BACKGROUND:Immunotherapy with autologous immune cel s has been developed as a major adjuvant therapy for malignant tumors, but its mechanism of action has not been elucidated. OBJECTIVE:To investigate the relationship between cytokine-induced kil er cel secretion and apoptosis in human liver cancer stem cel s. METHODS:Human liver cancer stem cel s, HepG2 cel s, were isolated and enriched using serum-free suspension method. The peripheral blood mononuclear cel s from patients with liver cancer were induced byγ-interferon, CD3 monoclonal antibody and recombinant human interleukin-2 to form kil er cel s. Passage 1 liver cancer stem cel s were divided into control group (culture alone) and experimental group (co-culture of cytokines-induced kil er cel s and human liver cancer stem cel s). At 48 hours after culture, apoptosis in human liver cancer stem cel s was detected using flow cytometry, and expression of caspase-3 mRNA and protein was detected using RT-PCR and western blot, respectively. RESULTS AND CONCLUSION:The apoptotic rate in the control group was significantly lower than that in the experimental group (P<0.05). The expressions of caspase-3 at mRNA and protein levels were both higher in the experimental group than the control group (P<0.05). Experimental findings show that cytokines-induced kil er cel s can significantly promote apoptosis in human liver cancer stem cel s, and up-regulate the caspase-3 mRNA and protein expressions dramatical y.

Objective To obtain information on the application value of percutaneous epididymal sperm aspiration(PESA) and percutaneous testicle sperm aspiration(PTSA) in the differentiating diagnosis of obstructive and nonobstructive azoospermia.Methods Sperm recovery procedures were done in infertile men with obstructive azoospermia(OA)(n=37) and nonobstructive azoospermia(NOA)(n=28) by PESA or PTSA.Cytological smears were analysed.Results Sperm was found in the 32 epididymides and 5 testicles of OA group and in the 7 epididymides and 11 testicles of NOA group.Sperm counts were significantly different in two groups.Conclusion PESA and PTSA are efficient methods in differentiating OA and NOA.

Objective To evaluate the effects of sevoflurane preconditioning on postoperative cognitive dysfunction in aged rats.Methods One hundred and twenty healthy male Sprague-Dawley rats,aged 18 months,weighing 400-450 g,were randomly divided into 3 groups (n =40 each) using a random number table:control group (group C),operation group (group O),and sevoflurane preconditioning group (group Sev).In group C,the rats were only anesthetized with pentobarbital sodium and did not undergo operation.In group O,the rats were anesthetized with pentobarbital sodium and underwent 30 min of exploratory laparotomy.In group Sev,the rats inhaled 2.4％ sevoflurane for 30 min and then inhaled air for 30 min,and the other procedures were similar to those previously described in group O.At 30 min before operation and on 1st,3rd,5th and 7th days after operation,Morris water maze test was performed to record the escape latency,time of staying at the original platform quadrant and frequency of crossing the original platform.At 30 min before operation and on 1st and 7th days after operation,10 rats in each group were sacrificed and hippocampi were isolated to detect the apoptotic rate and intracellular [Ca2 +] i (using flow cytometry) and the ultrastructure of hippocampal neurons was observed with transmission electron microscope.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the original platform quadrant was shortened,the frequency of crossing the original platform was decreased,and the apoptotic rate and intracellular [Ca2 +] i were increased after operation in O and Sev groups (P ＜0.05).Compared with group O,the escape latency was significantly shorten,the time of staying at the original platform quadrant was prolonged,the frequency of crossing the original platform was increased,and the apoptotic rate and intracellular [Ca2+]i were decreased after operation in group Sev (P ＜ 0.05).Microscopic examination showed no abnormality in the ultrastructure of hippocampal neurons in group C,and the pathological changes of the ultrastructure of hippocampal neurons were obvious in group O,and were significantly attenuated in group Sev.Conclusion 2.4％ sevoflurane preconditioning can reduce postoperative cognitive dysfunction in aged rats,and regulation of imbalance of calcium homeostasis and reduction of cell apoptosis are involved in the mechanism.

