To synthesise the evidence for the effectiveness of inpatient rehabilitation treatment ingredients (versus any comparison) on functioning, quality of life, length of stay, discharge destination, and mortality among older adults with an unplanned hospital admission.2).A systematic search of Cochrane Library, MEDLINE, Embase, PsychInfo, PEDro, BASE, and OpenGrey for published and unpublished systematic reviews of inpatient rehabilitation interventions for older adults following an unplanned admission to hospital from database inception to December 2020. Duplicate screening for eligibility, quality assessment, and data extraction including extraction of treatment components and their respective ingredients employing the Treatment Theory framework. Random effects meta-analyses were completed overall and by treatment ingredient. Statistical heterogeneity was assessed with the inconsistency-value (In = 12) of moderate to low quality, including 44 non-overlapping relevant RCTs were included. When incorporated in a rehabilitation intervention, there was a large effect of endurance exercise, early intervention and shaping knowledge on walking endurance after the inpatient stay versus comparison. Early intervention, repeated practice activities, goals and planning, increased medical care and/or discharge planning increased the likelihood of discharge home versus comparison. The evidence for activities of daily living (ADL) was conflicting. Rehabilitation interventions were not effective for functional mobility, strength, or quality of life, or reduce length of stay or mortality. Therefore, we did not explore the potential role of treatment ingredients for these outcomes.Systematic reviews (endurance exercise was effective for endurance in older adults with chronic obstructive pulmonary disease, and early intervention was effective for endurance for those with hip fracture. Future research should determine whether the effectiveness of these treatment ingredients observed in subgroups, are generalisable to older adults more broadly. There is a need for more transparent reporting of intervention components and ingredients according to established frameworks to enable future synthesis and/or replication.Benefits observed were often for subgroups of the older adult population e.g., CRD42018114323.PROSPERO Registration The online version contains supplementary material available at 10.1186/s12877-022-03169-2. The world’s population is ageing, reflecting advances in economic and social development, public health, sanitation, and medicine . Althougset of measures aimed at individuals who have experienced or are likely to experience disability to assist them in achieving and maintaining optimal functioning  when innments.” . Treatmeeatment” –12. TreaThere is a plethora of studies evidencing the effectiveness of inpatient rehabilitation for older adults admitted to hospital with an unplanned episode of injury or illness, summarised in systematic reviews and meta-analyses –17. ThisIt would be of value to both clinicians and researchers to determine which treatment ingredient(s) contribute to the effectiveness of rehabilitation . We propThe aims of this overview review were to inform evidence-based inpatient rehabilitation for older adults following an unplanned hospital admission, and to identify gaps in the evidence to inform future research. More specifically, the primary objective was to synthesise the evidence for the effectiveness of inpatient rehabilitation treatment ingredients (versus any comparison) on functioning  among older adults with an unplanned hospital admission. Secondary objectives included synthesizing the evidence for additional outcomes of quality of life, length of stay, discharge destination, and mortality.We registered the protocol on the international prospective register of systematic reviews (PROSPERO: CRD42018114323). We reported this review in adherence to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) statement [Eligibility criteria are outlined in Table We developed structured search strategies, in consultation with a librarian using thesaurus terms for intervention, setting and study design for each database  and free text, targeting the “title” and “abstract” fields . A third researcher resolved any discrepancies. We quantified inter-rater reliability using Cohen’s Kappa statistic . We avoiWe assessed the methodological quality of each included review in duplicate using the Assessment of Multiple Systematic Reviews (AMSTAR 2) tool  . AMSTAR We extracted data onto Microsoft Excel table templates defined a priori in duplicate . A third researcher resolved any discrepancies. We contacted authors to supplement missing or incomplete data.Organ Functions (example more specific treatment ingredient: strengthening exercise) [Skills and Habits (example more specific treatment ingredient: repeated practice of activities +/− increasing demands) [Changing Behaviour  [, increased medical care), we extracted them under Other Components. All treatment ingredients cited were assigned to a component in this review. For our outcomes, we extracted mean and standard deviation in each treatment arm for continuous outcome measures and proportions for categorical outcomes after inpatient rehabilitation and on longest follow-up (up to 1 year). We contacted all authors who presented data as medians, ranges, or 95% confidence intervals for means and standard deviations. If no response was received, we converted data presented as medians and ranges to means and standard deviations using methods as described by Hozo et al. [We extracted the following data items for the systematic reviews: review author, review year, population, intervention, comparators, outcome, number of studies eligible for the current overview, number of patients from eligibility studies. We extracted the following data items for eligible RCTs within the systematic reviews: RCT author, RCT year, country, sample size (intervention and comparator), characteristics of the population where available -age, gender, target group, and preadmission residence, comparison/s, interventions, outcomes and follow-up relevant to the current overview. For the interventions, we extracted three main treatment components and their more specific and measurable treatment ingredients specified by Treatment Theory –12. Compowledge) . Where to et al. . We convo et al.  and subsAll systematic reviews met the eligibility criteria for inclusion; however, 1) not all RCTs within reviews were relevant, and 2) there was considerable primary RCT overlap between reviews. Therefore, we re-analysed the data by performing random-effects meta-analyses within the subgroup of relevant RCTs for each outcome across the systematic reviews . We estiendurance exercise [2 and followed the Cochrane convention of 0–40% heterogeneity as may not be important, 30–60% as moderate, 50–90% as substantial, and 75–100% as considerable heterogeneity [We stratified meta-analyses which indicated the effectiveness of interventions on outcomes by individual treatment ingredients, e.g. exercise . For metexercise . We asseogeneity . Where aogeneity  and stran = 104), intervention (n = 9), outcome (n = 5), study design (n = 17), setting (n = 21). Cohen’s Kappa statistic following full text review was 0.73 indicating substantial agreement between assessors. Following generation of a citation matrix ordered by publication date, we excluded a further 10 reviews [We included 12 systematic reviews in this overview review. Initial searches identified 2677 systematic reviews, of which 583 were duplicates. On the title and abstract screening, a further 1916 were excluded. Of the 178 reviews assessed at full text screening, 155 were ineligible for the following reasons: population ( reviews –39 whichn = 12) [n = 11) [n = 11) [n = 11) [n = 9) [n = 12) [n = 11) [n = 11) [n = 5) [The results of the quality assessment are presented in Table n = 12) , 40–49, [n = 11) , 45–49, [n = 11) –49, risk[n = 11) , 45–49,  [n = 9) , 48, 49,[n = 12) , 40–49. [n = 11) , 44–49, [n = 11) , 40–49 a [n = 5) , 46, 47.n = 5) [n = 3) [n = 2) [n = 1) [n = 1) [n = 11) [n = 4) [n = 9) [n = 2) [n = 7) [The 12 systematic reviews included 41 unique RCTs and 10,444 older adults with an unplanned hospital admission relevant to this overview (mean (min - max) sample size per RCT: 261 12–1531)) (Table 1) (Tablen = 5) , 45, 48, [n = 3) , 49, wit [n = 2) , 47, ort [n = 1) . Outcome[n = 11) , 48, 49; [n = 4) , 48, 49; [n = 9) , 45–48;  [n = 7) , 48, 49.Organ Functions, treatment ingredients included: endurance exercise (n = 13), strengthening (n = 12), energy applied to soft tissue (n = 7), and/or breathing related exercises/training (n = 6). For component 2 Skills and Habits, treatment ingredients included: repeated practice activities (n = 15), functions (n = 8), and/or ‘exercise rehabilitation’ (n = 6). For component 3 Changing Behaviour, treatment ingredients included: shaping knowledge (n = 16), feedback and monitoring (n = 14), goals and planning (n = 11), antecedents (n = 12), natural consequences (n = 5), social support (n = 2), and/or comparison of behaviour (n = 1). For Other Components, treatment ingredients included: increased medical care for e.g., avoidance of complications and/or pain management (n = 14), early intervention (n = 12), team meetings and care planning (n = 11), discharge planning (n = 9), nutritional intervention (n = 8), home visits during inpatient stay (n = 5), and/or cognitive orientation exercise (n = 2).Characteristics (as well as their treatment components and more specific and measurable ingredients) of the 44 RCTs included in the 12 systematic reviews are detailed in Supplementary Files n = 42, 95.5%) identified from the systematic reviews. Physiotherapy/occupational therapy was a core component of usual care for 21 RCTs (50.0%), provided following a physician referral for 5 RCTs (11.9%), not a component of usual care for 13 RCTs (30.9%), or not specified for 1 RCT (2.4%). Two RCTs (4.8%) included education and usual care  as the comparator. The comparator was an alternative intervention - delayed ambulation or delayed weight bearing for 2 RCTs (4.6%).The comparator was usual care for the majority of RCTs , health-related quality of life, length of stay, discharge destination, and mortality Table . Details2 = 94.40; Change score: 3 RCTs including 139 participants; Log OR = 1.23, 95% CI: 0.68, 1.78. I2 = 54.96) supported by results of RCTs from one systematic review not included in the meta-analysis [endurance exercise  and shaping knowledge  had a large effect, while early intervention had a moderate effect  on walking endurance versus comparison after inpatient stay.Rehabilitation had a large effect on walking endurance versus comparison after inpatient stay . One systematic review reported on one RCT which noted no effect at follow-up [strengthening exercise or repeated exercise rehabilitation did not increase walking speed.Rehabilitation had a small effect on walking speed versus comparison after inpatient stay . The effect was similar but non-significant for ADL change score . The effect was not sustained at 1–12 month follow-up . The absence of an effect was supported by results of RCTs from six systematic reviews not included in the meta-analyses [p = 0.01). For estimates of total ADL after inpatient stay, the total effect of rehabilitation interventions adjusted for age, target population, RCT geography, and publication year was not significant (p = 0.12) in meta-regression.Rehabilitation had a small effect on ADL versus comparison after inpatient stay . There was no effect of endurance exercise, strengthening exercise, repeated practice activities, repeated exercise rehabilitation, goals and planning, feedback and monitoring, shaping knowledge, antecedents, increased medical