ObjectiveTo analyze the pharmacodynamic substance basis of Shangkeling Spray and its potential mechanisms in intervening knee osteoarthritis （KOA） using ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS）， network pharmacology， and molecular docking technology. MethodsUPLC-MS was used to identify the chemical components of Shangkeling Spray. Pharmacokinetic properties were employed to screen potential active ingredients. Network pharmacology methods were utilized to collect potential targets of these ingredients and the pathological gene set of KOA. An "active ingredient-disease" target network was constructed using databases such as STRING. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） functional enrichment analyses were performed using clusterProfiler. Libraries including NumPy were employed to calculate shortest path lengths to identify dominant pharmacodynamic links. Core gene clusters were identified using MCODE， validated through the Gene Expression Omnibus （GEO） database， and molecular docking was performed between key active ingredients and core targets. ResultsA total of 322 and 314 chemical components were identified under positive and negative ion modes， respectively， with 410 components in total after de-duplication， mainly including flavonoids， coumarins， terpenoids， organic acids， and alkaloids. Analysis of the "active ingredient-disease" network identified "development and regeneration"， "cell growth and death"， "immune system"， and "nervous system" as the dominant pharmacodynamic links of Shangkeling Spray in the treatment of KOA. Molecular docking showed that key active ingredients， such as bletillin A， formononetin， morin， oxymatrine， aconitine， gallic acid， curdione， apigenin， naringenin， and oleanolic acid， tightly bound to functional domains of 10 key targets including Jun proteins（JUN）， interleukin-6 （IL-6）， protein kinase B1 （Akt1）， Caspase-3， nuclear transcription factor-κB subunit p65（RELA）， nuclear factor-kappaB1（NF-κB1）， Cyclin D₁， mammalian target of rapamycin（mTOR）， tumor necrosis factor （TNF）， and Fos proto-oncogene protein （FOS）. These interactions synergistically regulated the phosphatidylinositol 3-kinase （PI3K）/Akt/mTOR-related signaling axis and nervous system-related pathways， mediating cartilage repair， reducing inflammation and pain， and improving KOA. ConclusionThis study preliminarily clarifies the pharmacodynamic substance basis of Shangkeling Spray and suggests that its main active ingredients may improve KOA by synergistically regulating the PI3K/Akt/mTOR-related pathways， providing a reference for subsequent exploration of its substance benchmark and mechanism of action.

OBJECTIVE: To explore the genetic etiology of 46 Chinese pedigrees affected with Hereditary multiple exostoses (HME) and provide genetic counseling and reproductive intervention. METHODS: Whole-exome sequencing and Sanger sequencing were carried out on 87 patients from the 46 pedigrees to analyze the variants of EXT1 and EXT2 genes. Pathogenicity of the variants was assessed based on the guidelines from the American College of Medical Genetics and Genomics and Association for Molecular Pathology (ACMG/AMP). Prenatal diagnosis and preimplantation genetic testing (PGT) were provided for couples with identified pathogenic mutations. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: LL-SC-SG-2014-010). RESULTS: In total 17 and 22 pathogenic variants were respectively identified in the EXT1 and EXT2 genes, among which 5 EXT1 and 12 EXT2 variants were unreported previously. Three patients with no family history were found to harbor de novo variants of the EXT1 gene. Twenty nine couples had opted for PGT or underwent prenatal diagnosis following natural conception, and 17 healthy babies were born. CONCLUSION This study has clarified the genetic etiology of 45 HME pedigrees and identified 17 novel variants, which has enriched the mutational spectrum of the EXT1 and EXT2 genes. Reproductive intervention through PGT and prenatal diagnosis have prevented the recurrence of HME in these families.
Humans ; Female ; Male ; Pedigree ; Exostoses, Multiple Hereditary/diagnosis* ; N-Acetylglucosaminyltransferases/genetics* ; Adult ; Exostosin 1 ; Asian People/genetics* ; Genetic Testing ; Exostosin 2 ; Mutation ; China ; Prenatal Diagnosis ; Pregnancy ; Genetic Counseling ; Preimplantation Diagnosis ; Exome Sequencing ; East Asian People