Objective To evaluate the effect of isoflurane on the apoptosis of SH-SYSY cells transfected with APPsw gene and the role of inositol 1,4,5-triphosphate (IP3) recepters.Methods The SH-SYSY ceils transfected with APPsw gene were seeded in culture flasks with the density of 1.2 × 104/cm2.The cells were randomly divided into 4 groups (n =6 each):control group (group C),IP3 receptor antagonist group (group Ⅹ),isoflurane group (group Ⅰ) and isoflurane + IP3 receptor antagonist group (group Ⅰ + Ⅹ).After the cells were cultured for 24 h and attached to the wall,the cells were cultured routinely in group C,and Xestospongin C 100 nmol/L (IP3 receptor antagonist) was added to DMEM culture medium in groups X and Ⅰ + X,and 30 min later the cells were exposed to 1.2 ％ sevoflurane for 8 h in groups Ⅰ and Ⅰ + X.The cells were collected for examination of the ultrastructure and for determination of cell apoptosis,intracellular free calcium ion concentration [Ca2 +] i (by flow cytometry) and expression of IP3 receptor protein (by Western blot).The apoptosis rate was calculated.Results Compared with group C,there was no significant change in the apoptosis rate,[Ca2 +]i or IP3 receptor protein expression in group Ⅹ (P ＞ 0.05),while the cell apoptosis rate and [Ca2 +] i were significantly increased and IP3 receptor protein expression was up-regulated in groups I and Ⅰ + Ⅹ (P ＜ 0.05 or 0.01).Compared with group Ⅰ,cell apoptosis rate and [Ca2+]i were significantly decreased and IP3 receptor protein expression was down-regulated in group Ⅰ + Ⅹ (P ＜ 0.01).The pathological changes of the cells happened in groups Ⅰ and Ⅰ + Ⅹ,and the pathological changes were severer in group Ⅰ than in group Ⅰ + Ⅹ.Conclusion Isoflurane can induce apoptosis of SH-SY5Y cells transfected with APPsw gene through increasing [Ca2+]i and up-regulating IP3 receptor protein expression.

Objective To explore the risk factors of urolithiasis incidence of migrant workers in Zhongshan,in order to provide the scientific evidence for preventing urolithiasis.Methods 1 630 workshop migrant workers and 1630 office white collar workers were selected as the subjects of this study.The questionnaire was conducted according to the age,gender,genetic factors,occupation,past illness factors,living environment,work environment factors,diet habit,the habit of drinking water survey,B ultrasound examination,the detection of blood uric acid.Results The incidence rate of urolithiasis was 6.53％,the incidence rate of workshop migrant workers was 8.28％,which was significantly higher than that of white collar 4.72％ (x2 =16.972,P ＜0.01).Conclusion Work environment,eating habits,drinking habits,smoking and sweating are related risk factors of urolithiasis.

Objective To investigate the effects of hypoxic conditions on expressions of hypoxia-inducible factor 1α (HIF-1α) and P-glycoprotein (P-gp),and the relationship between HIF-1α and P-gp.Methods Tumor tissues from 54 cases of patients with colonic neoplasm in Xingtai People's Hospital were selected after operation from June 2013 to June 2015.The expressions of HIF-1α and P-gp in colonic tumor tissues were detected by using immunohistochemistry,and their correlations to clinical and pathologic features were analysed.The expressions of HIF-1α and P-gp in colonic tumor cell lines under normoxia and hypoxia conditions were detected by using cell smear method,and correlation between HIF-1α expression and P-gp expression was analysed.Results Among tumor tissues from 58 cases of patients with colonic neoplasm,the positive rate of HIF-1α expression was 58.62％,and that of P-gp expression was 46.55％.The positive rates of HIF-1α and P-gp expressions of patients on the Dukes stage C+D were significantly higher than those of patients on A+B phase (P＜0.05).Additionally,the positive rates of HIF-1α and P-gp expressions of patients with lymphatic metastasis were significantly higher than those of patients without lymphatic metastasis (P＜0.05).The HIF-1α expression was positively correlated with the P-gp expression (r=0.574,P＜0.01).For the same cell lines,the expression levels of HIF-1α and P-gp under hypoxia condition were significantly higher than those under normoxia condition,there were statistically significant differences (P＜0.01).While,under the same oxygen conditions,no statistically significant difference was found in expression levels of HIF-1 and P-gp among different colonic tumor cell lines (P＞0.05).Conclusion Overexpression and coexpression of HIF-1α and P-gp exist in colon cancer.The expression levels of HIF-1α and P-gp in patients with different Dukes stages and patients with or without lymph node metastasis are significantly different,and positive correlation is observed between the expression of HIF-1α and P-gp.Hypoxia condition can induce an increase in expressions of HIF-1α and P-gp in colonic tumor cells.

