Objective : To investigate the brain microvascular tissue block culture method for extracting primary rat brain microvascular endothelial cells and identify its effect. Methods : Brain tissue from 4-week-old Sprague Dawley rats was screened, pre-digested and solidified to obtain brain microvascular segments. These segments were subsequently placed in a CO2incubator for primary culture. The target cells were identified by cell morphology and immunocytochemical staining for factor Ⅷ-related antigen. Results : After a 48-hour culture periodin vitro, the short spindle cells crawled out from around the brain microvascular segments. After 72 hours, island-like cell culsters formed. After 96 hours the clusters fused and the cells formed a typical monolayer, cobble stone-like, and mosaic arrangement. Factor Ⅷ-related antigen immunocytochemical staining showed that the cytoplasm of the cells appeared brown-red, indicating positive expression; DAB stained the nucleus, showing blue-dark. Conclusion The brain microvascular tissue block culture method can isolate and culture primary rat brain microvascular endothelial cells.

On the premise of respecting the objective law of the occurrence and development of traditional Chinese medicine（TCM） dispensing granules， relevant national departments have gradually formed the research and formulation ideas of national drug standards for dispensing granules based on the experiences and lessons learned in the development process of quality standards， as well as the formation mechanism of national standards for dispensing granules. This has certain reference significance for the formulation path of TCM quality standards. Combined with the general situation of the published standards and specific cases， the research concepts of the national standards for dispensing granules were analyzed and summarized in this paper， and the analysis of the technical characteristics of the issued national standards was focused， including the introduction of standard decoction， the overall quality control of TCM， the whole process quality control and other research ideas. At the same time， it summarized the industry common problems in the research and development process of national standards for dispensing granules， such as the source and process control of medicinal materials， and strived to solve them together， encouraging the demonstration and application of new technological means in the field of TCM dispensing granules. Finally， based on the literature analysis， the shortcomings of the current national standards were discussed， and relevant suggestions were put forward to further improve the national standards for dispensing granules. Through the overall analysis， it is helpful to comprehensively understand the technical characteristics of the national standards for TCM dispensing granules， and provide reference for the scientific exploration and practice of quality control methods for TCM.

OBJECTIVE To establish the characteristic chromatogram of the volatile oil in the standard decoction of Qingshang juantong decoction， and preliminarily infer the main active components of volatile oil that affect the clinical therapeutic effect. METHODS The volatile oil in the standard decoction of Qingshang juantong decoction was extracted by steam distillation. The ultra-high performance liquid chromatography （UPLC） characteristic chromatograms of 15 batches of samples were established by the Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）， and the similarity evaluation was carried out. The volatile oil of standard decoction was identified by UPLC coupled with quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF/MS）. Then the volatile oil components were analyzed by GC-MS. RESULTS The similarities of UPLC characteristic chromatograms for volatile oil of 15 batches of Qingshang juantong decoction were between 0.949 and 0.997. A total of 12 common peaks were obtained. According to the UPLC-Q-TOF/MS， the main components were methyl eugenol， E-ligustilide， E-butylidenephthalide and so on. A total of 23 components were identified by GC-MS， which were mainly 3，4，5-trimethoxy- methylbenzene， patchouli alcohol， Z-ligustilide and so on. CONCLUSIONS The characteristic chromatograms of the volatile oil in the standard decoction of Qingshang juantong decoction is established， and it is inferred that methyl eugenol， ligustilide， E- butylidenephthalide， patchouli alcohol， 3，4，5-trimethoxy-methylbenzene might be the main active components affecting the clinical therapeutic effect of the volatile oil of Qingshang juantong decoction.

Objective:To explore genes related to costimulatory molecule related to the prognosis of bladder cancer,and to construct and evaluate prognosis model based on costimulatory molecule-based signature(CMS).Methods:Gene expression matrix and clinical information of bladder cancer patients were downloaded from TCGA database and GEO database(GSE31684),and costimulatory molecule-related genes were retrieved from the literature.The univariate and multivariate Cox analysis were used to screened prognostic-related genes and constructed prognostic model.Forecast accuracy of model was verified in TCGA training group,TCGA validation data group and GEO group by Kaplan-Meier survival analysis and receiver operating characteristic curve(ROC).Considering risk score and clinical characteristics,we constructed a nomogram and evaluated its performance by consistency analysis and ROC.CIBERSORT algorithm was used to analyze immune cell composition of tumor microenvironment infiltration,and gene set enrichment analysis(GSEA)was performed to explore the potential mechanism.Results:Four prognostic-related CMSs were found:TNFRSF14,CD276,ICOS and TMIGD2,of which three were included in the risk score construction.Multivariate Cox regression results showed that the risk score based on CMS was an independent prognostic factor for bladder cancer patients.Consistency analysis and ROC results showed that the nomogram had ideal prognosis prediction accuracy.Immune infiltration analysis showed that the high risk group was likely to be in immunosuppressive state.GSEA results suggested that genes in high risk group were enriched in extracel-lular matrix(ECM)receptors interaction,cell cycle and other pathways.Conclusion:TNFRSF14,CD276 and ICOS may be potential prognostic biomarkers for bladder cancer patients.CMS-based risk score and nomogram could contribute to early prognosis and choice of personalized treatment.

