Ointments have been widely used as an efficient means of transdermal drug application for centuries. In order to create ointments suitable for various new medicinal drugs, the creation of ointment base materials, such as gels, has attracted much research attention in this decade. On the other hand, the chemical tuning of low-molecular-weight gelators (LMWGs) has been increasingly studied for two decades because LMWGs can be tailored for different purposes by molecular design and modification. In this review, several series of studies related to the creation of ointment base materials with enhanced properties using existing and newly-created LMWGs are summarized. Ointments are used as a method for delivering medicinal agents by direct application to the affected area of the skin or by application to the surface of the skin near the affected area, depending upon whether the wound or condition is internal or external. This is termed transdermal drug delivery  1,2]. I,2. I1,2]An ointment is composed of a medicinal agent and a base material that together form and maintain the structure of an ointment jelly. Thus, a base material must exhibit chemical stability and consistency of appearance in the presence of the formulated medicinal agent . It mustWhile the use of polymer gels is well-known and thoroughly investigated, molecular gels, which are constructed by the self-assembly of low-molecular-weight gelators (LMWGs), have attracted much attention as a new soft materials in the past two decades ,16,17,18In this review, the author presents a summary of studies on the application of molecular gels (mainly organogels for oil gels) used as base materials for ointments. These gels exhibit unique properties because they are composed of LMWGs; however, they exhibit some similar properties to polymer gels because of the formation of networks of fibers composed of LMWGs. First, ordinary ointments and molecular gels with thixotropy, which is a required property for ointment applications, are discussed. Then, recent studies on several organogelators for ointments are summarized. Finally, recent developments of new low-molecular-weight organogelators for ointments are described with respect to their required mechanical properties, including thixotropy.It should be noted that application studies of molecular hydrogels and organogels for drug delivery have been summarized in the literature ,20,21,22There are many representative medicinal drugs applied as an ointment to the skin as topical agents, such as anti-inflammatory agents , local anesthetics , immunosuppressive agents , and vasodilators  ,2. Low tOintments can be used as the active ingredient in dermal and transdermal patches. Owing to the development of many types of patches, ointments will become increasingly applied in this form. According to the wide range of future uses of ointments and the potential demand for ointment base materials with improved properties suitable to various types of medicinal drugs, the re-examination of both traditional and newly-developed base materials has been performed. Generally, ointments are produced by mixing and dissolving a medicinal agent, a base material, and a non-volatile oil. Typical non-volatile oils used in ointments are edible fats and oils, such as vegetable oil and animal fat, and synthetic oils and esters, such as silicones and isopropyl myristate, which are generally used in cosmetics. These fats and oils must be safe for human use.in vitro. In this apparatus, an ointment containing a medicinal drug is placed on an appropriate membrane, e.g., a cellulose acetate membrane or pig skin as an analog of human skin, and a buffer or aqueous solution near body temperature is used as a model for a physiological aqueous solution for receiving the permeated or diffused medicinal drug from the ointment through the membrane [Franz-type vertical diffusion cells are generally used to obtain data on the transdermal absorption of ointments membrane ,25. Contmembrane ,27,28,29Liposomes, microgels, and LMWGs have been examined as newly-developed base materials . On the Molecular gels are defined as crystalline fiber networks constructed from LMWGs in self-assembled manner that incorporate solvents within the networks, resulting in a gel-like appearance, much like that of polymer gels  5,6,7,8,16,17,18Using molecular gels as base materials for ointments is one of the most promising applications of LMWGs as soft materials. Molecular gels composed of LMWGs for use in ointment usage have several advantages, such as easy preparation and the possibility of improving their properties compared with those of polymeric compounds by molecular design. Preparation of LMWGs can be conducted inexpensively, with commercially available ingredients and a practical synthesis that results in a product suitable for use as an ointment. In general, LMWGs are easy to synthesize and prepare because of their low molecular weights and well-defined structures.Non-toxicity to humans is absolutely necessary. The design of non-toxic LMWGs that retain their required properties can be attained by tuning their chemical structure precisely using the results of toxicity tests. In terms of their molecular properties, molecular gels have poor mechanical strength, low durability, and poor long-term stability of appearance compared with polymer gels, which contain polymer networks resulting from multiple interactions between polymer chains. Despite the inferior properties of molecular gels compared with polymer gels, good stability and thixotropic behavior can be observed in molecular gels composed of molecularly designed LMWGs.Thixotropy is defined as mechanically induced reversible sol-to-gel or gel-to-sol changes and is generally observed in polymeric or inorganic nanosheet solution systems ,32,33,34Since medicinal drug molecules commonly have lipophilic properties, molecular organogels, prepared from low-molecular-weight organogelators, are used to retain the medicinal molecules within a gel network, and molecular hydrogels, prepared from low-molecular-weight hydrogelators, are used to support micelles containing the medicinal molecules within a gel network. In the case of ointments composed of molecular gels, there is a tendency to use the latter micelle-dispersed hydrogels owing to the frequent presence of aromatic rings in organogelators, leading to their use being avoided by medical personnel . Taking 1)) has been used by Pénzesa et al. in vitro and in vivo (rats) in the molecular gel state with the anti-inflammatory drug piroxicam, with white petrolatum and liquid paraffin being used as references [in vitro and in vivo more effectively than the reference materials.Glycerol esters, which are compounds commonly found in vegetable oils and animal fats, have been used as LMWGs for oils. The resulting organogels have similar feeling on skin application as soft paraffin (petroleum jelly), which is a mixture of hydrocarbons that forms a gel-like material. As an organogelator for use in ointments, Miglyol 812® ) is a fatty acid derivative well-known for its ability to form molecular organogels. 12-HSA functions as LMWG for vegetable oils and is often used as an additive to assist in the disposal of cooking oils. 12-HSA is a candidate for an ointment base material, and its use in oral controlled-release organogels has been studied. Iwanaga et al. reported that an organogel composed of 12-HSA, soybean oil, and the lipophilic medicinal drug ibuprofen formed a controlled drug release material in vitro and in vivo [in vitro and in vivo [12-Hydroxystearic acid ). They studied organogels composed of LMWGs containing a tyrosine unit and the parasympathomimetic drug rivastigmine and found that the drug was released slower in stiff organogels with higher Young’s moduli, suggesting the importance of evaluating the mechanical properties of molecular gels for drug delivery and ointments [et al. reported that the difference between the results observed in in vivo and in vitro experiments, which were inferior in vitro, was ascribable to the existence of in vivo enzymes, which led to enzyme-related degradation and erosion of the gel matrix, thereby affecting the diffusion mechanism of the drug [in vitro with the same performance as that in vivo. This strategy established a methodology for the fast creation of drug delivery materials in vitro without the need to change the gel formulation responsible for inferior results obtained in vivo. These results, and the concept developed, are important for the design of ointments using LMWGs because they show that the effect of biomolecules such as enzymes on the efficacy of ointments needs to be considered.Amino acid derivatives have been studied as a promising family of LMWGs. Using amino acid derivatives, Leroux intments ,55. Lerod-sorbitol ) is an old type of LMWG for organic solvents that forms molecular organogels known for over 100 years, and it has been used as an additive in cosmetics and dental materials [et al. have created novel DBS derivatives containing hydrazide units ) and examined two-component hybrid molecular hydrogel formations composed of DBS-CONHNH2 and medicinal drugs, such as mesalazine, naproxen, and ibuprofen ). Related to this studies, the basic material properties involving thixotropic behavior of toluene organogel composed of simple gelator with an amide was also evaluated (9)) [In recent years, 12-HSA has been considered as one of the most important core chemical structures for new high-performance organogelators. The relationship between the material properties of molecular gels composed of 12-HSA derivatives with different chemical structures was thoroughly studied ,59,60. Tted (9)) .10a–c)), stearohydrazide , palmitohydrazide , and octanohydrazide , exhibit organogelation abilities with organic solvents having various dielectric constantsfrom 2 (n-hexane) to 66 (propylene carbonate) [Recently, the author’s group found that simple alkylhydrazide derivatives (rbonate) . Previourbonate) . Furtherrbonate) . Better 11a–c)) [12a,b)) [Although several molecular gels showed the mixing-induced enhancement of molecular gel properties ,64,65, o11a–c))  and alky[12a,b)) , respectTo observe microstructures of mixed xerogels by scanning electron microscopy showed increase of network density by changes of shape of the network component from sheet-like or tape-like crystals to fiber-like crystals in increasing the ratio of gelator with shorter alkyl chain . The incFurthermore, multicomponent alkylamide organogels composed of C18Am, C16Am, and C8Am containing non-volatile oils, such as olive oil and squalane, were prepared for examining its properties of controlled drug release. Even in the presence of these non-volatile oils, a mixed alkylamide molecular gels showed thixotropic behavior. This organogel showed controlled drug (antipyrine) release in diffusion kinetics and will become one of candidates of spreadable host material for ointment base material .13a) and erucamide  as organogelatos in order to create new thixotropic non-volatile oil gels for controlled drug release (Mixed alkylamides with long-chain and short-chain alkyl groups formed thixotropic gels even in non-volatile oils. However, thixotropic multicomponent alkylamide organogels containing non-volatile oils showed drug release ability, according to its MSDS data, C8Am will bring about eye and skin irritation . Therefo release . These a release . It was In this review, the author has summarized several studies that aimed to create better molecular organogels applicable to ointment base materials by chemical approaches using LMWGs. The selection of suitable molecular gels for combination with new medicinal drugs and other additives needed to produce an ointment is challenging. Of course, the reexamination of existing molecular gels as ointment base materials is important; however, new molecular designs and new strategies for creating LMWGs that are suitable for ointments are required for the quick development of LMWGs for use with various medicinal drugs. The development of better treatments through the use of ointments will enhance our quality of life. Thus, more research in order to accumulate chemical libraries of different LMWGs with required properties such as thixotropy is required.