care, nutritional intervention, or early intervention, on ADL versus comparison.When included in a rehabilitation intervention, the treatment ingredient Rehabilitation did not improve functional mobility or lower limb strength versus comparison after inpatient stay or functional mobility at follow-up evidenced by meta-analysis. Two systematic reviews identified RCTs reporting a between group difference in functional mobility when measured with the Physical Performance and Mobility Examination after inpatient rehabilitation  or the S2 = 45.95) and at 3–12-month follow-up . When included in a rehabilitation intervention, the treatment ingredients repeated practice activities , goals and planning , increased medical care  early intervention , and discharge planning  increased the odds of living at home versus comparison after inpatient rehabilitation. When included in a rehabilitation intervention, the rehabilitation ingredients repeated exercise rehabilitation, antecedents, team meetings and care planning, and nutritional intervention had no effect on the odds of living at home after the period of inpatient rehabilitation. There was no evidence of small study sample bias. For total estimates after inpatient stay, the total effect of age, target population, RCT geography, and publication year was not significant (p = 0.14) in meta-regression suggesting these variables do not explain the observed heterogeneity. Subsequent meta-analysis was not carried out.Rehabilitation was effective at increasing the odds of living at home versus comparison after inpatient rehabilitation , or on 12-month follow-up . Three systematic reviews reported on RCTs not incorporated in the meta-analysis which favoured rehabilitation intervention versus comparison after inpatient stay [Rehabilitation did not increase health-related quality of life versus comparison after inpatient stay ; however, evidence was detected for small study sample bias (p < 0.001). For estimates of length of stay, the total effect of rehabilitation interventions adjusted for age, target population, RCT geography, and publication year was significant (p < 0.001) in meta-regression. A subsequent stratified meta-analysis by RCT geography was conducted. The absence of an effect of rehabilitation on length of stay persisted across regions with substantial heterogeneity for Australia (I2 = 86.26) and Europe (I2 = 76.47), and heterogeneity which may not be important for the United States of America (I2 = 18.10%).Rehabilitation did not reduce the length of stay versus comparison after inpatient stay  or 1–12 month follow-up , further supported by an RCT from 1 systematic review not included in the meta-analysis [Rehabilitation did not reduce mortality among older adults with unplanned hospital admission versus comparison after inpatient rehabilitation , early intervention (predominantly from RCTs of older adults after hip fracture) and shaping knowledge (exclusively from RCTs of older adults with COPD) on walking endurance after the inpatient stay versus comparison. We also reported beneficial effects of early intervention, repeated practice activities, goals and planning, increased medical care and/or discharge planning on discharge home. The evidence for effectiveness of treatment ingredients that improve ADL was conflicting. Rehabilitation interventions were not found to be effective for functional mobility, strength, or quality of life, or reduce length of stay or mortality. Therefore, we did not explore the potential role of treatment ingredients for these outcomes.We identified 12 systematic reviews of moderate to low quality which included 44 unique RCTs relevant to the current overview. When incorporated in a rehabilitation intervention, we report a large effect of the treatment ingredients Given ceaseless drives to decrease inpatient lengths of stay, it is important for clinicians to preferentially select treatment ingredients most likely to improve outcomes at discharge . Howeverendurance exercise had a positive effect on walking endurance. This is important as objective quantitative data indicate adults over the age of 65 years take a median of just 468 steps per day during their inpatient stay (no difference by admitting reason or illness severity) [endurance exercise; all included patients with COPD exacerbations and these favoured the intervention group. The treatment ingredient was comprised of pedal ergometry daily with increased resistance [The treatment ingredient everity) . Given teverity) , 52. Thrsistance , treadmisistance , or walksistance . The larsistance , 57 as wsistance  during tearly intervention as a treatment ingredient to be effective at increasing endurance and the likelihood of a home discharge when incorporated into inpatient rehabilitation for older adults after an unplanned hospital admission. This is unsurprising given potential for rehabilitation to mitigate hospital-associated deconditioning [n = 7, 78%) with early intervention defined by mobilisation from bed within the first two days of surgery. This evidence has informed wide acceptance older adults with hip fracture should receive early mobilisation after surgery with early mobilisation a key performance indicator in national audits [With bedrest, muscle strength is lost rapidly at a rate of 5% per day [itioning  and previtioning . Most RCl audits .goals and planning, repeated practice of activities, increased medical care, and/or discharge planning versus comparison. More specific detail for these treatment ingredients was limited. For example, repeated practice of activities often reflected ‘ADL training’ with no further detail related to the frequency, duration, or type of activities. One RCT specified transfers were practiced twice daily for 30 minutes [A discharge destination of home was more likely among participants who received interventions which incorporated treatment ingredients of  minutes . Another minutes .The findings of the current overview are consistent with those of the underlying systematic reviews which conclude that inpatient rehabilitation can improve functioning , 48, 49 repeated exercise rehabilitation where exercise rehabilitation was mentioned but not detailed, shaping knowledge where education was specified but not detailed, or increased medical care where examples of what ‘increased care’ may entail were provided but not explicitly measured. This may have led to an underestimation of more specific treatment ingredients. Third, we noted moderate to substantial heterogeneity for several outcomes overall and by treatment ingredient. It was not possible to complete meta-regression across all analyses due to the low number of RCTs [There are several limitations to this overview review. First, we needed to make two protocol changes a) outcome data were extracted at ‘end of inpatient rehabilitation’, which was a change from our protocol which specified ‘on discharge’ due to lack of clarity in published data, and b) we excluded systematic reviews exclusively addressing post-stroke rehabilitation at full text selection due to their often impairment focus  that would not be potentially translatable to other admitting diagnoses. Second, where intervention detail was limited, we termed treatment ingredients such as  of RCTs . For eac of RCTs . Nonethe of RCTs . Fifth,  of RCTs . Sixth,  of RCTs . Finally of RCTs .endurance exercise on endurance was reflective of three RCTs of older adults with an unplanned admission due to an exacerbation of COPD while the findings from early intervention predominantly reflected older adults with hip fracture. These treatment ingredients should be prioritised for implementation for these patient groups. It may be reasonable to generalise the recommendations to similar groups of older adults with an unplanned admission to hospital. For example, early intervention may be generalised to other non-hip fragility fractures [endurance exercise to patients admitted with exacerbations of other chronic lung diseases [Endurance exercise is a key component of most cardiac rehabilitation programmes offered to older adults with heart failure [early intervention (mobilisation) is recommended for older adults admitted with an exacerbation of heart failure [The effect of ractures , and enddiseases . Whether failure . However failure  a walkinIt was possible to assign treatment ingredients to inpatient rehabilitation interventions. However, for many, the interventions were poorly described limiting exploration of more specific treatment ingredients and/or the ingredient dose. Moreover, the description of usual care comparator groups was limited and those inclusive of rehabilitation could attenuate the between group comparisons for effectiveness. These are not new findings with several previous systematic reviews highlighting the challenges in synthesizing the evidence for rehabilitation interventions , 16. Thiearly intervention and endurance exercise were effective at improving endurance, and early intervention, goals and planning, repeated practice of activities, increased medical care, and/or discharge planning effectively increased the likelihood of discharge to home for older adults following an unplanned admission to hospital. Benefits observed were often for subgroups of the older adult population e.g., endurance exercise was effective for endurance in older adults with COPD, and early intervention was effective for endurance for those with hip fracture. Future research should seek to determine whether the benefits observed from these treatment ingredients are generalisable to older adults more broadly. Further, there is a need for more transparent reporting of rehabilitation intervention treatment ingredients to enable future synthesis and/or replication. Finally, the challenge of making meaningful change during a short period of inpatient rehabilitation emphasizes the importance of comprehensive post-discharge rehabilitation.The designation of treatment ingredients to interventions was challenging due to a paucity of detail specified by published interventions. Despite this, we reported the treatment ingredients Additional file 1: Supplementary File 1. Search strategies. Search strategies for electronic databases of published and unpublished evidence.Additional file 2: Supplementary File 2. Citation matrix. Citation matrix detailing the identification of unique (non-overlapping) randomized controlled trials from systematic reviews included in this overview review.Additional file 3: Supplementary File 3. Characteristics of randomized controlled trials. Characteristics of eligible randomized controlled trials identified from systematic reviews included in this overview review.Additional file 4: Supplementary File 4. Treatment ingredients. Treatment ingredients employed by eligible randomized controlled trials identified from systematic reviews included in this overview review.Additional file 5: Supplementary File 5. Meta-analyses results. Results of meta-analyses (forest plot) of randomized controlled trials identified from systematic reviews included in this overview review for functioning, quality of life, length of stay, discharge destination and mortality. Each meta-analysis is accompanied by a table which describes the characteristics of each randomized controlled trial included in each meta-analysis.Additional file 6: Supplementary File 6. Treatment ingredient meta-analyses results. Results of meta-analyses (forest plot) of randomized controlled trials identified from systematic reviews included in this overview review by treatment ingredient.Additional file 7: Supplementary File 7. Narrative results. Results of randomized controlled trials identified from systematic reviews included in this overview review which were not incorporated into the meta-analyses and reasons why they were not incorporated.


