Objective To construct adenoviral Ad-pki-67-E1 A-GFP containing the promoter of the ki-67gene to achieve replication specifically in ki-67-positive glioma cells.Methods The promoter sequence of the ki-67gene was cloned into the pGL3-basic vector using molecular biology methods.The ki-67 promoter activity was detected by the dual luciferase reporter gene as-say.The ki-67 promoter was constructed into a shuttle plasmid and co-transfected with a helper plasmid in 293 T cells to recombinant adenovirus Ad-pki-67-E1A-GFP.The Ad-pki-67-E1A-GFP was added to glioma cells,and the expression of GFP was ob-served by fluorescence microscopy.Meanwhile,the expression of ki-67gene,the expression of E1A gene,which was essential for adeno-virus replication,and the number of virus copies were detected by real-time fluorescence quantitative polymerase chain reaction(PCR).Results The cloned ki-67sequence was able to activate reporter gene expression in glioma cells;enzymatic digestion and sequencing demonstrated that the ki-67 promoter was constructed into a shuttle plasmid and recombined to obtain the adenovirus Ad-pki-67-E1A-GFP.Ad-pki-67-E1A-GFP could infect glioma cells,and the expression of E1A and the number of virus copies were posi-tively correlated with the expression of ki-67 in the cells.Conclusion The ki-67 promoter-modified adenovirus replicated in glioma cells,providing a basis for further modification for gene therapy of malignant gliomas.

AIM:To explore the core target and mechanism of α-Linolenic acid(ALA)in improving neuroinflammation through network pharmacology combined with in vitro experiments.METHODS:Pharmacological studies have shown that ALA has anti-inflammatory,antioxidant,and neuroprotec-tive properties.The targets of α-Linolenic acid were obtained from PharmMapper and Swiss Tar-get Prediction databases,the targets of neuroin-flammation were searched from GeneCards,TTD and OMIM databases,and the potential targets of ALA and neuroinflammation were obtained from Wayne diagram.Protein interaction network(pro-tein-protein interaction,PPI)of potential targets was constructed by STRING website,and the core targets in PPI were screened by Cytoscape 3.8.0 software.At the same time,potential targets are imported into DAVID database,GO and KEGG data were obtained and the results were visualized.Autodock vina and Pymol software were used to dock the selected core targets with ALA and visual-ize the results.An in vitro model of neuroinflamma-tion was constructed,and cell growth status,oxida-tive stress,and migration or repairing capacity were determined by CCK-8 analysis,SOD,MDA and cell scratches,and the expression of IL-6,iba 1,COX-2(PTGS2),and iNOS proteins was determined by ELISA or Western blot experiments.RESULTS:Network pharmacology analysis revealed 46 poten-tial targets of ALA for neuroinflammation,and 10 core targets,including IL-6 and PTGS 2.With 232 entries enriched by GO enrichment analysis and 70 signaling pathways enriched by KEGG enrichment analysis,molecular docking showed that ALA can form hydrogen bonding with COX-2.Experiments showed that ALA could improve cell viability,allevi-ate cell oxidative stress levels,and promote cell mi-gration and motor repair in an in vitro model of neuroinflammation.CONCLUSIONS:ALA may im-prove neuroinflammation by alleviating oxidative stress and inhibiting IL-6 and COX-2 protein expres-sion.

