ObjectiveTo verify the association between the Ddit3-Trib3-Akt signaling pathway and rat spermatogenesis by constructing an in vitro co-culture system of testis. MethodsTesticular tissue blocks from 20-25-day-old male rats were placed in an in vitro culture system， and the culture medium was replaced every 2 to 3 days. PCR was used to verify the expression of marker genes of various spermatogenic cells. RNA interference technology was employed to verify the correlation between the Ddit3-Trib3-Akt signaling pathway and rat spermatogenesis. ResultsThe co-culture system could be continuously cultured for more than 2.5 months in vitro. RT-PCR showed that specific marker genes of spermatogonia， spermatocyte and spermoblast were expressed. The RNA and protein expression of Trib3 and Akt changed after the knocking down of Ddit3 and Trib3， respectively. It demonstrated the existence of Ddit3-Trib3-Akt signaling pathway in rat spermatogenesis. ConclusionThe culture time of more than 2.5 months indicates that the culture system can temporarily maintain the proliferation and differentiation of stem cells， and simultaneously maintain and stabilize spermatogenesis in a simple system. The successful validation of the Ddit3-Trib3-Akt signaling pathway also confirms that this culture system can be used to study possible molecular mechanisms of spermatogenesis in vitro.

OBJECTIVE To explore the ameliorative effect and potential mechanism of vitexin on inflammation in ulcerative colitis （UC） mice. METHODS The UC mice model was established by continuous administration of 3% dextran sulfate sodium solution for 5 days. Mice with successful modeling were randomly divided into UC group， vitexin low- and high-dose groups （vitexin-L and vitexin-H groups， 40， 80 mg/kg）， mesalazine group （400 mg/kg）， and vitexin-H+recombinant Jagged canonical Notch ligand 1 （rJagged-1） group （vitexin-H+rJagged-1 group， 80 mg/kg vitexin+1 mg/kg rJagged-1）， with 12 mice in each group. Another 12 normal mice were used as the control （CK） group. Mice in each group were administered the corresponding drugs or the corresponding drugs and normal saline by gavage and intraperitoneal injection once daily for 7 consecutive days. General conditions were observed during the experiment. At 24 h after the last administration， the disease activity index （DAI） score was evaluated. Colonic histopathological morphology was observed and scored. Macrophage polarization levels in the spleen and colon tissues were measured. The protein expressions of interleukin-6 （IL-6）， IL-10， tumor necrosis factor-α （TNF-α）， transforming growth factor-β 1 （TGF-β 1 ）， Jagged-1， Notch1 and Notch intracellular domain （NICD） in colonic tissues were determined. RESULTS Compared with the UC group， the symptoms （reduced food and water intake， dull fur， etc.） and pathological changes （epithelial cell shedding， inflammatory cell infiltration， etc.） were significantly improved in the vitexin-L， vitexin-H and mesalazine groups. DAI scores， colonic histopathological scores， M1 macrophage contents in spleen tissue， M1/M2 macrophage ratios， M1 macrophage proportions in colon tissue， and protein expressions of IL-6， TNF-α， Jagged-1， Notch1 and NICD in colon tissue were significantly decreased （ P ＜0.05）. Meanwhile， the M2 macrophage contents in spleen tissue， M2 macrophage proportions in colon tissue， and protein expressions of IL-10 and TGF-β 1 in colon tissue were significantly increased （ P ＜0.05）. Moreover， the improvement effects in the vitexin-H and mesalazine groups were significantly superior to those in the vitexin-L group （ P ＜0.05）. Compared with the vitexin-H group， the above symptoms and pathological changes were aggravated， and all quantitative indicators were significantly reversed in the vitexin-H+rJagged-1 group （ P ＜0.05）. CONCLUSIONS Vitexin can ameliorate the inflammation of UC mice， which is associated with its inhibition of the Jagged-1/Notch1 pathway and regulation of macrophage polarization （inhibition of M1-type polarization and promotion of M2-type polarization）.

The Expert Consensus on Clinical Application of Qinbaohong Zhike Oral Liquid in Treatment of Acute Bronchitis and Acute Attack of Chronic Bronchitis （GS/CACM 337-2023） was released by the China Association of Chinese Medicine on December 13^th， 2023. This expert consensus was developed by experts in methodology， pharmacy， and Chinese medicine in strict accordance with the development requirements of the China Association of Chinese Medicine （CACM） and based on the latest medical evidence and the clinical medication experience of well-known experts in the fields of respiratory medicine （pulmonary diseases） and pediatrics. This expert consensus defines the application of Qinbaohong Zhike oral liquid in the treatment of cough and excessive sputum caused by phlegm-heat obstructing lung， acute bronchitis， and acute attack of chronic bronchitis from the aspects of applicable populations， efficacy evaluation， usage， dosage， drug combination， and safety. It is expected to guide the rational drug use in medical and health institutions， give full play to the unique value of Qinbaohong Zhike oral liquid， and vigorously promote the inheritance and innovation of Chinese patent medicines.

