OBJECTIVE To establish a content determination method for two specific C21‑steroid saponins namely 3- O - β -allopyranosyl-（1→4）- β -oleandropyranosyl-11- O -isobutyryl-12- O -acetyl tenacigenin B（compound T） and 17 β -tenacigenin B in Marsdenia tenacissima decoction pieces.METHODS Ultra‑high‑performance liquid chromatography‑tandem mass spectrometry （UPLC‑MS/MS） was adopted. An XSelect HSS T3 XP chromatographic column was used. Gradient elution was performed with 0.1% formic acid aqueous solution-methanol as the mobile phase. The flow rate was 0.3 mL/min， the injection volume was 5 μL， and the column temperature was 40 ℃. An electrospray ionization source was applied for mass spectrometry， and multiple reaction monitoring mode was used for positive‑ and negative‑ion acquisition.RESULTS The linear relationships of compound T and 17 β -tenacigenin B were good within the concentration ranges of 10-200 ng/mL and 20-200 ng/mL， with r values of 0.993 1 and 0.998 6， respectively； the RSD values of precision， repeatability， accuracy， and stability were all below 4.5% （ n ＝5-6）. Their contents in three batches of decoction pieces were 94.0-102.8 μg/g and 1 705.4-1 736.2 μg/g， respectively.CONCLUSIONS The established method can be applied for the content determination of the two specific C21‑steroid saponins mentioned above in M. tenacissima decoction pieces.

Objective To analyze the problems in the operation and management of drug clinical trials and put forward targeted suggestion. Methods An electronic questionnaire survey was conducted among medical staff in about 80 hospitals in the yangtze river delta region. Results 606 valid questionnaires were received. 71% of the respondents expressed their willingness to study and participate in drug clinical trials. There were significant differences in the cognitive demands, willingness and motivation of the respondents with different occupations and educational backgrounds about the drug clinical trial work （P<0.05）. During the operation of drug clinical trials, respondents reported the main factors affecting the quality of clinical trials which including good clinical practice (GCP) awareness and subjective enthusiasm of investigators （response rate 27%）, job stability of supervisors and research coordinators （27%）, compliance of subjects （45%）, quality control of the whole process of the circulation of test drugs, medical devices and biological samples （52%）, and the informatization level of clinical trial institutions （30%）. Conclusion Hospitals, institutions and project teams could take measures to cultivate and stabilize the drug clinical trial talent team, improve the quality management system of drug clinical trials, improve work efficiency, and promote the high-quality development of drug clinical trials in medical institutions.

ObjectiveTo systematically evaluate the application of narrative education in undergraduate nursing teaching， to understand the current application status of narrative education， and to provide a theoretical basis for the subsequent establishment of a sound narrative education system. MethodsA systematic search was conducted for studies published in Chinese and English databases on applying narrative education to undergraduate nursing teaching， with the search period ranging from database inception to February 23， 2025. Literature was screened， and relevant information was extracted. A rigorous quality evaluation was conducted on the included studies， and a descriptive analysis was performed on their content. ResultsA total of 20 papers were included， involving 3，180 research subjects， all of whom were undergraduate nursing students. The results of descriptive analysis showed that the teaching model of narrative education primarily encompassed reading narrative works， watching films and videos， performing narrative scenarios， and writing reflective journals. The course setting and content covered pre-teaching preparation and in-teaching implementation. The evaluation of teaching effectiveness included the evaluation of teachers’ teaching methods （student evaluation/self-evaluation） and the evaluation of students’ learning effectiveness （course grade evaluation/humanistic care scale/empathy scale assessment， and others）. ConclusionNarrative education combines abstract concepts with concrete clinical situations， which not only enriches students’ learning experiences but also enhances their humanistic literacy. Meanwhile， it provides teachers with opportunities to develop their narrative teaching skills， which requires them to possess profound professional knowledge and employ narrative techniques to guide students in reflection and critical thinking， thereby improving teaching quality and learning outcomes. Future efforts should consistently deepen the connotation research of narrative education and build a systematic nursing education system.

