OBJECTIVETo investigate the role of phospholipase C(PLC) in cytoskeleton rearrangement of mouse dendritic cells invaded by Mycobacterium tuberculosis.

METHODSMouse dendritic DC2.4 cells were co-cultured with Mycobacterium tuberculosis H37Rv. F-actin of DC2.4 cells were strained with phalloidin-TRITC, the microtubule was stained with anti-β-tubulin monoclonal antibody and FITC-conjugated AffiniPure anti-mouse IgG. The changes of cytoskeleton in DC2.4 cells induced by Mycobacterium tuberculosis H37Rv were determined by fluorescence microscopy and the rates of F-actin rearrangements were calculated. The expressions of PLC in cytoplasm and cytomembrane of DC2.4 cells were measured by ELISA. DC2.4 cells were pretreated with PLC inhibitor U73122, then F-actin rearrangements induced by invasion of Mycobacterium tuberculosis were observed.

RESULTSBacterial invasion was observed while DC2.4 cells were co-incubated with Mycobacterium tuberculosis H37Rv for 2 h. The rates of invasion were (26.1 ± 4.5)%, (39.9 ± 5.6)%, (51.2 ±5.9)%, (57.9 ± 6.1)% and (63.9 ± 6.8)% at 4, 6, 8, 10 and 12 h of co-culture, respectively; while those were (13.6 ± 3.1)%, (14.2 ± 3.9)%, (15.1 ± 4.3)%, (16.8 ± 4.0)% and (18.3 ± 5.2)% after blocked by PLC, respectively. The rates of the F-actin rearrangements at 2, 4, 6, 8, 10 and 12 h after DC2.4 cells were invaded by H37Rv were (26.9 ± 1.5)%, (59.3 ± 2.8)%, (72.7 ± 4.8)%, (78.2 ± 5.9)%, (63.3 ± 2.9)% and (43.2 ± 2.6)%, respectively; while those were (18.5 ± 1.2)%, (22.3 ± 1.7)%, (3.6 ± 2.5)%, (24.8 ± 2.3)%, (22.3 ± 1.3)% and (23.8 ± 1.8)% after blocked by PLC, respectively. There were no changes of the microtubule observed in DC2.4 cells at the same time points. The rates of the F-actin rearrangements before blocked by PLC were higher than those after PLC blockade at 4, 6, 8 and 10 h (P <0.05). The expressions of PLC in cytomembrane in DC2.4 cells increased after 2 h and reached its highest level at 8 h. The PLC inhibitor U73122 inhibited the expressions of PCL in cytomembrane of DC2.4 cells, but not in cytoplasm.

CONCLUSIONMycobacterium tuberculosis can provoke to F-actin rearrangements through PLC molecule, which would further lead to Mycobacterium tuberculosis invasion of DC2.4 cells.

Actins ; metabolism ; Animals ; Cell Line ; Coculture Techniques ; Cytoskeleton ; metabolism ; Dendritic Cells ; cytology ; microbiology ; Mice ; Microtubules ; metabolism ; Mycobacterium tuberculosis ; pathogenicity ; Type C Phospholipases ; metabolism

Objective:To analyze the feasibility and efficacy of ultrasound-guided percutaneous biopsy and radiofrequency ablation of gallbladder polyps.Methods:From April 2019 to January 2021, 25 patients with gallbladder polyps underwent ultrasound-guided biopsy and radiofrequency ablation in the ultrasound department of the First Affiliated Hospital of Zhengzhou University were collected, the maximum diameter of the lesion was 11.00(10.00, 12.50)mm. Under general anesthesia, ultrasound-guided injection of water into the gallbladder serosa layer was performed to make the gallbladder edema thicker than or equal to 10 mm. Percutaneous biopsy and radiofrequency ablation of gallbladder polyps were performed to ablate the gallbladder mucosa layer of polyps and polyp attachment.Intraoperative contrast-enhanced ultrasound was used to evaluate the need for supplementary ablation.The operation time, intraoperative and postoperative conditions were recorded. The complete ablation rate of gallbladder polyps, the reduction rate of lesion volume after ablation, the incidence of complications and the effect of operation on gallbladder wall thickness and gallbladder contraction rate were evaluated.Results:Biopsy and radiofrequency ablation of gallbladder polyps were successfully performed in 25 patients.There were 14 cases of cholesterol polyp, 5 cases of adenoma, 5 cases of inflammatory polyp and 1 case of gallbladder adenomyosis. Twenty-nine gallbladder polyps showed changes after ablation, and the lesion volume was reduced to varying degrees. In the first, third and sixth months, the volume reduction rates of ablation focus were 70.74%(58.55%, 77.56%), 89.47%(85.04%, 96.87%) and 100%(95.68%, 100%) respectively, and the differences were statistically significant ( P<0.05). There were no significant differences in gallbladder wall thickness and gallbladder contraction rate before and 1 month after operation ( P>0.05). The operation time was 14-39(23.32±6.68)min. During the operation, 3 patients(12.0%) had a decrease in heart rate, 2 patients(8.0%) had mild abdominal pain and 1 patient(4.0%) had obvious abdominal pain, which was relieved after treatment. No bleeding, gallbladder perforation, abdominal infection and other complications occurred. All patients were followed up for 1 to 22 months, with a median of 6 (3, 7) months. No patients were lost or follow-up, polyp recurrence, or new polyps, or secondary gallstone. Conclusions:Ultrasound-guided biopsy and radiofrequency ablation of gallbladder polyps is a feasible choice for gallbladder preserving treatment of gallbladder polyps with low complication rate.

