1.Effect of high mobility group box-1 protein on immune function of peritoneal macrophages of BALB/c mice
Feng LIU ; Yongming YAO ; Ning DONG
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
Objective To investigate the effect of high mobility group box-1 protein (HMGB1) on immune function of peritoneal macrophages in mice. Methods The peritoneal macrophages were isolated from female BALBc mice in vitro. The macrophages were stimulated with different doses of HMGB1 (1, 10, 100, 1 000ng/ml) for 6 hours, or with 100ng/ml HMGB1 for different duration (0, 6, 12, 24, 48, 72 hours). The phagocytosis ability of macrophages was determined by the neutral red dye uptake assay and optical densities of samples were read at 540nm. The cytotoxicity to L1210 cells was measured with MTT assay and was expressed by absorbance at 570nm. The chemotaxis was assayed by using the Costar Transwell cell culture chamber inserts and the cells on the lower surface of the transwell membrane were fixed and counted. The cells were harvested at the indicated time points, fixed, and stained for indirect immunofluorescence, then the expressions of I-Ak MHC class Ⅱ alloantigen on macrophages were determined with flow cytometry. Results The effects of HMGB1 on phagocytic activities, cytotoxicity, chemotaxis and I-Ak antigen expression of macrophages were enhanced in a dose-dependent manner, starting at doses as low as 1ng/ml and with a maximal response at 100ng/ml (P
2.Current situation of healthcare-associated infection management systems in Shaanxi Province
Fangfei LIU ; Hongliang DONG ; Shanhong FAN ; Hongmei WANG ; Yao SUO
Chinese Journal of Infection Control 2016;15(9):702-705
Objective To investigate the current situation of healthcare-associated infection(HAI)management in secondary and above medical institutions in Shaanxi Province,analyze development trend,and put forward sugges-tions for improvement.Methods In May-June,2016,170 secondary and above hospitals in 10 cities were selected for surveying through stratified random sampling method.Survey content included basic situation of hospitals,HAI management,HAI monitoring,and so on.Results Available questionnaires were obtained from 165 hospitals (43 tertiary hospitals,and 122 secondary hospitals).Of 165 hospitals,more than 90% have established HAI manage-ment organizations and regulations,but hospital risk management should be paid more attention,only 63.03% of hospitals perfected the risk management system and 66.06% conducted risk assessment.99.09% of hospitals im-plement training on HAI to all staff regularly and 88.41% conducted effective feedback.In the aspect of staff alloca-tion,88.48% of the hospitals assigned enough professionals for HAI management,but only 34.55% have specific training programme for these personnel.Only 33.94% of hospitals have special funds for HAI control;in the aspect of monitoring on HAI,21.21% of hospital installed and used HAI monitoring software;In the aspect of implemen-tation of monitoring programme,about 90% of hospitals developed monitoring on HAI cases and environmental hy-giene,but only 34.55% and 23.64% of hospitals conducted targeted monitoring on intensive care unit and neonatal intensive care unit respectively.Conclusion Organizational structure of HAI management in Shaanxi Province is perfect,relevant rules and regulations are basically established,basic monitoring projects are universal,but the awareness of risk management needs to be strengthened,professional allocation and professional quality develop-ment are both imbalance,informational monitoring is inadequate.
