Objective:To discuss the effect of zanubrutinib(BGB-3111)combined with bortezomib(Btz)on the apoptosis of the human multiple myeloma(MM)cells,and to clarify its possible mechanism.Methods:The human MM cell lines U266,PS-R,RPMI8226,KMS28-PE,KMS28-BM,and H929 were cultured in vitro.Western blotting method was used to detect the expression level of Bruton's tyrosine kinase(BTK)protein in various cells;cell counting kit-8(CCK-8)method was used to detect the survival rates of the RPMI8226,U266,and KMS28-BM cells after treated with 0,10,20,30,40,and 50 μmol·L?1 BGB-3111.The RPMI8226,U266,and KMS28-BM cells at the logarithmic growth phase were selected and divided into control group,BGB-3111 group,Btz group,and BGB-3111+Btz group.Flow cytometry was used to detect the apoptotic rates of the cells in various groups;Western blotting method was used to detect the expression levels of myeloid cell leukemia 1(MCL-1),B-cell lymphoma-2(Bcl-2),Bcl-2-interacting mediator of cell death(Bim),phosphorylated Bim(p-Bim),P65,phosphorylated P65(p-P65),tumor necrosis factor receptor-associated factor(TRAF)2,and tumor necrosis factor alpha-induced protein 3(A20)in different kinds of cells.The U266 cells were divided into A20 overexpression group(A20-OE group)and empty vector control group(EV group).Each group was further divided into control group,BGB-3111 group,Btz group,and BGB-3111+Btz group.The corresponding plasmids were transfected;Western blotting method was used to detect the transfection efficiency of the cells in various groups;flow cytometry was used to detect the apoptotic rates of the cells in various groups after over-expression of A20.Results:The Western blotting results showed that compared with KMS28-BM cells,the expression levels of BTK protein in the U266,RPMI8226,and H929 cells were significantly increased(P<0.05 or P<0.01).The CCK-8 results showed that compared with 0 μmol·L?1 BGB-3111 group,the survival rates of the RPMI8226 and U266 cells in 10,20,30,40,and 50 μmol·L?1 BGB-3111 groups were significantly decreased(P<0.05 or P<0.01),and the survival rates of the KMS28-BM cells in 20,30,40,and 50 μmol·L?1 BGB-3111 groups were significantly decreased(P<0.05).Compared with RPMI8226 and U266 cells,the survival rates of the KMS28-BM cells in 20,30,and 40 μmol·L?1 BGB-3111 groups were significantly increased(P<0.05).Therefore,10 μmol·L?1 BGB-3111 was selected for subsequent experiments.The flow cytometry results showed that compared with control group,the apoptotic rates of the RPMI8226 and U266 cells in BGB-3111 group,Btz group,and BGB-3111+Btz group were significantly increased(P<0.05 or P<0.01);compared with BGB-3111 group and Btz group,the apoptotic rates of the RPMI8226 and U266 cells in BGB-3111+Btz group were significantly increased(P<0.01);compared with control group,the apoptotic rates of the KMS28-BM cells in Btz group and BGB-3111+Btz group were significantly increased(P<0.01);compared with BGB-3111 group,the apoptotic rate of the KMS28-BM cells in BGB-3111+Btz group was significantly increased(P<0.01);compared with EV group,the apoptotic rates of the cells in A20-OE group in Btz group and BGB-3111+Btz group were significantly increased(P<0.05).The Western blotting results showed that compared with control group,the expression levels of Bim protein in the RPMI8226 and U266 cells in BGB-3111 group,Btz group,and BGB-3111+Btz group were significantly increased(P<0.05),while the expression levels of MCL-1,p-Bim,and Bcl-2 proteins in the RPMI8226 and U266 cells in Btz group and BGB-3111+Btz group were significantly decreased(P<0.05);compared with BGB-3111 group and Btz group,the expression levels of Bim protein in the RPMI8226 and U266 cells in BGB-3111+Btz group were significantly increased(P<0.05),while the expression levels of MCL-1,p-Bim,and Bcl-2 proteins were significantly decreased(P<0.05).Compared with control group,the expression levels of p-P65 protein in the RPMI8226 and U266 cells in Btz group and BGB-3111+Btz group were significantly increased(P<0.05),while the expression levels of TRAF2 and A20 proteins were significantly decreased(P<0.05);compared with BGB-3111 group and Btz group,the expression levels of p-P65 protein in the RPMI8226 and U266 cells in BGB-3111+Btz group were significantly increased(P<0.05),while the expression levels of TRAF2 and A20 proteins were significantly decreased(P<0.05).The flow cytometry results showed that compared with EV group,the expression level of A20 protein in A20-OE group cells was significantly increased(P<0.01).Conclusion:BGB-3111 induces apoptosis in the MM cells by inhibiting BTK activity and enhances the pro-apoptotic effect of Btz.Over-expression of A20 increases the sensitivity of the MM cells to the combined treatment.The antitumor effect may be related to the inhibition of the nuclear factor kappa B(NF-κB)signaling pathway.