Antibody-drug conjugates(ADCs)are a new type of targeting antibodies that conjugate with highly toxic anticancer drugs via chemical linkers to exert high specificity and efficient killing of tumor cells,thereby attracting considerable attention in precise oncology therapy.Cetuximab(Cet)is a typical antibody that offers the benefits of good targeting and safety for individuals with advanced and inoperable cutaneous squamous cell carcinoma(cSCC);however,its anti-tumor activity is limited to a single use.Cisplatin(CisPt)shows good curative effects;however,its adverse effects and non-tumor-targeting ability are major drawbacks.In this study,we designed and developed a new ADC based on a new cytotoxic platinum(Ⅳ)prodrug(C8Pt(Ⅳ))and Cet.The so-called antibody-platinum(Ⅳ)prodrugs conjugates,named Cet-C8Pt(Ⅳ),showed excellent tumor targeting in cSCC.Specifically,it accurately delivered C8Pt(Ⅳ)into tumor cells to exert the combined anti-tumor effect of Cet and CisPt.Herein,metabolomic analysis showed that Cet-C8Pt(Ⅳ)promoted cellular apoptosis and increased DNA damage in cSCC cells by affecting the vitamin B6 metabolic pathway in tumor cells,thereby further enhancing the tumor-killing ability and providing a new strategy for clinical cancer treatment using antibody-platinum(Ⅳ)prodrugs conjugates.

Objective:To conduct a scoping review of the characteristics, functions, risk of bias and applications of home-based self-management behavior assessment tools for lung transplant patients, so as to provide references for clinical medical staff to conduct further related research.Methods:The relevant literatures were systematically searched in China National Knowledge Infrastructure, Wanfang Database, VIP, China Biology Medicine disc, PubMed, Web of Science Core Collection, Embase, Cochrane Library and CINAHL databases. The search period was from the establishment of the databases to June 30, 2023. The scoping review method framework was used to screen the literature, extract information and standardize the report.Results:A total of 10 167 Chinese and English literatures were searched, and after deduplication and screening, 23 articles that met the criteria were finally included, involving 13 home-based self-management behavior assessment tools for lung transplant patients, including nine specific tools and four universal tools. There were eight kinds of lung transplantation patients whose reliability and validity were not verified. Of the 13 tools, those with single-dimensional assessments and single measures predominate. The level of home self-management behavior of lung transplant patients was reflected mainly through compliance, and medication management was the most frequently assessed content.Conclusions:At present, the measurement of home-based self-management behavior of lung transplant patients is mainly based on questionnaire surveys, and the evaluation tools are mostly single dimensional. The reliability and validity of most evaluation tools in the population of lung transplant patients are not yet clear, and there is a certain risk of bias. It is recommended that researchers pay attention to the reliability and validity verification report of the evaluation tool in the target population when selecting it, and integrate multiple measurement methods to reduce measurement errors.

Objective:To extract rat brain microvascular endothelial cells(BMECs)using thin-layer cell culture method.Methods:The brain cortex of four-week-old SD rats was obtained,which was chopped,sieved,and digested with type II collagenase to obtain microvascular segments.The amount of culture medium was strictly controlled,and it was inoculated into culture flasks for primary culture.The target cells were morphologically observed by using an invert-ed phase contrast microscope,and the factor Ⅷ related antigen(FⅧRag)was identified by using immunocytochemi-cal staining.Results:The target cells cultured in the inverted phase contrast microscope showed typical endothelial-like monolayer cobble stone-like mosaic growth;FⅧRag positive cells accounted for more than 99％of the total cells.Conclusion:Thin-layer cell culture method can successfully isolate and cultivate high-purity rat BMECs.