Objective To explore the value of analysis on the incidence of healthcare-associated infection(HAI)based on disease diagnosis-related grouping(DRG),case mix index(CMI),and relative weight(RW).Methods All discharged cases,DRG and HAI status in a tertiary first-class general hospital from January 1 to December 31,2023 were analyzed retrospectively.Incidences of HAI in different departments were adjusted and compared by CMI.Incidences of HAI in different DRG groups were adjusted by RW.Results Among the 47 695 cases included in the analysis,757 were HAI cases,including 225 DRG groups.The department of critical care medicine had the highest incidence of HAI(11.98％).After CMI adjustment,departments with higher incidence of HAI were main-ly the department of respiratory and critical care medicine(3.96％),department of critical care medicine(3.04％),and department of neurology(2.85％),et al.DRG groups with the top five high incidence of HAI were AH11(tracheotomy and with ventilator support ≥96 hours or extracorporeal membrane oxygenation[ECMO],accompa-nied by major complications and comorbidity[MCC],50.00％),BC29(ventricular shunt and revision surgery,31.43％),BB21(craniotomy other than trauma,accompanied by MCC,27.56％),BB11(craniotomy of brain trauma,accompanied by MCC,26.32％),and GB1A(major surgery of esophagus,stomach,and duodenum,accompanied by major or moderate complications and comorbidity,16.00％).After RW adjustment,the DRG groups with the top five high incidence of HAI were ES21(respiratory system infection/inflammation,accompanied by MCC,5.89％),BR21(cerebral ischemic disease,accompanied by MCC,5.17％),FR11(heart failure,shock,accompanied by MCC,4.80％),BC29(4.57％)and AH11(3.57％).Conclusion Analyzing the incidence of HAI based on CMI and RW can help to identify key departments and disease groups for infection prevention and control,and provide reference for precise prevention and control of HAI in the new era.

Objective To investigate the role and mechanism of microRNA-21-5p(miR-21)in cisplatin(CDDP)resistance in non-small cell lung cancer.Methods The expression of miR-21 in cancer tissues and paracancerous tissues of non-small cell lung cancer patients was detected by real-time fluorescence quantitative PCR.Non-small cell lung cancer CDDP-resistant cells H1299/CR and H1975/CR were constructed using non-small cell lung cancer cell lines H1299 and H1975.CCK-8 was used to detect the cell activity of each group of cells under CDDP treatment,and apoptosis was analyzed by flow cytometry.Dual luciferase was used to detect the role of miR-21 in relation to PTEN in H1299 cells,RT-qPCR was used to detect the miR-21 level,and Western blotting was used to detect the protein expression level of PTEN and PTEN downstream Wnt/β-catenin signaling pathway.H1299 and H1299/CR were used to construct a hormonal nude mouse model to verify the effect of miR-21 on the sensitivity of CDDP treatment in non-small cell lung cancer.Results The expression of miR-21 was significantly higher in cancer tissues than in adjacent normal tissues(P＜0.001).The expression of miR-21 in drug-resistant cells H1299/CR and H1975/CR was significantly elevated compared to that in H1299 and H1975 cells(P＜0.05).After transfection with miR-21 inhibitor,cell viability in the inhibitor group was significantly lower than in the inhibitor NC group when treated with CDDP ≥12 pmol/L(P＜0.001).Flow cytometry analysis showed that while the apoptosis rate in the CDDP and miR-21 inhibitor groups did not significantly differ from that in the untreated group,the apoptosis rate in the CDDP+miR-21 inhibitor group was significantly higher(P＜0.001).In contrast,after transfection with miR-21 mimic,the cell viability of the mimic-NC group was significantly reduced compared to the miR-21 mimic group when treated with CDDP ≥12 pmol/L(P＜0.05).TargetScan predicted that miR-21 could bind to the 3-UTR region of PTEN.Dual-luciferase reporter assays confirmed that miR-21 directly targeted PTEN.Overexpression of PTEN(PTEN-OE)together with miR-21 mimic co-transfection in H1299 cells resulted in decreased PTEN expression and increased levels of p-β-catenin(Ser552,Ser675)and β-catenin,while the PTEN expression in the miR-21 mimic+PTEN-OE group was elevated,with a corresponding decrease in p-β-catenin andβ-catenin levels.Flow cytometry showed that apoptosis was significantly increased in the mimic-NC group after CDDP treatment(P＜0.05).In H1299 xenograft models,after treatment with miR-21 mimic and CDDP,tumor growth was slower in the control+CDDP group than in the control group;tumors were larger in the miR-21+CDDP group than in the control+CDDP group at all time points.TUNEL staining revealed that apoptosis in the tumor tissues of the control+CDDP group was higher than in the control group,while apoptosis was reduced in the miR-21+CDDP group compared to the control+CDDP group.In the H1299/CR xenograft model,after 5 days of treatment,the tumors were smaller in the H1299/CR+CDDP group than in the H1299/CR group,and those in the H1299/CR+CDDP+inhibitor group were smaller than in the H1299/CR+CDDP group.TUNEL staining showed that apoptosis was increased in the H1299/CR+CDDP group compared to the H1299/CR group,and further increased in the H1299/CR+CDDP+inhibitor group.Conclusion In non-small cell lung cancer,miR-21 overexpression inhibits PTEN level and activates the Wnt/β-catenin pathway involved in the development of CDDP resistance in non-small cell lung cancer.Inhibition of miR-21 expression in tumor cells will enhance the sensitivity of tumor cells to CDDP.