Objective:To investigate the influence and mechanism of extracellular vesicles (EVs) derived from human adipose-derived mesenchymal stem cells (hADMSCs), i.e. hADMSC-EVs on pyroptosis of human umbilical vein endothelial cells (HUVECs) induced by high glucose, with the aim of providing evidence for improving vascular function in diabetic wounds.Methods:This study was an experimental research. The umbilical cords from 5 women aged 25 to 40 years were collected who had normal vaginal delivery at the Department of Obstetrics and Gynecology of the First Affiliated Hospital of Nanchang University from June to September in 2023, and HUVECs were isolated and successfully identified. Adipose tissue was obtained from 6 healthy women aged 25 to 35 years who underwent abdomen liposuction at the Department of Plastic Surgery of the above-mentioned hospital in the same period. After hADMSCs were isolated, hADMSC-EVs were extracted and successfully identified. The fourth passage of HUVECs were cultured in endothelial cell medium containing glucose in a molarity of 33 mmol/L and divided into phosphate buffered solution (PBS) group cultured with PBS, EV group cultured with hADMSC-EVs, and EV+LY294002 group cultured with hADMSC-EVs and the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway inhibitor LY294002. Western blotting was used to detect the expressions of PI3K/Akt signaling pathway-related proteins PI3K and Akt, and pyroptosis-related proteins nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), cysteinyl aspartate specific protease-1 (caspase-1), gasdermin D, interleukin-1β (IL-1β), and IL-18 of cells after 48 hours of culture. A cell counting kit-8 was used to test the proliferation levels of cells at 0 (immediately), 12, 24, 36, 48, 60, and 72 hours of culture. After 48 hours of culture, the cell scratch test was performed and the cell migration rates at 12 and 24 hours after scratching were calculated; the cell Transwell assay was conducted and the number of cells migrating in 24 hours was calculated; the cell tube formation experiment was performed, and the total length of tube formation and the number of branch nodes were measured and counted. The sample size was 3.Results:After 48 hours of culture, the protein expressions of PI3K and Akt of cells in EV group were significantly higher than those in PBS group ( P<0.05), and the protein expressions of PI3K and Akt of cells in EV+LY294002 group were significantly lower than those in EV group ( P<0.05). After 48 hours of culture, the protein expressions of NLRP3, caspase-1, gasdermin D, IL-1β, and IL-18 of cells in EV group were 0.54±0.08, 0.96±0.11, 0.525±0.061, 1.216±0.039, and 1.317±0.023, respectively, which were significantly lower than 2.32±0.11, 1.86±0.07, 1.256±0.113, 2.589±0.084, and 2.042±0.132 in PBS group ( P<0.05); the protein expressions of NLRP3, caspase-1, gasdermin D, IL-1β, and IL-18 of cells in EV+LY294002 group were 1.16±0.05, 1.37±0.06, 0.962±0.028, 1.834±0.017, and 1.803±0.065, respectively, which were significantly higher than those in EV group ( P<0.05). At 12, 24, 36, 48, 60, and 72 hours of culture, the proliferation levels of cells in EV group were significantly higher than those in PBS group ( P<0.05), and the proliferation levels of cells in EV+LY294002 group were significantly lower than those in EV group ( P<0.05). After 48 hours of culture, the cell migration rates at 12 and 24 hours after scratching in EV group were significantly higher than those in PBS group ( P<0.05), and the cell migration rates at 12 and 24 hours after scratching in EV+LY294002 group were significantly lower than those in EV group ( P<0.05); the number of cells migrating in 24 hours in EV group was significantly greater than that in PBS group ( P<0.05), and the number of cells migrating in 24 hours in EV+LY294002 group was significantly less than that in EV group ( P<0.05). After 48 hours of culture, compared with those in PBS group, the total length of tube formation of cells in EV group was significantly prolonged ( P<0.05), and the number of branch nodes was significantly increased ( P<0.05); compared with those in EV group, the total length of tube formation in EV+LY294002 group was significantly shortened ( P<0.05), and the number of branch nodes was significantly decreased ( P<0.05). Conclusions:hADMSC-EVs can inhibit the expression of pyroptosis-related proteins in HUVECs induced by high glucose through the PI3K/Akt signaling pathway and improve the proliferation, migration, and angiogenesis capabilities of HUVECs.

To meet the domestic clinical demand timely,the national health commission has released three batches of encourage generic drug catalogues,which plays a good guiding role in improving the supply level and accessibility of generic drugs.Based on literature investigation,the typical cases of novel pharmaceutical preparations were analyzed,and the pharmaceutical considerations were put forward in terms formulation,manufacturing process and quality control,aimed to provide scientific reference for research and development of such drugs.