3.Four-tube strategy for the treatment of thoracic cavity fistula following esophagus carcinoma resection
Sheng YAO ; Canhui LIU ; Kang WANG ; Demin LI ; Guohua DONG
Journal of Medical Postgraduates 2016;29(8):845-848
Objective Thoracic cavity fistula following esophagus carcinoma resection is a serious complication with a high mortality.This study aims at a better therapy for thoracic cavity fistula following esophagus carcinoma resection by summarizing the ex-perience with the four-tube strategy ( jejunal fistula tube, stomach tube, chest drainage tube, and nasal fistula tube) in the treatment of the complication. Methods We retrospectively analyzed the clinical data about 62 cases of thoracic cavity fistula following esopha-gus carcinoma resection, 35 treated with the four-tube strategy ( treatment group) and the other 27 with the three-tube ( stomach tube, chest drainage tube, and nasal fistula tube) method ( control group) .We compared the hospital days, wound healing time, mortality, and incidence of anastomotic stenosis at 6 months after operation between the two groups of patients. Results Compared with the controls, the treatment group showed remarkable decreases in the hospital days (P<0.05), wound healing time (P<0.05), and mortality (P<0.05), but no statistically significant difference was observed in the incidence rate of anastomotic stenosis at 6 months after operation between the two groups of patients ( P >0.05 ) . Conclusion Compared with the three-tube method, the four-tube strategy has the advantages of shorter healing time and lower mortali-ty, and therefore is preferable for the treatment of thoracic cavity fis-tula following esophagus carcinoma resection.
6.Study on the Transmission of Toxoplasma gondii by Semen in Rabbits
Shiguo LIU ; Chuan QIN ; Zhijun YAO ; Dong WANG
Chinese Journal of Parasitology and Parasitic Diseases 2006;24(3):166-170
Objective To confirm the transmission of Toxoplasma gondii by semen and to investigate the impact of vaginal status on the transmission of T. gondii in female rabbits. Methods Sixteen male rabbits were infected with T. gondii by intraperitoneal injection each with 1 ×105 RH tachyzoites. Eight rabbits died in 8-14 d after infection.Artificial vagina was used to collect semen from male rabbits weekly before and after infection for 8 weeks. If more than 2 portions of semen from 8 survived male rabbits were collected after infection, the collected semen was mixed weekly for later use. Twenty-seven female rabbits were divided into 4 groups: group 1 with normal vagina (7 rabbits), group 2with wounded vagina (7), group 3 with trichomonas vaginitis (7) and group 4 with colpomycosis infection (6). Tachyzoites were found in mixed semen digested by trypsinase, and were used for endovaginal artificial insemination to female rabbits by uterine cavity tube once a week for 8 consecutive weeks. 2-3 d after every insemination, 2 ml blood was collected from helix vein of each rabbit, and stored at -40 ℃ for use. Anti-T. gondii antibody was examined by ELISA and the B1 gene of T. gondii was detected by PCR. Results Anti-T. gondii antibody was detected in some rabbits (2, 3, 1, and 1 rabbits from each of the groups respectively) on the 16th day after the first insemination. The positive rate of ELISA was 25.9%. The amplification of B1 gene (200 bp) by PCR appeared positive from the blood samples on the 3rd day after the first insemination and the last positive one was proved on the 51th day after the first insemination.Number of positive samples was 2, 1, 3 and 1 in the 4 groups respectively, with an overall PCR positive rate of 18.5%.Only 3 of the 27 rabbits were positive by both ELISA and PCR. Conclusions T. gondii can be transmitted by semen and the health status of vagina shows no impact on it.