CDNA[η]. Furthermore, to complete previously reported experimental data, solutions of a large molecular weight DNA from calf-thymus were studied in a high concentration range (up to 40 mg/mL). The rheological behavior is discussed in terms of the generalized master curve obtained from the variation of the specific viscosity at zero shear rate  as a function of CDNA[η].Molecular weight, stiffness, temperature, and polymer and ionic concentrations are known to widely influence the viscosity of polymer solutions. Additionally, polymer molecular weight—which is related to its dimensions in solution—is one of its most important characteristics. In this communication, low molecular weight DNA from salmon sperm was purified and then studied in solutions in a wide concentration range (between 0.5 and 1600 mg/mL). The intrinsic viscosity of this low molecular weight DNA sample was firstly determined and the evidence of the overlap concentration was detected around the concentration of 125 mg/mL. The chain characteristics of these short molecules were studied in terms of the influence of their molecular weight on the solution viscosities and on the overlap parameter  Up to now, two of the most-studied fundamental properties of biomacromolecules such as DNA, proteins, and polysaccharides have been the hydrodynamic and conformational properties ,2. For tC/C* (where C is the polymer mass concentration and C* is the overlap concentration), with an effective exponent depending on the solvent quality parameter z. In their work, they determined the θ-temperature of dilute DNA solutions in the presence of salt excess, but they never considered the stiffness of the DNA molecule. Recently, a detailed study of the rheological behavior of high molecular weight DNA  solutions was reported, as well as evidence of the two critical concentrations of the system—i.e., the overlap and the entanglement concentrations  ) in a range of CDNA[η] values up to 40.Recently, Pan et al.  reportedctively) . Furthersp,0 vs. C[η] relationship based on the expression previously reported by Kwei et al. [C** and CDNA[η] values up to 160. Additionally, chain characteristics of very short molecules (only some tens of base pairs), which are of great importance to molecular or cell biology,  relationship was then found to be applicable to DNA chains with a molecular weight lower than the one corresponding to the persistence length of 50 nm, where DNA molecules behave as semi-flexible rods.In this paper, a novel representation of the ηi et al.  is propobiology, ,14,15 arMw = 6,559,500 g/mol  is the intrinsic viscosity (mL/g) and k’ is the Huggins constant.Then, since the DNA molecule bears one formal negative charge per nucleotide, its conformation is certainly sensitive to changes in the ionic strength, in the number of base pairs, and in the molecular weight ,21,22. Irelation .ηred =The intrinsic viscosity, (8.49 mL/g) was obtained from extrapolation to zero concentration. It is worth mentioning that low values of intrinsic viscosities between 7.5 mL/g and 160 mL/g were previously reported for oligonucleotides and oligosaccharides ,24,25. AThus, the calculated viscometric-average molecular weight for this sample is equal to 15,000 g/mol, equivalent to around 23 bp .C*) was estimated through the deviation from linear behavior in the dilute regime , which is also in good agreement with the value obtained from the relation to C*~[η]−1.Finally, the overlap concentration  by means of the generalized master curve, initially proposed for hyaluronans with various molecular weights. The importance of this approach is recognized since the intrinsic viscosity values ([η]) also contain information regarding the stiffness of the polymer −1) and the entanglement concentration .It should be pointed out that knowing the intrinsic viscosity of the polymer, it becomes possible to calculate the specific viscosity at zero shear rate 2/2! = 0.08 and k3 = 0.0213.Equation (3) presents a novel representation of the relationship between the specific viscosity and the overlap parameter, which is based on the expression previously proposed by Kwei et al. for hyaluronan samples .ηsp = C[η] > 10), instead of the value proposed by Kwei et al.  is then valid for DNA chains with molecular weights lower than 1 × 105 g/mol, where chains are in an almost rod-like state, and for DNA chains having higher molecular weights, where molecules behave as worm-like chains.All of the data for the specific viscosity at zero shear rate obtained for this LMW DNA were plotted together as a function of the overlap parameter, and collapse on the curve obtained for the HMW DNA. The proposed relationship between ηCDNA[η] = 160, the zero shear-rate viscosity determined through the Cross model follows the improved master curve. Up to C[η] = 160, the slope value remains at 4.3, with a behavior corresponding to that of the semi-dilute regime with entanglements.For high molecular weight DNA solutions from calf-thymus with concentrations up to


Klinefelter syndrome can present as a wide spectrum of clinical manifestations at various stages in life, making it a chromosomal disorder with no standardized set of guidelines for appropriate management. Understanding the genetic and hormonal causes of this syndrome can allow physicians to treat each patient on a more individualized basis. The timing of diagnosis and degree of symptoms can guide management. This report will provide an updated review of the clinical presentation at various stages in life and the implications for management. It is a subject of interest because of its wide spectrum of clinical manifestations, which include certain physical features, cognitive delays, and azoospermia.Klinefelter syndrome (KS) is a common male chromosomal disorder  that has been a topic of intrigue and inquiry since the 1940s, when it was first described2.The chromosomal aberration found in KS is due to either meiotic or mitotic non-disjunction, leading to sex-chromosomal aneuploidy. Two genetic variations exist. The majority (90%) of cases present as a pure form with a 47, XXY karyotype, whereas the remaining 10% include the following sex-chromosomal abnormalities: mosaicism , higher-grade aneuploidy , and structurally abnormal X chromosomes4. However, KS may be recognized during the prenatal, prepubertal, adolescent, or adult period. A patient’s management is guided by the extent and severity of clinical features as well as timing of presentation.KS has been reported to occur in 1 out of 600 male births, approximately 64% of which remain undiagnosed throughout life5. The severity of the presentation is also strongly correlated with the severity of the sex-chromosome aneuploidy . A prepubertal boy may be diagnosed during the workup of cryptorchidism or mild developmental delay, whereas KS may be discovered initially in the older individual during an infertility evaluation. Some physicians believe that delay in diagnosis can increase the morbidity of the patient; therefore, it is important for practitioners to become familiar with this spectrum of features.There is no single, classic phenotype that can describe KS. The familiar description of tall stature with a thin, eunuchoid body habitus is not only inaccurate but also inadequate. There are no distinct dysmorphic features, and presentation may vary according to the degree of gonadal dysfunction6. However, Nieschlag, Ferlin, Gravholtet al. provided a summary of the Second International Workshop on Klinefelter Syndrome , and one of the topics addressed was the advisability of neonatal screening and diagnosis7. Whereas Rogol argued that there are recognizable benefits to neonatal detection, Gravholt cautioned that there is yet no proof that early detection necessarily results in a reduction in morbidity and mortality with an improvement in outcomes when large populations are screened. Therefore, even expert opinion diverges on this issue at this time. The mini-puberty, namely activated luteinizing hormone (LH) and follicle-stimulating hormone (FSH) release in the first few months of life, matches that of 46, XY infants. Less than 3% to 10% of patients are diagnosed during the prepubertal period4. Tall stature may be seen, but not always, because of increased dosage of the sex chromosome-related short stature homeobox-containing gene (SHOX), which resides in the pseudoautosomal region of both X and Y and is not subject to X-chromosomal inactivation8. Additional physical features include cryptorchidism, gynecomastia, hypotonia, hypertelorism, clinodactyly, elbow dysplasia, pes planus, and a high arched palate9.There are no telltale physical signs of KS at birth. An increasingly important means of diagnosis is through prenatal testing. There is a growing utilization of non-invasive prenatal screening by cell-free fetal DNA testing. This test can identify the presence of extra chromosomes—autosomal and sex10. Though these features are more apparent in the severe chromosomal aneuploidies , KS males may exhibit “language-based learning disabilities, decreased fine motor skills, and discrepancies between nonverbal and verbal cognitive abilities” in the sense that visuoperceptual and nonverbal skills appear to be strengths11. In a cross-sectional study examining a prepubertal KS cohort, the presenting characteristic that ultimately led to diagnosis was developmental delay (11.6%)12. Gropman and Samango-Sprouse13 describe how young KS boys may have resultant academic difficulties in school, which broaches the topic of early diagnosis and whether testosterone replacement can prove advantageous. Although it is clear that androgens play a role in neurodevelopment, the progressive and more severe presentations of cognitive and motor delays seen with each additional X chromosome suggest a gene dosage effect of the X chromosome. In their double-blind randomized controlled trial, Rosset al. showed that 2 years of low-dose androgen treatment “did not have significant effects on most aspects of cognition ”14. Although the question of whether testosterone replacement would have a positive impact remains, early intervention with educational supports such as tutoring and counseling may be beneficial.These characteristics may be accompanied by learning difficulties that may manifest as attention disorders and speech or language impediments. Early literature has also suggested a higher prevalence of autism spectrum disorder, attention-deficit/hyperactivity disorder, and schizophrenia; however, results are somewhat incomplete, as those studies did not take into account family history of mental illness or learning disabilities15. However, in a minority