Objective To investigate the distribution and drug resistance characteristics of pathogens in donors and recipients undergoing simultaneous pancreas-kidney transplantation (SPK). Methods Clinical data of 231 pairs of donors and recipients undergoing SPK were analyzed retrospectively. The pathogens of samples from donors and recipients were identified by VITEK-2 analyzer, and drug sensitivity test was performed by K-B method. The source distribution and composition ratio of pathogens in donor and recipient samples, distribution characteristics of multi-drug resistant organism, infection of recipients and drug resistance characteristics of pathogens were analyzed. Results A total of 395 strains of pathogens were cultured from 1 294 donor samples, and the detection rate was 30.53%. Gram-negative bacteria mainly consisted of klebsiella pneumoniae, Gram-positive bacteria mainly comprised staphylococcus aureus, and fungi primarily included candida albicans, respectively. In total, 2 690 strains of pathogens were cultured from 10 507 recipient samples, and the detection rate was 25.60%. Gram-negative bacteria mainly consisted of pseudomonas maltophilia, Gram-positive bacteria primarily comprised enterococcus faecalis, and fungi mainly included candida albicans, respectively. Among 395 pathogens of donors, 15 strains of methicillin-resistant staphylococcus aureus (MRSA), 16 strains of extended-spectrum β-lactamase (ESBL) positive drug-resistant bacteria, 8 strains of carbapenem-resistant pseudomonas aeruginosa (CR-PA), 21 strains of carbapenem-resistant acinetobacter baumannii (CR-AB), 2 strains of carbapenem-resistant enterobacteriaceae (CRE) and 1 strain of multiple-drug/pan-drug resistant pseudomonas aeruginosa (MDR/PDR-PA) were identified. Among 2 690 strains of recipient pathogens, 73 strains of ESBL positive drug-resistant bacteria, 44 strains of CR-PA, 31 strains of CR-AB and 3 strains of MDR/PDR-PA were detected. One recipient developed donor-derived infection, 69 cases of pneumonia, 52 cases of urinary tract infection, 35 cases of abdominal infection and 2 cases of hematogenous infection were reported within postoperative 1 year. Gram-negative bacteria were resistant to certain antibiotics. Gram-positive bacteria were sensitive to vancomycin. Fungi were sensitive to amphotericin B. Conclusions Gram-negative bacteria are the main pathogens of SPK recipients, which are resistant to certain antibiotics. Empirical use of antibiotics can be delivered before culture results are obtained. Subsequently, sensitive antibiotics should be chosen according to the culture results to improve the survival rate of SPK recipients.

BACKGROUND: Rheumatoid arthritis (RA), a chronic systemic autoimmune disease, is characterized by synovitis and progressive damage to the bone and cartilage of the joints, leading to disability and reduced quality of life. This study was a randomized clinical trial comparing the outcomes between withdrawal and dose reduction of tofacitinib in patients with RA who achieved sustained disease control. METHODS: The study was designed as a multicenter, open-label, randomized controlled trial. Eligible patients who were taking tofacitinib (5 mg twice daily) and had achieved sustained RA remission or low disease activity (disease activity score in 28 joints [DAS28] ≤3.2) for at least 3 months were enrolled at six centers in Shanghai, China. Patients were randomly assigned (1:1:1) to one of three treatment groups: continuation of tofacitinib (5 mg twice daily); reduction in tofacitinib dose (5 mg daily); and withdrawal of tofacitinib. Efficacy and safety were assessed up to 6 months. RESULTS: Overall, 122 eligible patients were enrolled, with 41 in the continuation group, 42 in the dose-reduction group, and 39 in the withdrawal group. After 6 months, the percentage of patients with a DAS28-erythrocyte sedimentation rate (ESR) of <3.2 was significantly lower in the withdrawal group than that in the reduction and continuation groups (20.5%, 64.3%, and 95.1%, respectively; P  < 0.0001 for both comparisons). The average flare-free time was 5.8 months for the continuation group, 4.7 months for the dose reduction group, and 2.4 months for the withdrawal group. CONCLUSION: Withdrawal of tofacitinib in patients with RA with stable disease control resulted in a rapid and significant loss of efficacy, while standard or reduced doses of tofacitinib maintained a favorable state. TRIAL REGISTRATION Chictr.org, ChiCTR2000039799.
Humans ; Quality of Life ; China ; Arthritis, Rheumatoid/drug therapy* ; Piperidines/therapeutic use* ; Treatment Outcome ; Antirheumatic Agents/therapeutic use* ; Pyrroles/therapeutic use*

Objective : To explore the effect of C2 ⁃ceramide , one of the sphingomyelin substances , on apoptosis of non⁃small cell lung cancer cells( A549 and PC9) . Methods : Non⁃small cell lung cancer cell lines ( A549 and PC9) were cultured , total proteins were extracted and Western blot was performed to detect the expression of apoptotic protein Caspase⁃3 and cleaved Caspase⁃3 in the two cells. CCK⁃8 colorimetric method was used to screen drug concentration. Hoechst 33258 apoptosis staining was used to detect apoptosis. The apoptosis rate was detected by flow cytometry , and the expression of apoptotic protein Caspase⁃3 was detected by RT⁃PCR. Results : The cell viability was about 70% when ceramide was treated at 50 μmol/L. Compared with the control group , the expression of apoptotic proteins increased in the ceramide group (P < 0. 05) . Flow cytometry and apoptosis staining showed that the rate of apoptosis increased in the ceramide treatment group compared with the control group (P < 0. 05) . mRNA detection at gene level showed increased the expression of apoptotic protein Caspase⁃3 (P < 0. 05) . Conclusion C2 ⁃ceramide can promote the apoptosis of non⁃small cell lung cancer cells , thus providing a new therapeutic tar⁃ get for clinical lung cancer chemotherapy.

