ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L^-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.

Objective To explore the incidence and risk factors of sarcopenia in non-dialysis patients with stage 5 chronic kidney disease (CKD). Methods Non-dialysis stage 5 CKD patients in Jiading District Central Hospital Affiliated Shanghai University of Medicine & Health Sciences from October 2023 to October 2024 were enrolled and divided into the non-sarcopenia group and the sarcopenia group according to lean tissue index (LTI) and handgrip strength (HGS). Demographic data of patients were collected and biochemical indicators were detected. Serum advanced glycation end products (AGEs) were detected by competitive ELISA. Multivariate logistic regression analysis was conducted to analyze the risk factors of sarcopenia in these patients. The diagnostic efficacy of the risk factors was evaluated by the receiver operating characteristic (ROC) curve. Results A total of 102 non-dialysis stage 5 CKD patients with (68.52±7.39) years, were enrolled, including 61 male (59.8%). Among them, 24 were diagnosed with sarcopenia (23.5%). Compared with patients in non-sarcopenia group, patients in sarcopenia group were older and had higher interleukin-6 (IL-6) and AGEs, lower body mass index (BMI) and lower serum albumin (sAlb; P＜0.05). Multivariate logistic regression showed that advanced age (OR＝1.153, 95%CI 1.045–1.273, P＝0.005), higher IL-6 (OR＝1.165, 95%CI 1.042–1.302, P＝0.007)、lower sAlb (OR＝0.675, 95%CI 0.542–0.840, P＝0.001), and higher serum AGEs (OR＝1.105, 95%CI 1.013–1.206, P＝0.024) were independent risk factors of sarcopenia in non-dialysis CKD 5 patients. The areas under the curve (AUC) for the age, IL-6, sAlb, and serum AGEs were 0.896, 0.643, 0.658, and 0.724, respectively. Conclusions Sarcopenia is common in non-dialysis stage 5 CKD patients, and the risk of sarcopenia is higher in non-dialysis stage 5 CKD patients with advanced age, higher IL-6, lower sAlb, and higher serum AGEs.

ObjectiveTo observe the effects of electroacupuncture with "tonifying the kidney and dispelling stasis" acupoint prescription on sexual function in diabetic erectile dysfunction （DMED）model rats， and to explore its possible mechanism of action. MethodsSPF male SD rats were randomly divided into 10 each in blank group， model group， Tadalafil group， and electroacupuncture group. DMED rat model was prepared by high glucose and high fat diet combined with intraperitoneal injection of streptozotocin. After successful modelling， rats in the electroacupuncture group were given electroacupuncture intervention of "tonifying the kidney and dispelling stasis" acupoint prescription once every other day； Tadalafil group was given Tadalafil solution 0.5 mg/kg·d by gavage， and the blank group， model group and electroacupuncture group were given 10 ml/kg pure water by gavage once a day. Each group was intervened for 30 days. The body mass and blood glucose level of the rats were detected on the 1st， 8th， 15th， 22nd days and at the end of the intervention， respectively. At the end of the intervention， the penile erection of the rats was observed by using the apomorphine test； the level of plasma endothelial cell microparticles （EMPs）was detected by flow cytometry； the histopathological morphology of the penile cavernous body was observed by HE staining， and the pathological morphology of the endothelial cells of the penile vasculature was observed by electron microscopy. Serum sex hormones including testosterone （T）， follicle stimulating hormone （FSH）， luteinising hormone （LH） and vascular endothelial function-related factors including vascular endothelial growth factor （VEGF）， endothelin-1 （ET-1）， lectin-like oxidized low-density lipoprotein receptor-1 （LOX-1）， soluble E-selectin （sE-selectin）， and soluble intercellular adhesion molecule-1 （sICAM-1） were measured by ELISA method. ResultsCompared with blank group， the model group， Tadalafil group， and electroacupuncture group all had lower body mass and higher blood glucose levels at each time of testing （P＜0.01）. Compared with the blank group， the number of penile erections reduced in the model group， the level of CD31⁺ EMPs increased， the levels of serum T， FSH， LH， and VEGF reduced， and the levels of serum ET-1， LOX-1， sE-selectin， and sICAM-1 increased （P＜0.01）. Compared with the model group， the Tadalafil group and the electroacupuncture group showed an increased number of penile erections decreased level of CD31⁺ EMPs， increased levels of serum T， FSH， LH， and VEGF， and decreased levels of serum ET-1， LOX-1， sE-selectin， and sICAM-1 （P＜0.01）. Compared with the Tadalafil group， serum T， FSH， LH， VEGF levels increased and ET-1， LOX-1 levels decreased in the electroacupuncture group （P＜0.05 or P＜0.01）. HE staining and electron microscopic observation revealed that there was severe pathological damage to the cavernous tissue of the penis and vascular endothelial cells of the rat in the model group， which was ameliorated to a certain degree in both Tadalafil group and electroacupuncture group. ConclusionThe electroacupuncture prescription of "tonifying the kidney and dispelling stasis" can improve the erectile dysfunction of DMED rats， which is comparable to the effect of Tadalafi. Its mechanism of action may be related to the regulation of vascular endothelial function.