Objective To understand the current situation of healthcare-associated infection(HAI)in China,pro-vide data support and decision-making basis for formulating scientific and effective strategies for HAI prevention and control.Methods A nationwide cross-sectional survey on HAI was conducted among various types and levels of medical institutions in China according to a unified protocol of bedside surveys and case investigations.Results In 2024,a total of 5 736 medical institutions and 2 751 765 patients were surveyed.Among them,34 889 HAI cases were identified,with a prevalence rate of 1.27％.The number of HAI episodes was 38 032,and case prevalence rate was 1.38％.The prevalence rate of HAI in medical institutions in different regions of China ranged from 0.66％to 2.35％.Among medical institutions of different scales,those with a bed capacity of ≥900 had the high-est incidence of HAI,reaching 1.65％.The most common infection site was the lower respiratory tract(44.66％),followed by the urinary tract(12.94％),surgical site(9.32％),upper respiratory tract(7.02％),and bloodstream infection(5.78％).The top 3 departments with the highest HAI rates were the general intensive care unit(10.02％),department of neurosurgery(5.51％),and department(group)of hematology(5.34％).A total of 23 238 strains of HAI pathogens were detected,with 10 714 strains(46.10％)from lower respiratory tract speci-mens.The top 5 detected strains were Klebsiella pneumoniae(14.76％),Pseudomonas aeruginosa(13.33％),Escherichia coli(12.79％),Acinetobacter baumannii(9.23％),and Staphylococcus aureus(7.88％).231 944 pa-tients underwent class Ⅰ incision surgery were monitored,with 1 647 cases experienced surgical site infection,and the prevalence rate of surgical site infection was 0.71％.The number of patients who should undergo pathogen de-tection(patients receiving therapeutic and therapeutic combined prophylactic antimicrobial agents)was 715 179,while the actual number was 480 492,with a pathogen detection rate of 67.18％.425 225 patients received patho-genic detection before treatment,with a detection rate of 59.46％.Conclusion The overall HAI prevalence in Chi-na is lower,showing disparities among medical institutions of different regions and scales.Therefore,precise imple-mentation of measures is necessary for HAI prevention and control,with a focus on high-risk institutions and high-risk departments,key areas,and critical procedures.All levels of medical institutions should continuously reduce the incidence of HAI by strengthening monitoring,standardizing the use of antimicrobial agents,and reinforcing basic HAI prevention and control measures.