7.Effects of interleukin-18 on transdifferentiation in renal tubular epithelial cells
Cuiwei YAO ; Dong LIANG ; Huafeng LIU ; Deshen TANG ; Xiaowen CHEN
Chinese Journal of Immunology 1986;0(04):-
Objective:To study the effect of interleukin-18 on transdifferentiation in renal tubular epithelial cells(TECs).Methods:Human proximal tubular epithelial cell line(HK-2) was cultured in vitro.TECs were exposed to different concentrations(0,0.1,1,10 and 100 ng/ml) of IL-18 for 24,48 and 72 hours.At the end of each incubation,the expressions of the ?-SMA and TGF-?_1 mRNA were assessed by RT-PCR,the rate of ?-SMA expressing TECs was assessed by immunocytochemistry,and the expression of the ?-SMA protein was assessed by Western blotting.Results:(1)The expressions of ?-SMA and TGF-?_1 mRNA were increased significantly by a dose- and time-dependent manner when TECs were exposed to IL-18(P
8.Construction of stable Cdc25C knockout HeLa cell strains using CRISPR/Cas9 gene-editing system
Yebao YAO ; Guangfei WANG ; Qingcai DONG ; Cheng CAO ; Xuan LIU
Military Medical Sciences 2017;41(5):359-362
Objective To knockout the cell division cycle 25 homolog C(Cdc25C) gene in HeLa human cervical cancer cells and to construct HeLa Cdc25C gene knockout stable strains using clustered regularly interspaced short palindromic repeats(CRISPR)/Cas9 gene-editing system.Methods The sequence of the small guide RNA(sgRNA),which could specifically recognize the first exon of Cdc25C,was designed according to the target-designing rules of CRISPR/Cas9 for construction of eukaryotic recombinant expressional plasmids.After sequencing,the plasmid was transfected into HeLa cells.The stable Cdc25C-knocking out strains were screened through the stress of puromycin,and the knockout effect was detected by Western blotting.The cell cycle was analyzed by flow cytometry.Results The stable Cdc25C-knocking out strains were obtained.Moreover,the gene′s knockout obviously delayed the progression of G2/M phase.Conclusion The HeLa Cdc25C gene knockout stable strain is successfully built using CRISPR/Cas9 system,facilitating studies on the function of Cdc25C and the mechanism of carcinogenesis.
9.Disinfection of Dental Hand Piece Contaminated with HBV in Practice
Guangliang NIU ; Dong ZENG ; Gang LIU ; Lei SHI ; Yao XIE
Chinese Journal of Nosocomiology 2006;0(06):-
OBJECTIVE To investigate the disinfection efficacy of dental high-speed hand-pieces contaminated by HBV with the method of pressure steam sterilizer.METHODS Taking 50 oral clinical patients randomly,and three group-samples which included saliva,hand-pieces contaminated by clinical operation and disinfected by pressure steam sterilizing were collected,then the samples were detected HBsAg and HBV DNA,respectively.RESULTS There were 95% of the saliva samples being HBsAg positive,the positive rate of the high-speed hand-pieces contaminated by clinical operation was lower than the saliva samples,and the positive rate of the hand-piece disinfected by pressure steam sterilizing was the lowest.HBV DNA was undetectable in the sample before or after disinfected hand-pieces used in patients′ saliva which HBsAg s/co value higher than 5.0.CONCLUSIONS Pressure steam sterilizing is effective to reduce the contaminated HBV on hand-pieces,but the biology test should be taken to demonstrate whether the complete sterilizing is achieved.
10.Effect of high mobility group box-1 protein on Foxp3 expression in spleen regulatory T cells in mice
Ying ZHANG ; Yongming YAO ; Ning DONG ; Feng LIU ; Zhiyong SHENG
Journal of Third Military Medical University 2003;0(21):-
Objective Intranuclear forkhead/winged helix transcription factor p3(Foxp3) plays a key role in T cell-mediated immunosuppression.The present study was performed to investigate the effects of high mobility group box-1 protein(HMGB1) on Foxp3 gene as well as protein expressions in splenic regulatory T cells(Tregs) and their potential regulating mechanisms in mice.Methods CD4+CD25+Tregs isolated from the spleens of male BABL/c mice by magnetic beads were seeded on 96-well(1?105 cells/well) cell culture plates coated with anti-CD3(1 ?g/ml) and soluble anti-CD28(1 ?g/ml),and the cells were stimulated with HMGB1 at various intervals or at different concentrations.After being stimulated,the Foxp3 mRNA/protein expressions in the Tregs were determined.The time-dependent and dose-dependent responses between HMGB1 and intranuclear Foxp3 expression were analyzed by flow cytometry,and the expressions of Foxp3 mRNA of Tregs were analyzed by quantitative PCR of SYBR GREEN.Results After stimulation with HMGB1,the intranuclear Foxp3 protein and mRNA expressions of splenic Tregs in mice were markedly down-regulated in 24 h to 72 h(P