Objective To investigate the weight loss efficacy and mechanisms of oligopeptide-herbal medicine composite,and to provide new approaches for obesity treatment.Methods Twenty-three SPF female SD rats were randomly divided into control group(n=3)and modeling group(n=20).The control group was fed an ordinary diet for 6 consecutive weeks,and the modeling group was fed a high-fat diet for 6 consecutive weeks to establish a simple obesity rat model.After successful modeling,the modeling group was randomly divided into model group(n=10)and treatment group(n=10).The treatment group started anoint-and-massage therapy with oligopeptide-herbal medicine composite(3 g per rat per time,once a day,20 min each time),which was recorded as day 1.The control group and model group were not treated with oligopeptide-herbal medicine composite but received the same massage,and continued to be fed ordinary feed and high-fat feed respectively.Ten rats in the model group and ten in the treatment group were each divided into 3 groups,with 3,3,and 4 rats in each group.On days 3,6,and 9 after treatment,the rats in the three groups were weighed and cardiac blood collection was performed after isoflurane respiratory anesthesia.After euthanasia by cardiac bloodletting,abdominal subcutaneous adipose tissue(aSAT)and perirenal white adipose tissue(pWAT)samples were collected and weighed.Serum triglycerides(TG)and high-density lipoprotein cholesterol(HDL-C)levels were measured.Image J software was used to measure aSAT thickness and the diameter and area of perirenal white adipocytes and abdominal subcutaneous adipocytes.Immunofluorescence technique was used to observe the number of telocytes(TCs),cell junctions and exosomes per unit area of aSAT.Transmission electron microscopy was used to measure the length of telopodes(Tps)of dermal and subcutaneous TCs in abdominal skin,and to observe the distribution of exosome vesicles,rough endoplasmic reticulum,mitochondria,and cell junctions.Results Compared with the control group,the body weight of rats in the modeling group increased significantly(P＜0.05).Compared with the control group,rats in the model group showed a significant increase in serum TG level,a significant decrease in HDL-C level,and a significant increase in pWAT mass(all P＜0.05).Compared with the model group,the treatment group showed a significantly increased rate of body weight reduction,significantly decreased pWAT mass,significantly decreased serum TG level,and significantly increased HDL-C level(all P＜0.05).Compared with the model group,the treatment group showed significant reductions in aSAT thickness,as well as cell diameter and area in both aSAT and pWAT(all P＜0.05).Compared with the model group,the treatment group showed significantly increased number of TCs per unit area of aSAT,number of exosomes from TCs in aSAT,and Tps length(all P＜0.05).The treatment group showed an increasing trend in the numbers of mitochondria,rough endoplasmic reticulum,and cell junctions in TCs.Conclusion Oligopeptide-herbal medicine composite applied via anoint-and-massage therapy effectively improves obesity-related symptoms such as dyslipidemia and fat accumulation in obese rats by regulating TCs and their intercellular communication.