Objective:To conduct stakeholder identification and problem analysis of inclusive health insurance participation in rare disease coverage based on the stakeholder perspective.Methods:A literature review and a Mitchell scale were used to identify,categorize,and analyze the relationship among the stakeholders,and qualitative interviews were conducted with identified stakeholders to explore their demands,positioning,and barriers to participation in inclusive health insurance.Results:The Healthcare Security Ad-ministration,Banking and Insurance Regulatory Commission,and the primary insurer were the most influential"identified stakehold-ers"in the development of inclusive health insurance.A total of 63 concepts were coded,and the stakeholders were analyzed and de-scribed in categories of claims,roles,strengths,difficulties,and behavioral strategies.Conclusion:There were differences in the de-mands of the stakeholders,lack of clarity of roles,and lack of effective synergy.

Objective:To observe the effects of virtual reality (VR) technology combined with scenario-based simulation training in obstetrics and gynecology internship teaching.Methods:Ninety-eight medical students interned in the department of obstetrics and gynecology from June 2021 to May 2022 were included. Among them, 49 students received conventional clinical internship teaching of obstetrics and gynecology in the control group, while the other 49 students received scenario-based simulation training with VR technology in the observation group. The two groups were compared in terms of test scores, learning ability assessed by the Self-Rating Scale of Self-Directed Learning, post competency, and the degree of satisfaction with teaching quality. The data were analyzed through the chi-squared test and t test with the use of SPSS 20.00. Results:The observation group had significantly higher scores than the control group in theoretical knowledge [(35.51±2.21) vs. (32.17±3.22)], case analysis[ (16.52±1.51) vs. (13.37±2.03)], and practical operation skills (all P<0.05). After teaching, the learning ability and post competency were significantly improved in both groups (both P<0.05), which were significantly better in the observation group than in the control group (both P<0.05). The degree of satisfaction with teaching of the observation group was significantly higher than that of the control group ( P<0.05). Conclusion:VR technology combined with scenario-based simulation training can help improve teaching quality and enhance medical students' learning ability and post competency, with a high degree of satisfaction with teaching quality, which is worth promotion and application.

【Objective】 To construct a prediction model of severe obstructive sleep apnea (OSA) risk in the general population by using nomogram in order to explore the independent risk factors of severe OSA and guide the early diagnosis and treatment. 【Methods】 We retrospectively enrolled patients who had been diagnosed by polysomnography and divided them into training and validation sets at the ratio of 7∶3. Patients were divided into severe OSA group and non-severe OSA group according to apnea hypopnea index (AHI)>30. Variables entering the model were identified by least absolute shrinkage and selection operator regression model (Lasso), and logistic regression (LR) method. Then, multivariable logistic regression analysis was used to establish the nomogram, and the area under the receiver operating characteristic curve (AUC) was used to evaluate the discriminative properties of the nomogram model. Finally, we conducted decision curve analysis (DCA) of nomogram model, STOP-Bang questionnaire and Berlin questionnaire to assess clinical utility. 【Results】 Through single factor and multiple factor logistic regression analyses, the independent risk factors for severe OSA were screened out, including moderate and severe sleepiness, family history of hypertension, history of smoking, drinking, snoring, history of suffocation, sedentary lifestyle, male, age, body mass index (BMI), waist and neck circumference. Lasso logistic regression identified smoke, suffocation time, snoring time, waistline, Epworth sleepiness scale (ESS) and BMI as predictive factors for inclusion in the nomogram. The AUC of the model was 0.795 [95% confidence interval (CI): 0.769-0.820] . Hosmer-Lemeshow test indicated that the model was well calibrated (χ²=3.942, P=0.862). The DCA results on the visual basis confirmed that the nomogram had superior overall net benefits within a wide, practical threshold probability range which displayed the nomogram was higher than that of STOP-Bang questionnaire and Berlin questionnaire, which is clinically useful. The Clinical Impact Curve (CIC) analysis showed the clinical effectiveness of the prediction model when the threshold probability was greater than 82% of the predicted score probability value. The prediction model determined that the high-risk population with severe OSA was highly matched with the actual population with severe OSA, which confirmed the high clinical effectiveness of the prediction model. 【Conclusion】 The model performed better than STOP-Bang questionnaire and Berlin questionnaire in predicting severe OSA and can be applied to screening. And it can be helpful to the early diagnosis and treatment of OSA in order to reduce social burden.