Objective To investigate the role of OX40 in the mechanisms of memory T cells in islet transplant tolerance.Methods The expression of OX40 on native, like memory and memory CD8+T cells was detected by RT-PCR. Splenic T cells from B6 mice were injected into Rag-/- mice via the tail vein, and the Rag-/- mice were divided into three groups (n=8 each): control group, given IgG; treatment group, given anti-OX40L; and OX40 knock-out group, given T cells from OX40 knock-out B6 mice spleen. All recipients were induced into diabetes mellitus model after adoptive transfer. Islet transplantation was performed on all Rag-/- mice as recipients. The mean survival time of islet was observed.Results The expression of OX40 in native T cells, like memory T cells and memory T cells was 2.87, 111.24 and 146.15 respectively. The expression of OX40 in like memory and memory T cells was higher than in native T cells (P<0.05). Comparison with control group , The mean survival time of the DBA/2 islet allografts in treatment group (130 days) and OX40 knock-out group (125 days) was significantly longer than in control group (21 days, P<0.05).Conclusion The OX40 expression is high in memory T cells. The mean survival time of the islet allografts can be prolonged by blocking OX40/OX40L pathway. OX40/OX40L pathway may be the key point of transplant tolerance.

AIM:To study the mechanism of oleanolic acid induced apoptosis and its influence on cell cycle in HL-60 cells in vitro.METHODS:The HL-60 cells were treated with different concentrations of oleanolic acid and then cultured for 12 h,24 h,48 h and 72 h,respectively.MTT assay was used to evaluate the inhibitory effect of oleanolic acid on HL-60 cells in vitro.The argarose gel electrophoresis was used to detect the chromatin DNA fragmentation.FACS was used to analyze the cell cycle of HL-60 cells.Western blotting was used to detect the activation of caspase-3 which has been confirmed the last execution of apoptosis pathway.RESULTS:MTT assay showed that oleanolic acid dramatically inhibited the growth of HL-60 cells in vitro,more than 50% HL-60 cells were inhibited when the cells were treated with 80 ?mol/L oleanolic acid for 48 h;the apparent DNA ladder was detected after exposure of HL-60 cells to oleanolic acid for 48 h.FACS analysis showed that cell cycle of HL-60 cells was arrested in G1 phase,the inhibition ratio of HL-60 cells achieved 63.24% and 67.90% after treated with oleanolic acid for 48 h and 72 h correspondingly.Western blotting detected the activation of caspase-3 after exposure of HL-60 cells to 80 ?mol/L oleanolic acid for 48 h.CONCLUSION:These results suggest that oleanolic acid induces apoptosis and the cell cycle of HL-60 cells is arrested in G1 phase.

Objective To investigate the effects of blockade of OX40/OX40L costimulation pathway on mice islet allograft tolerance in CD40/CD154 costimulation pathway blockade mice.Methods C57BL/6 mice were induced into diabetes mellitus as recipients,and were transplanted with DBA/2 mice islets.The recipients were divided into four groups,(1) treated with IgG as controls,(2) anti-OX40L mAb,(3) anti-CD154,(4) combined treatment of anti-OX40L mAb and anti CD154mAb.The mean survival time (MST) of islet allograft was observed.The expression of OX40 in activated T cells of CD154 deficient mice was detected.Effector T cells were obtained from the spleen of CD154 deficient mice cultured with or without anti-OX40L mAb for 3 days.The proliferation of T cells was assayed.Results The MST in the control group,anti-OX40L mAb group,anti-CD154 mAb group and anti OX40L mAb + anti-CD154 mAb group was 19,22,48,and ＞150 days respectively (P ＜0.05).The OX40 expression was readily induced in the 66％ activated T effector cells.CD154 deficient T effector cells proliferation was inhibited by the addition of anti-OX40L mAb in the culture in a dose-dependent fashion.Conclusion The blockade of OX40/OX40L costimulation pathway can promote islet allograft tolerance in CD40/CD154 costimulation pathway blockade mice by inhibiting the proliferation of T cells.

Objective To investigate the effect of one-lung ventilation (OLV) on the occurrence of subcutanous emphysema during retroperitoneal laparoscopic urologic surgery (RPLUS).Methods Twenty-seven ASA Ⅰor Ⅱ patients,aged 29-64 yr,with body mass index 19-25 kg/m2,scheduled for elective RPLUS,were randomly divided into 2 groups:two-lung ventilation (TLV) group (group Ⅰ,n =15) and OLV group (group Ⅱ,n =12).In group Ⅰ,the patients were tracheal intubated and TLV was performed.In group Ⅱ,the left-sided double lumen endobronchial tube was inserted and TLV was performed,OLV on the non-operated side was performed starting from 10-15 min before pneumoperitoneum and TLV resumed at the end of pneumoperitoneum.The end-tidal CO2 partial pressure and minute ventilation volume were measured before pneumoperitoneum (T1),at 30 and 60 min of pneumoperitoneum (T2,3),and at 30 min after the end of pneumoperitoneum (T4).The CO2 absorption capacity was calculated.The degree of pneumoderma was assessed and the occurance of pneumoderma was recorded at the end of pneumoperitoneum.Results Compared with group Ⅰ,the CO2 absorption capacity was significantly reduced,and the degree and incidence of pneumoderma were significantly decreased in group Ⅱ (P ＜ 0.05).Conclusion OLV on the non-operated side can reduce the CO2 absorption capacity,decrease the degree of subcutaneous emphysema and reduce the occurrence of subcutanous emphysema during pneumoperitoneum in patients undergoing RPLUS.

