Finding a compatible donor for kidney transplant candidates requires overcoming immunological barriers such as human leukocyte antigens (HLA) compatibility and ABO compatibility. Emerging data suggest a role for red blood cell antigens (RCA) in renal transplant outcomes. The incidence of RCA alloimmunization is high in chronically transfused individuals, such as end stage renal disease patients, but whether antibodies to RCA can mediate renal graft rejection remains debatable. The Duffy blood group antigens (Fy) has been shown to be expressed in the kidney, among other tissues. There are some data to suggest that donor-recipient Fy mismatches may increase the risk for chronic allograft damage and that anti-Fy antibodies may be involved in renal graft rejection, however, while it is routine to screen renal transplant candidates for ABO antigens, detailed RCA phenotyping of the donor kidney is not routinely tested. In this paper, we review the current data on the role of Fy in renal transplantation and discuss the potential mechanisms of its biological function. Finding a compatible donor for kidney transplant candidates requires overcoming immunological barriers such as human leukocyte antigens (HLA) compatibility and ABO compatibility, which may prolong time to transplant. Emerging data suggest a role for red blood cell antigens (RCA) in renal transplant outcomes –3. As ofWhether antibodies to RCA can mediate renal graft rejections remains debatable. Various studies produced mixed results when looking at retrospective data and trying to correlate presence of antibodies with kidney graft survival , 3. TherFY*A and FY*B). These two alleles differ by a single nucleotide polymorphism at position 125 (G/A) resulting in the presence either of glycine or aspartic acid in position 42 of the polypeptide chain that gives rise to the two antigens, Fya and Fyb  and IgG2 (18%) . , 9. FY*APlasmodium vivax. RBC lacking Fy are more resistant to invasion from this parasite. The lack of expression of Fy in AA is caused by a point mutation in the GATA box promoter of the Fyb allele , which prevents expression of Duffy protein only on RBC [FY*02N.02) which introduces a stop codon for the Duffy gene in all tissues  express Fy on their RBC . Varianty on RBC , 7. ThisThe Duffy antigen was identified as a transmembrane glycoprotein coupled receptor and was renamed the Duffy antigen receptor for chemokines (DARC) , 15. DARn = 239) versus mismatched (n = 131) donor-recipient pairs and correlated Fy mismatch status with histologic findings of kidney biopsies and overall survival [FY*A mismatches and FY*B mismatches were associated with chronic lesions. However, the FY*B mismatched group showed only increased intimal fibrosis which may have been attributed to older age in this group compared with the Fy matched group. The FY*A mismatched group had no age or other demographic differences compared with the Duffy matched group, yet this group showed significantly more chronic histologic changes. The difference between FY*A and FY*B mismatches was attributed to the higher immunogenicity of FY*A compared to FY*B [One way in which DARC can affect renal transplant outcomes is by acting as a minor histocompatibility antigen. A Lerut et al. did a retrospective study of 370 renal transplant recipients comparing Fy matched , ischemia reperfusion injury (IRI), and acute renal allograft rejection . A studyThe functional role of DARC in recruitment of inflammatory cells is still not fully understood, and several mechanisms have been suggested: 1) DARC helps presenting chemokines on the endothelial cells to leukocytes expressing corresponding receptors; 2) The binding of chemokines to DARC helps to create a gradient flow to keep an influx of chemokines coming to the site; 3) Neutralization of bound chemokines , 20. Alta antibodies detected 2 years prior to her transplant, although at time of transplant her anti-Fya antibodies were undetectable. Her post-transplant hospital course was complicated by DGF and a biopsy at 26 days post-transplant revealed acute rejection, both of which are associated with upregulation of DARC expression in the kidney [a antibodies and a repeat biopsy at 3 months showed acute and C4d+ antibody-mediated rejection in the absence of HLA donor-specific antibodies (DSA). The authors concluded that the unfavorable outcome of her transplant is a result of the presence of antibodies to Fya [The idea that anti-Fy antibodies may participate in renal allograft rejection is supported by a case report by Watorek et al. The authors reported a 41 years-old Caucasian woman, Fy(a− b+) phenotype, who had anti-Fye kidney , 19. Twos to Fya . At the s to Fya –25.At our center, we recently evaluated a 30 years-old African female with a history of multiple RBC transfusions due to anemia for a kidney transplant. Her HLA panel reactive antibody (PRA) was 0%. An ABO type and screen identified antibodies to Fy3, JkB, E, C, and K. The finding of the anti-Fy3 suggested that this patient carries the very rare true Fy null phenotype. Despite her ability  to receive transfusions due to the absence of Fy on RBC in most AA, her ability to find a compatible kidney donor is virtually impossible due to the presence of Fy on nearly every donor kidney. Even with a perfectly HLA and ABO matched donor, there is a potential for rejection due to anti-Fy3 reactivity and therefore this patient was deemed not suitable for transplantation at our center. Currently, there are no guidelines for donor-recipient RCA matching, mostly since more data is needed and since this will narrow down the donor pool for many transplant candidates. However, a more detailed screening process for RBC antibodies and RCA phenotyping of the donor may be warranted in patients experiencing antibody-mediated rejection (ABMR) with no HLA-DSA, especially if a recent post-transfusion reaction was observed prior to the diagnosis of ABMR. RCA antibody characterization might also be helpful prior to re-transplant following an ABMR involving RCA antibodies, and in cases where the recipient has a rare null phenotype. Differentiating between the true null and the erythrocyte silent mutations can be achieved by utilizing DNA-based typing instead of serologic phenotyping.In conclusion, most of the emphasis in overcoming immunological barriers in solid organ transplantation is rightfully put on matching donors and recipients for HLA and ABO, however, the Duffy blood group system can present a unique and rare barrier to transplantation and potentially impact transplant outcomes.


Of these, 1.4 billion (18%) lack soap or water, while 1.6 billion (22%) have neither. This analysis explores the link between living conditions and the use of essential agents in sub-Saharan Africa. This secondary data analysis examines potential associations between the domiciliary environment and the use of essential agents in sub-Saharan Africa.Eighteen demographic and health surveys were used to analyze the association between household environmental factors and handwashing with essential agents. STATA version 16 was used to analyze data from 203,311 households across weighted samples. Using a multivariable multilevel mixed effect logistic regression analysis, it was possible to determine how each independent factor affected the outcome while taking the data clustering into account. The adjusted odds ratio and its associated 95% confidence interval were used to assess the independent factors’ statistical significance.Only one in three households 34.84%, practiced handwashing with essential agents, with the highest prevalence in Angola (70.2%) and the lowest in Malawi (6.5%). Educational status , female headship, household wealth, not sharing toilets with other homes, having a fixed place for hand washing, not having regular access to water and being a rural resident  were associated with handwashing.sub-Saharan nations are failing to demonstrate advancements in handwashing practices. There are still a lot of homes without access to basic infrastructure for handwashing and household water sources. For essential agent adoption programs to be successful in an environment with limited resources, Water, Sanitation, and Hygiene measures must be implemented. Furthermore, it is critical to include contextual factors from the current study as well as socio-cultural and psychological characteristics that dissuade people from using essential agents in intervention strategies. Handwashing is the act of cleaning hands for 10–15 seconds by using essential agents , 3.The WHO advises washing hands frequently, but especially before and after caring for anyone who may be ill; before, during, and after preparing food; before eating; after using the restroom; after assisting someone who has just done so; after blowing one’s nose; after coughing or sneezing; after touching an animal, animal feed, or animal waste; and after touching garbage .Regular handwashing has a significant impact on one’s capacity to maintain good health. Maintaining hand cleanliness promotes good nutrition, improves educational possibilities by reducing absences from class, and prevents disease, which promotes child development and, in turn, increases economic potential , 6. HandHowever, developing the practice of consistent handwashing remains difficult, particularly in developing world contexts where it has been noted that only 3–35% of people wash their hands at crucial times , 13. In The current state of knowledge on the household factors associated with handwashing with essential agents in sub-Saharan Africa is still evolving. Recent studies have highlighted several important factors that influence handwashing practices in households, including access to water and sanitation facilities, education and awareness, socio-economic status, and cultural beliefs and practices –18. HoweThe scant literature on the multilevel factors that determine the practice of handwashing with essential agents reveals that we still lack scalable models for hygiene and sanitation that can ensure desired practices in circumstances where risks are common, compliance costs are high, and enforcement capacity is weak. This study thus aimed to examine the fixed effects (measures of association) and the random effects (measures of variation) of handwashing behavior with essential agents in 18 countries in sub-Saharan Africa.Cross-sectional data from 18 nationally representative Demographic and Health Surveys conducted between 2016 and 2021 are used in this report. The Demographic and Health Survey(DHS) Program offers extensive secondary data collected through surveys using probability sampling techniques and adhering to recognized international standards . The surA total of 203,311 households were included in the study, which drew a weighted sample from 18 countries in sub-Saharan Africa across four regions, namely Western, Eastern, Central, and Southern Africa refer to . Prior twww.measuredhs.com was accessed for the sake of this investigation after permission was acquired via an online form that outlined the study’s objectives.The Demography Health Survey (DHS) program’s official database, The dependent variable for this study was the prevalence of "handwashing practice with essential agents in households." During interviews conducted by the Demographic and Health Survey, participants were requested to show the area within their household where handwashing was predominantly performed. This was done to gather data on variables related to handwashing responses. The sources of water used for handwashing were classified as either unimproved or improved. Furthermore, households were asked about the availability of hand cleaning agents, with responses ranging from soap and detergent to ash, mud, and sand. The availability of the two fundamental handwashing supplies, soap, and improved water, was examined in homes.The dependent variable was then split into two categories: "Yes" for households that had engaged in necessary handwashing practices during the study period and "No" for households that had