of cases, the degree of hypogonadism can be so severe that there may be minimal or no signs of pubertal development, prompting further workup. Testosterone supplementation to boost pubertal advancement may be of use in these cases.Diagnosis of the adolescent KS male can be more challenging. The degree of virilization is dependent upon the level of testicular testosterone production. The initial rise in testosterone levels during puberty may mirror that of a 46, XY male but typically plateau at low to low normal values. Although external secondary sexual characteristics may appear Tanner stage-appropriate as puberty progresses, reduced testicular volume is almost universally present. The average testicular volume for a KS postpubertal male is 2 to 5 mL16. Typically, LH and FSH will be elevated, testosterone will be low normal, and semen volume will be adequate. Erectile function is not commonly impacted.KS discovered in adulthood is often under the circumstances of evaluation for primary infertility. Although 8% of the KS population may have detectable sperm in the ejaculate, the majority will have non-obstructive azoospermia17 and Gravholtet al.18, cardiovascular, cerebrovascular, and metabolic syndrome and other comorbidities are increased in patients with KS. Cardiovascular mortality is higher in this population and this is usually due to aortic valve anomalies, pulmonary embolism, peripheral vascular disease, and deep vein thrombosis19. For example, as Zölleret al. detailed in their Swedish cohort study, the cumulative incidences of venous thromboembolism in men with KS were 8.6% at 50 years of age and 20.8% at 70 years of age20. In efforts to explain this association, Valaseket al.21 reported on a pediatric case in which the boy also had a type 1 protein C deficiency whereas Erkalet al.22 did not find a difference in PAI-1 gene polymorphisms when comparing KS patients and cohorts. Finally, Gluecket al. caution that perhaps prothrombotic long-term testosterone replacement therapy (TRT) in conjunction with undiagnosed familial thrombophilia may be the combination that leads to thromboembolism in the patient with KS23. They advocate for thrombophilia screening prior to initiation of TRT23. Cardiac structural abnormalities include left ventricular diastolic dysfunction, mitral valve prolapse, and increased thickness of inner tunica of carotid arteries24. Decreased exercise tolerance is also common. It is reasonable for practitioners to consider a diagnostic echocardiogram.As reviewed by Kanakis and Nieschlag25. The KS population is also noted to have a higher prevalence of metabolic syndrome and dyslipidemia. In one cross-sectional study, 47% of subjects with KS were noted to have metabolic syndrome, including increased total body fat, waist circumference, insulin resistance, triglycerides, and low-density lipoproteins26.Cerebrovascular mortality may be secondary to subarachnoid hemorrhage, rupture of an intracranial saccular aneurysm, or a thrombotic event27. As Tahaniet al. state, “bone loss in KS might, at least in part, be independent of hypogonadism”27.Men with KS may have reduced bone mineral density in the lumbar spine, femoral neck, and total hip, but TRT does not ameliorate all of these findingsAlthough the overall incidence of malignancy is not higher in the KS population, there is an increase in multiple types of cancer, including breast cancer and extragonadal germ cell tumors (GCTs). However, the overall incidence is so low that routine screening of an asymptomatic KS male is not recommended.et al., 3% of GCTs diagnosed in males under the age of 19 occur in boys with a 47, XXY karyotype28. Conversely, the risk of developing a GCT in KS patients of this age group is 1 in 4,000 (risk ratio of 18.8) compared with the normal 46, XY male population. In their study, the parent of origin of the supernumerary X chromosome was split equally between mother and father. Most GCTs in males with KS are mediastinal in location and teratomatous in histology. In fact, among KS males aged 14 to 19, GCTs are almost exclusively mediastinal. The exact biological etiology driving mediastinal GCT development is unknown at this point. These tumors may present at a young age as precocious puberty because of human chorionic gonadotropin secretion by the tumor cells. Older boys may present with chest pain or respiratory symptoms due to tumor location. Although today’s literature describes primarily pediatric cases of mediastinal GCTs in KS, adult males with KS have also been shown to develop these masses29. Males with mediastinal GCTs should be screened for KS.According to Williams30. With the absolute incidence still being quite low, routine screening mammography is not a formal recommendation30.In her literature review of the data to date, Brinton concluded that the risk of breast cancer in the male with KS is 20- to 30-fold higher than expected and that there is a 60-fold increase in mortality31. To give a brief embryologic overview, germ cells migrate to the genital ridges and populate the nascent seminiferous tubules. The Sertoli cells and Leydig cells form within and around the seminiferous tubules, respectively. Beginning just after birth, peaking in the 4th to 10th week after delivery and nadiring to prepubertal levels by 6 months of age, there is a surge in pituitary gonadotropins, also known as “mini puberty”, which stimulates the expansion and limited differentiation of the spermatogonial stem cell pool32. The seminiferous tubules lengthen, testis volume enlarges, and Sertoli cell number increases33. Cell cycle activity in the testes remains fairly quiescent until the initiation of puberty as a result of hypothalamic gonadotropin-releasing hormone (GnRH) stimulation of pituitary LH and FSH production and release. LH triggers intratesticular testosterone manufacture while FSH sparks Sertoli cell maturation and spermatogenic induction. In the KS male specifically, peripheral serum testosterone values rise but typically level off in the low normal range while the vast majority of 47, XXY spermatogonia become apoptotic as they fail to complete meiosis.KS is found to be the etiology of infertility in 10% of azoospermic males and accounts for up to 2% of infertility in the general male population34. However, it cannot be simply that an inability to sequence through the meiotic cascade leads invariably to such rapid and dramatic cell senescence and demise, as we would then expect the same events to unfold in spermatocytic maturation arrest resulting from genetic causes. In this latter anomaly of spermatogenesis, viable spermatogonia and spermatocytes stack up like cars on a highway behind an impenetrable roadblock—remaining alive but prevented from progressing further. However, the presence of spermatozoa in about 50% of KS testes is best explained by postulating a low level of gonosomal mosaicism. Escaping death and proceeding through meiosis I and II, as well as spermiogenesis, are scattered 46, XY spermatogonia which, through a second quirk of nature, have lost the additional X chromosome, resorted to a normal diploid state, and come to reside in a seminiferous tubule here or a seminiferous tubule there. The distribution is most likely random, explaining the patternless and haphazard location within the tiny testes of spermatogenically healthy tubules.Classic puberty in the adolescent heralds the start of intratesticular testosterone production, necessary for spermatogenesis. Although testosterone levels do increase on schedule, males with KS experience a blunting of both Sertoli and Leydig cell function after the initiation of puberty. There is progressive apoptosis of the 47, XXY spermatogonia, ultimately resulting in azoospermia in adulthood37.Until the advent of assisted reproductive techniques, males with KS were considered to be sterile. However, with further understanding that these men may have focal areas of spermatogenesis within the testes, testicular sperm extraction (TESE) has become a mainstay treatment option for sperm harvesting and future reproduction38. Subsequently, the resultant 46, XY daughter cells have the genetic opportunity to replicate. The extent of spermatogenesis in a male with KS appears then to depend on the presence and quantity of these 46, XY germ cells. Some argue that TESE procedures for sperm cryopreservation should be pursued during the adolescent period, or possibly earlier, prior to the progression of spermatogonial apoptosis. The opposite school of thought believes that TESE should be delayed until adulthood. As reviewed recently by Oates, many studies have examined this relationship between age at time of TESE and rate of successful surgical sperm retrieval (SSR)39. Although some of the evidence is contradictory, the majority have found no difference in TESE outcomes among different age groups40. In fact, Franiket al. demonstrated that SSR was lower in patients below the age of 16 who underwent TESE41.As described in the preceding paragraph, during mitotic renewal or differentiation of an occasional 47, XXY spermatogonial stem cell, the extra X chromosome may be lost because of anaphase laget al. found that only 30% of peripubertal patients from 12 to 16 years had any germ cells on testicular biopsy, suggesting that germ cell loss begins well before puberty40. The authors’ study beautifully addressed the timing of germ cell loss and the rate of TESE success in a number of different age ranges. They examined the testes of five fetuses with a 47, XXY karyotype and concluded that the number of germ cells identified “was not significantly different from controls”. There was very little fibrosis noted. However, Wingeet al. observed that, in eight KS fetuses they studied, germ cell loss may already be occurring, specifically failure of gonocytes to differentiate into the slightly more advanced stage of pre-spermatogonia42. The authors suggested an upregulation of X-chromosomal transcripts and enrichment of certain long non-coding RNAs (ncRNAs) as a possible etiologic explanation. Wingeet al., in a further study, concluded that the mechanisms underlying germ cell loss may be different in the fetal, prepubertal, and adult stages and that aberrant maturation of Sertoli and Leydig cells may also be contributory43.To elaborate on and investigate spermatogenic potential in 47, XXY testes, van Saenet al. also documented a paucity of detectable germ cells in prepubertal patients, no ongoing spermatogenesis in 20 biopsies from peripubertal boys, and sperm detection in only 1 out of 20 adolescent boys undergoing TESE. However, in the older adult patients with KS, TESE recovered fully formed