Objective:To observe the effects of Qizhi Tongluo Prescription on renal interstitial fibrosis in rats with membranous nephropathy based on the Shh/Gli1 signaling pathway; To explore its intervention mechanism.Methods:Totally 60 male SD rats were divided into blank group ( n=10) and model group ( n=50) using random number table method. The model of membranous nephropathy was established according to the modified Border method. The successfully modeling rats were divided into model group, benazepril group and Qizhi Tongluo Prescription low-, medium- and high-dosage groups using random number table method. Benazepril group was gavaged with benazepril hydrochloride 10 mg/kg, Qizhi Tongluo Prescription low-, medium- and high-dosage groups were gavaged with Qizhi Tongluo Prescription solution 1.22 g/kg, 2.43 g/kg and 4.86 g/kg, and blank group and model group were gavaged with equal volume of normal saline, once a day, for 4 weeks. The 24-hour urine was collected to detect the 24-hour urinary protein quantification, and the blood was taken from the abdominal aorta to detect the levels of total cholesterol (TC), triglyceride(TG), and serum albumin(ALB); the pathological changes of rat kidney were observed by light microscope and transmission electron microscope; the protein expressions of sonic hedgehog factor (Shh), zinc finger protein 1 (Gli1) and α-smooth muscle actin (SMA) in renal tissues were detected by immunohistochemistry; the protein expressions of Shh, Gli1, α-SMA, TGF-β1, Collagen Ⅳ and plasminogen activator inhibitor-1 (PAl-1) in renal tissues were detected by Western blot. Results:Compared with the model group, the quantitative level of 24-hour urinary protein of rats in each administration group decreased ( P<0.05), serum TC and TG levels increased ( P<0.05), ALB level decreased ( P<0.05), the positive expressions of Shh, Gli1, α-SMA protein in renal tissue decreased ( P<0.05), and the protein expressions of Shh, Gli1, α-SMA, Collagen Ⅳ, TGF-β1, PAI-1 in renal tissue decreased ( P<0.05). Conclusion:Qizhi Tongluo Prescription can improve renal interstitial fibrosis in membranous nephropathy rats, possibly by inhibiting the Shh/Gli1 signaling pathway to delay renal interstitial fibrosis.

Objective To investigate the spatio-temporal expression of nuclear receptor subfamily 0 group B member 1(NR0B1)in synovial tissues of rheumatoid arthritis(RA),and to uncover the potential upstream signalling pathway that may regulate NR0B1 expression in human fibroblast-like synovial cells,thereby clarifying the possible factors that induce the disorder of NR0B1 expression in RA.Methods Patients with 25 RA and 18 osteoarthritis(OA)who did not receive any immunotherapy were recruited to obtain the synovial tissues of knee joint,as well as their related clinical test information,and the expression characteristics of NR0B1 in synovial tissue were investigated using Real-time PCR,Western blotting,immunohistochemistry and double immunofluorescent staining.Meanwhile,Pearson Chi-square test was used to evaluate the correlation between NR0B1 mRNA expression level and clinical parameters in RA patients.Furthermore,the MH7ANR0B1-/-cells that were stably knocked down of endogenous NR0B1 were generated,and the effects of NR0B1 deficiency on the phenotype of rheumatoid synovial cells enhanced by vascular endothelial growth factor(VEGF)were then evaluated by means of cell viability and apoptosis assays,colony formation assay,wound healing assay,and cell invasion assay.Last,we studied the molecular mechanism underlying interleuikin 6(IL-6)-mediated regulation of NR0B1 expression at the transcriptional level by using the STAT3-specific inhibitor intervention,siRNA,and dual luciferase reporter gene detection.Results The expression level of NR0B1 mRNA in the RA synovial tissues was significantly higher than that in synovial tissues from OA patients(P＜0.05).This trend of the increased NR0B1 mRNA expression correlated to clinical parameters,including C-reactive protein(CRP),rheumatoid factor(RF)and erythrocyte sedimentation rate(ESR),in RA patients.NR0B1 protein was predominantly immunolocalized in fibroblast-like synoviocytes of RA synovial tissues.Functionally,knockdown of NR0B1 expression markedly neutralize the VEGF-mediated enhancement of cell viability,resistance to apoptosis,clonogenicity,as well as migration and invasion in the MH7A cells.Additionally,IL-6 could specifically regulate NR0B1 expression in the MH7A synovial cells.IL-6 upregulated the transcriptional expression of NR0B1 mainly through induction of the phosphorylation of its downstream STAT3 at the Tyr-705 site.Conclusion The upregulated NR0B1 expression in fibroblast-like synoviocytes is positively correlated with the progression of RA.The proinflammatory cytokine IL-6 potentiates the transactivation of NR0B1 by inducing the phosphorylation of Tyr-705 site of its downstream effector STAT3.NR0B1 may be a significant breakthrough point for understanding the interaction between IL-6 and VEGF signaling pathways and the progression of RA.