no .After reviewing recent literature, potential independent predictors of essential agent use were taken from the data set. Household-level variables were defined as respondent variables that were unique to each household , 12, 13.The factors applied at the community level were those that affected every home within the same community (cluster), such as residence, sub-region, community (cluster) poverty, and community education . The cluUsing STATA version 14, variables were organized, cleaned, and recoded before analysis. Households that had missing or unavailable outcome variables—those whose values couldn’t be used—were not included. These data were entered into the database using a unique code called "na," which either denoted "Don’t know" or replies that were deemed to be inconsistent with other entries on the questionnaires and hence likely to be erroneous.First, frequencies and proportions were computed using analytical and descriptive statistics. A weighted analysis was conducted to take into consideration the unequal likelihood of selection between the strata brought on by the non-proportional distribution of samples to various sub-regions, residences, and non-response rates.Due to the Demographic and Health Survey data’s hierarchical structure, where individuals are nested within households and households are nested inside clusters, multilevel (two-level) regression was used. The mixed-effect multilevel regression (MEMR) was the preferred statistical technique because of its exceptional flexibility and power in analyzing complex data that has a hierarchical structure, such as clustered data . This meFollowing the bi-variable multilevel logistic regression analysis, the variables with a p-value less than 0.25 were included in the multivariable analysis. To demonstrate the strength of the association, an adjusted odds ratio with a corresponding 95% confidence level was produced. For independent factors having a p-value of 0.05 or below, it was determined that the dependent variable was significantly associated with them. Multicollinearity between the household and community-level variables was examined using the Variance Inflation Factor (VIF) <10.Four random intercept models were fitted .Model 1 (null model): This model is based solely on the intercept and does not include any other independent variables. Statistics for measures of variance (random effects) were computed using the intra-class correlation coefficient (ICC) and median odds ratio (MOR). MOR can quantify unexplained cluster variability, whereas ICC explains cluster variability (heterogeneity) , 27. TheHousehold and community-level factors were both taken into consideration in Model 4. This model indicated that 11.7% of the unexplained variation might be explained by unobserved community- and individual-level factors. All models were compared and the fourth model with the lowest deviance was selected as the best-fit model. Additionally, were used to evaluate the goodness of fit. Better explanatory models are considered to have lowered AIC or BIC values  Table 4Table 4.All methods and procedures were carried out per the relevant guidelines and regulations of the Declaration of Helsinki. The Demographic and Health Surveys (DHS) collected written informed consent from all subjects and/or their legal guardian during the primary data collection. The retrieved data were only used for the registered research, and data were not shared with anyone other than the co-authors. Data were also fully anonymized by the Demographic and Health Survey program before the author team accessed them. No attempt was made to identify any household or individual, and the information was kept confidential. All data collected by the Demographic and Health Survey (DHS) are Institutional Review Board (IRB) certified and this secondary analysis received further IRB approval from the Wachemo University College of Medicine and Health Science Research Determination Committee.The weighted sample for this study included 203, 311 households. 36.67% of the study participants were household heads between the ages of 31 and 45. Likewise, 50.7% of the homes had four to seven people in residence. In the present study, 32% of the household heads had no formal education. 26.56% of the homes were female-headed households. 35.8% of households fell into the two quintiles with the lowest levels of wealth. In terms of the participants’ characteristics at the community level, 62.59% of the participants were rural residents. In addition, East Africa was home to 46.91% of the households.Only 49.36% of the homes had access to improved sanitation, while 58.04% had access to improved water sources. 67.74% of households accessed water from places other than their residences. In 37.1% of the studied households, there were shared toilets. In 16.6% of the houses, the distance to the water source was greater than 30 minutes. Mobile handwashing stations were available in 70% of the homes. Comparably, only over a quarter of the households (24.49%) treated their water at the point of use before utilizing it. In the 14 days before the study, water service interruption was reported in 57.20% of the homes as lasting a full day or longer. In the handwashing stations, only half of the examined households had running water.Interviewers were asked to inspect the location where family members wash their hands most frequently to collect information for the survey on handwashing. At the handwashing stations, soap and water were seen to be present. The dependent variable was then split into two categories: "Yes" for homes with handwashing stations equipped with essential agents and "No" for those without. Only 34.84% of the assessed homes had access to necessary amenities where family members could wash their hands .We performed a subgroup analysis based on the region of the survey in this analysis. As a result, the Eastern Africa and Central Africa regions had the highest and lowest prevalence of handwashing with essential agents respectively .Similar to this, the bivariate multilevel logistic regression established an association between hand washing with essential agents and the type of toilet facility, type of water source, location of the water source, sharing a toilet with another home or homes, water collection time, the place where members wash their hands, and water treatment .The geographic location of residence, sub-region, community poverty, and community education were all found to be significantly associated with hand washing using essential agents on a community level .After controlling for other variables, families with highly educated heads had a 77% higher likelihood of washing their hands with soap and water . The sex of the household heads had a substantial impact on the likelihood of using essential agents for handwashing, with female-headed households being more likely to do so .In comparison to households in the lowest quintile, households in the highest wealth quintile were four times more likely to follow hand washing recommendations. A higher likelihood of utilizing essential agents was likewise associated with not using a shared toilet facility inside a household or compound . Regarding water supply, not having regular access to water at hand washing stations and households with a fixed place for hand washing were both significantly associated with handwashing with essential agents and . Rural residents were 15% less likely to follow handwashing recommendations than urban residents  .Poverty is perpetuated by a lack of access to proper hygiene. For both people and society as a whole, having access to running water and soap reduces obstacles to economic development. It could result in fewer lost workdays, lower healthcare expenses, and greater access to educational opportunities . The preA disease must be seen as a significant community public health concern to be managed sustainably . PreviouIn this study, households in the highest wealth quintile had a fourfold higher likelihood of washing their hands as per recommended standards. It is well known that acquiring and cleaning water, as well as the cost of soap, consume a sizable portion of a poor family’s resources . In the In Africa, female-headed homes are typically poorer than male-headed homes, yet research also demonstrates that the children from these homes fare much better . ChildreNot sharing toilets with other homes was positively associated with washing hands with soap and water. The refusal of homes to share essential handwashing supplies with other households and toilet users is a significant contributor to the lack of water and/or soap in shared spaces. This act is a reaction to the "free riding" concern, which arises when it is believed that individuals profit from something without contributing to its provision . Future In the current study, contextual factors like regular access to water and the technology’s physical placement were significant predictors of handwashing with essential agents. A rising corpus of research, backed by behavioural plausibility, shows that if the aim is to promote habit formation, an interventional method that is much more automatic, unconscious, and cue-driven needs to be given more consideration . For exaIn the current study, urbanicity and handwashing with essential agents were associated. 57% of the water supply in sub-Saharan Africa’s rural households is unimproved . Little Using nationally representative, population-level, multi-country data, this study is one of the first of its kind to examine the relationships between domestic environmental conditions and handwashing with essential agents. The relatively high sample sizes that were examined in this study are one of its strengths since they improve the accuracy of the results. Additionally, this study serves as a resource for program managers and policymakers that are implementing multi-sectoral planning to scale up hand-washing practices. However this study is not without limitations, similar to any cross-sectional study, associations can be established but not causal relationships. Because the family head of household frequently responds to questions about water and sanitation, this study is limited by its reliance on him or her. The head of the home could be a man or woman from one of the various age groups found in the households. As a result, there may be variations in the reporting of WASH questions depending on who completes the household questionnaire.For many people, washing their hands with soap and water is more or less a fluid practice. Being aware of such variation in practice may be especially helpful for program developers when preparing for potential interventions and evaluation studies. To ensure that different types of users are taken into account and that ongoing training messages are tailored to each user, program designers should be aware of the various categories that households may fall into . Further, examining sub-national data for countries would be important for all of the countries to design effective and focused interventions.In terms of showcasing improvements in handwashing practices, sub-Saharan countries are off-course. There are still many nations without access to household water sources and fundamental sanitation infrastructure. The implementation of WASH initiatives will be crucial for the success of essential agent adoption programs in resource-limited settings. This includes educating the public on how to put up basic handwashing stations using affordable, locally accessible materials. Additionally, beyond everyday routines, the social context as well as the wider environment may have an impact on continuous handwashing behavior. Therefore, it is crucial to understand the factors that discourage people from using essential agents, including socio-cultural and psychosocial ones like educational status, female headship, household wealth, not using a shared toilet facility as well as contextual factors like regular access to water, handwashing technology’s physical placement, and urbanicity.