Adult ; Aged ; Aged, 80 and over ; Antiviral Agents ; therapeutic use ; Carcinoma, Hepatocellular ; epidemiology ; virology ; Female ; Hepatitis B ; drug therapy ; epidemiology ; Humans ; Liver Neoplasms ; epidemiology ; virology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

The abnormal glycans expressing on the surface of tumor cells are good targets to develop carbohydrate-based anti-cancer vaccines. However, one of the major problems is that carbohydrate antigens possess weak immunogenicity. This review summarizes the recent efforts to overcome this problem: glycoconjugates produced by coupling the carbohydrate antigens and proper carrier proteins improve their immunogenicity, many glycoconjugates have entered clinical trials; the vaccines become chemically well-defined when coupling the carbohydrate antigens with a T-cell peptide epitope and an immunostimulant to form fully synthetic multi-component glycoconjugate vaccines; the modification of carbohydrate antigens in combination with the technology of metabolic oligosaccharide engineering of tumor cells induces a strong immune response; and the fact that the antibodies elicited against the unnatural carbohydrate antigens can recognize the native carbohydrate antigens on tumor cells provides a new promising strategy for the development of anti-cancer vaccines.
Animals ; Antigens, Tumor-Associated, Carbohydrate ; chemistry ; immunology ; Cancer Vaccines ; chemical synthesis ; chemistry ; immunology ; therapeutic use ; Carbohydrates ; chemistry ; immunology ; Epitopes, T-Lymphocyte ; chemistry ; immunology ; Glycoconjugates ; chemistry ; immunology ; Humans ; Immune Tolerance ; Metabolic Engineering ; methods ; Neoplasms ; prevention & control ; therapy ; Oligosaccharides ; chemistry

To obtain the tervalent fusion toxin gene (named FT),three toxin gene fragments from three species of Vibrio parahaemolyticus,Vibrio vulnificus and Vibrio mimicus were connected with the flexible linker (GGGGS) using overla Pextension PCR. The three toxin gene fragments respectively encode the mature proteins of the thermostable direct hemolysin (TDH) of V. parahaemolyticus,the cytotoxin (VVC) of V. vulnificus and the heat-labile hemolysin (VMH) of V. mimicus. The identity of FT nucleic acid sequence was 99.6% with the corresponding toxin gene fragments. The open reading frame of FT was 3225 bp,encoding 1074 amino acid residues with the predicted molecular weight (MW) of 120.4 kDa. Then,FT was subcloned into the expression vector pET-22b(+). The construction of recombinant expression vector pET-22b-FT was followed by transforming into E. coli BL21(DE3) for expression. The SDS-PAGE electrophoresis results indicated that the MW of the fusion toxin protein was matched to the predicted MW. After induction by 1 mmol/L IPTG at 37℃,the fusion toxin protein was effectively expressed in E. coli BL21(DE3) with the amount of 11.49% through thin layer chromatography scanning (TLCS) analysis. Cavia cobaya was immunized using the purified cytorrhyctes to produce the anti-serum. Through the determination of the optimum working conditions,the sensitivity test,the specificity test,repeatability test and sample simulation test,the indirect ELISA method was established,which is a broad-spectrum,rapid and specific to detect various of food-poisoning Vibrio simultaneously.

OBJECTIVETo explore the crew and passengers' comfort on the Shijiazhuang-Taiyuan passenger dedicated line and physical factors, such as air pressure, noise, wind speed.

METHODSComfort investigation of all the crew (n = 244) and passengers (n = 377) on the Shijiazhuang-Taiyuan passenger dedicated line at speed of 250 km/h and 200 km/h and the detection of the air pressure, noise and wind speed were performed in 2011.

RESULTSSignificantly higher ratio of comfortable feeling, lower ratio of seriously discomfortable feeling were observed in crew and passengers at 200 km/h compared with those at 250 km/h (P < 0.05), as well as rapid disappearance of discomfortable feeling in crew (P < 0.05) and significantly higher ratio of lightly discomfortable feeling and lower ratios of tinnitus and eardrum discomfort induced by air pressure and noise in passengers at 200 km/h. No significant difference was observed in ear discomfort induced by air pressure and noise among crew, and the duration of disappearance of discomfortable feeling among passengers between 200 km/h and 250 km/h. The noise in carriages exceeded the related standard when the high-speed train passing through the tunnels.

CONCLUSIONThe individuals feel more comfortable at 200 km/h than 250 km/h in this line., which may be related with rapid variation of wind speed and noise when the train passes through the tunnels with high speed.

Adolescent ; Adult ; Atmospheric Pressure ; Female ; Humans ; Male ; Noise, Occupational ; Noise, Transportation ; Occupational Health ; Railroads ; Sensation ; Surveys and Questionnaires ; Wind ; Workplace ; Young Adult