spermatozoa in 48% with no difference noted between the various age groups , a finding supported by the meta-analysis by Coronaet al.35. To summarize, van Saenet al. concluded that “sperm recovery by TESE at early adolescent age does not appear to result in higher sperm retrieval efficiency compared to TESE at adult age”. Riveset al. also cautioned that “no predictive factors can actually demonstrate that early diagnosis of KS would allow increasing the chance of sperm retrieval”44. Taking a slightly different and novel focus, Ragabet al. recently reported on the microsurgical TESE (mTESE) SSR in two overall groups of patients with KS: 31% in those with a history of unilateral or bilateral cryptorchidism and 38% in those with eutopic testes at birth—no statistical difference was seen. SSRs also did not differ between those KS men born with unilateral or bilateral undescended testes45. Embedded within the excellent data of their study is the fact that, in the group of KS males with eutopic testes, the median age at which mTESE found sperm was comparable to the age at which no sperm were retrieved: 23 (range 15–26) and 20.5 (range 17–48), respectively45.In their study mentioned above, van Saenet al., which compares techniques of TESE in patients with KS specifically, demonstrates similar sperm retrieval rates with both conventional TESE and mTESE35. However, studies looking more generally at men with non-obstructive azoospermia do suggest that there is an increased sperm retrieval rate in mTESE46. With regard to the ultimate goal of reproduction, the couple must proceed with intracytoplasmic sperm injection (ICSI) to use the spermatozoa that may be retrieved in either the freshly harvested or the frozen-thawed state. Recent studies report an average sperm retrieval rate per TESE procedure of 34% to 44% and an average live birth rate per ICSI cycle of 29% to 43%35. In their latest report, aptly titled “Is genetic fatherhood within reach for all azoospermic Klinefelter men?”, Vloeberghset al. caution that a multiplicity of factors conspire to actually determine the ultimate chance of biological fatherhood, not just the probability of finding sperm on TESE or mTESE31. To wit, only 14 out of 138 (10.1%) KS men and their partners starting treatment succeeded in having a live birth. Hormonal parameters have not yet been shown to predict the success of TESE31.In a discussion of preferred surgical techniques, a large systematic review by Coronaet al. demonstrated “211 differentially expressed transcripts in the fetal KS testis”, a significant enrichment of X-chromosomal transcripts and long ncRNAs42. This suggested to them that the failure of gonocyte differentiation into pre-spermatogonia in the fetal testis led directly to the extremely poor (or absent) spermatogenesis in the adult KS testis. In their “companion” study on adult KS testes, they conclude that disturbed maturation of Sertoli and Leydig cells may further inhibit, and directly impair, spermatogenesis as well43.Sex-chromosome aneuploidies are also fascinating from a purely genomic perspective. Although the supernumerary X chromosome undergoes inactivation, there are clearly genes that escape inactivation that are either directly or indirectly responsible for the KS phenotype. Via RNA sequencing (RNA-seq), Wingeet al. further defined the genetic underpinnings of the phenotypic findings in Klinefelter males47. That is, they asked how the additional X chromosome is influencing form and function. They performed both genome-wide DNA methylation and genome-wide RNA-seq of peripheral blood leukocytes in 67 and 9 patients with KS, respectively. They had similar numbers of 46, XY male and 46, XX female controls for both groups. They built on previous work linking changes in the methylome and transcriptome of what we see in KS males49. They found 11 differentially methylated positions (DMPs) on the two X chromosomes when comparing KS and female controls (corresponding to eight genes) and 168 autosomal DMPs between KS and control males (corresponding to 85 genes). In their RNA-seq expression profiling studies, they found 31 differentially expressed genes between KS and male controls. Perhaps most importantly was the discovery that there were 23 differentially expressed autosomal and seven differentially expressed X-chromosomal ncRNA genes between the KS and control males. ncRNA genes may be involved in X inactivation as well as neurodevelopment and cognition51. Their conclusion forecasts future research and exploration efforts to causally link exact gene expression/regulation of the altered phenotype of KS males: “in conclusion, our results demonstrate a unique epigenetic and genetic landscape in KS involving both the X chromosome and the autosomal chromosomes, with few correlations between methylation values and gene expression”47. Raznahanet al. added to this growing body of data by also reporting on sex-chromosome dosage effects on gene expression in hopes of teasing out why 47, XXY; 47, XXX; 47, XYY; 45, XO; and 48, XXYY are all phenotypically distinct52.In their beautiful 2018 report, Skakkebæk54. Some practitioners may employ replacement therapy just after the onset of puberty, typically in the form of gel, as compliance is higher54. Aromatase inhibitors may be used in males with gynecomastia.A fair amount of focus has been placed on the role of androgen replacement therapy in KS, as testosterone levels are known to be low or low normal. Although this may be an important component of management with regard to pubertal advances, it will not have any positive effects on spermatogenesis or fertility. However, some studies suggest that TRT does not appear to impact sperm retrieval rates in patients with KSAs a genetic condition that affects 1 in 600 live male births, KS is being seen commonly enough that it is important for practitioners to understand the wide spectrum of issues and the multidisciplinary approach that is required to appropriately treat these patients. It is also essential to take note of distinct features that are known to be associated with KS, as this allows providers the opportunity to recognize undiagnosed cases. Along these lines, there is increasing momentum in genomic studies that are beginning to shed light on the differential gene expression that may explain the KS phenotypes and the variability within.Despite ongoing investigations, there are still many essential questions, including whether hormone replacement therapy plays a beneficial role and what timing is appropriate for sperm retrieval for those interested in reproduction. The evidence to date suggests that hormone replacement therapy with testosterone may be valuable for peripubertal KS boys who appear to have delayed initiation of puberty and distinctly lack virilization. There is also a lack of data to support the advantage of TESE in the adolescent compared with the adult patient with KS. In fact, the counterargument, that optimal timing for TESE in the patient with KS is during early adulthood, is becoming more apparent.


The development of novel drugs against the influenza virus with high efficiencyand low toxicity is an urgent and important task. Previous reports havedemonstrated that compounds based on sulfo derivatives of oligo- andpolysaccharides possess high antiviral activity. In this study, we haveexamined the ability of a novel sulfonated derivative of β-cyclodextrin(KS-6469) to inhibit the influenza virus A/WSN/33 (H1N1) infection in vitro andin vivo. The antiviral potential of KS-6469 against the influenza virus wasevaluated in Madin-Darby Canine Kidney epithelial cells treated with seriallydiluted KS-6469. We found out that KS-6469 completely inhibited viralreproduction after treatment of the infected cells with the compound for 48 h.Our data show that double intranasal treatment of mice with KS-6469 fullyprotected the animals from a lethal infection and significantly decreased theviral titers in the lungs of the infected animals. Thus, the novel sulfonatedβ-cyclodextrin derivative KS-6469 is a promising candidate for thedevelopment of antiviral drugs for preventing and treating the influenzainfection. The influenza virus causes respiratory diseases that are responsible for thedeath of up to 650,000 people each year worldwide. The emeIn this study, we investigated the antiviral properties of a sulfonated(C-sulfate) oligosaccharide (a bisulfite derivative of oxidizedβ-cyclodextrin KS-6469) against the influenza A/WSN/33 (H1N1) (IAV) virusin vitro and in vivo. The results revealed that KS-6469 effectively inhibitsthe replication of IAV in MDCK cells by acting in the late stages of the viralinfection, and that it possesses virucidal properties. The results obtained ona mouse model of lethal influenza infection in vivo confirmed the highantiviral potential of this compound.Synthesis of the KS-6469 compoundThe KS-6469 compound was developed and synthesized at the A.E. Favorsky IrkutskInstitute of Chemistry (Siberian Branch of the Russian Academy of Sciences)based on commercially available β-cyclodextrin .A detailed description of the synthetic procedure and the physicochemicalproperties of KS-6469 will be published separately. Oseltamivir  was prepared as a solution in phosphate-buffered saline (PBS) fororal gavage administration.Cytotoxicity of KS-6469 in MDCK cellsThe MTT assay. The cytotoxicity of KS-6469 was evaluated usingthe 3--2,5-diphenyltetrazolium bromide (MTT) test asdescribed previously [14]. Madin-Darby Canine Kidney (MDCK) cells were receivedfrom the Bank of Cell Cultures .The cells were maintained in Iscove’s Modified Dulbecco’s Medium supplemented with 5% fetal bovine serum , 100units/L penicillin, 100 mg/mL streptomycin, and 0.25 mg/mL amphotericin at 37°C in a humidifiedatmosphere containing 5% CO2 (from here on referred to as“standard conditions”). Briefly, MDCK cells were seeded into96-well plates and grown to confluence under standard conditions. The mediumwas replaced with a fresh medium containing serially diluted KS-6469, and thecells were incubated for another 24 h under standard conditions. Aliquots ofthe MTT solution were added to each well, and incubation was continued for anadditional 3 h. The dark blue formazan crystals formed within live cells weresolubilized with dimethyl sulfoxide (DMSO); absorbance was measured at 570 nmin a MultiskanTM FC plate reader . TheIC50 was determined as the compound concentration required todecrease the A570 to 50% of the control  and wasdetermined by interpolation from the dose-response curves.Real-time cell analysis. The cytotoxic effects of KS-6469 onthe MDCK cells were measured on an xCELLigence real-time