Bacteria account for nearly one third of the causes of community-acquired central nervous system infections, and traditional diagnostic methods are based on culture results, which are time-consuming and have a low detection rate leading to delayed diagnosis and treatment. Since metagenomic next-generation sequencing (mNGS) has the advantages of high timeliness and only detecting microbial trace gene fragments, it has been used more widely in recent years. Based on this, we explored whether the application of cerebrospinal fluid (CSF) mNGS is advantageous in patients with community-acquired purulent meningitis, especially in people who have already used antibiotics.This was a retrospective study of 63 patients with community-acquired purulent meningitis admitted to the Department of Neurology of Shanxi Bethune Hospital from March 2018 to November 2022. Data were systematically collected and classified into CSF culture group, blood culture group and CSF mNGS group according to different detection methods, and the total detection rate of each method was calculated. Each group of patients was divided into two subgroups according to whether antibiotics were used before sampling. The detection rates of the three groups were compared within and between groups to explore whether mNGS has advantages over traditional methods and the influence of antibiotic use on detection rates of the three methods.p < 0.05),suggesting that the detection rate of mNGS was higher than CSF culture (p < 0.05) and blood culture (p < 0.05),and the detection rate of blood culture higher than CSF culture (p < 0.05). Further grouping found that without antibiotics, the detection rates of CSF culture, blood culture and CSF mNGS were 28.57%, 56.25% and 88.89%, with statistical differences (p < 0.05), and the detection rate of CSF mNGS was higher than that of CSF culture (p < 0.05), but there was no statistical difference between CSF and blood culture (p > 0.05), nor between blood culture and CSF mNGS (p > 0.05). The detection rates of the three groups with antibiotics were 14.29%, 26.67% and 80.00%, with statistical differences (p < 0.05), and the detection rate of CSF mNGS was still higher than CSF culture (p < 0.05) and blood culture (p < 0.05). However, the detection rate of CSF mNGS also decreased after antibiotics were used for more than 3 days.Among the 63 patients, the cases of CSF culture, blood culture and CSF mNGS were 56, 46, 44, respectively. The total detection rates of the three methods were 17.86%, 36.96%, 81.82%, with statistical differences (The detection rate of CSF mNGS in patients with purulent meningitis is higher than traditional methods, especially in patients who have been given antibiotics, but the detection rate will decrease with the extension of antibiotic use. Infectious diseases are one of the leading causes of morbidity and mortality worldwide . As a coA total of 92 patients diagnosed with purulent meningitis/meningoencephalitis admitted to the Department of Neurology, Bethune Hospital, Shanxi Province from March 2018 to November 2022 since mNGS has been widely used in clinical practice in our hospital. According to Flow chart . Informed consent was waived because this was a retrospective study. We obtained patient data from the Medical Records and Statistics Room. We anonymously analyzed the data. The use of raw data was permitted by the Medical Ethics Committee of Shanxi Bethune Hospital.Inclusion criteria : 1. Patients present with clinical manifestations or signs of meningitis or encephalitis; 2. CSF characteristics suggest inflammatory response; 3. Patients whose CSF culture and/or blood culture results are bacterial infection according to traditional etiological examination and conform to clinical manifestations; 4. Traditional etiological examination was negative, but clinical manifestations and CSF manifestations were typical, and CSF characteristics were consistent with at least one of the following: ①CSF glucose < 1.9 mmol/L; ②CSF glucose/serum glucose < 0.23; ③CSF protein > 2.2 g/L;④CSF white blood cell count > 2000/μL or CSF multiple nuclear cell count > 1180/μL [ 1180/μL .Exclusion criteria: 1. Patients with incomplete data; 2. Patients with tuberculosis, virus, fungus and other types of infection were considered; 3. Nosocomial infection, craniocerebral postoperative infection, tumor, autoimmune encephalitis, rheumatic immune disease and other conditions are excluded. 4. Blood culture results were negative, but refuse to accept lumbar puncture or have contraindications for lumbar puncture; 5. Pregnant patients are excluded.Confirmed cases in this study include etiological confirmed cases  and clinical confirmed cases . (II) Positive rate refers to the proportion of the number of positive tests  to the number of tests; Detection rate refers to the proportion of the number of people who are positive and the result is consistent with the clinical diagnosis (true positive) to the number of people tested.(I) The sequencing depth of the CSF mNGS data is usually 20 M. Considering the difference in the microbial genome, the detected sequences are first normalized to reads per million (RPM), then the ratio of the RPM of clinical to the negative quality control (NTC) was calculated. When RPM-r10, the report result is positive .P < 0.05 was considered statistically significant.SPSS 23.0 was used to statistically analyze the data. Measurement data were expressed as median  or mean ± standard deviation(SD), counting data were expressed as rate, and rank sum test, and chi-square test or Fisher's exact probability method were used for comparison between groups, and Forty-four of the 63 patients completed CSF mNGS, and a total of 13 pathogens were identified. Among the 13 pathogens, 9 were true pathogenic bacteria, and 4 were polluting microorganisms. As traditional methods do not additionally identify other pathogens, the true pathogenic bacteria detected by mNGS also represents the pathogenic bacteria detected the pathogens detected by all methods.2 samples were both infected with two pathogens . However, combined with clinical manifestations and CSF characteristics of patients, among them, Torque teno virus 18 and Candida glabrata were interpreted as false positive, Nocardia brasiliensis and Klebsiella pneumoniae were interpreted as pathogenic bacteria, the results of 2 samples were ultimately interpreted as false positive (1 case was Timon Prevos and 1 case was Epstein-Barr virus), the remaining results were interpreted as true positive, and the pathogenic bacteria detected mainly were Streptococcus pneumoniae (11 cases) and Listeria glabella (8 cases). Klebsiella pneumoniae (6 cases), Brucella (4 cases), Staphylococcus aureus (2 cases), Streptococcus suis (2 cases). All pathogens detected are shown in Fig. Depending on the detection method, patients were divided into the CSF culture (CC) group(56cases), blood culture (BC) group(46cases) and CSF mNGS group (44cases), shown in the inside ring. The subgroups were divided according to whether antibiotics were used before sampling, shown in the middle ring. Light colors indicate antibiotics unused (Un) and dark colors indicate antibiotics already used (Used) before sampling. The outside ring shows number of positive and negative result. Color means positive(P), and gray means negative(N). The specific results above are shown in Fig. Among the 63 patients, 29 patients completed all three methods, as shown in Fig. Of all 63 patients, 56 underwent CSF culture, of which 10 were detected and all were true positive; 46 underwent blood culture, of which 17 were detected and all were true positive; 44 underwent CSF mNGS, of which 36 were true positive and 2 were false positive. Two additional pathogenic microorganisms were found in 36 true positive cases, which were finally interpreted as polluting microorganisms.. The overall detection rate of each group was calculated, and the three groups were further divided into 2 subgroups according to whether antibiotics were used before sampling. The detection rates of each subgroup were compared and statistically analyzed, and the results are shown in Fig. p = 0.247). The detection rate decreased significantly after 1 day of antibiotic use in the blood culture group (p = 0.000) and in the CSF mNGS group after 3 days of antibiotic use (p = 0.025).Calculate the cumulative detection rate of each time node (in days) of each group, that is, the detection rate of antibiotics used several days before sampling, make a line chart as shown in Fig. CNS infectious diseases, especially purulent meningitis, are rapidly progressive and fatal diseases that neurologists often have to face. Timely and correct diagnosis is the premise of effective treatment. The traditional diagnostic paradigm is often based on positive diagnosis of CSF smear, culture or blood culture, not only the detection rate is low but the time delay. Therefore, how to detect pathogens quickly and efficiently is still an urgent problem to be solved . EmerginIn further analysis of each subgroup, when antibiotics were not used before sampling, the detection rates of blood culture and mNGS were comparable, and both were higher than that of CSF culture. However, after antibiotic use, the detection rate of the blood culture group was significantly reduced and the detection rate of mNGS was significantly higher than that of the two traditional methods, indicating that the blood culture results were susceptible to antibiotics, but the detection rate of mNGS was less affected by antibiotics, that is, the detection rate before and after antibiotics was relatively stable, which is consistent with previous findings . LookingAlthough the detection rate of CSF mNGS is high, it is also affected by the length of antibiotic intervention before sampling. Studies  have indThis retrospective study also has some shortcomings: 1. Not all cases have completed the initial CSF culture and blood culture according to the standard diagnostic procedures. Of course, because of this, this result can more truly reflect the diversity and complexity of clinical diagnosis and treatment process, as well as the advantages of the CSF mNGS in the diagnosis and treatment of such cases. 2. According to the diagnostic criteria referred to in this paper, a part of patients with negative etiology and CSF traits that do not meet the clinical diagnostic criteria were missed, so it is not possible to evaluate whether CSF mNGS has an advantage after antibiotic use in this population.Currently, CSF mNGS has become an important and practical technique for the identification of the etiology of CNS infection. For patients who have already used antibiotics, the detection rate of mNGS is significantly better than that of traditional methods, which may be the direction for accurate etiological detection of CNS infection in the future. However, as the detection rate of mNGS decreases with the extension of antibiotic use, we still need to send samples for examination as early as possible.