Objective To analyze the diagnostic value of fat content measurement based on magnetic resonance multi-echo Dixon(ME-Dixon)technique combined with serological indicators for early liver fibrosis.Methods A total of 78 patients with early liver fibrosis who underwent liver biopsy at the First Affiliated Hospital of Army Medical University from August 2019 to October 2021 were included.According to the pathological results,the patients with early liver fibrosis were divided into the S0 group(n=21),the S1 group(n=32),and the S2 group(n=25).Meanwhile,the complete imaging examination results(including ME-Dixon sequence)and laboratory examination results[including hyaluronic acid(HA),procollagen Ⅲ(PCⅢ),collagen Ⅳ(CⅣ),laminin(LN),aspartate transaminase(AST),alanine amino-transferase(ALT),gamma-glutamyl transferase(GGT),alkaline phosphatase(ALP)]were collected.And the hepatic fat fraction(HFF)was measured by ME-Dixon technique.One-way ANOVA was used to analyze the differences in HFF and serological indicators among the three groups,and then factors with statistical differences in the univariate analysis were selected to draw receiver operating characteristic(ROC)curve,and the area under the curve(AUC)was calculated to evaluate their value in the diagnosis of liver fibrosis and the degree of early liver fibrosis.Results There was no statistically significant difference in the PCⅢ,LN,AST,or ALT among the S0,S1,and S2 groups(P>0.05),while there were statistically significant differences in the HFF,HA,CⅣ,GGT,and ALP(P<0.05),and the levels of HA,CⅣ,GGT,and ALP in the S2 group were significantly higher than those in the S0 and S1 groups(P<0.05).ROC curve analysis showed that the above indicators for diagnosing liver fibrosis and the degree of early liver fibrosis based on AUC values were in the following order:HFF+GGT(0.72)>HFF(0.68)>GGT(0.61)>CⅣ(0.54)>ALP(0.53)>HA(0.48)and HA+CⅣ(0.71)>CⅣ(0.68)>HA(0.66)>GGT(0.61)>ALP(0.59)>HFF(0.46).Conclusion The fat content measurement based on magnetic resonance ME-Dixon tech-nique combined with serological indicators has significant reference value for the diagnosis of early liver fibrosis,and it plays a certain guid-ing role in clinical diagnosis and treatment.

Bovine rotavirus(BRV)is an important pathogen causing diarrhea in calves.To understand the genomic charac-teristics and genetic variations in bovine rotavirus,and to further enrich data on the biological characteristics of rotavirus,we aimed to amplify 11 gene segments of the isolated and cultured G6P[1]bovine rotavirus BLL strain,perform whole genome se-quencing,and analyze the molecular characteristics.MEGA7.0 and DNAMAN software were used for homology and typing a-nalysis,and the whole genome phylogenetic tree was constructed to analyze genetic evolution relationships.The complete geno-type of the BLL strain was G6-P[1]-I2-R2-C2-M2-A3-N2-T6-E2-H3.Phylogenetic analysis of the VP7 and VP4 genes of the BLL strain showed that the VP7 gene had the highest homology with RVA/Cow-wt/HB01/China/2021,and the VP4 gene of the BLL strain was in the same branch as RVA/Human-tc/ISR/Ro8059/1995.From the sequence alignment of VP8*amino acids,the sialic acid domain of the BLL strain was found to be similar to that in other P[1]strains,but different from those in other types of strains,except for residue 189,which was the same as that in Ro8059 but different from that in other strains.The results suggested that the BLL strain might potentially infect humans.Therefore,continued monitoring and study of the biological characteristics of this strain are necessary to provide more information and evidence supporting further research on the cross-species transmission of group A rotavirus in China.