cell analyzer , which is based on the microelectronic biosensortechnology. The electrode impedance displayed as the cell index (CI) value wasmeasured to compare the status of treated and untreated cells. The cells wereplated in 16-well plates  in IMDM supplemented with10% FBS and a antibiotic-antimycotic solution and incubated overnight (20 h)under standard conditions. The growth medium was then removed, the cells werewashed with PBS, and 150 μL of IMDM either containing differentconcentrations of KS-6469 or without the compound was added to each well. Thecells were then incubated under standard conditions for 65 h with cellviability monitored every 15 min using the xCELLigence real-time cell analyzer.The points were run in duplicate, and the IC50 value was calculatedusing the xCELLigence real-time cell analyzer software. The concentration ofKS-6469 at which the diagram of CI for the treated cells coincided with that ofCI for the untreated cells was defined as the maximum tolerated concentration(MTC).Determination of the antiviral activity of KS-6469 in MDCK cellsThe influenza virus strain A/WSN/33 (H1N1) was obtained from the IvanovskyInstitute of Virology . MDCK cells were grown to confluence in24-well plates under standard conditions. The cells were then infected with IAVat MOI 0.1 in a medium supplemented with KS-6469 (70–600 μg/mL) andincubated at 37°C for 24 or 48 h under standard conditions. Twenty-four or48 h post-infection (p.i.), the cells were subjected to one freeze/thaw cycle(-20/20°C) and then the viral titer was determined by FFA as describedpreviously . The indc – To)/Tc × 100%,KI =  for 1 h under standard conditions. The KS-6469compound was added at a concentration of 5 mg/mL before, during, or after theIAV infection. After each period of incubation with the virus, the cells werewashed with PBS and incubated with a fresh infection medium at 37°C.Forty-eight h p.i., the cells were subjected to one freeze/thaw cycle and theviral titer was determined by FFA.The NA-Fluor™ Influenza Neuraminidase Assay Neuraminidase (NA) activity was measured using the NA-Fluor™ InfluenzaNeuraminidase Assay kit , according to themanufacturer’s protocol. The assay is based on the NA enzyme cleaving the2’-(4-methylumbelliferyl)-α- D-N-acetylneuraminic acid (MUNANA)substrate to release the fluorescent product, 4-methylumbelliferone. Thefluorescent signal was measured using a CLARIOstar® fluorescence platereader . The assays were performed in triplicate.Hemagglutination assayThe antiviral activity of KS-64649 was estimated using hemagglutination assay(HA). Viral suspensions were incubated with an equal volume of the medium withor without KS-6469. In this assay, a 0.5% suspension of chicken erythrocyteswas used. The viral titer of the sample was calculated as the inverse value ofthe dilution at which the last agglutinated appearance was detected.Analysis of the cholesterol level in the viral envelopeThe cholesterol level in the viral envelopes was determined using the AmplexRed Cholesterol assay kit , according to themanufacturer’s instructions. Briefly, the viruses were incubated withdifferent concentrations of KS-646 for 6 h at 37°C then pelleted andre-suspended in Amplex Red reaction buffer. Methyl-β-cyclodextrin was usedas a positive control. Fluorescence was then analyzed on a CLARIOstar®plate reader at an excitation wavelength of 550 nm and an emission wavelengthof 590 nm. The assays were performed in triplicate.Virucidal activity of KS-6469Viral suspensions were incubated with an equal volume of the medium with orwithout KS-6469 for 3 or 6 h at 4, 20 or 37°C. The viral titer wasestimated using the FFA. The assays were performed in triplicate.Enzyme-linked immunosorbent assay Enzyme-linked immunosorbent assays were performed using commercially availablemonoclonal antibodies against influenza virus hemagglutinin [IVC102], [1.B.408]and [B219M] (Abcam) as described previously [18].Antiviral activity of KS-6469 on the mouse model of lethal influenzainfection Animals. Female 4- to 6-week-old BALB/c mice werepurchased from the State Research Center of Virology and Biotechnology Vector. The animals were kept in the vivarium of theInstitute of Chemical Biology and Fundamental Medicine, SB RAS, with a naturallight regime on a standard diet for laboratory animals (GOST [State Standard] R5025892) in compliance with the international recommendations of the EuropeanConvention for the Protection of Vertebrate Animals used for Experimental andOther Scientific Purposes [Purposes , as wellIn vivo toxicity analysis. To evaluate thetoxicity of KS-6469, the mice were treated by intraperitoneal or intranasaladministration of varying amounts of the compound. Each of the experimentalgroups contained six BALB/c mice. The animals received KS-6469 in differentdoses intraperitoneally in 0.2 mL of PBS, or 250 mg/kg twice daily intranasallyin 40 μL of PBS. The mice from the control groups received the same volumeof PBS. After the treatment, signs of intoxication including general condition,weight, and depression of the central nervous system were assessed daily for 14days.50 being equal to ~1.2 × 104± 0.7 × 104 FFU of IAV immediately after intranasaladministration of KS-6469 at a dose of 250 mg/kg. The following day, the micereceived a second dose of KS-6469 intranasally. The study included a positivecontrol group that received oseltamivir daily (7.5 mg/ kg) for 5 days. Thecontrol mice received PBS instead of KS-6469. The animals were inspected dailyand weighed for 14 days after infection. The degree of protection was estimatedaccording to the reduction in the mortality rate. To assess the viral titer inthe lungs, BALB/c mice were divided into groups of six animals. The mice wereinfected intranasally with 3 LD50 of IAV immediately afterintranasal administration of KS-6469 at various doses. The following day, themice received a second dose of KS-6469 intranasally. The control mice receivedPBS instead of KS-6469. The mice in each group were euthanized on day 3 p.i.,and the lung tissue was harvested. The lungs were weighed and the medium wasadded at a ratio between the lung tissue and the medium of 1 : 10 (v/v). Thehomogenates were prepared using a Sonopuls HD 2070 ultrasonic homogenizer. The viral titers of the lung homogenates were determinedusing FFA.Protective efficacy in mice. To test the protective efficacy of KS-6469, weevaluated weight changes, the survival rate, and viral titer in the lungs ofthe infected mice. BALB/c mice with an average weight of 14 to 16 g weredivided into groups of six animals. The mice were anesthetized byintraperitoneal injection of tribromoethanol (Avertin®) and infectedintranasally with 3 LDStatistical analysisThe data are expressed as the mean ± SD. The statistical analysis wasperformed using the two-tailed unpaired t-test. P-values of less than 0.05 weredeemed statistically significant.Characterization of KS-6469The compound KS-6469 is a bisulfite derivative of oxidized β-cyclodextrin.The sulfonate moieties of bisulfite derivatives are bound directly to a carbonatom with the general formula (Gluox)7-Cx-(SO3Na)x, where (Gluox)7-Cx isoxidized β-cyclodextrin, x is the number of sulfonated carbon atoms, andSO3R is the bisulfite (sulfonate) group. The detailed structure andphysicochemical properties of KS-6469 are the subject of ongoing research andwill be published later.Cytotoxicity of KS-646950) value  was 15 ± 3.3 mg/mL, as determinedby MTT assay after 24 h incubation with the compound. An analysis ofthe cytotoxic effect of KS-6469 on the cells following 65 h exposure to KS-6469was measured using real-time cell analysis. The IC50 value obtainedby this method was found to be 8.9 ± 1.3 mg/mL after 65 h exposure.These results confirmed the low cytotoxicity of KS-6469 toward eukaryoticcells, which reached 15 and 8.9 mg/mL for the 24 and 65 h exposure durations,respectively. Based on the data from these experiments, we can conclude thatKS-6469 possesses low toxicity toward MDCK cells.The cytotoxicity of KS-6469 toward the MDCK cells was examined. The halfmaximal inhibitory concentration . Virus quantificationshowed tTo gain insight into the mechanism of KS-6469 inhibition of IAV replication, wedetermined the step of the viral life cycle that was affected by the compoundusing time-of-addition assay. First, we investigated whether KS-6469 added tothe cells prior to virus adsorption could protect the cells against infection.The cells were treated with KS-6469 2 h prior to IAV infection ((-3) to (-1) h).The virEffect of KS-6469 on the functional activity of influenza virus surfaceproteins.Functional balance between influenza virus hemagglutinin (HA) and neuraminidase(NA) was found to be very important for successful viral replication andfitness. In order to determine whether KS-6469 affected the structure of viralhemagglutinin, we investigated virus attachment to erythrocytes. The influenzavirus was treated with KS-6469 (5 mg/mL) for 1 or 6 h at 37 or 4°C, andthe viral HA titer was evaluated using HA assayThe ability of KS-6469 to affect the structure of the surface epitope of viralhemagglutinin was analyzed using a set of monoclonal antibodies specific tohemagglutinin of IAV. The treatment of IAV with KS-6469 (5 mg/mL) at 37°C,which resulted in a 16-fold decrease in HA titer, changed neither the structureof virus hemagglutinin nor the efficacy of its binding to specific monoclonalantibodies .It shouIn order to gain an understanding of the mechanism by which K-6469 inhibitsviral replication, we examined the inhibition of viral neuraminidase (NA),which mediates the release of viral progeny from infected cells and promotesviral transmission. The resultsindicateVirucidal properties of KS-6469It has been previously shown that sulfated polysaccharides possess virucidal activity, 22.In Antiviral activity of KS-6469 in the mouse model of influenzaIn vivo cytotoxicity of KS-6469. BALB/c mice weresubjected to a single intraperitoneal injection of different doses of KS-6469(250 mg/kg or 500 mg/kg) in PBS. The physical parameters and body weights ofthe animals were monitored daily,starting 30 min prior to viral challenge. On day 3 p.i., six animals per groupwere sacrificed and lung tissue extracts were analyzed by FFA and NA inhibitors, 24.VacOur study has shown that KS-6469 is a low-toxicity and safe agent thateffectively inhibits the development of the infectious disease caused by theinfluenza virus. Intranasal application of KS-6469 had anti-IAV effectscomparable to those of oseltamivir. Hence, KS- 6469 is a promising candidatefor developing an effective antiviral drug for the prevention and treatment ofinfluenza infections.