Objective To understand the current situation of healthcare-associated infection(HAI)in China,pro-vide data support and decision-making basis for formulating scientific and effective strategies for HAI prevention and control.Methods A nationwide cross-sectional survey on HAI was conducted among various types and levels of medical institutions in China according to a unified protocol of bedside surveys and case investigations.Results In 2024,a total of 5 736 medical institutions and 2 751 765 patients were surveyed.Among them,34 889 HAI cases were identified,with a prevalence rate of 1.27％.The number of HAI episodes was 38 032,and case prevalence rate was 1.38％.The prevalence rate of HAI in medical institutions in different regions of China ranged from 0.66％to 2.35％.Among medical institutions of different scales,those with a bed capacity of ≥900 had the high-est incidence of HAI,reaching 1.65％.The most common infection site was the lower respiratory tract(44.66％),followed by the urinary tract(12.94％),surgical site(9.32％),upper respiratory tract(7.02％),and bloodstream infection(5.78％).The top 3 departments with the highest HAI rates were the general intensive care unit(10.02％),department of neurosurgery(5.51％),and department(group)of hematology(5.34％).A total of 23 238 strains of HAI pathogens were detected,with 10 714 strains(46.10％)from lower respiratory tract speci-mens.The top 5 detected strains were Klebsiella pneumoniae(14.76％),Pseudomonas aeruginosa(13.33％),Escherichia coli(12.79％),Acinetobacter baumannii(9.23％),and Staphylococcus aureus(7.88％).231 944 pa-tients underwent class Ⅰ incision surgery were monitored,with 1 647 cases experienced surgical site infection,and the prevalence rate of surgical site infection was 0.71％.The number of patients who should undergo pathogen de-tection(patients receiving therapeutic and therapeutic combined prophylactic antimicrobial agents)was 715 179,while the actual number was 480 492,with a pathogen detection rate of 67.18％.425 225 patients received patho-genic detection before treatment,with a detection rate of 59.46％.Conclusion The overall HAI prevalence in Chi-na is lower,showing disparities among medical institutions of different regions and scales.Therefore,precise imple-mentation of measures is necessary for HAI prevention and control,with a focus on high-risk institutions and high-risk departments,key areas,and critical procedures.All levels of medical institutions should continuously reduce the incidence of HAI by strengthening monitoring,standardizing the use of antimicrobial agents,and reinforcing basic HAI prevention and control measures.

Objective:To explore the mechanism of follicular helper T (Tfh) cells, i.e. CD4 +CXCR5 +T cells, and the secreted cytokine programmed death factor 1 (PD-1) in the pathogenesis of tuberculosis, and to explore the significance of Tfh cells and PD-1 in the treatment of tuberculosis. Methods:Flow cytometry was used to detect the changes of Tfh cells and PD-1 in mononuclear cells during the treatment cycle of tuberculosis.Results:Before treatments, the ratio of Tfh cells/CD4 +T cells in peripheral blood mononuclear cells in the pulmonary tuberculosis group was 3.37%±0.45%, which was significantly higher than 2.21%±0.47% of the healthy control group ( P<0.01), and significantly higher than 2.39%±0.38% after treatments ( P<0.01). Before treatments, the ratio of CD4 +CXCR5 +PD-1 +T cells/Tfh cells in the peripheral blood of the tuberculosis group was 25.33%±10.08%, which was significantly higher than 8.42%±2.31% of the healthy control group ( P<0.01), and significantly higher than 11.35%±2.65% after treatments ( P<0.01). After treatments, the levels of Tfh cells and PD-1 in the sputum smear-negative group and the sputum smear-negative group were lower than that before treatments, and the difference between the groups was statistically significant (all P<0.05). Conclusions:The levels of Tfh cells and PD-1 in patients with tuberculosis are significantly higher than those in healthy people, and after drug treatment, the levels of both can be reduced. With the prolongation of the treatment cycle, the sputum smear-transforming group and the non-negative group began to show significant differences. In the course of pulmonary tuberculosis, monitoring changes in Tfh cells and PD-1 levels is helpful for the diagnosis of tuberculosis, and has certain guiding significance for its treatment and outcome.