Objective To analyze the risk factors for the occurrence of post transplantation diabetes mellitus (PTDM) in renal transplant recipients, establish a prediction model for PTDM and evaluate its prediction value. Methods Clinical data of 915 renal transplant recipients were retrospectively analyzed. According to the occurrence of PTDM, all recipients were divided into the PTDM group (n=78) and non-PTDM group (n=837). The main indexes of recipients were collected. The risk factors for the occurrence of PTDM in renal transplant recipients were analyzed by univariate and multivariate analysis. The prediction model for PTDM was established and its prediction value was evaluated. Results Family history of diabetes mellitus, body mass index (BMI), preoperative 2 h postprandial blood glucose and preoperative glycosylated hemoglobin were the independent risk factors for the occurrence of PTDM in renal transplant recipients. The prediction model for PTDM was logit (P)=2.199×family history of diabetes (yes=1, no=0)+0.109×BMI+0.151×2 h postprandial blood glucose (mmol/L)+0.508×glycosylated hemoglobin (%)-9.123. The results of receiver operating characteristic (ROC) curve showed that the area under the curve (AUC) of these 4 predictors combined for predicting PTDM in renal transplant recipients was 0.830 [95% confidence interval (CI) 0.786-0.873], the cut-off value was 0.0608, the sensitivity was 0.821, the specificity was 0.700, and the Youden index was 0.521 (P < 0.05). Conclusions Family history of diabetes mellitus, BMI, preoperative 2 h postprandial blood glucose and preoperative glycosylated hemoglobin are the independent risk factors for the occurrence of PTDM in renal transplant recipients. The prediction model for PTDM combined with4 predictors yield relatively high prediction value for PTDM.

Objective:To explore the clinical efficacy of aspirin plus low molecule heparin for pancreatic thrombosis during simultaneous pancreas and kidney transplantation (SPK).Methods:A total of 129 patients aged 18 years or higher underwent SPK between September 2016 and March 2020.They were divided retrospectively into two groups of aspirin ( n=60) and heparin ( n=69) according to different anticoagulant regimens.The aspirin group received only aspirin 100 mg/d at Day 1 post-operation.The heparin group received subcutaneous injection of enoxaparin 2 000 AxaIU daily for 7 days and followed by aspirin and clopidogrel.Outcomes and complication rates were compared between two groups. Results:All operations were successful without any mortality.In aspirin group, there were 5 cases of pancreatic thrombosis and one patient underwent pancreatectomy.There was no pancreatic thrombosis in heparin group ( P=0.014). There were 8 cases of intestinal anastomotic bleeding in aspirin group and 19 cases in heparin group.Statistically significant inter-group difference existed ( P=0.048). However, no significant inter-group difference existed in delayed recovery or rejection. Conclusions:Heparin anticoagulation can significantly lower the incidence of pancreatic thrombosis after SPK.Despite a higher incidence of intestinal anastomotic bleeding, no serious complication occurs after conservative meaures.

Objective:To explore the feasibility of preparing an iodine-coated titanium plate with potassium iodide and verify its antibacterial performance.Methods:Iodine was coated onto the surface of a titanium plate in electrolyte of potassium iodide using the electrophoretic deposition method. The signs and composition of the surface of the iodine-coated titanium plate were observed by scanning electron microscopy and energy dispersive spectroscopy. The experiment was conducted in a control group and 3 antibacterial test groups. The control group consisted of 10 titanium plates which had been pretreated but not loaded with iodine; the 3 experimental groups also had in each 10 titanium plates which had been pretreated and loaded with iodine in the electrolytes of concentrations of 1,000 mg/L, 2,000 mg/L and 4,000 mg/L, respectively. The antimicrobial tests in vitro were conducted with standard strains of staphylococcus aureus [1×10 6 Colony-Forming Units (CFU)/mL ATCC25923] to determine the antibacterial property of the plates. Results:The iodine-coated titanium plates appeared grey and their surface was evenly covered with a flat coating with no collapse. The scanning electron microscopy observed on the surface of the iodine-coated titanium plates an iodine coating with scattered irregular collapses in different sizes. The iodine content was 0 mass%, 5.10 mass%, 10.32 mass% and 15.05 mass%, respectively, in the control, 1,000 mg/L, 2,000 mg/L and 4 000 mg/L groups under the energy dispersive spectroscopy. Their counts of in vitro antibacterial colony were 56.00±5.09, 21.40±2.76, 9.10±2.51, and 2.00±1.88, respectively, showing significant differences between groups ( P< 0.05). Conclusions:A titanium plate with a steady and even iodine coating can be prepared by virtue of the electrophoretic deposition method in electrolyte of potassium iodide. The antibacterial property of an iodine-coated titanium plate is superior to that of a titanium plate without iodine coating.

