Cytotoxic effects of γδT cells on bladder cancer cells and expression of MICA/B in bladder cancer / 北京大学学报(医学版)

OBJECTIVE@#To explore the role of γδT cells against bladder cancer and to detect the expression of stress proteins MICA/B recognized by γδT cells in bladder cancer.@*METHODS@#γδT cells from peripheral blood drawn from 6 bladder cancer patients with pamidronate stimulating were expanded. Flow cytometry was used to detect the purity and expansion folds of γδT cells, and the expression of CD107a on γδT cells after PMA/ionomycin stimulated. The cytotoxicity assay was carried out to test the cytotoxicity of γδT cells against human bladder cancer cell lines. The expression of MICA/B on bladder cancer cell lines and in bladder cancer tissues were detected through flow cytometry and immunohistochemistry respectively.@*RESULTS@#γδT cells from peripheral blood drawn from 6 bladder cancer patients were successfully expanded. The purity was 75%-94% and the expansion folds were 109-371 times. After being stimulated by PMA/ionomycin, the proportion of CD107a+ γδT cells increased significantly, reaching 40%-82%. γδT cells from the 6 bladder cancer patients showed obvious cytotoxic effects on 3 human bladder cancer cell lines which was enhanced as the effector: the target ratio increased. MICA/B were detected both in 3 bladder cancer cell lines and in 26 bladder cancer tissues. The staining score of MICA/B in invasive bladder cancer was slightly higher than that in non-invasive bladder cancer, and in advanced bladder cancer was higher than that in low grade bladder cancer, but the statistical analysis showed that the staining score of MICA/B was no significant correlation between the tissue and the tumor stages and grades.@*CONCLUSION@#γδT cells from the peripheral blood of the bladder cancer patients could be successfully expanded in vitro, and showed significant anti-bladder cancer effect. MICA/B were detected both in bladder cancer cell lines and in bladder cancer tissues. The statistical analysis showed that there was no significant correlation between the staining scores of MICA/B in the tissue and the tumor stages and grades.

Clinical significance of high mobility group protein B1 in patients with severe pneumonia complicated with sepsis / 新乡医学院学报

Objective To investigate the clinical significance of the changes of serum level of high mobility group protein B1 (HMGB1) in patients with severe pneumonia complicated with sepsis.Methods Fifty patients with severe pneumonia complicated with sepsisin in Respiratory Intensive Care Unit(RICU) of Xinxiang Central Hospital from April 2014 to March 2017 were selected as observation group;while 50 healthy individuals were selected as control group.The patients in the observation group were divided into death group(n =32) and survival group(n =18) according to the prognosis.The serum levels of procalcitonin(PCT) and HMGB1 of patients in the observation group were detected on the 1st,3rd,7th day of patients hospitalized in the RICU,while the acute physiology and chronic health evaluation Ⅱ (APACHE lⅡ)scores of the patients were evaluated.The serum levels of PCT and HMGB1 of subjects in the control group were detected during physical examination.Results There was no statistic difference in the mean arterial pressure,oxygenation index,body temperature and total white cell count of patients between the death group and survival group(P ＞0.05).On the first day of patients hospitalized in the RICU,the serum levels of PCT and HMGB1 of patients in the observation group were significantly higher than those in the control group (P ＜0.05).The serum levels of HMGB1 and the APACHEⅡ scores of patients in the death group were significantly higher than those in the survival group at each time point(P ＜0.05).On the first day of patients hospitalized in the RICU,there was no statistic difference in the serum level of PCT of patients between the death group and survival group (P ＞ 0.05);the serum level of PCT of patients in the death group was significantly higher than that in the survival group at another time point (P ＜ 0.05).The serum level of HMGB1 of patients in the observation group was positively correlated with the PCT and APACHE Ⅱ score (r =0.562,0.460;P ＜0.05).Conclusion The serum level of HMGB1 in patients with severe pneumonia complicated with sepsis is increased;and the increase of serum level of HMGB1 in the death cases is more obvious than that in the survival cases.So it can be used to evaluate the patient's condition and judge the prognosis.