Researchers commonly use cyclization recombination enzyme/locus of X-over P1 (Cre/loxP) technology-based conditional gene knockouts of model mice to investigate the functional roles of genes of interest in Sertoli and Leydig cells within the testis. However, the shortcomings of these genetic tools include high costs, lengthy experimental periods, and limited accessibility for researchers. Therefore, exploring alternative gene silencing techniques is of great practical value. In this study, we employed adeno-associated virus (AAV) as a vector for gene silencing in Sertoli and Leydig cells. Our findings demonstrated that AAV serotypes 1, 8, and 9 exhibited high infection efficiency in both types of testis cells. Importantly, we discovered that all three AAV serotypes exhibited exquisite specificity in targeting Sertoli cells via tubular injection while demonstrating remarkable selectivity in targeting Leydig cells via interstitial injection. We achieved cell-specific knockouts of the steroidogenic acute regulatory ( Star ) and luteinizing hormone/human chorionic gonadotropin receptor (Lhcgr) genes in Leydig cells, but not in Sertoli cells, using AAV9-single guide RNA (sgRNA)-mediated gene editing in Rosa26-LSL-Cas9 mice. Knockdown of androgen receptor ( Ar ) gene expression in Sertoli cells of wild-type mice was achieved via tubular injection of AAV9-short hairpin RNA (shRNA)-mediated targeting. Our findings offer technical approaches for investigating gene function in Sertoli and Leydig cells through AAV9-mediated gene silencing.
Animals ; Male ; Leydig Cells/metabolism* ; Mice ; Dependovirus/genetics* ; Sertoli Cells/metabolism* ; Gene Silencing ; Genetic Vectors ; Testis/cytology*

OBJECTIVE: Exposure to polycyclic aromatic hydrocarbons (PAHs) or metal(loid)s individually has been associated with neural tube defects (NTDs). However, the impacts of PAH and metal(loid) co-exposure and potential interaction effects on NTD risk remain unclear. We conducted a case-control study in China among population with a high prevalence of NTDs to investigate the combined effects of PAH and metal(loid) exposures on the risk of NTD. METHODS: Cases included 80 women who gave birth to offspring with NTDs, whereas controls were 50 women who delivered infants with no congenital malformations. We analyzed the levels of placental PAHs using gas chromatography and mass spectrometry, PAH-DNA adducts with ³²P-post-labeling method, and metal(loid)s with an inductively coupled plasma mass spectrometer. Unconditional logistic regression was employed to estimate the associations between individual exposures and NTDs. Least absolute shrinkage and selection operator (LASSO) penalized regression models were used to select a subset of exposures, while additive interaction models were used to identify interaction effects. RESULTS: In the single-exposure models, we found that eight PAHs, PAH-DNA adducts, and 28 metal(loid)s were associated with NTDs. Pyrene, selenium, molybdenum, cadmium, uranium, and rubidium were selected through LASSO regression and were statistically associated with NTDs in the multiple-exposure models. Women with high levels of pyrene and molybdenum or pyrene and selenium exhibited significantly increased risk of having offspring with NTDs, indicating that these combinations may have synergistic effects on the risk of NTDs. CONCLUSION Our findings suggest that individual PAHs and metal(loid)s, as well as their interactions, may be associated with the risk of NTDs, which warrants further investigation.
Humans ; Neural Tube Defects/chemically induced* ; Polycyclic Aromatic Hydrocarbons/adverse effects* ; Female ; Case-Control Studies ; China/epidemiology* ; Adult ; Pregnancy ; Environmental Pollutants ; Maternal Exposure/adverse effects* ; Metals/toxicity* ; Young Adult ; Risk Factors