Objective To compare the protective effects of pharmacological batch RC28-E1 and pilot batch RC28-E2 on retinal vascular endothelial cells (RF/6A) under the stimulation of vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF).Methods RF/6A cells were divided into normal control group,VEGF + FGF group and RC28-E1 groups with different concentrations.The optimal concentration of RC28-E1 was determined by cell counting kit-8 (CCK-8) method.Cells were divided into normal control group,VEGF+FGF group,RC28-E1 group,RC28-E2 group,conbercept group and FGF trap group,and cultured with serum-free culture medium,serum-free culture medium containing VEGF+FGF,serum-free culture medium containing VEGF+FGF+RC28-E1,serum-free culture medium containing VEGF+FGF+RC28-E2,and serum-free culture medium containing VEGF+FGF+ conbercept,serum-free medium containing VEGF+FGF+FGF trap,respectively.Cell proliferation rate was measured by CCK-8 method,cell migration ability was detected by Transwell test,and tube formation ability was detected by Matrigel assay.Results The cell proliferation rate of 0.080 mg/ml RC28-E1 group was significantly lower than that of VEGF+FGF group (P＜0.05).The cell proliferation rate of RC28-E1 group,RC28-E2 group and FGF trap group were significantly lower than that of VEGF+FGF group (P＜0.05).The number of migrated cells in RC28-E1 group,RC28-E2 group,conbercept group and FGF trap group were significantly lower than that in VEGF+FGF group (P=0.000).The numbers of meshes formed by retinal vascular endothelial cells in RC28-E1 group,RC28-E2 group,conbercept group and FGF trap group were significantly lower than that in VEGF+FGF group (P =0.003,0.001,0.009,0.018).The number of tube formation in FGF trap group was significantly higher than those in RC28-E1 group,RC28-E2 group,conbercept group and normal control group (P =0.014,0.000,0.008,0.014).Conclusions Under the stimulation of VEGF + FGF,the inhibitory effect of RC28-E on the proliferation of retinal vascular endothelial cells is greater than that of conbercept,and its inhibitory effect on the tube formation is superior to that of FGF trap.There is no significant difference in the effects of different batches of recombinant decoy receptor innovative drugs on retinal vascular endothelial cells.

OBJECTIVE： To promote rational use of proton pump inhibitors （PPIs） during perioperative period. METHODS： PDCA （Plan， Do， Check， Action） cycle management was used， the irrational use of PPIs of 300 medical records in neurosurgery department of our hospital were collected. The reasons were analyzed， management target was formulated and measures were implemented. The effects of management were evaluated through comparing the rate of irrational drug use and ratio of irrational type of PPIs in 300 medical records of neurosurgery department during perioperative period after management. RESULTS： Through collecting related data to confirm risk factors of stress ulcer， establishing rationality evaluation criteria for perioperative prophylactic use of PPIs， conducting rational drug use training among medical staff， drawing up various management systems and strengthening supervision and management， the rate of irrational use of PPIs was decreased significantly in our hospital； the number of irrational drug use cases decreased from 240 before management to 156 after management， among which the rate of prophylactic drug use without indication decreased from 37.33％ to 29.00％ （P＜0.05）； the irrational dosage rate decreased from 11.33％ to 6.33％ （P＜0.05）； the rate of irrational dosing frequency dropped from 12.67％ to 5.00％ （P＜0.01）. CONCLUSIONS： PDCA cycle management of our hospital can standardize the prophylactic use of PPIs in neurosurgery department during perioperative period and promote rational use of PPIs.