Objective The aim of this study was to explore the acceptance and influencing factors of self-collected urine samples for human papillomavirus(HPV)testing in cervical cancer screening among eligible women,and to provide scientific evidence for promoting this testing in low resource areas.Methods A population-based cross-sectional study was conducted from 2022 to 2023 at the Maternal and Child Health Hospital in Shuangliu district,Chengdu City Sichuan Province.The study subjects were women aged 21 to 69 years old,and a customized questionnaire was used to conduct general information and acceptance surveys on the partic-ipants.Results A total of 2,062 women were included,with an average age of 51.58±9.34 years.Among them,1,501(72.79％)women believed that self-sampling urine was very easy.However,although 1,333(64.65％)women were still willing to accept doctor sampling as a cervical cancer screening method,only 729(35.35％)were more willing to accept self-sampling urine HPV testing.Age,educational level,annual household income,awareness of HPV,HPV vaccination status,and a sense of shame about the doctor's sampling process were all associated with the acceptance of self-collected urine HPV testing among women undergoing cervical cancer screening(P＜0.001).The results of multivariate logistic regression analysis indicated that older women(OR=0.965,95％CI:0.951-0.979)and those who were not familiar with HPV(OR=0.760,95％CI:0.602-0.961)were more likely to undergo self sampling urine HPV testing,while those with junior high school education(OR=1.330,95％CI:1.053-1.682),high school education or a-bove(OR=1.990,95％CI:1.401-2.827),and a sense of shame towards the doctor's sampling process(OR=2.314,95％CI:1.706-3.142)were more likely to undergo self sampling urine for HPV testing.Conclusions Most women believe that self sampling urine for HPV testing is very easy,but compared to doctor sampling,only some women choose to self sample urine for HPV testing.Key health education interventions should be carried out for older and lower educated populations to promote acceptance of urine HPV testing.

The effective integration of ideological and political education into medical courses is one of the most important aspects of medical teaching reform and a key approach to fostering students'moral character.Based on the Outcomes-Based Education(OBE)philosophy,this article focuses on the learning outcomes of Medical Immunology ideological and political education,integrating relevant ideological and political elements with modular teaching content to form a themed"Medical Immunology Ideological and Politi-cal"teaching goal with distinctive features of Medical Immunology.Through a blended learning model of online and offline teaching methods,students are guided to learn modular teaching content.This study creatively integrates Medical Immunology theory with ideo-logical and political elements,carries out"Medical Immunology Ideological and Political"through professional knowledge,and achieves the goal of subtle and deep-seated ideological and political education.Moreover,it effectively improves students'comprehensive learning and application abilities,enabling them to apply what they have learned.

OBJECTIVE T o assess the in vitro antimicrobial activity of aztreonam-avibactam against metallo-β—lacta-mase(MBL)-producing carbapenem-resistant Enterobacteriaceae(CRE)using different antimicrobial susceptibility testing methods,evaluate the consistency of results based on the latest breakpoints recommended by European Committee on Antimicrobial Susceptibility Testing(EUCAST),and analyze the clinical applicability.METHODS The imipenem-or meropenem-resistant Enterobacteriaceae strains were isolated from the clinical specimens of Bei-jing Chaoyang Hospital from Jan.2019 to Mar.2023,MBL-producing CRE were selected after they were con-firmed by colloidal gold immunoassay and Sanger sequencing as the study subjects.The minimum inhibitory con-centrations(MICs)of single aztreonam and aztreonam-avibactam compounds were determined by using broth mi-crodilution method.In addition,the disk diffusion method and the gradient diffusion method were employed to further detect the in vitro susceptibility to aztreonam-avibactam.RESULTS Among the 87 strains of MBL-producing CRE that were included in the study,Escherichia coli was the most common species,accounting for 44.83％.NDM-5 was the predominant carbapenemase type,detected in 48.28％of the isolates.According to the latest EUCAST break-points,6 isolates were aztreonam-avibactam-resistant strains based on broth microdilution method,all of which were E.coli,and the resistance rate was 6.90％(6/87).However,the resistance rate that was determined by the disk diffu-sion method was significantly higher(22.99％,20/87).Among these,14 strains was within the area of technical uncer-tainty/resistant,making result interpretation difficult.In addition,the categorical agreement between the gradient diffu-sion method and the broth microdilution method reached up to 98.85％.CONCLUSIONS Aztreonam-avibactam has high antimicrobial activity against MBL-producing CRE.However,based on the latest EUCAST breakpoints,the antimicrobial susceptibility testing by the disk diffusion method results of some strains are hard to interpret.It is necessary to integrate with other methods for further validation.