Hepatocellular carcinoma (HCC) is one of the most frequent malignancies. Alpha-fetoprotein (AFP) has some limitations in diagnosing early HCC. Recently, long noncoding RNAs (lncRNAs) showed great potential as tumor diagnostic biomarkers, and lnc-MyD88 was previously identified as a carcinogen in HCC. Here, we explored its diagnostic value as a plasma biomarker.Quantitative real-time PCR was adopted to detect lnc-MyD88 expression in plasma samples of 98 HCC patients, 52 liver cirrhosis (LC) patients, and 105 healthy people. The correlation between lnc-MyD88 and clinicopathological factors was analyzed through chi-square test. The receiver operating characteristic (ROC) curve was used to analyze the sensitivity, specificity, Youden index, and area under the curve (AUC) of lnc-MyD88 and AFP alone and in combination for the diagnosis of HCC. The relationship between MyD88 and immune infiltration was analyzed by single sample gene set enrichment analysis (ssGSEA) algorithm.Lnc-MyD88 was highly expressed in plasma samples of HCC and hepatitis B virus (HBV)-associated HCC patients. Lnc-MyD88 had better diagnostic value than AFP in HCC patients using healthy people or LC patients as control . The multivariate analysis showed that lnc-MyD88 had great diagnostic value for distinguishing HCC from LC and healthy people. Lnc-MyD88 had no correlation with AFP. Lnc-MyD88 and AFP were independent diagnostic factors for HBV-associated HCC. The AUC, sensitivity, and Youden index of the combined diagnosis of lnc-MyD88 and AFP combined were higher than those of lnc-MyD88 and AFP alone. The ROC curve of lnc-MyD88 for the diagnosis of AFP-negative HCC was plotted with a sensitivity of 80.95%, a specificity of 79.59%, and an AUC value of 0.812 using healthy people as control. The ROC curve also presented its great diagnostic value using LC patients as control . Lnc-MyD88 expression was correlated with microvascular invasion in HBV-associated HCC patients. MyD88 was positively correlated with infiltrating immune cells and immune-related genes.The high expression of plasma lnc-MyD88 in HCC is distinct and could be utilized as a promising diagnostic biomarker. Lnc-MyD88 had great diagnostic value for HBV-associated HCC and AFP-negative HCC, and it had higher efficacy in combination with AFP. Hepatocellular carcinoma (HCC) is ranked as the sixth most common cancer and also ranked third on the leading causes of cancer-induced mortality across the globe . What isAt present, the diagnosis of HCC mainly depends on alpha-fetoprotein (AFP) and imaging technology, but in the early diagnosis of HCC, the missed diagnosis rate (AFP <20 ng/ml) is as high as 30%–40% . Even inNoncoding RNAs (ncRNAs) have been attracting the attention of many scholars in recent years. A large amount of research data indicate that ncRNAs partake in cell proliferation, differentiation, apoptosis, migration, and invasion –12. NcRNBecause lncRNA has a regulatory role, it may have significant implications in a variety of diseases through complex mechanisms. Therefore, looking for lncRNAs related to HCC may be very important for discovering and treating HCC. In addition, past literature has elucidated that certain lncRNAs are associated with the occurrence and development of cancer and can be detected in plasma . For exavia Toll-like receptor 4 (TLR4)-MyD88-Nuclear Factor Kappa-beta (NF-κB) pathway. MyD88 had been considered as a treatment and prognostic evaluation target for HCC  was defined as a carcinogenic gene that promoted the occurrence and development of HCC  for HCC –22. Prev for HCC . In the The present research was conducted in collaboration with the Affiliated Changzhou No. 2 People’s Hospital of Nanjing Medical University and Changzhou No. 3 People’s Hospital. In total, plasma samples of 98 patients with HCC, 52 patients with liver cirrhosis (LC), and 105 healthy subjects were acquired from the aforementioned hospitals. Prior to surgery, no treatment including chemotherapy, radiotherapy, and antitumor treatment using biological products was performed on patients. According to relevant guidelines, the diagnosis of LC or HCC is based on typical morphological findings and pathological observations using CT or ultrasound , 25. TumVenous blood samples were collected from participants within 2 h, and they were centrifuged at 4,000 rpm for 10 min; this was to detect the level of plasma lnc-MyD88. The supernatant plasma was centrifuged at 12,000 rpm for a further 15 min to eliminate any leftover debris. To avoid hemolysis, the entire procedure was meticulously monitored and controlled, and the plasma was kept at −80°C until RNA was extracted.−ΔΔCt method. The primer sequences were synthesized by RiboBio . Lnc-MyD88 primer sequences: forward, 5′-TACCACAGGGGCTGGAGTTAT-3′; reverse, 5′-AGTCTGTCCCCCTCCAGGTT-3′; β-actin primer sequences: forward, 5′-CGGAGTCAACGGATTTGGTCGTATTGG-3′; reverse, 5′-CCATGGTGTCTGAGCGATGT-3′.Total RNA extraction was achieved through the TRIzol LS reagent  (Lot no. 223306) following the guide supplied by the manufacturer. NanoDrop spectrophotometer was used to assess the quality and quantify RNA. The reverse transcription (RT) reactions took place using the Prime Script™ RT Reagent Kit  (Lot no. AK4802). The 20-μl solution contained five samples of 4-μl Prime Script Buffer Mix, 1 μg of template RNA and 1 μg of Prime Script RT Enzyme Mix I, 1 μl of Oligo dT Primer and RNase-free H2O; it was then incubated firstly at 37°C for 30 min, followed by 85°C for 5 s, and finally 4°C for 60 min. RT-qPCR was completed using a CFX96™ real-time system . The cDNA solution generated was diluted, and 2 μl of this solution was then combined with 12.5 μl of SYBR Premix Ex TaqTM, 0.5 μl of Dye II  (Lot no. 7E480I0), 1 μl forward and reverse primers (10 μM), and 9 μl of nuclease-free water, so that the total volume was 25 μl following the guideline by the manufacturer . Then, the solution was incubated at 95°C for 30 s, after that, 45 cycles of 95°C and 60°C alternating for 5 s and 34 s, respectively. Three repetitions were the minimum standard for each test. The specificity of the RT-qPCR products was examined with the melting curve analysis. The relative gene expression level was normalized by the endogenous control-β-actin, then assessed using the 22) tests were used for count data. Nonparametric tests including Mann–Whitney and Kruskal–Wallis were also utilized to contrast skewed RNA data (2−ΔΔCt) from independent samples. The diagnostic accuracies of the parameters examined were assessed by the receiver operating characteristic (ROC) curve analysis. In addition, the logistic regression analysis was performed in order to recognize the predictor RNA that is correlated with the predisposition of developing HCC. The association between the expression of individual RNA subtype and the occurrence of HCC was established by conducting a univariate analysis. Spearman rank correlation test determined the relationship between the two quantitative nonparametric variables. Data input and statistical analyses were performed using the Statistical Package for the Social Science  version 22.0 software. Significance level was set at the P < 0.05 with a 95% confidence interval (CI). The association between MyD88 and immune cells was analyzed through the ssGSEA algorithm of Gene Set Variation Analysis (GSVA) package (1.34.0) in R software (3.6.3). The immune cells incorporated cytotoxic cells, eosinophils, dendritic cells (DCs), immature DCs (iDCs), activated DCs (aDC), CD8 T cells, B cells, natural killer (NK) CD56bright cells, neutrophils, NK cells, NK CD56dim cells, mast cells, macrophages, T cells, plasmacytoid DCs (pDCs), T helper cells, T effector memory (Tem), T follicular helper (Tfh), T central memory (Tcm), T gamma delta (Tgd), Th1 cells, Th2 cells, Th17 cells, and Treg25. The correlation in respect of the immune-related gene expression and MyD88 expression was analyzed and visualized by employing R software (3.6.3) and ggplot2 (3.3.3) package.The results were illustrated as a ranking of means yielded or percentages. Among the three groups of clinical data, one-way ANOVA was used for measurement data, and chi-square (xP < 0.05) among HCC patients. By comparing the clinicopathological data of 255 individuals in the three groups, we found that the subjects in the three groups had significant differences in Hepatitis B surface antigen (HBsAg), Hepatitis B e-antigen (HBeAg), and Child grades , 52 patients with LC (38 men and 14 women), and 105 healthy subjects (40 men and 65 women). In the LC group, 67.3% of the patients were classified as Child A, 26.9% were classified as Child B, and 5.8% were classified as Child C. Whereas none was scored as Child C, 96.9% were scored as Child A, and 3.1% were scored as Child B , and ALT  had potential to distinguish HCC patients from healthy subjects (P = 0.022), DBIL , AFP , and HBsAg  had potential to distinguish HCC patients from LC patients  could distinguish HCC patients from healthy people, with areas under the curves (AUCs) being 0.776 and 0.725, respectively. The sensitivities were 74.5% and 62.2% at the cutoff values of 1.46 and 6.63, respectively; and the specificities were 80.0% and 92.4%  and low lnc-MyD88 expressed group (n = 49) based on the median value. The result showed that the expression of lnc-MyD88 in the plasma of HCC patients has no significant correlation with various clinicopathological data  , T helper cells , and Tcm . MyD88 had a negative correlation with pDCs .Our previous research determined that lnc-MyD88 expression had a positive correlation with MyD88 expression by utilizing Pearson analysis  . Emergin< 0.001) . The reland CCL18XCL1, CXCL9, CXCL8, CXCL6 CXCL3, CXCL13, CXCL12, CXCL11, CXCL10, CX3CL1, CCL8, CCL7, CCL4, CCL3, CCL28, CCL24, CCL23, CCL22, CCL2,   were positively correlated with the expression of MyD88  were all associated with MyD88 expression  RNA sequencing data were downloaded from TCGA online platform, and the immune-related analysis was conducted by R software. The outcome indicated that MyD88 expression was related to several chemokine genes including HCC is a highly invasive, highly