Additionally, the proportion of neutrophils, lymphocytes, and monocytes did not exceed 75%, 75%, and 15%, respectively, in 95% of the population. Furthermore, in 95% of the specimens, the concentration of chloride, glucose, and protein was >115 mmol/L, >2.2 mmol/L, and <2.3 g/L, respectively. Compared to the normal reference values, the CSF indexes after spontaneous SAH showed significant changes, especially in the leukocyte count, chloride concentration, and glucose concentration. Using “white blood cell count < 880/mm3, glucose > 2.2 mmol/L, chloride > 115” as the reference values for SAH pathological status is more meaningful for reference purposes.The cerebrospinal fluid (CSF) analysis in ruptured aneurysms can be greatly affected by subarachnoid hemorrhage (SAH), making the diagnosis of intracranial infection more difficult after surgery. This study aimed to identify the reference value range of CSF in the pathological state following spontaneous SAH. A retrospective analysis of demographic and CSF data of all spontaneous SAH patients treated between January 2018 and January 2023 was conducted. A total of 101 valid CSF specimen data were collected for analysis. Our results indicate that in 95% of patients after spontaneous SAH, the leukocyte count in CSF was less than 880 × 10 Cerebrospinal fluid (CSF) laboratory testing is necessary to aid in the diagnosis of infection. However, the number of CSF cells and range of biochemical indicators in patients with subarachnoid hemorrhage (SAH) can change significantly due to bloody CSF, making the diagnosis of intracranial infection more challenging. To date, no studies have been conducted to determine the fluctuation range of CSF indicators in the pathological state following spontaneous SAH. In theory, the probability of intracranial infection caused by intravascular intervention treatment for aneurysms is extremely low because it does not result in a breach of the blood-brain barrier. This study collected routine and biochemical results of CSF from SAH patients who received interventional embolization or conservative treatment and analyzed the fluctuation interval to identify the reference value range of CSF in the pathological state following spontaneous SAH.Cases of spontaneous SAH were continuously enrolled at the Third People’s Hospital of Hubei Province from January 2018 to January 2023. Patients or their legal representatives signed both the informed consent form for diagnosis and treatment, as well as the informed consent form for this study. The inclusion criteria were as follows: (1) patients with spontaneous SAH; (2) patients who underwent interventional therapy or conservative treatment after admission; (3) patients without pre-existing nervous system diseases prior to hospitalization; and (4) patients who underwent lumbar puncture after SAH. The exclusion criteria comprised the following: (1) Patients who underwent craniotomy after admission; (2) patients who underwent other invasive neurological procedures besides aneurysm embolization prior to CSF collection; (3) patients whose CSF culture indicated bacterial infection; and (4) patients who experienced increased SAH during or after interventional therapy. All procedures were conducted in compliance with relevant guidelines and regulations. This study received approval from the local ethics committee.A retrospective study was conducted to collect all cases of spontaneous SAH from January 2018 to January 2023. Patient data, including age, gender, time of SAH, time of lumbar puncture, and various CSF indicator results, were recorded. Cases were screened according to inclusion and exclusion criteria, and data from all eligible cases were analyzed.p < 0.05 was considered statistically significant.Statistical analyses were conducted using IBM SPSS Statistics version 26 . The normality of the measurement data was tested. Continuous variables were presented as medians with interquartile ranges (IQR) and compared using the Mann-Whitney U test. Categorical variables were reported as numbers with percentages. A total of 118 cases of spontaneous SAH treated with either interventional or conservative methods were included in this study. After exclusion criteria were applied, a total of 101 eligible CSF specimens were collected. Of these, 86 cases (85.1%) underwent endovascular embolization, while 15 cases (14.9%) were treated conservatively . The pri6 cells/L, the 5–95th percentiles were (8–880) × 106 cells/L, and the 10–90th percentiles were (10–496) × 106 cells/L  of all specimens showed a skewed distribution A. The me cells/L . In term cells/L . The pro cells/L B–D. The  cells/L . The pro cells/L . Similar cells/L . After s6 cells/L, with 5–95th percentiles ranging from  × 106 cells/L and 10–90th percentiles ranging from  × 106 cells/L (The red blood cell (RBC or erythrocyte) count in CSF increased and exhibited a skewed distribution A. The me cells/L . The rat cells/L B. The me cells/L . These fThe distribution of chloride concentration was found to be skewed A, with t6 cells/L, with a median value of 100 × 106 cells/L, which was much higher than the normal value range of (0–8) × 106 cells/L  × 10 cells/L A. The lo cells/L B. Simila cells/L C. These The incidence rate of central nervous system (CNS) infections after neurosurgery ranges from 2.5% to 4.6% . CNS infThe routine and biochemical examination of cerebrospinal fluid (CSF) plays a crucial role in the diagnosis and treatment of neurological and neurosurgical diseases, and its reference values are essential for guiding clinical decision-making. However, the CSF results can be significantly altered after aneurysmal subarachnoid hemorrhage (SAH), rendering the normal reference values unreliable. Patients with ruptured cerebral aneurysms often require craniotomy and aneurysm clipping and may experience postoperative symptoms such as fever, headache, cervical rigidity, and impaired consciousness, which could be caused by pulmonary infections, meningeal irritation, or the absorption of bloody CSF. It is challenging for clinicians to differentiate whether these patients have concomitant postoperative intracranial infections based solely on their symptoms. CSF examination is often necessary to aid in diagnosis, but the positive rate of CSF bacterial culture for CNS infection is generally low ,7. Moreo3 and a proportion of multinucleated cells >0.7. If the number of leukocytes in CSF is less than 1000 cells/mm3, bacterial infection cannot be ruled out. A comprehensive evaluation of risk factors, clinical signs, and other biochemical indicators is necessary to make an accurate diagnosis [At present, early diagnosis of CNS infection in neurosurgery still poses a significant challenge ,9. The t6/L is suggestive of a central nervous system infection [6 cells/L, and the 5–95th percentiles were (8–880) × 106 cells/L. Therefore, 50% of the SAH population would have a CSF WBC count that exceeds the clinical diagnostic criteria for intracranial infection. The median proportion of neutrophils was 46.0%, and 13% of patients had a neutrophil ratio >0.7. The reference standard for CSF routine in current guidelines for the diagnosis of CNS infections is not applicable for judging whether patients with SAH have an intracranial infection. In this study, the CSF WBC count was <880 cells/mm3, and the neutrophil ratio was <0.75 in 95% of the population after spontaneous SAH. Using a “WBC count > 880 cells/mm3” as a reference indicator for the presence or absence of intracranial infection would be more instructive in patients with spontaneous SAH. However, bacterial infections cannot be ruled out in patients with white blood cell counts in the range of 100–880 cells/mm3. After a subarachnoid hemorrhage (SAH), there is a significant increase in the red blood cell (RBC) count in the cerebrospinal fluid (CSF). This study demonstrates that the 10% to 90% percentile interval for the CSF RBC count in patients with SAH is  × 106 cells/L, while the percentile interval for the ratio of RBC count to white blood cell (WBC) count is (9–1670), which fluctuates widely and is not suitable for clinical use. Despite this, the authors observed that some clinicians still try to convert the RBC/WBC ratio to estimate the actual WBC count in the CSF. However, the authors caution against this method for patients with spontaneous SAH due to two reasons. Firstly, patients are typically bedridden for a prolonged period after the hemorrhage and the RBCs deposit in a low position. Thus, the CSF samples collected through lumbar puncture may not accurately reflect the total RBC content in the CSF. Secondly, the premise of using the RBC count to correct the WBC count is that fresh bleeding occurs during the lumbar puncture procedure. Nonetheless, in the long-term bed rest process for SAH patients, cell separation occurs due to the sedimentation of blood cells, making the conventional correction method of little practical significance.According to the literature, a CSF WBC count greater than 100 × 10nfection . The resThe normal concentration of glucose in CSF is 2.5–4.5 mmol/L, which is two-fifths of the serum glucose level due to its dependence on serum glucose . In thisPatients with aneurysmal SAH may develop pulmonary infections, which can cause fever and hyperthermia even without invasive neurological treatment. Additionally, most patients will experience symptoms such as impaired consciousness, headache, neck stiffness, and increased intracranial pressure stimulated by bloody CSF. These symptoms can be similar to those of CNS infections, making it difficult to determine whether intracranial infection is present after surgery . Moreove6/L and less than 880 × 106/L in 95% of patients. The proportion of neutrophils, lymphocytes, and monocytes did not exceed 75%, 75%, and 15%, respectively, in 95% of the population. The red blood cell (RBC) count to WBC count ratio in the CSF exhibited significant variability after SAH. Furthermore, the concentration of chloride, glucose, and protein was >115 mmol/L, >2.2 mmol/L, and <2.3 g/L, respectively, in 95% of the specimens. If the 5–95% quartile interval is taken as the reference range, a “white blood cell count > 880 mm3, glucose < 2.2 mmol/L, chloride < 115” can be used as the reference index for detecting the presence of intracranial infection, which may have greater guiding significance for patients with spontaneous SAH. These results are useful for the identification of intracranial infection in patients with SAH, which can reduce the abuse of antibiotics. However, further prospective studies are necessary to validate the reference range.In this study, we conducted a statistical analysis of the pathological changes in cerebrospinal fluid (CSF) indexes following spontaneous subarachnoid hemorrhage (SAH) and established a reference range. Our findings indicate that in 90% of patients with spontaneous SAH, the white blood cell (WBC) count in the CSF was less than 500 × 10Our study has several limitations that should be acknowledged. Firstly, it is a retrospective study, and the patients included in the study had severe clinical symptoms and a high degree of SAH. This could have resulted in elevated levels of erythrocytes and leukocytes in the CSF. Future prospective studies are needed to compare CSF indicators in patients with different degrees of hemorrhage. Secondly, the sample size of this study was small. Therefore, the range of CSF index fluctuation in SAH patients needs to be confirmed through larger studies conducted at multiple centers.3, glucose > 2.2 mmol/L, chloride > 115” as the reference values for SAH pathological status is more meaningful for reference purposes.Compared to the normal reference values, the CSF indexes after spontaneous SAH showed significant changes, especially in the leukocyte count, chloride concentration, and glucose concentration. Using “white blood cell count < 880 mm