The VFA-to-SFA ratio is the most predictable body composition model for use in predicting the incidence of SSI after gastrectomy.Recent studies have shown that body composition is an important factor that affects surgical site infection (SSI). However, each study has utilized different body composition criteria. Therefore, in this study, we aim to determine the most predictable body composition criteria for the prediction of SSI after gastrectomy. The visceral fat area (VFA), subcutaneous fat area (SFA), and muscle area were assessed by a preoperative-stage computed tomographic (CT) scan. To compare the predictive performance of body composition for SSI, logistic regression models were used, and the models were compared using the receiver operation characteristic (ROC) curve and the area under the curve (AUC) value. Of the 1038 eligible patients, 58 patients (5.6%) developed SSI. The VFA-to-SFA ratio showed the best predictive performance (mean AUC 75.11). The cutoff value for the SSI of the VFA-to-SFA ratio was 0.94, and the sensitivity and specificity were 67.86% and 77.65%, respectively. A multivariate logistic analysis indicated that a total gastrectomy (OR, 2.13;  Gastric cancer is the fifth most common cancer in the world and the third most common cause of cancer-related death . SurgerySurgical site infection (SSI) is one of the most common postoperative complications of gastrectomy. SSI is associated with prolonged hospitalization and increased mortality . TherefoRecently, the effects of different body compositions on SSI have been attracting attention. Especially, the effects of fat distribution and loss of skeletal muscle, called sarcopenia, have been focused on. Some studies have shown that visceral fat is a risk factor for postoperative complications in digestive surgery ,7,8. SarBetween January 2015 and December 2015, 1067 patients underwent gastrectomy for gastric cancer in the Severance Hospital in Seoul, Korea. We the excluded patients who required emergency surgery, those with active infections, and those for whom a preoperative CT scan was unavailable. Consequently, we enrolled 1038 patients who underwent gastrectomy and who did not qualify for exclusion. A schematic flow of the study is shown in 2/m2 as the visceral fat index and muscle index, respectively. Examples of the areas of VFA and SFA are depicted in The visceral fat area (VFA), subcutaneous fat area (SFA), and total abdominal muscle area (TAMA) of the patients were assessed using the latest preoperative CT scans. The cross-sectional VFA, SFA, and TAMA values were measured at the level of the third lumbar vertebra, as previously described . SpecifiSurgeons checked for SSI every day during the patients' in-hospital stays, and at every outpatient clinic or emergency room visit until 30 days after surgery . PatientTypical surgical procedures for open, laparoscopic, and robotic gastrectomy have been previously described in detail ,18,19,20p < 0.05 in the univariate analyses, we selected variables for multivariate logistic regression based on the clinical significance known by previous studies [p-value of 0.004. In order to compare the predictive performance of body composition for the SSI, logistic regression models were used, and the models were compared using the receiver operation characteristic (ROC) curve and the area under the curve (AUC) value. The AUC value was extracted by using the standard error estimated from the bootstrap samples. As a method of comparing the predictive performance of the models, we used the bootstrap samples to show the confidence interval for the difference, and we used the DeLong method to compare with the control. The consistency between the predicted and the observed probabilities of the actual data was assessed using the Hosmer–Lemeshow test. The cutoff value, the AUC value, and the sensitivity and specificity of the selected model were determined using the Youden method, and the odds ratio (OR) was obtained by multivariate logistic regression analysis. p-values of <0.05 were considered statistically significant. All of the statistical analyses were performed using the R package, version 3.4.4 .To verify the normality of the continuous variables, the Kolmogorov–Smirnov test and Shapiro–Wilk test were conducted. The independent t-test and the Mann–Whitney test were used to compare the continuous variables of the two groups. The Chi-square test or the Fisher exact test was used to compare the categorical variables. Among variables with  studies ,24,25,26The clinical and surgical characteristics of the 1038 patients who had undergone gastrectomy are shown in p = 0.017), stage III or IV cancer , and a high VFA-to-SFA ratio  were independent risk factors for SSI after gastrectomy. The effects of the interacting variables are presented separately in In the univariate logistic regression analysis, male sex, smoking, an ASA score ≥3, neoadjuvant chemotherapy, stage III or IV cancer, total gastrectomy, open surgery, D2 or more lymph node dissection, and longer operation times were significantly associated with SSI. After checking the multicollinearity, gender, smoking history, surgical method, extent of surgery, and staging, and one of the body composition criteria were selected for multivariate logistic regression analysis based on the clinical significance. The results of the logistic regression analysis, including the VFA-to-SFA ratio, are shown in p-value of 0.3812. The superiority of the VFA-to-SFA ratio to the control was identified using the DeLong method using a p-value of 0.022. These results were also applied to the organ/space SSI, and the results about organ/space SSI are shown in Using the ROC curve and the AUC value, the body composition criteria variable with the highest predictive power for SSI was determined. As shown in The cutoff point of the VFA-to-SFA ratio was 0.94, using the Youden method to improve the predictive power of the SSI. To compare the performance of Model M1-6-1 using the VFA-to-SFA ratio as a continuous variable, and Model M1-6-2 using the VFA-to-SFA ratio as a binary variable with a cutoff point of 0.94, we used 1000 bootstrap samples, and the mean AUC value was obtained and the results are shown in p < 0.001). This was mainly due to the organ/space SSI . In the high VFA-to-SFA ratio group, the incidence rate of a Clavien-Dindo score of IIIa or a higher for postoperative complication, mean days of postoperative hospital stay, and the incidence rate of re-admission within 30 days were significantly higher than in the low VFA-to-SFA ratio group. However, there was no difference in the mortality rate between the two groups . This study demonstrated that the VFA-to-SFA ratio had the highest predictive power for SSI after gastrectomy among the body composition criteria assessed, especially the organ/space SSI. Moreover, the VFA-to-SFA ratio could be widely applicable, because it is a value defined by a ratio, not an absolute value. The large number of patients who underwent gastrectomy in our study increased the clinical significance of this result. 2, and reported that SSI occurred more frequently in the high VFA group [A high VFA-to-SFA ratio indicates either an increased VFA or a decreased SFA. As the area of the abdominal fat and muscle is obtained through the CT scan, several studies have shown that VFA is better than BMI at predicting postoperative complications, including SSI. One reason patients with visceral obesity are more likely to develop an SSI could be due to the surgical difficulty associated with the surgeon having a deeper and poorer view of the surgical field, as well as the fragile, easily bleeding tissue in high-visceral obesity patients . In addiFA group . However2/m2 for men and ≤38.5 cm2/m2 for women, and a visceral fat area of ≥100 cm2 in both sexes, is an independent risk factor for the development of SSI after laparoscopic total gastrectomy [Conversely, SFA improves insulin sensitivity. Thus, it can work as a buffer against the lipid accumulation of VFA. Consequently, SFA insufficiency causes increased lipid accumulation in visceral fat, and therefore the VFA-to-SFA ratio is effective for evaluating the risk of diseases affected by the adipose tissue . For thitrectomy . HoweverRecently, some studies have been published that address the risk of postoperative complications, such as SSI, by values calculated by a ratio rather than a specific value based on body composition criteria. According to Pecorelli et al., the high VFA-to-TAMA ratio was a major risk factor for postoperative complications, especially in pancreatic cancer patients ,34. HoweIn addition to the individual fat distribution, total gastrectomy and stage III or IV cancer also influenced the occurrence of SSI. Total gastrectomy is one of the most invasive gastrointestinal surgeries, and the risk of SSI after a total gastrectomy is known to be higher than after a partial gastrectomy ,36. ThusThis study had several limitations. First, this study was a nonrandomized retrospective study of a single country with a homogenous ethnic background. Second, we excluded 10 patients with follow-up loss or incomplete data review of the 1067 patients who underwent a gastrectomy in 2015. Although these patients did not develop SSI during the hospitalization, and the rate of SSI occurrence after discharge was low , we could not completely rule out the possibility that SSI occurred after discharge in these patients. Third, active post-discharge surveillance using phone calls or family physicians has not been conducted. Thus, there was a potential for some SSI incidents to be missed. Fourth, as a result of the model comparison, the 95% confidence interval of the AUC value of each model overlapped. Fifth, in the present study, the incidence of SSI after gastrectomy was 5.6%, which is not high. Therefore, this raises the issue that the sensitivity and PPV were low for predicting the increased risk of SSI with a specific cutoff value. However, SSI is a complication that cannot be accurately predicted before surgery, so it is meaningful to find patients who are at high risk for SSI before surgery. Surgeons may consider minimal invasive surgical methods and try to reduce the operation time so as to reduce SSI in patients with a high VFA-to-SFA ratio. In this regard, even though the sensitivity and PPV were low, the specific cutoff value we determined would be beneficial. In addition, the high NPV of the VFA-to-SFA ratio cutoff value allows surgeons to identify patients at a very low risk of developing SSI before surgery. Further research is needed to determine if it will be applicable in other abdominal surgeries and other countries.A high VFA-to-SFA ratio is an independent risk factor for the development of SSI after gastrectomy. Also, the results of this study suggest that the VFA-to-SFA ratio is the most predictable body composition model to predict the occurrence of SSI after gastrectomy. It would be beneficial for surgeons to pay greater attention to SSI when operating on patients with high VFA-to-SFA ratios.