metastatic cancer, and it has a high rate of recurrence and low rate of survival . The maiAt present, many studies have established a range of biomarkers for diagnosing and predicting the prognosis of HCC, including miRNAs, lncRNAs, and some specific genes. Shaker et al.  mentioneIn our prior research, we found that lnc-MyD88 is significantly expressed in liver cancer tissues and can affect the growth and metastasis of HCC by affecting MyD88 . In the Our previous research verified that lnc-MyD88 had a positive correlation with MyD88, and it can influence the progression by mediating MyD88 expression. In the present study, the results showed that MyD88 expression was correlated with the infiltrations of eosinophils, T helper cells, Tcm, and pDCs. Moreover, the immune-related genes including chemokine genes, chemokine receptor genes, immunostimulatory genes, and immunoinhibitory genes all positively correlated with the MyD88 expression. Recent studies mentioned that eosinophils may contribute to the regulation of the tumor microenvironment , 34. YanOur study also had many limitations. The major limitation of this study is that it neither investigated the prognosis of HCC patients nor explored whether lnc-MyD88 in plasma has a guiding role in the prognosis assessment of HCC. Secondly, the diagnostic value of lnc-MyD88 should be proven again by utilizing a new patient population. Especially, we need to enroll more patients with other types of virus infection to validate the diagnostic value of lnc-MyD88. These will be performed in our future work.The distinct expression of lnc-MyD88 is high in HCC and may be used as an innovative but promising diagnostic biomarker. It also had great potential to be an immunotherapeutic target for HCC patients.The original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding authors.The project was approved by the Ethics Committee of the hospital and written informed consent was obtained from each patient who enrolled in the study.ZW and PG contributions to the conception and work design; JJ and SX gathered the clinical samples and literature; ZW and AR created the diagrams and tables; PG and CZ completed the writing and editing of this manuscript. ZW, PG, WS, AR, JJ, SX, CX, CZ, and XQ completed the review and revision of this manuscript. All authors contributed to the article and approved the submitted version.


Augmented reality(AR) is a mixed technology that superimposesthree-dimensional (3D) digital data onto an image of reality. Thistechnology enables users to represent and manipulate 3D chemical structures.In spite of its potential, the use of these tools in chemistry isstill scarce. The aim of this work is to identify the real situationof AR developments and its potential for 3D visualization of molecules.A descriptive analysis of a selection of 143 research publications(extracted from Web of Science between 2018 and 2020) highlights somesignificant AR examples that had been implemented in chemistry, inboth education and research environments. Although the traditional2D screen visualization is still preferred when teaching chemistry,the application of AR in early education has shown potential to facilitatethe understanding and visualization of chemical structures. The increasingconnectivity of the AR technology to web platforms and scientificnetworks should translate into new opportunities for teaching andlearning strategies. For instance, relevant featurestypically covered by stereochemistry, crystallography, or molecularsymmetry are normally approached only in the 2D form. As a result,making viable and graphically accurate reproductions of complex chemicalstructures4 In this sense, descriptive resources ranging from electron microscopyimages to drawings, scale models, or even analog videos6 have been deployed with different success. Nevertheless, these resourcesstill require considerable capacity for visual and spatial thinking.7 Thus, it is common to find presentations of 3Dinformation using 2D projections in chemistry research even thoughthis undermines true representation of its characteristics  is a mixed technology thatsuperimposesa 3D digital image onto reality. Normally, the representation of 3Dmodels using AR is done with a video camera on a computer screen,smartphone, or head-mounted display (HMD) with semitransparent glasses,such as HoloLens, Microsoft.14 This technology seemsto be a powerful tool to facilitate the acquisition of competencein spatial visualization skills16 and could also serveas a source of motivation for learning, although little informationon its impact is available. This review collects and analyzes theuse of AR in chemistry with the aim of triggering the implementationof this appealing technology. 3D representations can favor the processof dissemination of the research results, and these images have animpact on other people interested in understanding the phenomenoninvestigated. The results of research are transferred to society throughdifferent ways, and education constitutes a suitable platform to applyinnovations systematically. In this sense, the use of AR in educationcould serve as an important tool to facilitate the incorporation ofinnovations in a more efficient manner. Specific features of AR, suchas visualization and 3D representation capabilities, and its simplicityare relevant to education because it serves as a motivation and resultedin better results in the assessment.AR can significantly enhance human performance,especially in theeducational field.17 In the first decade of the 21st century, thistool was applied in training within laboratories working effectivelywith virtual instruments.18 There are reviewsof scientific literature that describe how AR technology itself works,20 and the speed of technological evolution means that the analysesof these topics become rapidly outdated.24 For instance, AR research delvesinto procedures to trigger digitally stored information and its superpositionon real images,25 detecting the spatialposition and pose of user’s fingers.19At the beginning of the 1990s, Rosenbergintroduced the term “AR”.26 For instance, the development of new interfaces called augmentedchemistry (AC), where elements can be composed into 3D molecular models,27 or the first experiences with HMD HoloLens glassesfor AR in inorganic chemistry were disclosed.28 Thus, AR could be an interesting solution to the problems relatedto 3D crystal structures in inorganic chemistry, and undergraduatestudents might understand these topics, facing symmetry-related problemssupported by 3D figures.29The first reports of AR regarding the teaching of chemistryappearin the 2010s. Most of these reports were published at conferences,introducing new technological designs and the advantages this interactioncould bring to the education of chemistry.24 In this sense, new hardware developments have also appeared, aswell as some proposals describing haptic prototypes developed to workwith augmented chemical reactions have also appeared.30 AR emerges with other technological advances and new educationalstrategies, such as gamification.31 AR and thesemethodologies seem to change the traditional teachings that introduceeducational and collaborative games in chemistry.32Previousreviews of AR in education by scientific literature articlesdescribed, in general, a positive impact of AR on academic outcomes.23 Indeed, several studies highlight how this toolwith computer-assistedlearning improved the results of low-achieving students, as well astriggered a positive attitude.33 Thereare hardly any studies that describe problems generated after theintroduction of these technologies. In this way, some students likedto manipulate AR by rotating markers, while many of them preferredto interact with physical models in order to get a feeling of physicalcontact.34AR tends to improve students’ motivation.36 In addition, most of the analyses of AR experiences were limitedto studies of a general nature, such as organic chemistry37 or, in contrast, were used in very specifictopics. For instance, various experiences in biochemistry have beendescribed based on these technological developments, such as AR metabolicpathways ARMET,38 which allows studentsto visualize 3D molecular structures and evaluate their own levelsof understanding.The implementation andincorporation of AR technology, especiallyat the university level has been growing at a slow pace.16 Along these lines,it could be a priority to collect the practices of teachers in theirexperimental laboratories.16 Thus, a reviewof the recent research results published in the scientific literatureis essential to provide an overall view of available tools, for implementationin further research and in various educational sectors. The aim ofthis study is to clarify the main findings from research on AR appliedto educational environments in the area of chemistry. This might serveto provide some guidelines to university lecturers for the incorporationof AR technology into chemistry studies. The methodological designof this study focuses on a qualitative descriptive analysis of theimpact of AR.More importantly, there is a lack of reviewsspecifically focusedon scientific literature related to AR and chemistry. This new technologyneeds to be further explored in order to agree on a solid theoreticaland practical approach.The scientificdatabase Web of Science Clarivate Analytics (WoS) provided us withrelevant research material of proven quality with objective indicators.WoS offers collected data with reliable measures of the impact ofthe research, including the amount and type of citations. As for typeof documents, research results published in books, articles, and congresscommunications were selected. The cases with the greatest scientificimpact are classified within the Journal Citation Report (JCR) level.Keywords were entered to locate the relevant documents in thisrepository by applying BIbExcel2016.2.20 software to a set of reiterativeterms in the JCR-classified articles; for example, “AR”,“3 dimension visualization”, “3D”, “stereoscopicinterface”, “molecular visualization software”,“scientific visualization”, and “immersive analytics”.The terms were linked to “chemistry”, and the searchwas run to determine which of these terms extracted the most registersrelated to “advanced atomic/molecular visualization”.The most representative terms generated by this analysis were “AR”and “chemistry”, and the categorization of registerswas considered if the terms appeared in the title, abstract, or keywords.WoS All Databases contain a universe of several million registersfrom all over the world. In the case of the population of descriptors,the terms “chemistry” and “AR” provideda total of 235 registers 1. Such a39 Since then, a significant number of researcharticles on teaching chemistry using AR have appeared, as indicatedby Analysis by date shows that AR had a reduced impactbefore 2017.Because the inventions from a few years ago seem to be obsolete today,new technology is being reconfigured. Actually, there are not manystudies on AR in Chemistry published in high impact journals beforethose dates. This technology might be renewed because of the augmentedcapacity of smartphones’ ability to play audio, take pictures,as well as viewing videos. Smartphones and tablets can now be usedto perform many tasks previously addressed by desktop computers.Contentanalysis was performed on complete documents in the text form downloadedfrom the WoS platform; this database analysis was coded using toolssuch as Aquad7 software. The texts were converted into *.rtf formatand the concepts were counted together, with the frequency of theirappearance and interrelations as the indicator of their strength relatedto the key terms “chemistry” and “augmented”.There were two stages: (1) verification of whether the title, descriptors,and abstract contained visualization experiences and, if so, the nextstep of the analysis continued. (2) Analysis and extraction of findingsin each document, prioritizing results, discussion, and conclusion.Additionally, a confirmation of whether the text reached a JCR categorydocument was accomplished.The analysis of research at the JCR level assumesa guarantee ofhigh scientific quality and relevance in these documents. Gatheringdata for qualitative analysis is difficult yet Aquad7 software allowedus to examine texts accurately. Two external professors have participatedin this part of the study. This analysis includes the evaluation ofthe articles, extracting the related fundamental terms and determiningtheir importance in each text. Thus, we measure its objective relevanceby checking its position in the title, within the list of keywordsor/and within the abstract. The most important findings were alsoquantified and these terms were grouped and quantified, giving themspecial value when they were repeated.Among the limitationsof this research is the fact that there werefew previous studies in chemistry of this nature to draw on, althoughthis could be seen as an opportunity to identify new lines in theliterature and investigation. Even though this research successfullyanalyzes all the relevant elements and achieves its goals, this articleis only a snapshot of existing research on atomic/molecular representation,and the area needs further and deeper investigation. The number ofpublications analyzed was limited in comparison with the broad scopeof AR technology. In this sense, the present work is certainly exploratoryand tries to shed some light on a complex scenario such as AR technology.Accordingly, only reports regarding application and implementationof AR in education, chemistry, have been analyzed.The performed analysis of the contents differentiatedand contrastedthe following themes: (a) two-dimensional atomic/molecular representationsoftware and (b) JCR research on AR.First, this study reviewed the softwareon internet based on asearch of the terms “molecular atomic visualization software”;this search yielded 175 applications that possessed a functionalityrelated to this type of 2D representation on a screen, including mostof the operative systems. The large number of 2D representation programsranges from desktop software to others working from a specific website or cloud software, including open source and licensed commercialsoftware or applications. There are tools for specific areas of Chemistryas well as multidisciplinary areas. This fact makes it difficult tocategorize these computer tools given their variability in time. Hence,we opted for an open sample of software available for the users 2.Supporting Information. The majority of those tools allow the user tobuild and/or analyze the geometry of the molecular systems. In additionto this, some of them  incorporatethe realization of MM (molecular mechanics) calculations, the visualizationof molecular orbitals and molecular surfaces, and the animation ofnormal modes of vibration or the possibility to perform conformationalanalysis. Programs providing chemical structures based on densityfunctional theory or ab initio calculations are also included.40Programs for the 2D visualization of chemical structures on screenscollected in 41 Applications based onWeb browsers such as Iview and Jmol, a Java-based molecular visualizationtool, are also employed to simulate 3D molecules on a 2D computerscreen.8 Moreover, interesting experiencesof 3D model generation with open-source optical structure recognitionapplication (OSRA) together with the simplified molecular input line-entrysystem and Jmol are commonly used. We observed that users handle variouscomputer tools in a complementary way. This combination of softwareis capable of narrowing the performance gap between students withdifferent levels of spatial perception abilities.42However, it is difficult to make a universalclassification, andwe found that users combine several programs simultaneously. For instance,there are 3D molecular simulation experiments that use software suchas PyMOL with Python language programing, and open-source programinglanguage libraries to create large multidimensional vectors and matrices,such as NumPy.43 These APPs have only emerged recently and arenormally free or low-cost, with underpowered molecular visualization.Researchers who have analyzed these APPs indicated that their applicationsare still mainly restricted to simple molecular structure visualization.44As already indicated, there is an abundance of specificsoftwarefor smartphones and tablets. APP is the generic name of these specializedprograms for mobile devices. Google Play repositories for Androidand the Apple APP store for IOs Apple have hundreds of such tools.Many of them are offered in both repositories, as is the case of MoleculAR.46 and users can view chemistryconcepts, interact with them by hand, and learn through a self-directedlearning experience.47There is a set ofinvestigations onthe 3D representation of the atomic/molecular structure using AR ormixed reality technology. For a basic operation, AR runs with computingequipment along with a device capable of capturing images, such asa webcam, and a specific program to superimpose digital objects onthe image of the reality across the user’s screen. Most recently,these APPs are increasing their presence on smartphones,48 Researchon AR also emphasizes the emergence of increasingly powerful hardwaresuch as HMD equipment with semitransparent glasses to allow the userto view 3D digital data overlapping reality , which makes an entire digital world aroundthe user in a stereoscopic vision. reality 2. These 51 Several researchers have compared these HoloLens AR glasses withvisualization experiences on 2D computer screens or laptops, whichshows that traditional equipment is faster and more precise.52 That is why some authors have demonstrated theshortcomings of the AR technology and why resources for visualizingchemical reactions are limited to static models and preprogramed animations.47 In this sense, the lack of user interactivitywith real experiences could be an important aspect of teaching, andthis cognitive skill is crucial to the understanding of chemistryas a subject.47Currently, the technological evolutionof AR allows interactionswith objects through digital monitoring of the user’s hands,in a similar way as it happens in the VR technology.53 Vuforia,55 and ARchemy.56 The specific use of ARwith positive repercussions on the field of chemical education isdescribed in various investigations,59 especially at lowerlevels of training.60 Indeed, they analyzedthe outcomes of understanding the phenomena in the short and mediumterm. Students were positive to this experience, with a willingnessto become active learners. In particular, application in experimentalcourses showed its utility to teach while minimizing the use of chemicals,reducing handling risks and environmental impact.54Other investigationshave examined the potential of programs, suchas ARChemEx, ARKimia Kit, AC,61 Having said that HMD headsets are evolving theirportability, and the improvements try to overcome the motion sicknessand the sense of isolation.48Research on AR stressed its simplicity and flexibilitywhen comparedto VR 3. BecausAR is linked to real elements of the reality and the uploadingof digital data for visualization is key. Normally, users can printa card-marker or employ a camera shooting pattern and launch theirown interactions 4. This a46 and provides an individual approach for enhanced learning.Importantly, undergraduate students enjoy using these methods.59These card-markers usually appear in printed form in books andmagazines or can be executed from websites and function on normalcomputers. For the fabrication of these self-made shooting patterns,undergraduate students download and print the marker. Then, pointinga smartphone or a tablet toward this pattern, a previously storeddigital image, like a molecule, is superimposed. A specific benefitlies in the illusion of moving simultaneously with the projection,and the possibility of interaction with the image, including rotationand translation of the chemical structures along 3-axis, as well asenlarging and reducing their size as required. The AR task ensuresa personal pace for understanding complicated systems61 The visualization software of 3D can be