Demand for donor hearts and lungs exceeds their supply. Extended Criteria Donor (ECD) organs are used to help meet this demand, but their impact on heart-lung transplantation outcomes is poorly characterized.n = 447) from 2005‒2021. Recipients were stratified based on whether they received ECD hearts and/or lungs. Morbidity was analyzed using Kruskal-Wallis, chi-square, and Fisher's exact tests. Mortality was analyzed using Kaplan-Meier estimation, log-rank tests and Cox regression. Sixty-five (14.5%) patients received two ECD organs, 134 (30.0%) received only an ECD lung, and 65 (14.5%) only an ECD heart. Recipients of two ECD organs were older, more likely to have diabetes, and more likely transplanted from 2015‒2021 (p < 0.05). Groups did not differ by pre-transplant diagnosis, intensive care unit disposition, life support use, or hemodynamics. Group five-year survival rates ranged from 54.5% to 63.2% (p = 0.428). Groups did not differ by 30-day mortality, strokes, graft rejection, or hospital length of stay.: The United Network for Organ Sharing was queried for data on adult heart-lung transplantation recipients (: Using ECD hearts and/or lungs for heart-lung transplantation is not associated with increased mortality and is a safe strategy for increasing donor organ supply in this complex patient population. Demand for donor hearts and lungs continues to exceed their supply. One strategy to increase supply is the use of Extended Criteria Donor (ECD) organs. While there is no consensus on what the extended criteria for hearts or lungs are, previously published lists have considerable overlap and tend to include increased donor age, ischemic time, and other risk factors for end-organ dysfunction ,2. SeverSimultaneous Heart-Lung Transplantation (HLTx) remains the preferred treatment for concomitant end-stage cardiac and pulmonary dysfunction. Yet, finding organs for these very ill patients can be challenging. While the benefit of using carefully selected ECD organs for single-organ heart and lung transplantations has been shown, it is unclear what effect ECD organs have on HLTx outcomes. Furthermore, the complexities of this patient subset, including the presence of Eisenmenger syndrome with an unrepairable cardiac defect, differ from those of single-organ heart or lung transplantation The authors undertook this study to 1) Assess the effect of ECD organs on morbidity and mortality in the setting of HLTx and 2) Assess the prevalence of ECD heart and lung use in HLTx over time.The United Network for Organ Sharing (UNOS) thoracic database was queried for all patients aged 18 or older who received HLTx from January 1, 2005, through July 19, 2021. 2005 was selected to exclude transplantations prior to the implementation of lung allocation scores. Patients with unknown mortality status at the last follow-up were excluded. Each HLTx recipient was assigned to one of four groups based on donor organ status. The four groups were recipients of either two Standard Criteria Donor (SCD) organs, two Extended Criteria Donor (ECD) organs, an ECD heart and SCD lung, or an ECD lung and SCD heart. Donor hearts and lungs were determined to be ECD organs if they satisfied two ECD criteria . ECD criBaseline characteristics, morbidity, and mortality data are reported for all donor recipients and compared by donor organ status. Predicted total lung capacity was calculated from height using previously described methods The University of Pennsylvania Institutional Review Board deemed the study to be not human subjects research and waived the need for patients’ written informed consent . The study was completed in conformity with STROBE Statement guidelines.n = 111, 24.9%). The largest cohort of recipients  received two SCD organs. 134 (30.0%) received only an ECD lung, 70 (15.7%) received only an ECD heart, and 65 (14.5%) received both an ECD lung and an ECD heart.447 adults undergoing simultaneous HLTx from 2005 to 2021 were analyzed . 232 (51p < 0.05) , inotrope need, serum creatinine, infiltrates on chest radiographs, purulent secretions on bronchoscopy, a ratio of arterial oxygen partial pressure to the fraction of inspired oxygen (P/F ratio), and circulatory death ( < 0.05) , Table 4For all adults undergoing HLTx from 2005‒2015, one-year survival was 81.6% and five-year survival was 58.5% . 120 (27p = 0.339), one-year survival , or five-year survival  (p = 0.529), 1-year , or 5-years . These groups also did not differ in terms of early postoperative outcomes including rates of intubation  and ECMO at 72 h , rates of reintubation , and median hospital length of stay . Additionally, the donor organ status groups had similar incidences of complications including rejection within one year of transplant , new dialysis requirement , and postoperative stroke . The groups did however differ in terms of postoperative permanent pacemaker placement with recipients of two ECD organs having the highest rate (4.7%) and recipients of two SCD organs having the lowest .The four donor organ status groups did not differ in terms of 30-day survival  . When lip < 0.001) and P/F ratio < 300   . No otheAmong adults undergoing simultaneous HLTx from 2005 to 2021, the use of an ECD heart and/or lung is not associated with increased mortality at five years, one year, or 30 days. Further, the use of one or more ECD organs is not associated with increased graft rejection, postoperative stroke, or hospital length of stay. The only measure of morbidity positively associated with ECD organ use is postoperative permanent pacemaker placement, which was more likely in recipients of both an ECD heart and an ECD lung. Of the 12 extended donor criteria identified in this study, only two – age ≥ 55 and P/F ratio < 300 – were associated with mortality at 5 years.The comparable mortality and morbidity between recipients of SCD and ECD organs are not easily explained by recipient characteristics. In fact, recipients of two ECD donor organs were less healthy than other recipients with respect to increased age and diabetes prevalence. The only reported characteristics that may have biased outcomes in favor of recipients of ECD organs were the relative increase in transplantation of ECD organs from 2015‒2021, the higher proportion of SCD organ donors on inotropes at the time of procurement, and the lower rates of cytomegalovirus positivity among recipients of ECD organs. Yet on Cox regression, year of transplant, donor inotrope use, and recipient cytomegalovirus positivity were not significantly associated with five-year mortality. Further, the donor organ status group itself was not associated with five-year survival. It is unclear why recipients of two ECD donor organs had increased pacemaker need, though this may be attributable to these recipients being older The relative safety of using ECD hearts and ECD lungs is consistent with existing literature showing that the use of ECD hearts or lungs in single-organ transplantation yields comparable outcomes to using SCD organs Limitations of this study are a function of its retrospective design and reliance on the UNOS database. While the UNOS database is invaluable for understanding trends in transplantation, it does not report all pre-transplant variables that are useful for assessing donor organ health nor does it investigate every significant post-transplant outcome. It fails, for example, to report the dose of donor inotrope support at the time of procurement, which is notable given that high-dose inotrope support has been identified as an extended donor criterion. It is thus possible that the present study underestimated the prevalence of ECD organs being used and/or mischaracterized some ECD organ recipients and SCD organ recipients. Additionally, the UNOS database does not report significant post-operative outcomes including primary graft dysfunction, pulmonary function test findings, or echocardiographic measures. Future studies could better characterize the impact of ECD organs on morbidity and mortality by using a prospective design and reporting these additional pre- and post-transplant variables.With appropriate patient selection, using one or two ECD organs in HLTx represents a viable strategy for increasing the supply of limited donor hearts and lungs.This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.Noah Weingarten: Conceptualization, Data curation, Formal analysis, Methodology, Writing – original draft, Writing – review & editing. Amit Iyengar: Conceptualization, Data curation, Formal analysis, Methodology, Supervision, Writing – original draft, Writing – review & editing. David Alan Herbst: Writing – original draft, Writing – review & editing. Mark Helmers: Writing – original draft, Writing – review & editing. Danika Meldrum: Writing – original draft, Writing – review & editing. Sara Guevara-Plunkett: Writing – original draft, Writing – review & editing. Jessica Dominic: Writing – original draft, Writing – review & editing. Pavan Atluri: Conceptualization, Resources, Supervision, Writing – original draft, Writing – review & editing.The authors declare no conflicts of interest.


Sp)-PB/PO chimeric oligouridylate had higher hybridization ability than the unmodified counterpart to an unmodified oligoadenylate. This is the first report that elucidates the effect of both stereochemistry and type of P-modification (PB and PS) on properties of oligoribonucleotides.This study describes the stereoselective synthesis of boranophosphate/phosphate (PB/PO) and phosphorothioate/phosphate (PS/PO) chimeric oligouridylates using the solid-phase method. Oxazaphospholidine monomer was used to construct the stereodefined PB and PS linkages. The study introduces modifications to oligouridylate derivatives in the intended positions with the intended stereochemistry of phosphorous atoms. Additionally, biophysical and biochemical properties of the synthesized oligomers were evaluated. Notably, it was found that a ( Since discovering the RNA interference (RNAi) mechanism , much efFive siRNA drugs, Patisiran , GivosirP-modified phosphate linkages into siRNAs. A phosphorothioate modification is the most prevalent strategy due to its easy access and reliable nuclease resistance .,26.25,26. 2. 