Objective To investigate the effect of Fas and FasL abnormal expression on immune escape mechanism of renal cell carcinoma(RCC). Methods The expression of Fas and FasL was detected in 44 cases of RCC and 10 cases of normal kidney tissue by immunohistochemical method. Tumor infiltrating T lymphocyte's apoptosis in 44 cases of RCC was examined by TUNEL assay. FCM was used to detect the expression of Fas and FasL as well as apoptosis in RCC cell line 786-0. In addition, the apoptosis of Jurkat T cell with Fas exprssion was also analysed with FCM. Results Fas expression rate (22.8%) in RCC was significantly lower than that (53.8%) in normal kidney tissue(P

Objective To test the effects of restuscitation with air or oxygen on the blood gas and cerebral superoxide dismutase (SOD) concentration in neonatal rats with experimental intrauterine asphyxia. Method Seventy-seven neonatal rats were randomly (random number) divided into three experimental groups: sham operation group (SHAM), air resuscitation group (AR), and oxygen resuscitation group (OR). In groups AR and OR, animal models of intrauterine asphyxia were established and then resuscitated with air (AR) or oxygen (OR) for 30min. Comparison was made between groups including the mortality 0 hour after resuscitation, and the levels of blood gas and cerebral SOD concentrations 0 h, 6 h and 24 h after resuscitation. Results Mortality of neonatal rats in SHAM group, AR group and OR group were 0 (0/24), 0 (0/26) and 3.7% (1/27), respectively (P ＞0.05). The average levels of blood PaO2 in OR group and AR group 0 h after resuscitation were (69.2 ± 8.2)mmHg and (55.5±10.3) mmHg,respectively (P=0.004). Blood pH and PaCO2 and BE levels in OR group 0 h after resuscitation were not significantly different from those in AR group (P＞0.05). Blood pH, PO2, PCO2and BE levels in OR group were also not significantly different from those in AR group 6 h and 24 hours after resuscitation. The average concentrations of cerebral SOD in OR group 0 h and 6 hours after resucitation were (38.3±9.8) U/mgprot and (8.6±3.6) U/mgprot, and those in AR group were (53.8± 10.6) U/mgprot and (13.0±4.6) U/mgprot, respectively (P = 0.003, 0.04). The cerebral SOD concentration in OR group 24 hours after resuscitation was not significantly different from that in AR group (P＞0.05). The cerebral SOD concentrations in SHAM group 0 h,6 h and 24 hours after resuscitation were much higher than those in OR group and AR group (P＜0.05). Conclusions Resuscitation with air is as good as pure oxygen in neonatal resuscitation, in respect of early mortality and improvement of acidosis in neonatal rats after intrauterine asphyxia. Resuscitation with air will generate less radical oxygen species than pure oxygen in neonatal rats after intrauterine asphyxia.

[Objective] To explore the in-vitro determination method for the recovery rate of sinomenine in microdialysis probe and to investigate the influencing factors. [Methods] The recovery rate of sinomenine was detected by concentration difference method (by gain and by loss) and zero-net flux method. [Results] The recovery rate detected by increment method was as the same as that by decrement method; the recovery rate had no correlation with the sinomenine concentration in the solvent. Sinomenine recovery assessed by concentration difference method had a good intra-day stability and intra-day reproducibility. Sinomenine concentration and recovery in the solvent could be determined accurately by zero-net flux method. [ Conclusion ] Microdialysis sampling can be used for the pharmacokinetic study of sinomenine and decrement method (ie. retrodialysis) can be used for the determination of recovery rate of sinomenine.

To understand the degree of local medical students' clinical practice skills, evaluate teaching achievements and reveal weaknesses by analyzing the test papers and grades, so as to provide scientific advices for reform in clinical education.

This study is to investigate the inhibitory effect of kaempferol on inflammatory response of lipopolysaccharide(LPS)-stimulated HMC-1 mast cells. The cytotoxicity of kaempferol to HMC-1 mast cells were analyzed by using MTT assay and then the administration concentrations of kaempferol were established. Histamine, IL-6, IL-8, IL-1β and TNF-α were measured using ELISA assay in activated HMC-1 mast cells after incubation with various concentrations of kaempferol (10, 20 and 40 µmol.L-1). Western blot was used to test the protein expression of p-IKKβ, IκBα, p-IκBα and nucleus NF-κB of LPS-induced HMC-1 mast cells after incubation with different concentrations of kaempferol. The optimal concentrations of kaempferol were defined as the range from 5 µmol.L-1 to 40 µmol.L-1. Kaempferol significantly decreased the release of histamine, IL-6, IL-8, IL-1β and TNF-α of activated HMC-1 mast cells (P<0.01). After incubation with kaempferol, the protein expression of p-IKKβ, p-IKBa and nucleus NF-κB (p65) markedly reduced in LPS-stimulated HMC-1 mast cells (P<0.01). Taken together, we concluded that kaempferol markedly inhibit mast cell-mediated inflammatory response. At the same time, kaempferol can inhibit the activation of IKKβ, block the phosphorylation of IκBα, prevent NF-KB entering into the nucleus, and then decrease the release of inflammatory mediators.