Our studies reveal a continuous morphology of hybrid nanofibers with the mean fiber diameter of 189 nm ± 86 nm, which demonstrate a high thermal stability. Finally, we describe a protocol for the neutralization of nanofibers, which enabled us to achieve their structural stability in phosphate-buffered saline (PBS) for more than six months, as confirmed by microscopy and FTIR measurements. The formed hybrid nanofibers exhibit unique physicochemical properties essential for the development of future tissue engineering platforms.We report a methodology for the fabrication of neutralized chitosan-based nanofiber matrices decorated with bare Au nanoparticles, which demonstrate stable characteristics even after prolonged contact with a biological environment. The methodology consists of electrospinning of a mixture of bare (ligand-free) laser-synthesized Au nanoparticles (AuNPs) and solutions of chitosan/polyethylene oxide (ratio 1/3) containing chitosan of a relatively high molecular weight (200 kDa) and concentration of 3% ( The elaboration of artificial scaffold platforms, capable of replacing and/or repairing failing tissues or full organs, is still challenging for the scientific community ,2,3,4. M2 groups on a chitosan nanofiber surface make it suitable for the immobilization of enzymes and negatively charged proteins. Moreover, a highly reactive chitosan surface can offer additional opportunities for its functionalization with nano-engineered particles to improve physicochemical characteristics  and enable biological  or other theranostic functionalities [Chitosan is one of the most extensively exploited naturally derived biopolymers, which is widely used to develop novel electrospun nanofibrous matrices for a variety of biomedical applications, including anti-microbial, antifungal, wound healing, drug, and gene delivery ,16,17,18nalities . Howevernalities ,21,22. Hnalities . The empnalities .w/v of chitosan, ratio 1:4), together with nanoparticles prepared by methods of laser ablation, as described in our previous studies [3+ [Various chitosan/PEO formulations have been tested at different ratios of chitosan/PEO to obtain homogenous electrospun nanofibers with different characteristics ,24,25,26 studies ,34,35,36 studies ,33,34,35 studies , and Sur studies , the incdies [3+ ,37,38.w/v) using 10% (v/v) acetic acid with a chitosan/PEO ratio of 1:3 in the electrospinning solution, which renders possible the solution of the dissolubility problem. Moreover, we describe a simple protocol of neutralization of NH3+ in the alkaline/alcoholic solution of 5 M NaOH or 1 M K2CO3 to ensure the structural stability of nanofiber membranes in biological media. Novel formulations of chitosan/PEO functionalized with bare laser-synthesized AuNPs look very promising for tissue engineering applications.In this article, we report a further advancement of the electrospinning procedure in order to improve the properties of the formed AuNPs-decorated hybrid chitosan/PEO nanofibers. Here, we show the possibility for increasing concentration of 200 kDa chitosan up to 3% , while NaOH was purchased from VWR International GmbH .A medium molecular weight chitosan (200 kDa) and a PEO powder with the molecular weight of 300 kDa were purchased from Sigma-Aldrich . A target (1 cm × 1 cm) of high purity Au (99.999%) was purchased from GoodFellow  to fabricate bare AuNPs. Chitosan was solubilized using acetic acid (99–100%) purchased from SCS GmbH . For the neutralization stage, methanol (99.9%), ethanol (99.5%), and KBare laser-synthesized AuNPs were prepared using methods of femtosecond ablation and fragmentation in deionized water ,30,31,33w/v) using 10% (v/v) acetic acid. In a separate cuvette, we prepared 4 mL of PEO with a concentration of 8% (w/v). The solutions were then mixed in the ratio of 1:3, respectively. A concentrated AuNPs solution was added to the polymer solution and ultra-sonicated for 2 h, which ensured the removal of bubbles and a homogenous dispersion of nanoparticles. Concentration of bare laser ablated AuNPs was measured to be 0.15 g L−1. The nanoparticle solution was concentrated before the addition to the electrospinning solution by heating it at 30 °C in an oven for several hours. Such a procedure led to 3-decreasing of the solution volume. A total of 2 mL of the concentrated solution 0.45 g L−1 was used to functionalize the electrospinning polymer solutions. This sample was denoted as Ch-AuNPs. A second sample of nanofibers without AuNPs (Ch-0) was prepared as a reference for comparative studies. In this case, 2 mL of AuNP solutions were replaced by 2 mL of PEO. The procedure was carried out in such a way that the final concentration of the polymer was the same as in the main sample . All steps of the preparation protocol are presented in A total of 2 mL of medium molecular weight chitosan was dissolved at a concentration of 3% (−1. The pump was connected via a Leur-Lock and PTFE (Polytetrafluoroethylene) tube with a blunt 0.8 mm-diameter needle at one end of a spinneret, where 14 kV was applied to the solution. The distance between the collector and spinneret was kept at 15 cm based on previous optimization studies. The collector had a negative voltage of −1 kV and was wrapped in aluminium foil to collect the fibers. Climate conditions were fixed for all experiments at 25 °C temperature and 50% of humidity.The fabrication of nanofibers was done using specialized equipment purchased from IME medical electrospinning technologies. A climate-controlled setup was employed to ensure constant environmental conditions and fixed parameters during the process. An electrospinning setup contained a rotating cylinder set at 2300 rpm. Electrospinning solutions were transferred into a 5 mL syringe, which was connected to a programmable pump providing the constant feed rate of 0.3 mL h® software. JSM-IT100 InTouchScope™ system  operating at 5−20 kV accelerating voltages was the main microscope used at lower magnifications. A DSM 982 Gemini Zeiss system  operating at accelerating voltage of 20 kV was used to produce higher magnification micrographs. Both SEM systems were coupled to energy dispersive X-ray analysis. Chemical characteristics were evaluated by Fourier transform infrared (FTIR) spectroscopy  equipped with a universal attenuated total reflection (ATR) sampling accessory. The measurement histogram was performed by analyzing 2 micrographs; one from center of the collector, where most of the fiber deposition takes place, and another from the edge of the collector. The frequency distribution, as obtained using integrating ImageJ results with the origin, ranged from 100 to 15,000.A high-resolution transmission electron microscopy (HR-TEM) system  was used to characterize laser-synthesized AuNPs. Thanks to ImageJ software , Bethesda, MA, USA), statistic measurements were performed on more than 200 AuNPs to determine diameter size distribution. A scanning electron microscope was used as a primary analysis method to verify if the electrospinning process led to the formation of nanofibers. Due to the limited charging effect on samples, it was unnecessary to proceed to the sputter coating step. Size distributions and morphology of fibers were characterized using scanning electron microscopy (SEM), and a subsequent treatment of images using ImageJ−1, preventing the combustion of the samples, and heated at the rate of 10 °C min−1. Similarly, DSC was carried out under a nitrogen environment with a constant flow rate of 21 mL min−1 and heating rate was kept at 5 °C min−1. The operation temperature was between 30 and 700 °C during TGA studies and between 30 and 180 °C during DSC analysis.Thermal properties of nanofibers were studied with the help of thermogravimetric analysis  and differential scanning calorimetry . A total of 7 mg and 5 mg of nanofibers were used for each TGA and DSC measurement, respectively. A TGA heating chamber was flushed with nitrogen at 20 mL min2CO3, dissolved in 15 mL of ethanol (70%), and 5M NaOH dissolved in 15 mL of methanol (70%). The nanofibers were immersed in the solutions for a total of 20 h at ambient conditions. The immersion process was repeated 3 times by changing solutions after every immersion step to ensure effective neutralization. The samples were then washed with distilled water and dried at room temperature for 24 h before SEM-EDX and FTIR analyses to assess their structural stability and chemical composition.A total of two protocols were investigated based on two solutions of 1M Kw/v) concentration of chitosan is used at variable chitosan/PEO ratios. Recently, chitosan/PEO nanofibers functionalized with bare laser synthesized NPs were successfully electrospun [w/v) to 3% (w/v) and the chitosan/PEO ratio was increased from 1:4 to 1:3. Moreover, the concentration of acetic acid used to dissolve chitosan was also decreased down to 10% (v/v), as compared to highly acidic 90% (v/v) solutions used in our previous study [As mentioned above, the electrospinning of pure chitosan is quite challenging due to its poor solubility and short chain length. In most cases, 1% . A typical HR-TEM image of AuNPs formed by laser ablation and corresponding AuNPs size distribution is given in Such initial composition of chitosan/PEO was thus used to prepare a novel solution containing highly concentrated AuNPs (0.09 mg mL® software and fitted with a Gaussian approximation (blue curve) (3+ groups and negatively charged (–23.1 ± 2.61 mV) Au nanoparticles [SEM-EDX examinations of chitosan/PEO nanofibers functionalized with AuNPs revealed a homogenous network of cylindrical nanofibers without any beads. All of these results are consistent with conclusions of our previous study , which ee curve) . AuNPs warticles ,31,32,33articles ,19. It iIn our tests, we used TGA and DSC analyses to examine thermal properties of formed nanofibers and the impact of AuNPs on them. A TGA curve a revealeSubsequently, DSC was carried out to record phase transition changes, which could take place e.g., at nanofiber melting points after their functionalization with gold nanoparticles. Here, both types of nanofibers displayed classical DSC curves. As shown in 2 and OH, stretching from 3500–3000 cm−1. A peak for amide absorption could also be observed at 1570 cm−1 for both functionalized and reference nanofibers [2 stretching in PEO could also be observed at 2885 cm−1. The spectra beyond 1500 seems to be dominated with intense PEO bands at 1145, 1095, and 1059 cm−1, complimentary to C-O-C stretching vibrations [The chemical composition of nanofibers was analyzed using FTIR before and after the functionalization. As shown in nofibers ,40,41. Abrations . The fun3+ group can lead to a rapid dissolution of chitosan/PEO complexes, making them hardly useful for applications in biological systems [3+ group into the NH2 insoluble form: (i) 1M K2CO3 dissolved in ethanol (70%) and (ii) 5M NaOH in methanol (70%). After the treatment, the samples were washed in distilled water and immersed in phosphate-buffered saline (PBS) for 24 h to test the neutralization efficacy. For the first method employing 1M K2CO3, the overall structure of the treated nanofibers had a non-woven mat-like morphology , and functionalized these nanofibers with bare laser-synthesized AuNPs. Thanks to systematic physicochemical analysis, improvements on the structural characteristics of the fibers were clearly demonstrated compared to previously reported nanofiber matrices [In summary, we prepared novel formulations of hybrid chitosan/PEO nanofibers, with the ratio of 1:3 at a relatively high concentration of chitosan 3% (matrices ,19. Firsmatrices ,19. Ther