Objective To discuss the clinical safety,feasibility and efficacy of transcatheter arterial infusion chemotherapy(TAI)combined with lipiodol chemoembolization in the treatment of advanced colorectal cancer(CRC).Methods The clinical data of 37 patients with advanced CRC,who received TAI combined with lipiodol chemoembolization at the First Affiliated Hospital of Zhengzhou University of China between June 2016 and December 2022,were retrospectively analyzed.The clinical efficacy was evaluated,the progression-free survival(PFS)and the serious complications were recorded.Results A total of 55 times of TAI combined with lipiodol chemoembolization procedures were successfully accomplished in the 37 patients.The mean used amount of lipiodol emulsion was 2.9 mL(0.8-10 mL).No serious complications such as bleeding and intestinal perforation occurred.The median follow-up time was 24 months(range of 3-48 months).The postoperative one-month,3-month,6-month and 12-month objective remission rates(ORR)were 67.6%(25/37),67.6%(25/37),64.9%(24/37)and 56.8%(21/37)respectively,and the postoperative one-month,3-month,6-month and 12-month disease control rates(DCR)were 91.9%(34/37),91.9%(34/37),89.2%(33/37)and 81.1%(30/37)respectively.The median PFS was 16 months(range of 2-47 months).As of the last follow-up,22 patients survived and 15 patients died of terminal stage of tumor.Conclusion Preliminary results of this study indicate that TAI combined with lipiodol chemoembolization is clinically safe and effective for advanced CRC,and it provide a new therapeutic method for patients with advanced CRC.

Objective: To construct the competitive endogenous RNA (ceRNA) network related to gastric cancer and explore the molecular mechanism. Methods: The expression profiles of lncRNA, miRNA and mRNA in gastric cancer and paracancer tissues were analyzed by biochip technology, edgeR package in R software was used to filtrate differential expression genes (multiple change of >1.5 times, P<0.05) and volcano map was drawn. Based on the online miRNA-lncRNA prediction tool lncBase database and the miRNA Target gene prediction database (miRTarBase, target-scan, miRDB, starBase), the relationship between miRNA, lncRNA and mRNA was predicted. Cytoscape software was used to construct lncRNA-miRNA-mRNA ceRNA network and key genes (hub genes) were identified based on cytohubba calculation of degree score of each node. Then Hub genes related to the prognosis of gastric cancer were verified in the TCGA database. The GO and KEGG enrichment analysis of differentially expressed mRNA was performed using the online biological information annotation database DAVID, P<0.05 and false discovery rate (FDR)<0.05 were used as cut-off criteria. R software was used to download the RNA sequencing data and mirna-seq data of gastric cancer and adjacent tissues in TCGA database, edgeR package was used to screen out differentially expressed mRNA, miRNA and lncRNA, and some differentially expressed genes in our data were verified. In OncoLnc database, STAD project of TCGA data was selected and hub gene was input. Patients were divided into two groups based on the median value for hub genes and Kaplan-meier analysis was performed. Results: The differentially expressed 766 mRNA, 110 lncRNA and 10 miRNA were screened out, among them 90 mRNA, 4 lncRNA and 6 miRNA were used to construct the ceRNA network, and 2 of the 20 hub genes were related to the prognosis of patients. MLK7-AS1, SPP1, SULF1, hsa-miR-1307-3p were upregulated in gastric cancer tissues from our biochip, while MT2A, MT1X were downregulated, which were consistent with the results of TCGA gastric cancer database. The differentially expressed mRNAs were significantly enriched in the biological process (BP) and the mineral absorption pathway. CHST1 was negatively correlated while miR-183-5p was positively corelated with the survival of patients. Conclusion The establishment of ceRNA network for gastric cancer is conducive to further understanding of the molecular biological mechanism. CHST1 and miR-183-5p can be used as prognostic factors of gastric cancer.