sophisticatedfor manipulating and visualizing a model in almost infinite ways;however, the procedure with QR codes is straightforward in representingstructural data. This can be used in lectures or seminars by projecting/presentingthese codes, and the audience can directly reproduce the 3D structureon their mobile device. The application of this strategy is not onlylimited to represent molecules, including large ones such as proteins,but also allows the elaboration of 3D data sets from the results fromspectrometry experiments to map interaction partners.62Work on AR visualization can also be drivenby the use of quickresponse (QR) codes 2, as exa63 Once again, AR wasfound to be motivating for learners,64 providingvaluable digital 3D information for visualizing discrete moleculesas well as their changes during chemical transformations.65All this research underlines the fact that AR techniquescan beused over a period of time longer than VR. Moreover, the AR technologyallows users to work in parallel by taking notes or handle other documentsin another screen.9 In this sense, students showed a marked preferencefor more tactile AR models, providing the user a greater control overmolecular manipulations.9 The majority of experienceswith AR have been focused on early education because of the simplicityfor its implementation. For instance, AR was employed to study redoxreactions using Vuforia SDK and Unity3d55 or in molecular geometry.67 Improvements in performancewere noted in preuniversity students when working with HP Reveal softwareto identify organic compound formulas and nomenclatures, chemicalbonds, and intermolecular forces.37 Atthe university level, studies on chemical kinetics, chromatography,or the Krebs cycle reported improvements in the understanding of complexconcepts, as long as they are visually presented.68 Noticeableimprovements of the assimilation of concepts such as the understandingof molecular symmetry with diverse structural geometries7 have been reported by using the molecular symmetryalternate conception test, MACT.As stated above, the documents analyzedin this review clearlyemphasized that this technology is not restricted to visualizing structuresbut enables these elements to be controlled and manipulated.69 This line includes the resultof the activity in virtual laboratories (VL), and these results weresimilar to those obtained in real laboratories with traditional resources.4 Other research showed improvements in learningoutcomes70 and understanding in practicalchemistry classes.71 In spite of thesediscrepancies, all these features together justify the growing useof AR in education and research.Nevertheless, AR research ineducation does not only describe successfulexperiences, some studies did not observe beneficial effects of thistechnology.42 Computer tools are commonly used, while APPsand software for smartphones are still used for more simple structures.44AR is a relatively simple technology with most of thedocumented experiences focused on an early education stage. This technologycan help to visualize 3D molecules in an effective and efficient way.72 who enjoy using AR.73AR is motivatingfor the students,54AR might be helpful to minimize costs and risks in experimentalcourses.69Improvements in the educational performance with ARare still not obvious.74 In this sense, these technologies are noveland lack a proven track record of effectiveness. Moreover, the requiredtools, such as HoloLens glasses, need a high-performance computingto display 3D motion images, and it can be expensive for students.3D simulations and interactive atomic/molecular representationsoftware are widely, effectively used on 2D screens of the traditionalcomputer terminal, in chemistry classes of all levels. AR has problemsfor its systematic incorporation in educational environments.In general, some key findingsof AR can be described so far:75 in spiteof the difficulties in developing molecular software fulfilling rigorousscientific criteria. Regarding educational environments, it seemsadvisable to incorporate gradually the AR at early stages of chemistrystudies according to a number of studies supporting its effectiveness.For instance, AR visualizations appear as a valuable complementarytool to learn key concepts such as chirality, stereochemistry, coordinationof metal complexes, and biomacromolecular structures6 or for more advanced topics such as exploration of molecularsurfaces, geometries, transition states, or intermolecular interactionsin biomolecules, among others. Additionally, AR might be an interestingcomplement to assist laboratory courses through the use of digitalreplicas. The trend observed suggests that many of the next learningscenarios will be virtual workspaces.4 Nevertheless,the technological literacy of chemistry teachers must be significantlyincreased,13 and teachers need to reviewthe most appropriate learning methodologies to apply AR effectivelyin education.16Current3D chemical representations and manipulations are growingacross all stages in chemistry,76 The emerging technologies still require rigorous academic and scientificanalysis,16 particularly in the field ofeducation.72AR technology makesuse of software for chemical visualization:online programs, PC software, and APPs for smartphones. Additionally,there is an AR markup language (ARML), which is being used in XMLgrammar and software development kits (SDK) for simple AR environments.The biggest change driven by AR will be a human interaction with reality,including digital component in this process. In this way, new gesturalpatterns are being incorporated to facilitate the interaction andrecognition of user gestures without the need of an intermediate interface,such as a camera making an eye-tracking.77People use their mobilephones in countless ways that are becominga part of the educational activity of both students and teachers.Currently, smartphones have some limitations to operate with AR software,but these APPs will have a more effective and complex use in chemistry.This will be combined with artificial intelligence, the big data,the networks, and global collaboration.77 The science needs increasingly collaborative work to solveglobal challenges. In this sense, many researchers are starting topursue open-source software development collaborative projects totackle a globally accessible and effective chemistry teaching.20Investigations that analyze the research cooperative networksareof particular interest, and some of these experiences are supportedby decentralized and interconnected databases.78 for instance,in complex networks of chemical reactions based on huge quantitiesof data and vast quantum and chemical robotized explorations. The3D scenes digitally recreated are becoming powerful tools for exploringmolecule and protein energy landscapes.78It is also interesting to know new sophisticated virtual environmentsthat simulate reality,80 and this augmentsconventional physics-based approaches in computational research.80 This new technology infuses robustness intothe molecular models, and it is bound to become a mainstream toolin chemical research.2 Hence, we encouragemore research work into how all these possibilities could have aneducational utility.The interaction of reality data with digital information isthefoundation of the machine learning, where computers can learn fromdata without being programed by people,The results providea snapshot of current research on advancedatomic/molecular representation. The AR technology helps to visualizecomplex 3D environments in chemistry that today are done only in thelaboratory, for example, to understand chemical reaction mechanismsand their evolution in an optimization or a reaction path. AR allowsanalyzing different molecular characteristics, such as structures,bond distances, dihedral angles, geometry of a system, calculationof electrostatic potentials and vibrational modes, and so forth. Thesetypes of visualizations are efficient showing 3D molecules with stereocenters,as they could display an amino acid, a chiral catalyst, or processeswhere stereochemistry is relevant.The main research findingsreveal discrepancies regarding the effectivenessof this technology, especially when applied to higher education. Thus,the representation on a bidimensional screen is used in chemistryclasses at all educational levels, whereas the AR technology has afurther practice among initial students. In this sense, smartphoneapplications focus the content primarily on the visualization of simplemolecular structures, while more advanced users prefer traditionalatomic representation. This fact is justified because the latter equipmentis faster and more precise, especially when they combine several programssimultaneously in order to see and to understand complex molecularinteractions.Simple AR software is now becoming widely available,and the studentsusually have smartphones capable of showing these pictures. AR iseffective when students are relatively new to the laboratory environment,and when the user is working on complex processes and need assistanceprovided by additional digital information. These graphic tools arenot critical to the analysis of chemical reactions, but might helpthe 3D perception, which makes it more interesting for students, increasingtheir stimulation to understand chemistry. Therefore, it seems advisableto incorporate this technology into early chemistry education.The publications analyzed concur in that implementation of thetechnologies that simulate this reality cannot replace in-person experiencesbut they can complement them. This fact is especially relevant whenreal experiments are impeded by physical obstacles. In this respect,these digital technologies should contribute to the development ofan effective online accessible education.To conclude, thisanalysis can contribute to the identificationof potential practical references for teachers and academics, as wellas for researchers, who are designing the future smart educationalenvironment. Finally, inclusion of scientific innovation and thesetechnologies as complementary tools might result in a symbiotic effectfor teaching chemistry.