Background and purpose:Cytokine-induced killer (CIK) has both the advantages of T lympho-cytes’ powerful anti-tumor activity and NK cells’ tumor killing capacity without MHC restriction. It could directly kill tumor cells, regulate and enhance immune function, without damaging the structure and functions of the immune sys-tem. Its effects on the treatment of malignant solid tumors has been widely recognized. This study aimed to evaluate the anti-proliferation effects of CIK cells obtained and cultivated from lung cancer patients’ lymph nodes. Meanwhile, the safety of clinical transfusion was observed.Methods:The peripheral blood and lymph nodes of 6 surgery patients with lung cancer from Shanghai Chest Hospital were used to cultivate CIK cells for 14 days. The phenotypes of CIK cells were detected by lfow cytometry. The anti-proliferation activities of CIK cells on A549 lung cancer cells were detected by CCK8 assay. The morphological changes of CIK cells were observed by invert microscope. The expression of CEA level and adverse events were evaluated after CIK transfusion.Results:The proportion of CD3+CD56+T lymphocyte in two groups were both more than 30%. The CCK8 assay showed that the suppression rate of lymph nodes group was higher than that of peripheral blood group at each effect/target ratio(P<0.05). The adverse effect of CIK transfusion was mild and tolerable. The expression of CEA level decreased in patients.Conclusion:Lymph nodes of surgery patients with lung cancer can be used for cultivation of CIK cells. The anti-tumor activity of CIK isolated from lymph nodes is better than that of CIK cells cultivated from peripheral blood. Preclinical experiments showed high safety.

[ ABSTRACT] AIM:To study the effects of anti-aging Klotho protein on neonatal rat myocardial cells with hypo-xia/reoxygenation ( H/R) injury.METHODS:The cardiomyocytes of neonatal SD rats were cultured to establish hypoxia/reoxygenation model.The myocardial cells were divided into normal control group, H/R model group, different concentra-tions of Klotho protein (0.1μmol/L, 1μmol/L and 10μmol/L) pretreatment groups.The myocardial cells pulse frequen-cy was observed before and after H/R.The cell viability was measured by MTT assay.The leakages of LDH, CK and AST, the content of MDA and the activity of SOD were detected.The apoptotic rate of the myocardial cells was analyzed by flow cytometry.The mRNA expression of endoplasmic reticulum stress markers and apoptosis-related molecules GRP78, CRT, CHOP and caspase-12 was measured by real-time PCR.The protein levels of CHOP, caspase-12 and phosphorylated Akt in the myocardial cells were determined by Western blot.RESULTS: Compared with normal control group, the pulse fre-quency, cell viability rate and SOD activity of myocardial cells were significantly decreased, the cell apoptotic rate as well as the contents of LDH, CK, AST and MDA were increased in H/R model group.The mRNA expressions of GRP78, CRT, CHOP and caspase-12 as well as the protein levels of CHOP and caspase-12 were increased, whereas p-Akt level was decreased obviously.Compared with H/R model group, the pulse frequency, cell viability rate and SOD activity were in-creased significantly, the cell apoptotic rate as well as the contents of LDH, CK, AST and MDA were decreased in Klotho pretreated group.The mRNA expression of GRP78, CRT, CHOP and caspase-12 as well as the protein levels of CHOP and
caspase-12 were decreased, while p-Akt level increased significantly.CONCLUSION:Anti-aging Klotho protein improves the myocardial cell survival and inhibits the apoptosis by increasing the resistance of the cells to oxidative stress and exces-sive endoplasmic reticulum stress response, which is related with the activation of Akt phosphorylation in H/R-injured my-cardial cells.