Objective To study the prognostic role of plasma neutrophil gelatinase-associated lipocalin (NGAL) early after renal transplantation.Methods A total of 37 kidney recipients were enrolled from Department of Organ Transplantation,Guangdong Second Provincial General Hospital within a 12-month period of time.Plasma NGAL was measured immediately before and at 6 and 12 h post-transplantation.Changes of serum creatinine were documented daily within the first week postoperation.Acute kidney injury (AKI)/graft rejection during the first week after transplantation was the outcome variable.Results The levels of serum NGAL in the 37 patients were (311.14 ± 102.69),(317.81 ± 107.28) and (312.16 ± 134.80) μg/L respectively immediately before and at 6 and 12 h post-transplantation.There was no significant difference in serum NGAL levels before and 6 h or 12 h after operation (P =0.70,and P =0.96).There were no significant differences in gender and age between the two groups (P =0.29,and P =0.20).There was significant difference in creatinine levels between the AKI group and the non-AKI group (P =0.002) and between pre-operation and 6 or 12 h postoperation.The preoperative levels of serum NGAL in AKI group and non-AKI group were (333.58 ± 116.30) and (300.36 ± 96.15) μg/L (P =0.36),and those were (383.3 ± 147.16) and (286.32 ± 65.97) μg/L (P＜0.01) at 6 h,and (437.33 ± 164.16) and (252.08 ± 57.53) μg/L (P＜ 0.001) at 12 h after operation.The sensitivity and specificity of serumNGAL (317μg/L at 12 h after operation as the cutoff value) predicting AKI was 100％ and 92％ respectively,which was much better than that of serum creatinine at the corresponding time point (sensitivity =66.7％,and specificity =61.9％).Conclusion Plasma NGAL,particularly at 12 h after transplantation,is a very sensitive and specific biomarker for predicting AKI.

AIM:To explore a novel method to isolate human nucleus pulposus mesenchymal stem cells (hNP-MSCs) in vitro and to identify their biological characteristics.METHODS:The explant culture method was employed to isolate hNP-MSCs from nucleus pulposus tissue obtained by percutaneous endoscopic lumbar discectomy (PELD).The isolated cells were passaged for purification and cultured in vitro followed by morphological observation.The cell proliferation ability was detected by CCK-8 assay.Growth curves of the cells were drawn and surface antigens were detected by flow cytometry.The cells at the 3rd~6th passages were induced for adipogenic, osteogenic and chondrogenic differentiation, and examined by oil red O staining, alizarin red staining and Alcian blue staining.RESULTS:The cells with self-renewal were obtained from nucleus pulposus tissue obtained by PELD.The results of flow cytometry analysis revealed that the cells were positive for CD29, CD44, CD90, CD73 and CD105, but negative for CD34 and CD45.The proliferative capacity was consistent with the growth characteristics of MSCs and multilineage differentiation potential was identified.CONCLUSION:A novel method to efficiently isolate and culture hNP-MSCs,PELD combined with explant culture method,was established, which would promote the study of regenerative medicine based on hNP-MSCs.

Objective To establish a diagnostic model of multiple cytokines for differential diagnosis of tuberculous pleural effusion , and compare its diagnostic accuracy with tuberculosis infected T cells detection ( T-SPOT.TB ) in order to evaluate its diagnostic performance.Methods Case-control study.Totally 147 patients with pleural fluid in Tianjin Haihe Hospital were enrolled and categorized as tuberculous pleural effusion group ( n=95 ) and malignant pleural effusion group ( n=52 ) from December 2011 to June 2013.Pleural effusion cytokines including interferon-γ( IFN-γ) , C-X-C motif chemokine 10 (CXCL-10), tumor necrosis factor-α(TNF-α), vascular endothelial growth factor (VEGF), IL-2, IL-16, IL-17, IL-27 and IL-33 were tested by liquid chip technology and analyzed by Binary Logistic regression and receiver operating characteristic curve (ROC), and the pleural effusion was also detected by tuberculosis infected T cells detection ( T-SPOT.TB) as a control.Results The comparison of the AUC of cytokines is:CXCL-10>IL-27>IFN-γ>IL-33 >IL-17>IL-16>TNF-α>VEGF>IL-2; After that, CXCL-10, IFN-γ, IL-27 and IL-33 were included the Binary Logistic regression model.The regression equation is P=1/1+e-( -16.851+0.390 ×IFN-γ+0.006 ×IL-27+0.020 ×IL-33).The AUC, sensitivity and specificity of the diagnostic model were 99.5%, 96.84%, and 98.08%, respectively.Both AUC and sensitivity of the diagnostic model were superior to those of any single index.Compared with T-SPOT.TB (0.995 ±0.003), the AUC of the diagnostic model (0.921 ±0.023) was significantly greater ( Z=3.235, P <0.01), but no significant difference was found when it comes to diagnostic results consistency (χ2 =0.062 5, P>0.05).The Kappa of the two methods was 0.795, which meant fine agreement of the evaluations of the two raters.Conclusion The application of liquid array technology of high sensitivity and repeatability with high throughput provided a novel insight and method in the clinical diagnosis , treatment and prevention for tuberculous pleural effusion scientifically and accurately.