Tripartite Motif 47 (TRIM47) protein plays a prominent role in many cancers. This study aimed to investigate the biological roles of TRIM47 in ovarian cancer.TRIM47 was knocked down and overexpressed in ovarian cancer cell lines SKOV3 and OVCAR3, and the effects on proliferation, clone formation, apoptosis, invasion, and growth of xenograft tumors in nude mice were determined. The expression levels of the selected candidates were tested by western blotting and quantitative real-time PCR.TRIM47 knockdown suppressed proliferation and encourages apoptosis of ovarian cancer cells. Similarly, TRIM47 knockdown suppressed ovarian cancer cell invasion, migration, and epithelial-mesenchymal transition. Ovarian cancer cell xenograft assays demonstrated that TRIM47 knockdown significantly inhibited tumor growth. Mechanistically, TRIM47 knockdown suppressed STAT3 phosphorylation and the expression of several downstream genes, including MCL-1, MMP2, and c-MYC. Silencing of STAT3 partially prevented TRIM47–induced tumor cell proliferation and invasion.The present study's findings demonstrate that by activating STAT3 signaling, TRIM47 functions as an oncogene in ovarian cancer. TRIM47, therefore, appears to be a potential target for ovarian cancer prevention and/or therapy. Ovarian cancer is one of the most detrimental gynecological malignancies. In 2020 alone, approximately 313,959 new patients and 207,252 ovarian cancer deaths were reported.The Tripartite Motif (TRIM) family of proteins are evolutionarily conserved proteins that share a common N-terminal ‘Really Interesting New Gene’ (RING) finger domain, followed by 1 or 2 B-box domains and coiled-coil sequences.The authors here report that TRIM47 knockdown suppressed proliferation and invasion and promoted apoptosis of ovarian cancer cells, whereas TRIM47 overexpression enhanced cell proliferation and invasion. In addition, TRIM47 enhanced STAT3 phosphorylation, and the knockdown of STAT3 attenuated the oncogenic role of TRIM47 in ovarian cancer. These observations shed light on the mechanism by which TRIM47 promotes ovarian cancer.2 at 37°C.Antibodies against TRIM47, β-actin, Bax, Bcl-2, E-cadherin, vimentin, and STAT3 were purchased from Proteintech . Antibodies against phospho-STAT3 (Tyr-705) were obtained from Cell Signaling Technology . The secondary antibody for western blotting was purchased from Servicebio . EdU proliferation supplies, CCK-8 kit, and enhanced chemiluminescent reagent were acquired from the Boyetime . TRIzol® reagent was purchased from Invitrogen . The human ovarian cancer cell lines SKOV3 and OVCAR3 were purchased from the Chinese Academy of Sciences Cell Bank  and grown in RPMI-1640 medium  supplemented with 10% FBS  in a humidified environment with 5% COshRNA sequences for TRIM47 (shTRIM47) and Negative Control shRNA (NC) were cloned into the lentiviral vector pLKO.1. Virus particles were harvested 48h after lentiviral vector transfection alongside the packaging plasmids in HEK293T cells using Lipofectamine 2000. Ovarian cancer cells were infected with lentivirus-transducing units in the presence of 5 mg/mL polybrene. The TRIM47 coding sequence was cloned into pcDNA3.1 (+) to generate a TRIM47 overexpression vector. STAT3 siRNA was procured from GeneChem . Cells were collected 48h after plasmid or siRNA transfection.Western blotting was performed as previously described.−△△CT method and individual data represent triplicate measurements.cDNA synthesis was performed using a PrimeScript™ reverse transcriptase reagent kit , and total RNA was isolated from tissues or cells using TRIzol® reagent. The mRNA levels of E-cadherin, vimentin, Mcl-1, MMP2, and c-Myc were determined by qPCR using SYBR Green  on an ABI 7900 instrument  with GAPDH as an internal control. The fold-change in gene expression normalized by the internal control was estimated by the 23 cells in 96-well plates and cultured for 24h, 48h, 72h, and 96h. Then, 20 μL of CCK-8 reagent was added to each well and cultured for a further two hours. A standard microplate reader was used to measure the absorbance at 450 nm. For the colony formation assay, 1000 cells were plated in a 6-well plate and maintained in RPMI-1640 medium containing 10% FBS for two weeks. After methanol fixation, the colonies were stained with 0.1% crystal violet for 15 min before being counted. All assays were performed in triplicate.Cell proliferation was measured using a 5-Ethynyl-2′-Deoxyuridine (EdU) proliferation detection kit according to the manufacturer's instructions. Briefly, the cells were pretreated with EdU for 2h before being stained with DAPI. Images of EdU-positive cells were captured using a fluorescence microscope . For the Cell Counting Kit-8 (CCK-8) test, transfected cells were plated at 2 × 105 cells in 200 μL of RPMI-1640 medium without serum were placed in Boyden chambers . Similar cell quantities were placed in Boyden chambers covered with 150 mg Matrigel®  for the invasion test. Respective sections were subsequently infused into 24-well plates before 24h of incubation in RPMI-1640 medium supplemented with 10% FBS. Cells that remained on the upper surface of the membrane were removed, while those adhered to the lower surface were fixed, stained for 1h in a solution containing 0.05% crystal violet, and then counted using an Olympus microscope to determine their respective relative numbers. All results are presented as the mean of three separate analyses.For the migration test, 1 × 105 cells were fixed in ice-cold 70% ethanol, followed by incubation with the testing reagents of the Annexin V-FITC apoptosis kit (Boyetime) according to the manufacturer's guidelines. The fluorescence signal was detected using a BD FACSCanto™ flow cytometer and analyzed using FlowJo™ software.To measure cell apoptosis, 5 × 10http://gepia.cancer-pku.cn/).The TCGA database outcomes were primarily investigated using web-based gene expression profiling interactive analysis  × 0.5, where A is the length and B is the width of the tumor. The wet weight of each tumor was also determined.All the animal experimental procedures were approved by the Institutional Animal Care and Use Committee of Jinshan Hospital . Six-week-old female BALB/c nude mice were purchased from the SLAC Laboratory Animal Co. Ltd. . Subcutaneous xenografts were established by subcutaneous injection of 2 × 10IHC was performed according to a standard method, as described previously.t-test, while one-way analysis of variance (ANOVA) was used for multiple groups. Statistical significance was set at p < 0.05. All estimations were performed using the GraphPad Prism software (version 5.0).All data are presented as means ± the Standard Deviation (SD). Data between two groups were compared quantitatively using Student's To explore the role of TRIM47 in ovarian cancer tumorigenesis, the authors first performed loss-of-function assays by silencing TRIM47 with lentiviruses expressing shRNA against TRIM47 in the ovarian cancer cell lines OVCAR3 and SKOV3. Western blotting verified the successful knockdown of TRIM47 in these cells A. EdU inNext, the authors examined the effect of TRIM47 knockdown on ovarian cancer cell migration and invasion. In keeping with the Transwell® migration test results, the numbers of migrating SKOV3 and OVCAR3 cells were reduced after TRIM47 knockdown A. In addThe STAT3 signaling pathway is generally thought to be involved in cell proliferation, survival, motility, and invasion.To further determine whether TRIM47 functions in ovarian cancer by activating STAT3 signaling, SKOV3 cells were transfected with TRIM47 plasmids. TRIM47 overexpression enhanced the levels of p-STAT3 and promoted cell proliferation, migration, and invasion A–D. Whenin vivo. When the tumors were harvested, the weights and volumes of the tumors in the TRIM47 knockdown group were lower than those in the control groups .This study was supported by the Animal experiments complied with the ARRIVE guidelines and were carried out in accordance with the National Research Council's Guide for the Care and Use of Laboratory Animals, which was approved by the Institutional Animal Care and Use Committee of Jinshan Hospital .The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.The authors declare no conflicts of interest.


A relevant study found that allergic rhinitis (AR) may be related to the imbalance of nasal flora. Therefore, if the nasal flora of AR patients can be detected quickly, it is of great significance to study the distribution law of nasal flora in AR patients and explore its correlation with AR.  To design a new and convenient nano-DNA sensor for quick screening of nasal flora in allergic rhinitis (AR) patients, so as to provide experimental basis for the prevention and treatment of AR. 2-DNA sensor which can be combined with samples from nasal mucosa or secretion with high efficiency and detect certain flora in situ without DNA extraction or RNA sequencing. In a physical property test, firstly, we tested the permeability, solubility, and storage temperature of nano-TiO2, so as to provide experimental basis for the synthesis of Nano-TiO2-DNA sensor. Subsequently, the permeability of Nano-TiO2-DNA sensor in Staphylococcus aureus was further tested. In a clinical experiment, we selected 60 AR patients treated in our hospital from September 2020 to September 2021 as the AR group and 60 healthy people who underwent physical examination at the same time as the control group. The Nano-TiO2-DNA sensor was used to detect typical nasal flora in AR patients, and Pearson's correlation analysis was used to explore the correlation between nasal flora with serum IgE and eosinophils.  We create a synthesized nanostructured DNA biosensor called Nano-TiOStaphylococcus aureus, Klebsiella pneumoniae, and viridans streptococci in AR patients compared with healthy people, which will help these patients in the prevention of acute sinusitis and acute or subacute pneumonia. Furthermore, we found Proteus had the strongest positive correlation with AR while Actinomyces had the biggest negative correlation.  