Objective To study the expression of human papillomavirus ( HPV) and ATM protein in laryngeal squamous cell carcinoma (LSCC), and discuss the correlation among the expression and its prognosis Method The expression of HPV16/18 mRNA was detected by PCR , and the expression of ATM proteins by immunohistochemical method in 63 LSCC specimens and 30 specimens of normal tissue adjacent to cancer. Results The positive expression rates of HPV16 /18 and ATM protein in LSCC group were 39.7% (25 /63) and 41. 3% (26 /63) respectively and those of HPV16 /18 and ATM protein in normal group were 9.5% ( 6/63 ) and 83.3% (25 /30) respectively. There was no correlation between the expression of ATM and HPV16/18 in LSCC. The accumulative 5-year survival rates of HPV16/18 positive group and HPV16/18 negative group in 63 patients were 69.8% and 52.6% respectively ,and there was no significant difference (P> 0.05. The accumulative 5-year survival rates of ATM positive group and ATM negative group in 63 patients were 65.4%and 45.9% respectively and there was no significant difference (P>0.05. Conclusion Both HPV16/18 and ATM are abnormally expressed in LSCC , but there is no correlation between the expression of HPV16/18 and ATM and the expression and its prognosis.

Objective To evaluate the necessity and practicability of optimizing the management of type 2 diabetes mellitus in community after acquiring the peer education and the music therapy to their physical and mental issues and sleep problems. Methods Totally 179 type 2 diabetes mellitus patients who were followed up in Ruijin 2nd community health service center, the random numbers table used to randomize the patients into 2 groups:control group ( 97 cases) and experiment group ( 96 cases), the conventional treatment was used in control group. Besides the conventional treatment measures, the peer support was used by patients in the experiment group. In the second step, 45 patients were met the inclusion and the exclusion criteria, the random numbers table used to randomize the patients into 2 groups:the multiple intervention group (22 cases) and the conventional treatment group (23 cases), the multiple intervention included the music therapy, the peer support and the sleep health education, the conventional treatment included the conventional treatment and the sleep health education. The t test was used to compare the patient's HbA1c and other quantitative data in two groups of patients after the intervention. Results In the first stage of research, compared with the control group patients, the patients 'HbA1c in intervention group was significantly improvement after 6 months(7.26%±1.37%vs.7.53%±1.63%,t=2.148, P<0.05),besides, the intervention group individuals achieved significant improvement in diabetic self-management behaviors and self-efficacy after 6 months, and the improvement in self-efficacy of peer support group was significant different compared with routinely educated patients(104.09±16.40 vs.110.96± 13.86,t=2.120,P<0.05), and the PHQ-9(5.95 ± 4.02 vs.2.55 ± 1.67,t=2.630,P<0.05)between the two group had significant difference, while no improvement was found in PSQI, BMI, and WHR between intervention group and control group. Conclusions Peer support could improve the blood glucose metabolism in patients with type 2 diabetes. With the effect of yoga music and physical exercises, peer support can improve the quality of sleep and decrease depression in T2DM patients, who also have sleep disorders and mild depression.

AIM:To study the protective effect of anti-aging Klotho protein on human umbilical vein endothe-lial cells (HUVECs) treated with high glucose (HG).METHODS:HUVECs were cultured in vitro, and divided into PBS control group, 5.5 mmol/L glucose group, 33.3 mmol/L glucose group, 0.1 μmol/L Klotho +33.3 mmol/L glucose group, 1 μmol/L Klotho+33.3 mmol/L glucose group , and 10μmol/L Klotho+33.3 mmol/L glucose group .The viabili-ty of the HUVECs was measured by MTT assay .The content of malondialdehyde ( MDA) , and the activities of lactate de-hydrogenase (LDH), superoxide dismutase (SOD) and glutathione (GSH) in cell culture supernatants were observed . The production of reactive oxygen species ( ROS) in HUVECs was analyzed by flow cytometry .The levels of nitric oxide ( NO) , endothelin ( ET-1 ) , intercellular adhesion molecule-1 ( ICAM-1 ) in HUVEC culture medium were detected by ELISA.The protein expression of nuclear factor-kappa B (NF-κB) in the HUVECs was determined by Western blot .RE-SULTS:Compared with PBS control group , 33.3 mmol/L glucose significantly decreased the HUVEC viability , increased ROS, LDH and MDA levels , reduced the activities of SOD and GSH , decreased the NO secretion , and induced the ET-1 and ICAM-1 secretion and the protein expression of NF-κB in HUVECs.When HUVECs were treated with Klotho protein at different concentrations combined with 33.3 mmol/L glucose, the cell viability was increased significantly , the ROS, LDH and MDA levels were decreased significantly , the antioxidant SOD and GSH activities were significantly increased , the se-cretion of NO was increased , but ET-1 and ICAM-1 releases and protein expression of NF-κB were significantly reduced . CONCLUSION:Anti-aging Klotho protein promotes the viability of HUVECs treated with HG , reduces the oxidative dam-age and ROS production , and restores the normal secretory function of HUVECs , thus playing a protective role in vascular endothelial cells through reducing the protein expression of NF-κB.