Objective To evaluate the clinical value of bedside index for severity in acute pancreatitis (BISAP) and APACHEⅡ score in predicting the severity and organ failure of acute pancreatitis (AP).Methods One hundred eighty-five patients of AP admitted to Department of Gastroenterology of First affiliated Hospital of Soochow University from January 2012 to December 2014 were studied retrospectively.According to BISAP score, patients who were ≥3 points were considered as high risk group, while ＜3 points were considered as low risk group.According to APACHEⅡ score, patients who were ≥ 8 points were considered as high risk group, while ＜ 8 points were considered as low risk group.According to the criteria of Pancreatic Diseases Group of Chinese Society of Gastroenterology of Chinese Medical Association, the patients were diagnosed as mild acute pancreatitis (MAP), moderately severe acute pancreatitis (MSAP), and severe acute pancreatitis (SAP).The BISAP, APACHEⅡ scores were calculated and compared between MAP group and MSAP + SAP group, respectively.The incidence of MSAP + SAP between high risk group and low risk group was also compared.The area of ROC curve (AUC) was used to evaluate the ability of BISAP and APACHEⅡ scoring system for predicting the severity of AP and the multiple organ dysfunction syndromes (MODS).Results Among 185 patients, MAP was identified in 101 patients, MSAP in 76 patients and SAP in 8 patients.Twenty-five MSAP patients developed organ dysfunction, and all the 8 SAP patients developed organ dysfunction.The BISAP scores of MSAP + SAP group and MAP group were (1.43 ± 0.89), (0.38 ± 0.61),andAPACHⅡ scores were (2.45± 1.36), (0.87± 0.62), the scores of MSAP+ SAP group were significantly higher than those in MAP group (P ＜0.01).In the 137 patients of low risk BISAP group, there were 47 MSAP + SAP patients (34.3％), while in the 48 patients of high risk BISAP group, there were 37 MSAP + SAP patients (77.0％);in the 153 patients of low risk APACHEⅡ group, there were 56 MSAP + SAP patients (36.6％), while in the 32 patients of high risk APACHEⅡ group, there were 28 MSAP + SAP patients (87.5％);the incidence of MSAP + SAP patients was significantly higher in high risk group than that in low risk group (P＜0.01).The AUC of BISAP, APACHEⅡ for MSAP+ MAP was 0.804 (95％ CI 0.738 ～ 0.870), 0.794 (95％ CI 0.725 ～ 0.863), and the AUC for organ dysfunction was 0.758 (95％ CI 0.686 ～0.830), 0.781 (95％ CI 0.710 ～0.852) , and the difference between BISAP and APACHE Ⅱ was not statistically significant (P ＞ 0.05).Conclusions The BISAP has the prediction ability for AP severity and prognosis similar to APACHEⅡ , and it consists of only 5 parameters and can be completed in the first 24 h of admission, therefore it is worth of clinical application.