Aim To study the role and mechanism of ML210 in glioma.Methods The cell viability was detected by CCK8 assay.The percentage of dead cells was detected by SYTOXstaining.The role of ferroptosis-signaling pathway in gliomas was detected bygenomics.Cell proliferation was observed by EdU staining and clone formation assay.Cell migration ability was detec-ted by scratch healing assay.The apoptosis was detec-ted by flow cytometry.Cell mitochondrial function was assesses by JC-1 staining.The mechanism of action of ML210 was detected by molecular docking coupled with immunoblotting assay(Western blot).The levels of ROS,MDA were observed by ELISA.Results Compared with the control group,ML210 treatment dose-dependently decreased glioma cell viability,in-hibited cell proliferation,migration,and increased cell apoptosis and mitochondrial dysfunction,which were reversed by ferroptosis antagonists.Gene microarray screening showed that 688 genes of the ferroptosissig-naling pathway were aberrant and 10 signaling path-ways were altered in gliomas.Molecular docking re-sults showed that ML210 binding to GPX4 significantly inhibited the protein expression level of GPX4 and pro-moted the elevation of ROS and MDA levels.Conclu-sions ML210 produces anti-glioma cells via GPX4-mediated ferroptosis pathway.

OBJECTIVE To observe the effects of silicone oil removal detergent and multi-enzyme detergent on re-moving the residual silicone oil from digestive endoscopes so as to provide bases for clinical cleaning of digestive endoscopes.METHODS Totally 60 colonoscopes that were used in the endoscopy center of Shenzhen Hospital of Southern Medical University in Jul.2024 were chosen as the research subjects and were randomly divided in to the experimental group with 30 colonoscopes(the silicone-removing multi-enzyme detergent group)and the control group with 30 colonoscopes(the multi-enzyme detergent group),the two groups were respectively treated with sil-icone-removing multi-enzyme detergent and multi-enzyme detergent during the cleaning process.The average bac-terial colony counts of rinsed endoscopes,residual water droplet counts of the cleaned endoscopes and absorbance values of residual methyl silicone oil in the biopsy forceps channels of the endoscopes were observed and compared between the two groups so as to evaluate the effect on removing silicone oil.RESULTS The median average bacteri-al colony counts were 3.05(1.62,4.68)× 103 CFU/piece in the experimental group,36.70(34.66,38.10)× 103 CFU/piece in the control group(Z=-6.654,P＜0.001).The median counts of residual water droplets on the en-doscopes were 7.00(5.00,16.50)in the experimental group,31.50(12.00,43.00)in the control group(Z=-3.940,P＜0.001).The median absorbance value of residual methyl silicone oil in the biopsy forceps chan-nels of the endoscopes was 6.85(5.58,7.85)× 10-3 in the experimental group,36.50(35.50,43.65)× 10-3 in the control group(Z=-6.655,P＜0.001).CONCLUSION The silicone-removing multi-enzyme detergent shows more remarkable effect on removing the residual silicone oil on the endoscopes than the multi-enzyme detergent,it improves the cleaning effect of the endoscopes and is worthy to be verified and promoted in the hospital.