Objective To investigate the weight loss efficacy and mechanisms of oligopeptide-herbal medicine composite,and to provide new approaches for obesity treatment.Methods Twenty-three SPF female SD rats were randomly divided into control group(n=3)and modeling group(n=20).The control group was fed an ordinary diet for 6 consecutive weeks,and the modeling group was fed a high-fat diet for 6 consecutive weeks to establish a simple obesity rat model.After successful modeling,the modeling group was randomly divided into model group(n=10)and treatment group(n=10).The treatment group started anoint-and-massage therapy with oligopeptide-herbal medicine composite(3 g per rat per time,once a day,20 min each time),which was recorded as day 1.The control group and model group were not treated with oligopeptide-herbal medicine composite but received the same massage,and continued to be fed ordinary feed and high-fat feed respectively.Ten rats in the model group and ten in the treatment group were each divided into 3 groups,with 3,3,and 4 rats in each group.On days 3,6,and 9 after treatment,the rats in the three groups were weighed and cardiac blood collection was performed after isoflurane respiratory anesthesia.After euthanasia by cardiac bloodletting,abdominal subcutaneous adipose tissue(aSAT)and perirenal white adipose tissue(pWAT)samples were collected and weighed.Serum triglycerides(TG)and high-density lipoprotein cholesterol(HDL-C)levels were measured.Image J software was used to measure aSAT thickness and the diameter and area of perirenal white adipocytes and abdominal subcutaneous adipocytes.Immunofluorescence technique was used to observe the number of telocytes(TCs),cell junctions and exosomes per unit area of aSAT.Transmission electron microscopy was used to measure the length of telopodes(Tps)of dermal and subcutaneous TCs in abdominal skin,and to observe the distribution of exosome vesicles,rough endoplasmic reticulum,mitochondria,and cell junctions.Results Compared with the control group,the body weight of rats in the modeling group increased significantly(P＜0.05).Compared with the control group,rats in the model group showed a significant increase in serum TG level,a significant decrease in HDL-C level,and a significant increase in pWAT mass(all P＜0.05).Compared with the model group,the treatment group showed a significantly increased rate of body weight reduction,significantly decreased pWAT mass,significantly decreased serum TG level,and significantly increased HDL-C level(all P＜0.05).Compared with the model group,the treatment group showed significant reductions in aSAT thickness,as well as cell diameter and area in both aSAT and pWAT(all P＜0.05).Compared with the model group,the treatment group showed significantly increased number of TCs per unit area of aSAT,number of exosomes from TCs in aSAT,and Tps length(all P＜0.05).The treatment group showed an increasing trend in the numbers of mitochondria,rough endoplasmic reticulum,and cell junctions in TCs.Conclusion Oligopeptide-herbal medicine composite applied via anoint-and-massage therapy effectively improves obesity-related symptoms such as dyslipidemia and fat accumulation in obese rats by regulating TCs and their intercellular communication.