Objective:To determine the detection rate of myopia among primary and secondary school students in Hunan Province in 2022 and to analyze the influencing factors at both the school and individual levels, thereby providing a scientific basis for developing myopia prevention and control strategies.Methods:From October to November 2022, a multi-stage stratified cluster random sampling method was employed to select students from Year 4 of primary school to Year 3 of senior high school across 14 prefecture-level (autonomous prefecture) cities in Hunan Province for vision screening and questionnaire surveys. A multilevel regression model was utilized to analyze the influencing factors of myopia at both the school and individual levels.Results:A total of 189 343 primary and secondary school students were included in this study. The overall myopia detection rate was 55.56%, with a significantly higher prevalence observed in female students (60.49%) compared to males (51.03%) and in urban students (59.12%) versus those from rural areas (53.50%). A marked upward trend in myopia prevalence was identified with advancing grade levels (trend test χ2=16 246.13, P<0.001). Multilevel regression analysis revealed that at the individual level, female gender, higher grade level, parental myopia history, daily homework duration ≥2 hours after school, improper reading/writing postures, and taking breaks only after more than 15 minutes of near work were associated with an increased risk of myopia. Conversely, adequate sleep duration, outdoor activity ≥2 hours, and outdoor breaks during recess demonstrated protective effects. At the school level, non-compliant blackboard illumination uniformity emerged as a significant risk factor for myopia development. Conclusions:The detection rate of myopia among primary and secondary school students in Hunan Province remains relatively high and is associated with multiple factors at both the school and individual levels. Targeted interventions should be implemented at different levels to mitigate the risk of myopia.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Objective:Analyze the imaging and clinical data of cases undergoing thoracoscopic subsegmental resection for non-small cell lung cancer(NSCLC) with a diameter≤2 cm, and explore the clinical outcomes of subsegmental resection.Methods:A retrospective analysis was conducted on the clinical data of 58 patients who underwent thoracoscopic subsegmentectomy in China-Japan Friendship Hospital from January 2020 to July 2024. Three-dimensional reconstruction technology was used for surgical planning before the operation, and thoracoscopic subsegmentectomy was performed, including single lung subsegmentectomy(Group 1), multiple lung subsegmentectomy(Group 2), and combined segmentectomy and subsegmentectomy(Group 3).Results:All patients successfully completed the surgery, with 23 cases of single lung subsegmentectomy, 6 cases of multiple lung subsegmentectomy, and 29 cases of combined segmentectomy and subsegmentectomy. The median intraoperative blood loss was 30.0(20.0, 30.0)ml, the average operation time was(2.03±0.68) h, the average pathological size of the nodules was(10.53±4.45) mm, and the average postoperative tube retention was(2.55±0.92) days. There were 6 cases of postoperative complications, including pulmonary air leakage in 2 cases, cerebral embolism in 1 case, pulmonary embolism in 1 case, pulmonary infection in 1 case, and atrial fibrillation in 1 case. All patients had negative surgical margins in the postoperative pathology. Group 1 had less average intraoperative blood loss than Group 2, with statistically significant differences( P=0.027). Surgical procedures for the upper lobe of the lung mainly involve the resection of combined segments and subsegments, while those for the lower lobe primarily consist of single segmentectomy. Conclusion:Subsegmentectomy is an effective surgical approach when the nodule is small and a clear margin can be ensured, allowing for better preservation of remaining lung tissue. Bleeding during multiple subsegmentectomies is greater than that in single subsegmentectomy and combined segmentectomy with subsegmentectomy, which may be related to the more complex vascular variations in multiple subsegmentectomies.

Objective:To analyze the human immunodeficiency virus (HIV) screening status and the resulting residual risk (RR) among blood donors across 17 provincial blood centers in China.Methods:This study used a cross-sectional study. Data on HIV infection markers per 100 000 first-time donors (FD) and repeat donors (RD) from January 2015 to December 2024 were extracted from the National Blood Establishment Performance Comparison Information Management System. Questionnaires were used to collect each center′s HIV screening strategy, algorithm, serological test (ST) kit manufacturers, gray-zone setting for ST, and nucleic acid test (NAT) modality, method, and platform. The incidence-window-period model was used to calculate the residual risk for first-time donors (RR FD), repeat donors (RR RD), and total donors (RR TD) at each center. Horizontal and vertical analysis of RR FD, RR RD, and RR TD across centers and years were performed. Results:All 17 centers applied the same HIV screening strategy which was two rounds of ST followed by one round of NAT. Eight of them operated a single screening algorithm, six employed two algorithms and three used three. Eleven centers used both imported and domestic ST kits, five relied on domestic ST kits only, and one used imported ST kits only, while four centers never set a grey zone for ST throughout the decade. For NAT modalities, eight centers adopted both individual nucleic acid test (ID-NAT) and minipool nucleic acid test (MP-NAT), eight used MP-NAT only and one used ID-NAT only. Seven centers combined transcription mediated amplification (TMA) and polymerase chain reaction (PCR), nine used PCR only and one used TMA only, and fourteen centers ran both imported and domestic NAT systems, two used imported systems only and one used a domestic system only. Over the ten-year period, the mean RR FD across the centers ranged from 2.22 to 12.33 per 10 6 person-years, RR RD from 0.83 to 3.29 per 10 6 person-years and RR TD from 1.59 to 9.29 per 10 6 person-years, with center Z4 consistently showing the lowest values for all three metrics and center U4 recording the highest RR FD and RR TD, while center D2 had the highest RR RD. In 2024 compared with 2015, eleven centers achieved a lower RR FD and ten centers achieved lower RR RD and RR TD. The RR FD and RR TD of centers W2 and U4 displayed pronounced fluctuations and an upward trend in recent years. Conclusions:The 17 provincial blood centers maintain consistent HIV screening strategies, while demonstrating variations in screening algorithm, ST kit manufacturers, NAT modalities, methods, and platform. And the RR FD, RR RD, and RR TD differ across centers. Although most centers show declining trend in RR over the ten-year period, some centers exhibite data fluctuations with a rising trend, suggesting potential for further optimization of HIV screening protocols.