Objective To analyze the characteristics of intestinal flora of personnel stationed on an island,so as to lay the foundation for maintaining the intestinal microecological balance of personnel stationed on island and provide accurate medical security.Methods Several subjects stationed on an island and several subjects from coastal areas were enrolled by random and sampling method,and their fecal samples were sequenced by 16S rRNA high-throughput sequencing.Diversity and composition of gut microbiota in 2 cohorts of personnel were compared.Results Alpha diversity analysis of intestinal flora showed that the abundance of intestinal flora in subjects stationed on the island was significantly higher than that of subjects from coastal areas.Beta diversity analysis indicated significant differences in the composition of intestinal microbial communities between the subjects stationed on the island and those from coastal areas(P=0.001).The abundance of the Bacteroidota in the intestinal tract of subjects stationed on the island was significantly lower than that of subjects from coastal areas(30.8%vs 48.3%,P＜0.001),while the abundance of the Proteobacteria was significantly higher than that of subjects from coastal areas(28.3%vs 10.2%,P＜0.001).After multiple hypothesis testing correction,it was found that the abundance of the Bacteroides,Roseburia,Alistipes,and Parabacteroides in the intestines of subjects stationed on the island decreased significantly,while the abundance of the Prevotella,Escherichia-Shigella,Citrobacter,and Eubacterium_coprostanoligenes increased significantly.Conclusion The special environment of islands affects the characteristics of intestinal flora of personnel,and the intestinal microecological health needs precise maintenance.

ObjectiveTo investigate and analyze an outbreak of rotavirus infectious diarrhea in a prison in Chongqing Municipality, to provide a basis for adult rotavirus surveillance and prevention, and to explore the public health problems in special settings. MethodsA retrospective survey was conducted to collect and analyze data on individual cases with diarrheal disease on-site. The clinical characteristics, as well as the temporal, spatial and geographical distribution patterns of the epidemic were described. Multi-pathogen detection tests were conducted both on diarrhea cases and environmental samples, with viral genotyping performed on positive samples. A case-control analysis was performed to identify the causes of the outbreak, and an SEIR model was adopted to predict the outbreak trend and evaluate the effectiveness of interventions. ResultsA total of 65 cases were found among the inmates, with an attack rate of 2.03%. The predominant clinical manifestations included diarrhea (89.23%), watery stool (73.85%), and dehydration (18.46%). The epidemic curve indicated a “human-to-human” transmission pattern, with an average incubation period of 5‒6 days. The attack rates among chefs in the main canteen (80.00%, 8/10) and caterers (28.33%, 17/60) were significantly higher than those of other inmates （P<0.05）. Multi-pathogen polymerase chain reaction (PCR) testing detected positive for group A rotavirus, with the viral genotyping identified as G9P [8] strain. Factors such as unprotected "bare-handed" food distribution among cases with diarrhea (OR=9.512, 95%CI: 4.261‒21.234) and close contact with diarrhea cases (OR=3.656, 95%CI: 1.719‒7.778) were the possible cause of the outbreak. The SEIR model (r₀=5, α=0.3, β₁=0.08, β₂=0.04) was constructed using prison inmates as susceptible population, aiming at fitting the initial transmission trend of the outbreak, and the epidemic rate declined rapidly after intervention measures were implemented (r_t≈0). ConclusionThis rare rotavirus infection diarrhea outbreak among adults in confined settings suggests that the construction of public health prevention and control systems in prison may be overlooked. Cross infection during meal processing and distribution in the canteens of such settings is likely to be the cause of the outbreak. Given the potential neglect of public heath system construction in special settings, it is imperative to enhance the surveillance and monitoring of rotavirus and other intestinal multi-pathogens among adults, as well as the construction of public health prevention and control systems in these special settings.

Laparoscopic subtotal cholecystectomy (LSC) has been a safe and viable alternative to conversion to laparotomy in cases of severe cholecystitis. The objective of this study is to determine the utility of intraoperative choledochoscopy in LSC for the exploration of the gallbladder, cyst duct, and subsequent stone clearance of the cystic duct in cases of severe cholecystitis. A total of 72 patients diagnosed with severe cholecystitis received choledochoscopy-assisted laparoscopic subtotal cholecystectomy (CALSC). A choledochoscopy was performed to explore the gallbladder cavity and/or cystic duct, and to extract stones using a range of techniques. The clinical records, including the operative records and outcomes, were subjected to analysis. No LSC was converted to open surgery, and no bile duct or vascular injuries were sustained. All stones within the cystic duct were removed by a combination of techniques, including high-frequency needle knife electrotomy, basket, and electrohydraulic lithotripsy. A follow-up examination revealed the absence of residual bile duct stones, with the exception of one common bile duct stone, which was extracted via endoscopic retrograde cholangiopancreatography. In certain special cases, CALSC may prove to be an efficacious treatment for the management of severe cholecystitis. This technique allows for optimal comprehension of the situation within the gallbladder cavity and cystic duct, facilitating the removal of stones from the cystic duct and reducing the residue of the non-functional gallbladder remnant.