Objective:To analyze the epidemiological characteristics of drug resistance genes of carbapenemase-producing Escherichia coli (CPECO) in Henan Province Hospital of Traditional Chinese Medicine from 2021 to 2023, providing data support and theoretical basis for controlling nosocomial infections of CPECO.Methods:Using a cross-sectional study, 30 carbapenem-resistant Escherichia coli (CRECO) strains confirmed by VITEK-2 Compact identification and drug sensitivity test in the Clinical Microbiology Laboratory of Henan Province Hospital of Traditional Chinese Medicine from 2021 to 2023 were tested, using carbapenemase inhibitor enhancement test to conduct preliminary screening of carbapenemases, and colloidal gold immunochromatography and polymerase chain reaction (PCR) were used to determine the phenotypes and genotypes of common carbapenemases ( blaKPC, blaNDM, blaVIM, blaIMP, blaOXA) respectively, and the genotypes ( blaSHV, blaTEM, blaCTX) of common extended Spectrum beta-lactamases (ESBL) were confirmed using PCR. The PCR amplification products of carbapenemase and ESBL positive strains were Sanger-sequenced, and the sequencing products were compared on the Blast website to determine the exact carbapenemase and ESBL genotypes. Sequence typing (ST) was performed on CPECO using the Achtman multi-locus sequence typing scheme to determine the cloning relationship between different strains. Results:A total of 21 CPECO strains were screened. Drug sensitivity test results showed that CPECO strains showed widespread drug resistance, with the resistance rate to monocyclic (aztreonam) and trimethoprim/sulfamethoxazole being over 60%(16/21, 14/21), and the resistance rate to other antibacterial drugs being 100%. Only the sensitivity to aminoglycosides and fosfomycin remained relatively high, and no strains resistant to tigecycline and colistin were found. Colloidal gold immunochromatography detected 18 blaNDM types, 2 blaKPC types, and 1 blaIMP type. Sequencing of drug resistance gene PCR products classified 17 blaNDM-5 strains, 1 blaNDM-4 strain, 2 blaKPC-2 strain, and 1 blaIMP-4 strain, which were completely consistent with the results of screening test and colloidal gold immunochromatography. ESBL resistance gene testing showed that the detection rate of blaTEM was 42.9%(9/21), blaCTX-M was 33.3%(7/21), and blaSHV was 4.8%(1/21). The rate of blaNDM producing CPECO carrying both ESBL resistance genes was 27.8%(5/18). The MLST typing results revealed 11 sequence types (STs), including one ST155 clonal complex and nine singleton STs. Among these, there were seven strains of ST167, five strains of ST410, and one strain each of ST58, ST68, ST69, ST93, ST131, ST155, ST648, ST1114, and ST3268. Conclusion:The main resistance mechanism identified in this study for CPECO was the production of blaNDM-5 carbapenemase, with a high proportion of strains also carrying blaTEM-1D and/or blaCTX-M-15 ESBLs. MLST typing found that the epidemic strain of CPECO showed certain polymorphism, but there were clonal transmission of multiple clonal complexes between ST167 and ST410.

Objective The aim of this study was to explore the acceptance and influencing factors of self-collected urine samples for human papillomavirus(HPV)testing in cervical cancer screening among eligible women,and to provide scientific evidence for promoting this testing in low resource areas.Methods A population-based cross-sectional study was conducted from 2022 to 2023 at the Maternal and Child Health Hospital in Shuangliu district,Chengdu City Sichuan Province.The study subjects were women aged 21 to 69 years old,and a customized questionnaire was used to conduct general information and acceptance surveys on the partic-ipants.Results A total of 2,062 women were included,with an average age of 51.58±9.34 years.Among them,1,501(72.79％)women believed that self-sampling urine was very easy.However,although 1,333(64.65％)women were still willing to accept doctor sampling as a cervical cancer screening method,only 729(35.35％)were more willing to accept self-sampling urine HPV testing.Age,educational level,annual household income,awareness of HPV,HPV vaccination status,and a sense of shame about the doctor's sampling process were all associated with the acceptance of self-collected urine HPV testing among women undergoing cervical cancer screening(P＜0.001).The results of multivariate logistic regression analysis indicated that older women(OR=0.965,95％CI:0.951-0.979)and those who were not familiar with HPV(OR=0.760,95％CI:0.602-0.961)were more likely to undergo self sampling urine HPV testing,while those with junior high school education(OR=1.330,95％CI:1.053-1.682),high school education or a-bove(OR=1.990,95％CI:1.401-2.827),and a sense of shame towards the doctor's sampling process(OR=2.314,95％CI:1.706-3.142)were more likely to undergo self sampling urine for HPV testing.Conclusions Most women believe that self sampling urine for HPV testing is very easy,but compared to doctor sampling,only some women choose to self sample urine for HPV testing.Key health education interventions should be carried out for older and lower educated populations to promote acceptance of urine HPV testing.