Since the rapid development of theoretical progress on the two-dimensional graphyne nanoribbons and nanojunctions, here we investigate the electronic band structures and transport properties for the junctions based on armchair-edged γ-graphyne nanoribbons (AγGYNRs) with asymmetrically nitrogen (N)-substituting in the central carbon hexagon. By employing first-principles calculation, our computational results imply that the number and the location of single or double N-doping can efficiently modulate the electronic energy band, and the N-doping hexagonal rings in the middle of the junction play a vital role in the charge transport. In specific, the effect of negative difference resistance (NDR) is observed, in which possesses the biggest peak to valley ratio reaching up to 36.8. Interestingly, the N-doped junction with longer molecular chain in the central scattering region can induce a more obvious NDR behavior. The explanation of the mechanism in the microscopic level has suggested that the asymmetrically N-doped junction by introducing a longer molecular chain can produce a more notable pulse-like current-voltage dependence due to the presence of a transporting channel within the bias window under a higher bias voltage. In addition, when the spin injection is considered, an intriguing rectifying effect in combination with NDR is available, which is expected to be applied in future spintronic devices. Several two-dimensional (2D) carbon materials have been demonstrated as the potential candidate for spintronic devices –5. RecenAs we know, research works show that the graphene nanoribbons can be tailored and cut into many structures as molecular devices in experiment , 30. Simn-doping or p-doping semiconductors. Thus, it is essential to consider single or double N-doping in our proposed junctions of AγGYNRs here.The γ-graphyne nanoribbon (γGYNR) , which c2 and double N-doping of M6 can induce the spin splitting of energy band. The transport current of 3-AγGYNR junction without N-doping is weakened as the number of repeated units in the scattering region increases; in contrast, the currents are intensified with a longer molecular chain for 3-AγGYNR junctions with single or double N-substituting positions. Interestingly, the rectification and obvious NDR effects are observed in the N-doping junctions of M2 and M6. Such behaviors generate from the different coupling between two electrodes and the scattering area. In order to explain the mechanism of NDR behavior in a microscopic level, the reason displayed that the longer molecular chain contained in the asymmetrically N-doped junctions can induce a more obvious pulse-like current-voltage dependence due to the existence of an opened transporting channel within the corresponding bias window under higher bias. Additionally, the hexagonal ring with N-substituting positions has a vital impact in the transport process.Motived to deeply understand the spin electronic and transport properties of several length-adjustable molecular chain sandwiched between two semi-infinite AγGYNRs with different N-substituting positions, we have finished the computational work by using first-principles calculation in combination with a Landauer-Büttiker approach in this paper. The results of theoretical simulation suggest that the N-doping can efficiently reduce the energy gap of 3-AγGYNR junctions, then the single N-doping of MThe paper is divided as follows: In the “1 in the second picture of the bottom panel in Fig. 6 ring are numbered as 1, 2, 3, 4, and 5 as pointed under the corresponding atoms of the red frame, respectively. Similarly, the pristine device without N-doping is called as M0, where the models with two-typical single N-substituting positions (replacing the atomic positions of 1 or 2) are M1 and M2, and the ones with five-typical double N-doping at different substituting positions  are named as M3–M7, respectively. The red shaded part enclosed by a dashed rectangular box in the main view of Fig. 1 with the single N-instituting position of 1 including a molecular chain of four repeated molecular units as D should be call for M1D.The molecular wires consisting of 1~4 repeated molecular units, which are made of one benzene and one acetenyl without or with N-doping, are shown in the middle panel of Fig. −1. To perform the calculations of electronic structure, a k-point grid of 1 × 1 × 15 Monkhorst-Pack in Brillouin zone is adopted. The Monkhorst-Pack mesh of reciprocal space sampling for the spin-dependent transport calculation is 1, 1, and 100 in the x, y, and z directions, respectively, and the cut-off energy is adopted to 150 Ry. The double-ζ polarized basis is set to all elements including C, H, and N. Furthermore, the convergence criterion for total energy is set to 10−5 eV. Since the transport direction is set to the z axis, an interlayer vacuum distance of ~ 25 Å is used to avoid interactions between the periodic images [E) and bias voltage (V) is defined asGR(A) is the retarded (advanced) Green’s function of the central scattering area and ГL(R) is the coupling matrix of the left (right) electrode. The spin transport current is calculated by using the Landauer-Büttiker formula [μL/R = EF ± eV/2 is the electrochemical potential in terms of the Fermi energy (EF) of the material common to both leads under an external V, and the Fermi-Dirac distribution function is We firstly optimize all the structures of unit cells and molecular junctions by implementing the density functional theory calculation in the Atomistix ToolKit (ATK) package –48, 58 Ac images , 64. The formula –68\docuz direction of the ribbon axis is considered. For the convenience of contrast observation, we show all the unit cells in the form of illustrations for M0–M7 in Fig. 6 rings with N-doping are enclosed in a blue dashed frame with a blue shaded area, in which the projected density of state has also been calculated and shown in the right panels of Fig. To perform the practical electronic band structure calculations, the periodic super unit cell with N-doping along the 0 displays that the AγGYNR is a semiconductor with a direct energy gap of 1.16 eV. The lowest conduction band and the highest valence band originate from π* and π sub-band, respectively [1 and M2 with single N-instituting position in Fig. 2 is spin splitting. The inclusion of single N-doping narrows the energy gap at the Brillouin zone boundaries. As a result, the band structures for M1 and M2 behave metal property. When the unit cell of system doped with double N-doping for M3–M7 in Figs. 3, M4, and M7 have been narrowed into 0.06, 0.04, and 0.44 eV due to the using of dopant in the pristine structure, which images that they are still semiconductors after double N-doping. However, we can find that the band structures of M5 and M6 perform metallicity with zero band gap in Fig. 6 in Fig. 6 ring of AγGYNR. As shown later, the central part of the C6 ring indeed influences the conduction properties of AγGYNRs reported in our present work.Firstly, we investigate the structural and electronic characteristics of AγGYNRs. As shown in Fig. ectively , 69. But6 ring in our proposal structure.To deeply illustrate the impact of N-instituting position, the spin-dependent electrons on N atoms can be identified from the spin density distribution of the AγGYNRs . When the applied voltage is larger than 1.2 V, we can find out that the longer the molecular chain is, the current is weaker, implying that the molecular chain could impede the hopping of electrons from the left to right electrodes. The corresponding explanation is displayed in Fig. EF since the current is largely contributed by the transmission peak [0A tracks several peaks around the Fermi level; on the contrary, the transmission peak becomes lower and lower from M0A to M0D with the increasing length of the molecular link. For clarity, the inset of Fig. 0C and M0D refers to account for the reduction of current. Indeed, the pristine AγGYNR is not a perfect electrode to construct a spin (electronic) junction; the issue of N-instituting position is needed to be considered here.To further understand the transport properties of these two-probe junctions, we compute the ion peak , 20. The1) or 2 (M2), respectively, the opposite situation occurs, and we notice that all the currents are enhanced in Fig. V ≤ 1.2 V, and it happens to decrease with the increase of bias for device M1A–M1D in Fig. I-V curve displayed that the NDR effect is also found for M2B in Fig. When the devices are doped with single N atom by position 1  and the blue line (for M2D) are larger than the ones of the black line (for M0D). Seen from Fig. 1D extends to the energy range of − 0.26 eV ≤ E ≤ 0.8 eV, contributing to the electron flowing through the central scattering region. There exists a sharp transport peak around the Fermi level for M2D (the blue line) which is little lower than the former one; as a result, a relative weaker current curve appears. Certainly, zero transport gap for M0D  in Fig. 6, we plot all the I-V characteristics from M6A to M6D, finding that the NDR effect begins to strengthen with the increase of length for molecular chain. The PVR can increase from 1.7 for M6A to 5.4 for M6B, then a PVR maximum of 24.5 can be reached for M6D from the value of 12.8 for M6C. Note that the length of the molecular chain can efficiently modulate the occurrence and intensity of NDR behavior.As explained before, all the transmission spectra of three junctions hold many transmission peaks near the Fermi level  behaves a crucial action in Fig. 6D with the double N-instituting positions. It could be an outstanding candidate for a spin-switch of the nanoelectronic device based on AγGYNRs in the future. Therefore, the generation and transport features of spin-polarized currents are still vital issues for spintronics devices [There are many NDR effects that have been observed in our proposed models; to better interpret the foundation of NDR, we draw the relative diagrams in Fig.  devices .Fig. 8(6 ring of nanoribbons. We can obtain some semiconducting nanoribbons with narrow band gap or conductors of AγGYNRs. With regard to the transport properties, the different lengths of molecular chains induce interesting negative difference resistance behavior which has been expected for nanoelectronic junctions. In particular, the hexagonal rings in the middle of nanoribbons hold a vital role in the transport properties. The longer the molecular chain is, the more obvious NDR effect can be observed in the junctions including N-instituting positions. For the crucial N-doping for junctions M2D and M6D, the spin-polarized currents with the maximums of rectification ratio and peak to valley ratio of 480% and 36.8 in spin-up direction have been found, respectively. We propose the distinct physical mechanisms notably suggesting that the molecular junctions of AγGYNRs endow potential applications for future nanoelectronic devices.In summary, the comprehensive ab initio calculations based on the density functional theory combined with non-equilibrium Green’s function formalism on the 2D armchair 3-γ-graphyne sheets and nanoribbons with the incorporation of nitrogen atoms possess many electronic and transport characteristics that are obviously different from those of well-known graphene and typical graphynes. By exploring the impact of single or double N-doping defects of AγGYNRs, our results confirm that band structures of super unit cells are highly dependent on the positions of the dopant in the central C