Clinical observation of exenatide for treatment of obesity type 2 diabetes mellitus complicated with obstructive sleep apnea hypopnea syndrome / 新乡医学院学报

Objective To observe the clinical effect of exenatide for treatment of obesity type 2 diabetes mellitus (T2DM) conplicated with obstructive sleep apnea hypopnea syndrome(OSAHS).Methods Eighty patients of obesity T2DM complicated with OSAHS in the Department of Endocrinology,the First People's Hospital of Xinxiang from August 2014 to August 2016 were chosen and randomly divided into observation group(n =40) and control group(n =40).The patients in control group taken orally mefformin 800 mg twice daily for 24 weeks;based on this,the patients in observation group were injected subcutaneously exenatide 10 μg at one hour before meal,twice daily for 24 weeks.The fasting blood glucose (FBG),2-hour postprandial blood glucose (2 hPBG),glycosylated hemoglobin (HbA1 C),body mass index (BMI),waistline,total cholesterol (TC),triglyceride (TG),apnea hypoventilation index (AHI),Epworth sleepiness scale (ESS) score,the lowest oxygen saturation in sleep (LSpO2) were compared between the two groups before and after treatment.Results There was no statistic difference in the HbA1 C,FPG,2 hPBG levels of patients between the two groups before treatment (P ＞ 0.05);the HbA1 C,FPG,2 hPBG levels of patients in the two groups after treatment were significantly lower than those before treatment (P ＜ 0.05);and the HbA1 C,FPG,2 hPBG levels of patients in observation group were significantly lower than those in the control group after treatment(P ＜ 0.05).There was no statistic difference in the BMI,waistline and TC,TG levels of patients between the two groups before treatment(P ＞ 0.05);the BMI,waistline and TC,TG levels of patients in the two groups after treatment were significantly lower than those before treatment (P ＜ 0.05);and the BMI,waistline and TC,TG levels of patients in observation group were significantly lower than those in the control group after treatment(P ＜ 0.05).There was no statistic difference in the AHI,ESS scores,SpO2 and LSpO2 levels of patients between the two groups before treatment(P ＞0.05);the AHI,ESS scores,SpO2 and LSpO2 levels of patients in the two groups after treatment were significantly lower than those before treatment(P ＜0.05);and the AHI,ESS scores,SpO2 and LSPO2 levels of patients in observation group were significantly lower than those in the control group after treatment(P ＜0.05).The incidence of adverse reaction of patients in observation group and control group was 7.5％ (3/40) and 5.0％ (2/40) respectively;there was no statistic difference in the incidence of adverse reaction between the two groups (P ＞ 0.05).Conclusion Exenatide can effectively control blood glucose,improve the function of beta cells and reduce insulin resistance.It can effectively reduce body weight,BMI and waistline,improve the quality of sleep breathing.

Oxidative stress and peripheral skeletal muscle dysfunction in rats with emphysema / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Peripheral skeletal muscle dysfunction in patients with chronic obstructive pulmonary disease (COPD) may be due to the disease per se or as a result of concomitant confounding factors. Although the mechanistic basis for this functional impairment is uncertain, oxidative stress may play a role. The purpose of this study was to investigate whether local oxidative stress is associated with the reduced peripheral skeletal muscle performance in rats with emphysema.</p><p><b>METHODS</b>In situ mechanical properties of gastrocnemius were measured in Sprague-Dawley rats 5 months after intratracheal instillation of either elastase (EMP, n = 10) or normal saline (CON, n = 10). Lipofuscin inclusions, myocyte apoptosis and antioxidant enzyme activities were examined in the gastrocnemius muscle.</p><p><b>RESULTS</b>Lipofuscin inclusions were significantly higher in the gastrocnemius muscle of EMP compared with CON (3.2 + or - 0.4 vs. 1.7 + or - 0.4, P < 0.01). The activities of antioxidant enzymes were significantly increased in muscle homogenates of EMP as compared to CON. No significant differences were found in myocyte apoptosis between EMP and CON (1.2 + or - 0.9 vs. 1.0 + or - 0.8, P > 0.05). EMP decreased the fatigue endurance of gastrocnemius muscle (half-time to fatigue recovery: (150.0 + or - 55.4) seconds vs. (55.2 + or - 29.3) seconds, P < 0.01) and had no effect on maximal tetanic force ((467.4 + or - 36.6) g vs. (493.2 + or - 30.5) g, P > 0.05). A significantly positive correlation was found between the level of lipofuscin inclusions and the half-time to fatigue recovery of gastrocnemius muscle in EMP (r = 0.664, P < 0.05).</p><p><b>CONCLUSION</b>Local oxidative stress may have important functional consequences for peripheral skeletal muscle in rats with EMP.</p>