The coil adverse events in the U.S.Food and Drug Administration Manufacturer and User Facility Device Experience(MAUDE)database from January 2021 to June 2024 were analyzed retrospectively.The risks of coils during the clinical application and their causes were explored with hospital survey and expert demonstration in Shandong Province.Some improving measures were put forward for the safe use of coils,including implementing the main responsibility of the registrant,enhancing the professional skills of the using institutions and strengthening the supervision of the supervisory authorities.[Chinese Medical Equipment Journal,2025,46(6):83-87]

OBJECTIVE To understand the disinfectant-resistant genes in the gram-negative bacteria isolated from the dirt retention on air recure filters of air conditioners and observe the drug resistance.METHODS The dirt re-tention samples were collected from the air return filters of air conditioners of some wards in 3 hospitals of Wuhan(Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology,Wuhan Central Hospital,and Hubei Provincial Maternal and Child Health Hospital)from 2018 to 2024.The gram-negative bac-teria were screened out,the disinfectant-resistant genes in the strains were detected by polymerase chain reaction(PCR),and the results of drug susceptibility test were analyzed.RESULTS Of 354 dirt retention samples that were collected from the air return filers of air conditioners,77 were detected with 138 strains of gram-negative bacteria,87 of which were Acinetobacter baumannii,50 were Enterobacteriaceae,and 1 was Pseudomonas aerug-inosa.The detection rates of qacEΔ1,qacEΔ1-sul1,aceI and qacA/B were 73.19％,82.61％,69.57％and 2.90％,respectively.None of the strains were detected with qacC,qacH or qacJ.The result of drug susceptibility test showed that 76.81％of the gram-negative bacteria were resistant to at least 1 type of antibiotic;93 strains of multidrug-resistant gram-negative bacteria were isolated,most of which were isolated from intensive care unit(ICU).The detection rates of qacEΔ1 and qacEΔ1-sul1 were higher in the drug-resistant strains than those in the non-drug-resistant strains;there were significant differences in the drug resistance rates to carbapenems,quin-olones and β-lactams between the qacEΔ1-sul 1-positive strains and the qacEΔ1-sul1-negative strains(P＜0.05).CONCLUSIONS There are drug-resistant gram-negative bacteria contaminations in some wards of the 3 hospitals in Wuhan.The carrying rates of disinfectant-resistant genes of the strains are high,and the strains show varying degree of resistance to the commonly used antibiotics;the strains carrying the qacEΔ1-sul1 have certain statistical association with the drug resistance.It is suggested that the hospital should take targeted disinfec-tion measures for the environment and reasonably use antibiotics.

Objective:To explore the activation pathway of CD4+CD28-T cells and their specific role in rheumatoid arthritis(RA).Methods:Peripheral blood mononuclear cells(PBMCs)were isolated from RA patients and healthy individuals.Flow cytome-try was used to detect the proportion and activation level of CD4+CD28-T cells,as well as the expressions of perforin and TNF-α,and to analyze the correlation between the proportion of CD4+CD28-T cells and age,C-reactive protein,anti-cyclic citrullinated peptide(CCP)antibodies,rheumatoid factors(RF)and erythrocyte sedimentation rate(ESR).The expression of Toll-like receptor(TLR)2 on CD4+CD28-T cells was detected by flow cytometry.Purified RA CD4+T cells were stimulated with anti-CD3 antibody and TLR2 agonist alone or in combination,and the activation level and cytokine secretion of CD4+CD28-T cells were detected by flow cytometry.Results:The proportion of CD4+CD28-T cells in RA patients was higher than that in healthy individuals(P<0.05),and was positively correlated with rheumatoid factors and erythrocyte sedimentation rate(r=0.554 1,P<0.01;r=0.363 4,P<0.01).Compared with CD4+CD28+T cells,CD25 expression was downregulate(P<0.01),but CD40L and TLR2 expressions were upregulated(P<0.01;P<0.000 1)in CD4+CD28-T cells from RA patients,and they secreted more perforin and TNF-α(P<0.001;P<0.001).After TLR2 ago-nist stimulation,there was no significant change in CD40L and CD25 expression,as well as perforin and TNF-α secretion in CD4+CD28-T cells from RA patients.Conclusion:The proportion of CD4+CD28-T cells is increased in RA patients,and they secrete a large amount of perforin and TNF-α,which may be involved in the inflammatory state and joint damage of RA,but TLR2 cannot re-place CD28 in activating CD4+CD28-T cells.