As for physicochemical characteristics, this sensor can permeate into certain bacteria directly and specifically. It has high affinity ability with a target, and the combination can be detected by evaluating the released fluorescence qualitatively and quantitatively. It can be stored at −20°C in ethyl alcohol stably. By this sensor, we have successfully detected 2-DNA sensor will help an outpatient doctor more for quick screening certain nasal flora in AR patients and improve the prevention of AR-related complications. The Nano-TiO Allergic rhinitis is a global health disease which has a profound impact on people's social work, daily life, and school events, although leading to no serious sequelae and death . The worAR is associated with many factors, such as irregular work and rest, unhealthy diet, and abused or overuse of antibiotics, which make the nasal mucosa more easily and frequently exposed to foreign microorganisms and antigens , 6. It h2-DNA sensor which can be combined with samples with high efficiency and detect certain flora in situ without DNA extraction or RNA sequencing. By this method, we can easily detect known flora by observing and quantifying the releasing fluorescence from the biosensor. So far as we know, there are few clinical studies on the specific detection of flora using nano-DNA sensors, and the emergence of this technology may bring new changes to the rapid detection of microbial flora.At present, the commonly used bacterial detection methods in clinic mainly include smear detection, bacterial culture, serological detection, bacterial nucleic acid detection, and second-generation sequencing, which plays a key role in the study of nasal flora distribution in AR patients, but there are also some deficiencies. For example, bacterial culture generally takes 3 days, and special flora even takes a week, which cannot meet the needs of early diagnosis. The specificity of serological antibody detection is relatively poor, which is prone to false positive or false negative results. In addition, second-generation sequencing technology and PCR amplification can go through almost all the abnormal flora distribution in nasal mucosa in detail via its high throughput detecting process , 16, but2 nanoparticles  were prepared. As these particles were used as the vector to enhance the penetration capacity of nano-DNA, here, we first packaged the particles to test its bind ability with bacteria. Hence, we first test its binding ability with bacteria by coupling fluorescein isothiocyanate (FITC). The TiO2-FITC was mixed with Staphylococcus aureus  which was stained by MycoLightRed . Second, we need an appropriate medium to carry our designed DNA sensor with maximum solubility and a stable storage temperature. So, we dissolve TiO2-FITC in different kinds of medium including distilled water, Phosphate-Buffered Saline (PBS), methyl alcohol, ethyl alcohol, and formic acid and test the fluorescence abortion of precipitated TiO2-FITC after standing for 24 h away from light. After confirming the best medium, dissolved TiO2-FITC was stored at different levels of temperature including −80°C, −20°C, 4°C, 37°C, 58°C, and 98°C for another 24 h, and precipitated TiO2-FITC was tested again via fluorescence abortion. Confocal microscopy  was applied for fluorescence image observation, and SpectraMax  was used for fluorescence quantitative analysis.TiO2 particles were conjugated with nano-DNA after stirring the mixture at 37°C for 12 h in borate buffer and removing free nano-DNA via centrifugation at 3000 rpm for 5 min. Dioleoyl-trimethy-lammonium-propane  was coated surrounding the TiO2-DNA to enhance the biocompatibility in samples with less immunological rejection, using distilled water to wash the sensor three times and store at 4°C for later use  Two separate Staphylococcus aureus-related oligonucleotides were designed. (2) FITC-labeled beacon was merged in the oligonucleotides via stirring at 60°C for 30 min and cooling down at 4°C for 5 min. (3) An fluorescence negative stop beacon tag was assembled in the opposite side of oligonucleotides after incubating at 37°C for 2 h and 60°C for 5 min with a mixture of glycerol to form the key part of the oligonucleotides which will bind to the target sequence of candidate bacteria into a ring structure, which will stop the fluorescence release. Then, TiOater use . Other f2-DNA sensor were separately mixed with Staphylococcus aureus to compare the infiltration ability. The reaction process was 95°C for 1 min, 62°C for 3 min, and 72°C for 5 min. Thus, the fluorescence intensity was detected 10 min later by fluorescence microscopy . As this sensor was designed for quick observation, no more waiting minute was needed. Besides, the fluorescence intensity detection was performed every 12 h and lasted for 72 h to test the fluorescence stability.Pure nano-DNA sensor and nano-TiO60 AR patients as well as the same number of healthy people who performed healthy examination in our hospital were included, all aging from 5 to 65 years. For AR patients, serum IgE and eosinophil percentage from blood were evaluated and relative data were collected from a clinical lab. All patients participating in this study voluntarily signed informed consent, and the study was approved by the medical ethics association of our college. As we plan to create a quick method to confirm the common or typical nasal flora in AR patient, here, we need to perform a conventional nasal flora examination to confirm these typical floras. Hence, nasal secretion was collected from AR patients and control people. After total DNA extraction , 16 s rRNA primers in V1 and V2 region including 27F and 338R were used for PCR amplification. 250 bp × 2 double-terminal sequencing of PCR products was performed using the Hiseq second-generation high-throughput sequencing platform (Illumina).After confirming the typical nasal flora in AR patients, we selected the top three kinds of flora in AR patients and healthy people separately as a target for testing. Before performing the test, the specific sensors related to certain floras will be synthesized based on the same protocol above and then mixed with the samples which come from nasal secretion of AR patients and healthy people quickly. The results can be evaluated after 10 min, and whether the positive result matched the conventional nasal flora date would be known.To support the testing result by DNA sensor and to provide some helpful information for AR prediction, we further analyzed the correlation of nasal flora with IgE and eosinophils, in order to obtain the most relevant flora with AR.n (%), and the quantitative data was described by x ± s. A t-test was employed for intergroup comparison. Pearson's correlation analysis was used to detect the correlation between nasal flora with IgE and eosinophils. P < 0.05 stands for striking difference.SPSS 22.0 and MedCalc software were adopted for statistical analysis. The quantitative data were described by 2 as the vector to enhance the infiltration ability of our sensor, here, we test its infiltration characteristics in flora. Obviously, TiO2 could permeate into Staphylococcus aureus directly (Figures 2 in some candidate media and find ethyl alcohol as the best medium to dissolve TiO2, with the least sediment after the solution standing by for 24 h (2 had the best stable status when being stored at both −80°C and−20°C, so the constructed sensor could be stored at −20°C to maintain its stability (Figures As we selected TiOfor 24 h . Besides2 and DOTMA outside of the DNA sensor to form a Nano-TiO2-DNA sensor, based on the better permeation capacity of TiO2 and good biocompatibility of DOTMA. It was proved that the TiO2-DNA sensor had better permeation capacity than the pure DNA sensor (Figures Although we can synthesize the pure nanostructured DNA, it was not able to permeate the flora with high efficiency; thus, we packaged the TiO Figures .Staphylococcus aureus, Klebsiella pneumoniae, and viridans streptococci, while Coagulase-negative Staphylococci, Corynebacterium, and Propionibacterium acnes were the top three of healthy people (Here, we presented the age distribution of AR patients and control people with no significant difference  and sex y people . Hence, 2-DNA sensor which had separate sequence information for each kind of flora to quickly test the nasal mucosa sample from AR patients and healthy people. Consistent with the result by a conventional sequencing method, our sensor could detect Coagulase-negative Staphylococci (Corynebacterium (Propionibacterium acnes (Klebsiella pneumoniae (viridans streptococci (Here, we applied our designed TiOylococci , Corynebacterium  and Propum acnes  with mucum acnes , Klebsieeumoniae , and virptococci  in healtNext, we analyzed the correlation of the above nasal flora with the level of IgE and eosinophils, aiming to find some cues for better prevention and therapy of AR. Finally, we found that Proteus had the strongest positive correlation with AR while Actinomyces had the biggest negative correlation, as shown in 2-DNA sensor. The working principle of this kind of synthesis nanostructured DNA is the “open and shut” process for fluorescence beacon. In a pretest situation, the fluorescence beacon was silenced by a stop beacon after forming a circle structure in the DNA sequence. This silent structure has no fluorescence release, so we cannot detect any signal before mixing the sensor with samples. During the testing process, the biosensors will be mixed with samples and find their complementary sequence if they exist under a certain reaction condition. In this process, the ring structure will be opened and the silent fluorescence will be released again. Hence, if there exists a certain kind of flora in the nasal sample, we can detect the fluorescence signal, and the fluorescence intensity will reflect the quantity of certain targets. As a result, we can confirm certain flora both qualitatively and quantitatively.In this study, we focus on the nasal flora in AR patients as this evidence will contribute to AR diagnosis and complication prevention caused by AR –19. To qThis kind of sensor has the following advantage. First, this sensor is very easy for use. It can be dropped on samples which were collected from the surface of nasal mucosa directly and under a simple reaction temperature cycle without DNA extraction, PCR process, and sequencing. Thus, the sensor screening method can be realized at an outpatient department by an assistant without sending to clinical lab or even a third party of inspection company. This will save work time for a physician and save money for patients obviously. Second, the result is easy to get. As the read out is the fluorescence signal, so it is easy for judgment without too many false positive or false negative results.2 biosensor to the detection of E. coli, which effectively improved the targeting and specificity of the E. coli detection, consistent with the results of the present study. To our knowledge, the above floras detected in AR patients have much correlation with body infection including acute sinusitis and acute or subacute pneumonia, so we guess this may be an explanation why some AR patents tend to suffer from these two kinds of infectious disease [By this sensor, we successfully detect some high level of flora in AR patients including Staphylococcus aureus, Klebsiella pneumoniae, and viridans streptococci. Yoo et al.  applied  disease , 22. TheThis study mainly discussed the sensitivity of nanosensor to the detection of typical nasal flora, but the sample size is relatively small, and the pathogenesis of AR caused by flora is not studied, which should be improved in the follow-up study.2-DNA sensor. This sensor was able to detect certain nasal flora in situ quickly without DNA extraction, PCR process, and conventional sequencing technology. We have successfully used this kind of sensor to detect Staphylococcus aureus, Klebsiella pneumoniae, and viridans streptococci in AR patients which will help these patients in the prevention of acute sinusitis and acute or subacute pneumonia. This method will help an outpatient doctor more for quick screening certain nasal flora in AR patients and improve prevention of AR-related complications.In this study, we developed a synthesized nanostructured DNA called TiO