Objective:To evaluate the feasibility and safety of laparoscopic jejunostomy with central venous catheterization set (CVC, Arrow International Inc., USA) during the operation of totally minimally invasive Ivor-Lewis esophagectomy (MIIE). Methods:The clinical data of 88 patients with esophageal squamous cell carcinoma who were admitted to the Fudan University Cancer Hospital from February 2013 to April 2014 were retrospectively analyzed. Among them, 48 patients with early mid-lower esophageal cancer un-derwent laparoscopic jejunostomy with CVC, and 40 patients accepted nasogastric tube nutrition. Short-term clinical outcomes were collected. Results:No significant difference in nutrition index was found between the two groups, but the rate of unplanned extubation in the laparoscopic jejunostomy with CVC group was less than that in the nasogastric tube nutrition group. Conclusion:Laparoscopic jejunostomy with CVC set is a safe and feasible technique. It is potentially accepted as an optional approach in MIIE for post-operative nutrition support.

Objective To analyze the metastatic rule of common hepatic artery lymph node of thoracic esophageal squamous cell carcinoma,and to investigate the strategies of common hepatic artery lymph node dissection.Methods The clinical data of 682 patients with esophageal squamous cell carcinoma who were admitted to the Cancer Hospital of Fudan University from May 2005 to December 2010 were retrospectively analyzed.The locoregional lymph node metastasis of thoracic esophageal squamous cell carcinoma,relationship between metastatic rates of common hepatic artery lymph node and clinicopathological factors and the postoperative complications were analyzed.The enumeration data were analyzed using the chi-square test.Results A total of 18 277 lymph nodes were dissected (27 lymph nodes per patient).The lymph node metastatic rate was 55.87％ (381/682),and the metastatic lymph node ratio was 7.87％ (1438/18 277).Lymph nodes adjacent to the cardia of stomach,laryngeal nerve,lesser curvature of stomach,cervical esophagus,left gastric artery had a higher metastatic rate,while common hepatic artery lymph node had a lower metastatic rate.All the common hepatic artery lymph node metastasis was accompanied with locoregional metastasis.A total of 1480 common hepatic artery lymph nodes were dissected (2 common hepatic artery lymph nodes per patient).Twenty-four patients had common hepatic artery lymph node metastasis,with the metastatic rate of 3.52％ (24/682) and the lymph node ratio of 2.16％ (32/1480).The common hepatic artery lymph node metastatic rates of upper,middle and lower esophageal squamous cell carcinoma were 2.33％ (1/43),3.76％ (16/425) and 3.27％ (7/217),with no significant difference (x2 =0.295,P ＞ 0.05).The common hepatic artery lymph node metastatic rates of patients in T1,T2 and T3 stages were 2.35％ (2/85),5.46％ (10/183) and 2.90％ (12/414),with no significant difference (x2 =2.850,P ＞ 0.05).The common hepatic artery lymph node metastatic rates of patients with high,moderate and poor differentiated esophageal squamous cell carcinoma were 0(0/63),3.50％ (16/457) and 4.94％ (8/162),with no significant difference (x2=3.259,P ＞ 0.05).The common hepatic artery lymph node metastatic rates of patients with diameter of tumor under 3 cm,3-5 cm and above 5 cm were 2.59％ (6/232),3.02％ (11/364) and 8.14％ (7/86),with significant difference (x2 =6.267,P ＜ 0.05).The common hepatic artery lymph node metastatic rates of patients in N0,N1,N2,N3 stages were 0(0/301),2.53％ (5/198),5.65％ (7/124) and 20.34％ (12/56),with significant difference (x2 =62.368,P ＜ 0.05).The common hepatic artery lymph node metastatic rates of patients in stage Ⅰ,Ⅱ,Ⅲ and Ⅳ were 0(0/62),1.78％ (6/337),5.06％ (13/257) and 19.23％ (5/26),with significant difference (x2=25.959,P ＜0.05).Two hundred and twenty-eight patients had postoperative complications with the complication rate of 33.43％ (228/682).The incidence of anastomotic fistula was the highest,which was 11.58％(79/682).Conclusions The metastatic rates of common hepatic artery lymph node in thoracic esophageal squamous cell carcinoma is the lowest.For patients suffered from esophageal cancer in stage I or the tumor diameter under 5 cm,the dissection of common hepatic lymph node can be ommitted in surgery.