2.11) was condensed with an oxazaphospholidine monomer 2 [3. Then, the resultant phosphite 3 was boronated. The 5′-O-DMTr group was removed under acidic conditions to give an intermediate 4 in the presence of Et3SiH as a cation scavenger to prevent the side reaction on borano groups with DMTr cation [3–EtOH  solution afforded a diuridine boranophosphate (CEM-on), and the mixture was analysed using reverse-phase high-performance liquid chromatography (RP-HPLC). The results are shown in Sp)−2 was used as a monomer and the boronation reaction was conducted using the BH3·SMe2 complex in DMAc solvent. However, there was a large peak just after the peak derived from the product in the RP-HPLC profile , indicating that the BH3·SMe2 complex caused a side reaction. Using 0.05 M BH3·THF complex in THF instead of BH3·SMe2 complex gave the UPBU (CEM-on) in a high yield with good diastereoselectivity (entry 2). The (Rp)-2 monomer also afforded (Rp)-UPBU (CEM-on) with a good HPLC yield and stereoselectivity (entry 3).First, we attempted the synthesis of PB/PO chimeric ORNs. The boronation reaction conditions were studied to convert a phosphite intermediate to a boranophosphate. Thus, solid-phase synthesis of stereocontrolled diuridine boraphosphates was investigated using an automated synthesizer, as shown in onomer 2 , whose 2r cation ,29. Subst-butylhydroperoxide (TBHP) as an oxidizing reagent, which is necessary to create phosphate linkages because boranophosphotriester was almost inert towards TBHP in liquid-phase synthesis, as we previously reported [3COIm is a favourable capping reagent for the stereocontrolled synthesis of PS-DNAs and RNAs using the oxazaphospholidine method [3COIm. However, the reduction of trifluoroacetamide moiety upon treatment with a boronation reagent would affect the synthesis of an oligomer with boranophosphate linkages. Thus, Unicap® phosphoramidite was chosen as a capping reagent for the synthesis. CEM-on octamers, (Sp)- and (Rp)-UPOUPOUPOUPBUPOUPOUPOU were synthesized by repeated cycles consisting of the condensation, capping, boronation or oxidation and detritylation, then the removal of auxiliary groups and release from the solid support was conducted by treatment with a concentrated NH3 aq–EtOH  solution. To construct the phosphate linkage, we used the phosphoramidite 5 and 5-(ethylthio)-tetrazole as a monomer and activator, respectively. For the stereodefined boranophosphate linkage formation, we used the oxazaphospholidine 2 and PhIMT (O-CEM group by treatment with tetra-n-butylammonium fluoride (TBAF) in dimethyl sulfoxide (DMSO) in the presence of 0.5% MeNO2 as a scavenger of acrylonitrile [Sp)- and (Rp)-octamers in 19% and 32% yields, respectively, based on UV absorbance value at 260 nm -2 or (Sp)-2 with a hydroxy group. After we conducted a capping step with CF3COIm and N-methylimidazole (NMI), we sulfurized the phosphite intermediate using 3-phenyl 1,2,4-dithiazoline-5-one (POS) [5 as a monomer, acetic anhydride (Ac2O) as a capping reagent and TBHP for the oxidation step. We followed the procedure for the oligomer release from a solid support and removal of the 2′-O-CEM group to synthesize PB/PO chimeric U12  . To formeric U12 . The PS/eric U12 . Isolate. 2.2P-modified linkages in their central region with an equal amount of dodecaadenylate in a buffer solution, evaluating the Tm value with a thermal denaturation test. 2HPO4–NaH2PO4, we could not determine the Tm values of all duplexes . Thus, Tm values were calculated from the results obtained from the experiments with a higher salt concentration -PS/PO and (Rp)-PB/PO-ORNs had reduced duplex-forming ability, whereas (Rp)-PS/PO-ORN showed comparable ability. Notably, (Sp)-PB/PO chimeric U12 formed a marked stable duplex. This result agrees well with our previous report that an all-(Sp)-PB 2′-O-Me decauridylate formed a more stable duplex with a complementary ORN than an unmodified counterpart, whereas the Tm value of an all-(Rp)-PB 2′-O-Me decauridylate and complementary ORN could not be determined owing to the instability of duplex [Rp)-PB/PO chimeric U12 and A12 had a small hyperchromicity compared to the other duplexes. We attributed the phenomenon to the low thermal stability of the duplex. A part of the duplex might be dissociated even at 0°C. From these results, the order of hybridization ability of dodecamers was found to be as follows: (Sp)-PB/PO > (Rp)-PS/PO ≈ unmodified > (Sp)-PS/PO > (Rp)-PB/PO.Next, we moved to the evaluation of the obtained oligomers. We annealed each stereocontrolled PB/PO and PS/PO chimeric dodecauridylate having f duplex . From thRp)-PS/PO and (Sp)-PB/PO chimeric dodecauridylates, which formed relatively stable duplexes, were used for the experiments and the results are shown in H and ΔS values of duplex formation of (Rp)-PS/PO chimeric dodecauridylate and the complementary strand was clearly lower. On the other hand, ΔH and ΔS values of (Sp)-PB/PO chimeric dodecauridylate and dodecaadenylaye were comparable and slightly higher than those of the unmodified duplex, respectively. These results suggested that introduction of (Rp)-PS modification led to an enthalpy gain upon duplex formation whereas (Sp)-PB modification resulted in a reduced entropy loss. The detailed study must be conducted with oligomers bearing four nucleobases.To gain further insights into the effect of chemical modifications on duplex formation, the thermodynamic parameters of duplex formation were calculated by denaturation tests at varied duplex concentrations. Unmodified, , (Rp)-UPBU with SVPDE for designated times and then divided and analysed a portion of the solution using RP-HPLC. The RP-HPLC profiles clearly showed that (Rp)-UPBU was stable under the reaction conditions, whereas (Sp)-UPBU was gradually cleaved (Rp)-counterpart had higher resistance to SVPDE digestion and did not contradict previous reports [Subsequently, we investigated the nuclease resistance of synthesized oligomers using snake venom phosphodiesterase (SVPDE) from nuclease . First,  cleaved . This re reports , suggestP-modified linkages at 3′-ends with SVPDE to investigate the effect of the type of modification and configuration of phosphorous atoms on nuclease resistance. Rp)-PS/PO U12 degraded the most rapidly, followed by its (Rp) and (Sp)-PB/PO counterparts. (Sp)-PS/PO U12 seemed to be the most resistant to the SVPDE digestion. The effect of phosphorous configuration on nuclease resistance was apparent for PS/PO chimeric U12, whereas (Rp) and (Sp)-PB/PO oligomers showed similar behaviour.Next, we treated PB/PO and PS/PO chimeric oligomers having . 3P-modified linkages. We used Unicap® phosphoramidite for the capping steps in the synthesis of PB/PO-ORNs to suppress side reactions. We also evaluated the biophysical and biochemical properties of the resultant oligomers. The following order was elucidated:We have developed an efficient method for synthesizing PB/PO and PS/PO chimeric oligouridylate derivatives in a stereoselective manner. Concurrent usage of phosphoramidite and oxazaphospholidine monomers enabled the construction of both phosphate and stereocontrolled Sp)-PB/PO > (Rp)-PS/PO ≈ unmodified > (Sp)-PS/PO > (Rp)-PB/POhybridization ability to complementary ORN: (Sp)-PS/PO > (Rp)-PB/PO ≈ (Sp)-PB/PO > (Rp)-PS/PO > unmodified.3′-exonuclease resistance: .Synthesized dimers were analysed using RP-HPLC. Synthesized oligomers (octamers and dodecamers) were analysed and purified using reversed-phase HPLC and identified by MALDI or ESI mass spectroscopy. Isolated yields of oligomers were estimated by measuring the UV–Vis spectra using the following molar absorption constant at 260 nm (octamer: . 4.2Sp) and (Rp)-UPBU was conducted following the procedure shown in O-DMTr-uridine loaded HCP via a succinyl linker . After the synthesis, we treated the HCP with concentrated aqueous NH3–EtOH  at 25°C for 3 h, filtered and washed with EtOH. The filtrate and washings were combined and concentrated under reduced pressure. The residue was analysed using RP-HPLC, conducted with a linear gradient of 0%–30% MeCN in a 0.1 M triethylammonium acetate (TEAA) buffer (pH 7.0) at 30°C for 60 min with a flow rate of 0.5 ml min−1 in a C18 column.An automated solid-phase synthesis of (. 4.3Sp) and (Rp)-octamers and dodecamers was conducted following the procedure shown in −1, 0.2 µmol). After the designed length was achieved, the HCP was treated with concentrated aqueous NH3–EtOH  at 25°C for 3–10 h, filtered and washed with EtOH. The filtrate and washings were combined and concentrated under reduced pressure. The residue was analysed and purified using RP-HPLC, conducted with a linear gradient of 0%–40% MeCN in a 0.1 M TEAA buffer (pH 7.0) at 55°C for 75 min with a flow rate of 0.5 ml min−1 in a C18 column.An automated solid-phase synthesis of . After 5–10 h, the reaction was quenched by the addition of 40 ml 0.1 M TEAB buffer (pH 7). The mixture was desalted by C18 cartridge. The cartridge was washed with 5% MeCN and the desired oligomer was eluted by 40% MeCN. The solution was lyophilized. The residue was analysed and purified by RP-HPLC, which was performed using a linear gradient of 0%–40% MeCN for 75 min in 0.1 M TEAA buffer (pH 7.0).The purified octamer or dodecamer bearing CEM group on the 2′ positions was treated with 0.5 M TBAF solution in dry DMSO containing 0.5% MeNO. 4.52PO4–Na2HPO4 buffer or 1 M NaCl in 100 mM NaH2PO4–Na2HPO4 buffer (pH 7.0) was heated for 10 min at 90°C and cooled to 0°C at a rate of 1.0°C/min and then left at 0°C for 30 min. Denaturation tests were conducted in a 1 cm path length quartz cell. The temperatures at the crossing points of the melting curves and the median lines of the low- and high-temperature baselines were calculated as Tm values. The three independent tests were conducted and the average values along with standard deviations were calculated.A solution containing pairs of complementary strands  and 100 mM NaCl in a 10 mM NaH. 4.62PO4–Na2HPO4 buffer (pH 7.0) was heated for 10 min at 90°C and cooled to 0°C at a rate of 1.0°C/min and then left at 0°C for 30 min. The diluted duplex solutions  were also prepared by diluting the solution by a 1 M NaCl aqueous solution in a 100 mM NaH2PO4–Na2HPO4 buffer (pH 7.0). Denaturation tests were conducted in a 1 cm path length quartz cell. The temperatures at the crossing points of the melting curves and the median lines of the low- and high-temperature baselines were calculated as Tm values. The ΔH° and ΔS° values were calculated from the following equation [A solution containing pairs of complementary strands (16 µM and 2.4 nmol each) and 1 M NaCl in a 100 mM NaHequation .1Tm=RΔHl. 4.72, 0.01 mM ZnCl2 and 0.1 mM spermidine at 37°C. The reactions were conducted for 0.5, 3, 6, 12 and 28 h. After the designated time, a part of the reation mixture was divided and a 0.1 M TEAA buffer (100 µl) was added to the mixture, and the mixture was heated to 100°C for 1 min to denature the enzyme. The mixture was analysed using RP-HPLC.In the enzymatic digestion experiments, 5.0 nmol (50 µM) of each dimer was treated with SVPDE from Crotalus adamanteus (0.3 U) in a 5 mM Tris-HCl buffer  containing 0.1 mM MgCl. 4.8–2 U) in a 5 mM Tris-HCl buffer  containing 0.1 mM MgCl2, 0.01 mM ZnCl2 and 0.1 mM spermidine at 37°C. The reactions were conducted for 0.5, 2, 4, 8 and 12 h. After the designated time, a part of the reation mixture was divided, a 0.1 M TEAA buffer (100 µl) was added to the mixture, and the mixture was heated to 100°C for 1 min to denature the enzyme. The mixture was analysed using RP-HPLC.In the enzymatic digestion experiments, 1.0 nmol (10 µM) of each oligomer was treated with SVPDE from Crotalus adamanteus (2 × 10