Ion channel mechanism of regulatory volume decrease in human epithelial cells / 中国应用生理学杂志

<p><b>AIM</b>To observe the regulatory volume decrease (RVD) process of human intestine cells and investigate its ion channel mechanism.</p><p><b>METHODS</b>Cultured human intestine cells were exposed to hypotonic solution and the cell volume was measured using Coulter Counter System. RT-PCR was explored to detect the mRNA expression of Ca(2+) -activated K+ channel.</p><p><b>RESULTS</b>Human intestine cells showed a RVD process and this process could be blocked by Cl- channel blocker NPPB and K+ channel blocker TEA. Further results demonstrated the subtype of K+ channel involved in RVD was an intermediate-conductance, Ca(2+) -activated K+ channel (IK) because of its high sensitivity to clotrimazole. RT-PCR results also showed the expression of IK in this cell line.</p><p><b>CONCLUSION</b>The RVD process of intestine cell was dependent on the parallel activation of Cl- channel and K+ channel. The subtype of K+ channel in volved in the RVD process was IK channel.</p>

Using oligonucleotide suspension arrays for laboratory identification of bacteria responsible for bacteremia / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The aim of this study was to develop and validate an oligonucleotide suspension array for rapid identification of 15 bacterial species responsible for bacteremia, particularly prevalent in Chinese hospitals. The multiplexed array, based on the QIAGEN LiquiChip Workstation, included 15 oligonucleotide probes which were covalently bound to different bead sets. PCR amplicons of a variable region of the bacterial 23S rRNA genes were hybridized to the bead-bound probes. Thirty-eight strains belonging to 15 species were correctly identified on the basis of their corresponding species-specific hybridization profiles. The results show that the suspension array, in a single assay, can differentiate isolates over a wide range of strains and species, and suggest the potential utility of suspension array system to clinical laboratory diagnosis.

Identification of closely related bacteria via phylogenetic methods / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To differentiate closely related pathogenic bacteria via phylogenetic method on the basis of gyrB gene sequences.</p><p><b>METHODS</b>GyrB sequences of 19 strains of E.coli, 13 Shigella spp. 2 Aeromonas caviae, 2 Aeromonas hydrophilia,1 Aeromonas veronii were determined and combined with sequences retrieved from public databases to construct phylogenetic trees. For each sequence tested, the identification deduced from the clustering relation of sequences was compared with the phenetic identification.</p><p><b>RESULTS</b>All the tested sequences, except those of Shigella boydii and Shigella dysenteriae, were corresponded with the 5 closest sequences on the tree at the species level. While the BLAST queries returned some other bacteria organisms or undetermined entries.</p><p><b>CONCLUSION</b>Phylogenetics displays good discriminative power in bacterial sequences differentiation.</p>

Inhibition of HBV core antigen gene expression in human embryonic kidney cell line AD293 by plasmid-based RNAi / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To inhibit HBV core antigen gene expression with plasmid-based RNAi.</p><p><b>METHODS</b>The shRNA expression vector targeting HBV core antigen gene was designed and constructed. Human embryonic kidney cell line AD293 was co-transfected with HBcAg-EGFP fusion protein expression vector and shRNA expression vector transiently, and the cells without shRNA-transfection and with non-specific shRNA transfection were used as controls. Inhibitory effect of RNAi was detected by fluorescence-activated cell sorting (FACS) and real-time fluorescence quantificational RT-PCR.</p><p><b>RESULTS</b>HBV core antigen gene expression in AD293 was inhibited by shRNA, with the maximal inhibition rate of 76 % measured by FACS and of 63.1 % by real-time PCR.</p><p><b>CONCLUSION</b>Effective inhibition of HBV core antigen gene expression by plasmid-based RNAi provides an alternative for anti-HBV study in vitro, which has potential clinical application.</p>