Surface electromyography (sEMG) is widely used in monitoring human health. Nonetheless, it is challenging to capture high-fidelity sEMG recordings in regions with intricate curved surfaces such as the larynx, because regular sEMG electrodes have stiff structures. In this study, we developed a stretchable, high-density sEMG electrode array via layer-by-layer printing and lamination. The electrode offered a series of excellent human‒machine interface features, including conformal adhesion to the skin, high electron-to-ion conductivity (and thus lower contact impedance), prolonged environmental adaptability to resist water evaporation, and epidermal biocompatibility. This made the electrode more appropriate than commercial electrodes for long-term wearable, high-fidelity sEMG recording devices at complicated skin interfaces. Systematic in vivo studies were used to investigate its ability to classify swallowing activities, which was accomplished with high accuracy by decoding the sEMG signals from the chin via integration with an ear-mounted wearable system and machine learning algorithms. The results demonstrated the clinical feasibility of the system for noninvasive and comfortable recognition of swallowing motions for comfortable dysphagia rehabilitation. The costs of hospital tests and treatments are frequently very high, making them uneconomical for protracted healing processes. An alternative strategy is to use portable monitoring devices to enable home disease monitoring. This model enables real-time monitoring of the patient’s health status, and the data are processed and saved via wireless transmission, and the results are presented with simple visualizations4. Since it is a noninvasive and easy-to-use technique, surface electromyography (sEMG) is often used for monitoring health and treating diseases. The measurement of the EMG signal depends mainly on the interface between the skin and the electronic acquisition circuit. However, the commercial Ag/AgCl electrodes used in existing sEMG acquisition devices are insufficient to establish a robust, conformal interface for long-term high-fidelity sEMG signal acquisition. The electrodes are typically made of nonstretchable materials and are electrically connected via metal buttons to ensure the stability of the device. When the skin is distorted by human activity, interface delamination occurs between the electrode and the skin because of the significant modulus difference. In addition, commercial electrodes add a gel serving as a binder and ionic medium, which leads to loss of signal quality due to dehydration over long periods of use6.With continuous advancements in medical technology, the prevention and rehabilitation processes for diseases with long treatment cycles, such as strokes, are becoming major concerns12. To eliminate motion artifacts arising from the different moduli for the skin and the electrode, stretchable electrodes provide soft conformal contact even during skin deformation. In general, there are two main design strategies for stretchable electrodes. One is based on an elaborate structure such as serpentine, where a nonstretchable conductor is spatially endowed with deformability19. The other category uses conductive fillers filled with intrinsically stretchable composites, such as hydrogels or Polydimethylsiloxane (PDMS) filled with silver nanosheets23. These methods are designed to improve the conformal contact of the flexible electrodes with skin exhibiting large curvature and complex textures. Curved skin interfaces frequently have complex muscle distributions. Examples include the larynx and the face, where there are many more small muscles than there are in the arm. The traditional bipolar disk sEMG acquisition mode can only acquire sEMG signals from one measured area, which may have multiple muscles or muscle groups superposed and does not accurately reflect the specific muscle to be analyzed. High-density surface electromyography (HD-sEMG) compensates for the shortcomings of conventional differential electrodes. It quantitatively analyzes the intensity and temporal sequence information from different muscles in space. Moreover, it enables accurate, comprehensive, and objective evaluation of the synergistic effects of different muscles during muscle activity. Kim et al. presented a reusable, multichannel sEMG sensor array that covered multiple muscles over relatively large areas. The stretchable structure and array design based on serpentine interconnections enabled it to acquire sEMG information from complex curved surfaces such as the back of the hand or the face16. Although dry electrodes can achieve stable, long-term monitoring, the lack of robust adhesion to the skin leads to inevitable relative slippage against the skin. Hydrogels can establish robust bioelectronic interfaces with properties close to those of human tissue as well as easy processability with which to engineer the mechanical strength, biological adhesion, or electrical conductivity26. For example, Gong et al. reported a neutral polyampholyte hydrogel (PA gel), which showed rapid, strong, and reversible adhesion to charged hydrogels or biological tissues through coulombic interactions with its self-adjustable surface27. For hydrogel-based on-skin electrodes, another important feature is the conductivity of the hydrogel. It can be enhanced by adding ions or by adding conductive fillers such as carbon nanotubes and conductive polymers31. In addition, to enable long-term monitoring of physiological signals, the resistance of the hydrogels to water loss can be enhanced by adding humectants in the network32. While it is a difficult task to accomplish mass production of the aforementioned dry electrodes or wet electrodes at an affordable price, they are meant to be used widely for physiological electrode detection as a replacement for commercial electrodes. This is due to the high cost and the need for sophisticated equipment for traditional MEMS-based manufacturing techniques. In this context, the continuous advancement in printing electronics with large-scale manufacturing methods, such as screen printing, offers a way to make highly adhesive flexible electrodes more widely available34.Fortunately, the rapid development of electronics based on flexible materials and structures offers new options for high-fidelity recording of bioelectrical signals. Flexible electron-based myoelectric electrodes have good compliance and biocompatibility and can build seamless and conformal skin-electrode interfaces, and some electrodes can also be stretched to a certain degree–1 adhesion to porcine skin, ultrastretchability to 960%, a skin-matched modulus of approximately 10 kPa, and a much lower skin interface impedance than commercial electrodes (due to the abundance of free ions in the gel). More importantly, this patch withstands a high strain of 80% without mechanical delamination between any of the layers or water loss during one week of EMG monitoring without deteriorating the signal quality (due to the introduction of glycerol in the gel network). To demonstrate its practical utility in the detection of EMG signals on subchinococcygeal and subglottic muscle groups, the patch was integrated with a 16 analog channel readout circuit and a WiFi module in an ear-mounted wearable device for the classification of swallowing activity. Using a deep learning algorithm, eleven different swallowing actions are recognized and categorized with 80% accuracy. This work holds promise for screening and treatment of dysphagia and offers a viable and affordable method for remote rehabilitation.In this work, we propose a stretchable and durable HD-sEMG electrode array patch to be worn on complicated skin interfaces, as shown in Fig. 2. Finally, the patch was trimmed into a v-shape that fit snugly against the submandibular to increase its wearability and obtain high-quality sEMG signals for swallowing activity recognition.Figure 35. Contact with positively or negatively charged objects causes charge redistribution on the gel surface. As a result, the charges in the gel flow to the opposing charged objects and create powerful electrostatic interactions27. The strong biological adhesion of the GW-PA gel was investigated with FTIR-ATR spectroscopy, and Fig. –1, the GW-PA gel exhibited this peak, which was attributed to a C-O stretching vibration of glycerin, indicating the presence of glycerin in the gel network. Additionally, the high viscosity of glycerol promoted strong interfacial interactions, which also enhanced the intermolecular interactions29. More excitingly, due to nonspecific adhesion by the gel, it adhered to a variety of different materials  presented an inverse pattern, i.e., a larger glycerol proportion increased the stretchability of the gel. The maximum stretchability approached 960% at 50% (w/w) glycerol, although it decreased as more glycerol was added. Given that it has the lowest hardness of 10 kPa, which was similar to that of human soft tissue37, and the highest stretchability of 960%, the GW-PA gel with 50% (w/w) glycerol was chosen for construction of the GW-PA-Ag electrode.The mechanical properties of the GW-PA gels were optimized by adjusting the glycerol weight content. As shown in Fig. 2 to 62 J/m2, while the adhesion strength of the shear force increased drastically from 10 kPa to 30 kPa. The increased adhesion seen when increasing the glycerol content was due to the dual effects of high glycerin viscosity and a lower Young’s modulus. The low Young’s modulus allowed the gel to stick to skin with a complicated texture, which increased the contact area and enhanced the adhesion strength. As a result, the gel exhibited strong adhesion and a low Young’s modulus, so it attached firmly to fingers and withstood uniaxial tension with up to 900% strain  acted as an ion conductive layer, which resulted in capacitive coupling of the ionic and electronic currents between the silver electrode and the skin24. As a result, the GW-PA gel generated a much larger interfacial capacitance Cd and a much lower impedance than a commercial Ag/AgCl electrode or a dry Ag electrode. Figure As described previously, the GW-PA gel was polymerized and bonded in situ on the surface of the silver electrode to form the GW-PA-Ag electrode and served as the on-skin sEMG interface. Figure sEMG signals from the swallowing muscles are often smaller than those from larger muscles, including those in the forearm. This poses a challenge for the acquisition of swallowing sEMG signals, as the sEMG signals may be drowned in noise when sEMG is used for activities with low muscle activity. We evaluated the performance of the GW-PA-Ag electrode by recording sEMG signals for different levels of submandibular force, since either mouth opening or swallowing requires the use of the targeted muscles in the submandibular region. As shown in Fig. 38. For this reason, we evaluated the robustness of the GW-PA-Ag electrode by examining the signal quality for motion artifacts, which were manually created by cyclically stretching and compressing the skin around the electrode when performing a grip strength exercise of 50% MVC. As shown in Fig. A good sEMG electrode should recognize motion artifacts, as they are the main sources of interference when monitoring muscle activity signals on the body surface. Conventional sEMG electrodes encounter relative slippage between the skin and electrodes during human muscle action as a result of the stress mismatch. This disrupts the charge distribution at the interface and consequently results in motion artifacts40. As shown in Fig. 4c). Among the three electrodes, the electrode with 50% glycerol had the highest SNR of 28 ± 1.1 dB lasting for 7 days, while the electrode with 0% glycerol had the highest SNR of 38 ± 0.5 dB on Day 0. This was because the latter had the most water in its network, which enabled faster ion migration and, consequently, gave the strongest sEMG signal. However, since it was nearly totally dried after one day and was unable to collect the sEMG signals, the scenario did not endure for more than one day. As a result, the sEMG SNR dropped significantly to 0 dB. This also occurred for the electrode with 25% glycerol. In addition, in decreasing or increasing the humidity to 40% or 80%, respectively, similar drying patterns were observed for all of the electrodes with variations of no more than 5%  cells that were in close contact with the on-skin electrodes. For this test, human HaCaTs were cultured directly on the materials for 24 h or 72 h. The cell survival rates are shown in Fig. 41, the subchinococcygeal and subglottic muscle groups are the active muscles that generate swallowing action, and their sEMG signals can, to some extent, reflect the ease of hyoid elevation movements42. To precisely cover these areas for submandibular sEMG measurements, a single electrode was engineered into a patch with a patterned array of 24 electrodes. Figure According to previous studies on the correlation between sEMG activities and swallowing43. The crosstalk rate represents the level of contamination of the tested muscle’s sEMG signal by adjacent muscles. In the test, two electrodes were fixed upon the flexor digitorum longus (FPL) muscle, while another two pairs of electrodes were placed on top of the flexor digitorum longus (FDS) and flexor digitorum ulnaris (FCU) muscles. By comparing the sEMG amplitude for the FDS or FCU to that for the FPL, it was possible to determine the crosstalk rate because only the FPL was activated when the thumb moved. Figure The electrode size is an important parameter allowing an HD-sEMG patch to obtain accurate, high-quality sEMG signals. This was evaluated by placing a pair of GW-PA-Ag electrodes on the participant’s forearm and testing their signal quality as a function of electrode size , as shown in Fig. 2/Hz of industrial frequency interference at 50 Hz. However, these interferences were successfully removed by denoising  FPC cable Fig. . The rea45. The EMG signals for the open mouth and cough were distinctly different from those for normal swallowing movements in terms of the amplitudes and durations. Meanwhile, the sEMG energy maps for swallowing with the three head positions revealed that when swallowing normally, the muscle activities on both sides of the lower jaw were nearly symmetrical, whereas when the head was turned to one side, the muscle activities on this side were inhibited  and bananas (5 g) were used to represent swallowing various types of foods. The viscous liquids were obtained by varying the proportion of edible cellulose dissolved in water. In this context, the first seven swallowing actions involved the physical difficulties of ingesting foods with various volumes or formations, which are usually employed as indicators of the severity of dysphagia. The remaining four of the eleven were chosen to simulate abnormal swallowing conditions, in which the left cephalic or right cephalic position was achieved by turning the head to the side at a 45° angle and swallowing 5 ml of water. Figure Five volunteers aged 22 to 27 and without recorded medical diseases were asked to perform the eleven swallowing actions for 2 s and repeat them 10 times. To position the electrode, the participant was required to sit comfortably to keep the neck relaxed and avoid interference from other muscle activities. Then, the electrode was placed on the lower jaw so that its edge coincided with the mandibular medial edge. Note that the target skin was cleaned carefully with alcohol before each measurement. A total of 550 groups of data were gathered, and 60% of them were used to train the model, 20% to validate it, and the remaining 20% to test it. Figure −1, an ultrastretchability of 1000%, a skin-matched modulus of approximately 10 kPa, and long-term stability of the SNR measuring no less than 30 dB over one week in an open environment. Compared to a commercial Ag/AgCl electrode, this electrode had a much lower contact impedance in the sEMG frequency range of 1 to 1000 Hz and half the noise baseline with large skin deformations. In the practical demonstration, this patch was applied with a trained CNN model for the recognition of eleven swallowing activities, three of which involved actual digestion of food, while four imitated abnormal swallowing motions. A high average classification accuracy of 80% was achieved, indicating the potential of this system for use in diagnosing dysphagia.In summary, this work developed a stretchable, durable HD-sEMG patch that was used for the classification of swallowing activities on complex epidermal surfaces. The patch was prepared with scalable, layer-by-layer printing and lamination to fabricate an array of GW-PA-Ag electrodes sandwiched between two thin PU films, which was used as a human‒machine interface to measure the sEMG data from targeted muscles. The fabricated electrode exhibited an excellent electrophysiological interface with an adhesion strength of 22.8 N mp-styrenesulfonate  was purchased from Sigma‒Aldrich, Shanghai, China. Dimethylaminoethylacrylate quaternized ammonium  and 3-(methacryloylamino)propyltrimethylammonium chloride  were purchased from J&K Chemical Ltd. Glycerol (≥99%) and α-ketoglutaric acid (α-keto) were purchased from Sinopharm Chemical Reagent Co. Ltd. Stretchable conductive silver pastes (EM-1046) were purchased from Guang Zhou City Silver Well Trading Co., Ltd. All reagents were of analytical grade and were used as received. Deionized water  was used in all experiments.Sodium co-DMAEA-Q) hydrogel was prepared via free radical polymerization. First, the anionic monomer (NaSS), cationic monomer (DMAEA-Q), and photoinitiator (α-keto) were mixed with the water-glycerol binary solvent to form a pregel solution. The total ionic monomer concentration (CM) and the molar fraction of the anionic monomer were fixed at 2.3 M and 0.487, respectively, and the mole fractions of the initiators were 0.10 mol% relative to the CM. Different GW-PA gels were prepared by changing the mass fraction of glycerol in the binary solvent. The gels used for testing were prepared as follows: the pregel solution was injected into a reaction cell consisting of a pair of glass plate walls and a silicone spacer (10 cm × 10 cm × 1 mm). The gel was then polymerized for 5 h with an ultraviolet lamp  at room temperature. The hydrogels for sEMG measurements were polymerized in situ on the surface of the silver electrode with a layered array of patterned silicone film, and the rest of the process was the same as above.The P to measure the tensile strength. Before the tests, the samples were cut into dumbbell shapes (gauge length l (12 mm) × width w (2 mm) × thickness t (1 mm)). The tests were performed at ambient temperature with a stretching velocity of 100 mm min−1. The Young’s modulus, E, was calculated from the initial slope of the stress−strain curve at a strain within 10%. Each type of experiment was performed in triplicate.The interfacial toughness of the GW-PA gel was measured against porcine skin with the standard 180-degree peel protocol (ASTM F2256). It was determined by multiplying the averaged platform forces at the stable peel state and dividing it by the adhesion width. To measure the shear strength, the adhered samples were tested with the standard lap-shear test (ASTMF2255). The shear strength was determined by dividing the maximum force by the adhesion area. All samples were prepared with dimensions of 50 mm × 15 mm × 1 mm (length × width × thickness). The hydrogels were bonded with poly (PET) films using a cyanoacrylate adhesive to prevent the gel from being stretched during adhesion testing. Before the tests, a 5 N preload was placed on the sample for 60 s. All tests were performed at ambient temperature with a universal testing machine  with a constant tensile speed of 50 mm/minThe electrode-skin impedance was measured with an electrochemical analyzer  with a three-electrode system. During the measurement, the reference and counter electrodes were placed at distances of 3 cm and 6 cm from the working electrode, i.e., the fabricated GW-PA-Ag electrode. To cover the range of the sEMG signals, the scanning frequency range was set from 1 to 1000 Hz. For the long-term stability tests, the hydrogel electrodes were stored in a programmable constant temperature and humidity chamber with a temperature of 25 °C and a humidity of 60% before the experiments were performed. The single-electrode sEMG data acquisition experiment was performed with a commercial Noraxon wireless sEMG acquisition system. The signal-to-noise ratio (SNR) was calculated as follows:3 cells/well, and the DMEM was supplemented with 10 vol % fetal bovine serum (FBS). A CCK-8 assay was performed to investigate the 24- or 72-h viability of the cells quantitatively. In addition, the absorbance was measured at 450 nm with a microplate reader .The GW-PA and GW-PA-Ag electrodes were cocultured with human immortalized keratinocyte (HACAT) cells to evaluate their cytotoxicities. Pristine Dulbecco’s modified Eagle’s medium (DMEM) served as the negative control. After sterilization via UV irradiation, the samples were added to the medium at a ratio of 10 mg/ml and extracted for 24 h. HACATs were seeded in 96-well plates at a density of 8 × 10Ethics approval and informed consent were obtained from all participants, and the protocol was approved by the Maoming People’s Hospital Department of Ethics Committee (PJ2020MI-K179-01). The study was carried out under the Helsinki Declaration.supporting informationsupporting video S1supporting video S2


Cerebrospinal fluid (CSF) Polymerase chain reaction (PCR) panel is routinely used for etiologic diagnosis of meningitis. While its role in this setting has been proven, there is no data on its benefits on duration of antibiotic use and hospital length of stay.Retrospective cohort study was performed to compare meningitis patients who received the CSF PCR panel versus those who did not in a large database of HCA hospitals across the United States.Data was collected from January 2014 to May 2022 on all patients diagnosed with meningitis with a CSF WBC greater than 10. Using logistic regression and negative binomial regression, the study compared utility of CSF PCR panel use versus non-use with regards to length of stay, antibiotic duration, resolution of fever and leukocytosis, and hospital readmission rate.Demographics by Panel UseA total of 4895 patients were included in the study, of which 379 received the CSF PCR Meningitis Panel.A total of 4895 distinct patient encounters were included, of whom 379 patients had the CSF PCR panel. CSF PCR use was associated with a significant decrease in readmission rates within 30 days  and had decreased log count days of antibiotic duration  with a mean duration of antibiotic use of 73.91 hours in CSF PCR panel patients versus 82.05 hours. Panel use was not significantly associated with length of stay . The likelihood of fever resolution and leukocytosis resolution was the same for those who received the CSF PCR Panel versus those who did not.Antimicrobial Medications Received by Panel UseAntimicrobial Duration and Clinical Presentation by Panel UseThere was a decrease in antibiotic use of 73.91 hours in CSF PCR panel patients versus 82.05 hours. There was also a significant decrease in hospital readmission rates within 30 daysCSF PCR panel benefits included reduced antibiotic duration of use and reduced 30-day readmission rates. Fever resolution and leukocytosis or leukopenia resolution was not impacted by CSF PCR panel use.All Authors: No reported disclosures