AIM: To investigate the role of reactive oxygen species ( ROS)-mediated mitochondrial oxidative injury in isonicotinyl hydrazide ( INH)-induced DNA damage and the protective effect of quercetin on L-02 cells.ME-THODS:The injury model of hepatocyte L-02cells in vitro induced by INH was established .The cells were divided into control group, INH group, low-dose quercetin group and high-dose quercetin group.The DNA damage of L-02 cells was evaluated by the comet test .The mitochondrion was prepared , and the level of mitochondrial ROS and the value of mitochondrial membrane potential (ΔΨm ) were detected by fluorescent probes DCFH-DA and rhodamine 123.The content of MDA was measured by TBA method .The activity of SOD was assessed with the xanthine oxidase method .The protein expression of Bcl-2 and Bax was determined by Western blotting , and the value of Bax/Bcl-2 was calculated .RESULTS:INH induced obvious DNA damage , increased the level of mitochondrial ROS , the content of MDA and the value of Bax/Bcl-2, and markedly reduced the value of ΔΨm and the activity of SOD in the L-02 cells.Quercetin attenuated DNA dam-age, reduced the level of mitochondrial ROS , elevated the value of ΔΨm , declined the content of MDA , increased the ac-tivity of SOD and decreased the value of Bax/Bcl-2 in the L-02 cells.CONCLUSION:INH induces DNA damage in L-02 cells by generation of mitochondrial oxidative stress .Quercetin has a protective effect on L-02 cells to attenuate the INH-in-duced DNA damage by inhibiting ROS-mediated mitochondrial oxidative damage .

Purpose To study the expression of HK2 in human prostate cancer (PCa) tissues and its effect on malignant phenotype of prostate cancer cells.Methods HK2 expression in PCa tissues was determined by microarray database and immunohistochemical staining.Subsequently,the change of cellular phenotype was detected by glycometabolism kit,CCK-8 kit,and flow cytometry after HK2 knockdown.Results HK2 expression was elevated followed by prostate cancer development.HK2 depletion inhibited cellular proliferation and aerobic glycolysis,and increased the ratio of early apoptosis.Conclusion HK2 expression increases in the process of PCa malignant progression.It plays a critical role in cellular proliferation,glycometabolism,and apoptosis,the mechanism of which needs further exploration.

OBJECTIVETo explore the incidence and genotypes of glucose-6-phosphate dehydrogenase (G6PD) gene mutations among infertile patients in Shenzhen.

METHODSDNA samples from 851 infertile patients were tested for 25 G6PD gene mutation sites using a multiplex SNaPshot assay.

RESULTSThe incidence of G6PD gene mutations among infertile patients in Shenzhen was 17.63%. Male and female abnormal rates were 15.13% and 20.09% respectively. Most of the female abnormal cases were heterozygotes. Mutations involved 11 haplotypes in 10 sites. 1311C> T/IVS-11 93T> C was the most common mutation, accounting for 72.00% (108/150) abnormal cases. Forty three cases of missense mutations were detected, including 19 cases of 1376G> T, 9 cases of 1388G> A, 5 cases of 95A> G and 871G> A/1311C> T/IVS-11 93T> C, 1 case of 202G> A, 835A> T, 1360C> T, 1376G> T and 392G> T/1311C> T/IVS-11 93T> C.

CONCLUSIONThe incidence of G6PD gene mutations among infertile patients in Shenzhen was high and the mutation types were various. Therefore, the G6PD deficiency genetic screening should be performed prior to assisted reproduction. This investigated results provided valuable basic data for genetic counseling, preimplantation genetic diagnosis and prenatal diagnosis.

Asian Continental Ancestry Group ; genetics ; China ; epidemiology ; DNA Mutational Analysis ; Female ; Gene Frequency ; Genotype ; Glucosephosphate Dehydrogenase ; genetics ; Haplotypes ; Humans ; Incidence ; Infertility, Female ; ethnology ; genetics ; Infertility, Male ; ethnology ; genetics ; Male ; Mutation

Objective To carry out a pilot investigation into fungus diversity in corn samples harvested from Keshan disease areas and obtain the information of dominant fungi,to predict possible secondary toxins in future study.Methods In Keshan disease areas (Heshui County and Zhengning County of Gansu Province,Yuanbao Town of Heilongjiang Province) and non-Keshan disease area (Harbin suburbs of Heilongjiang Province),5 samples of newly harvested corn from local plants were collected via the simple random sampling method,and the samples (n =20) were put in a sterile bags and sealed,numbered and transported to laboratory,placed in the incubator for cultivating mould fungi,cuhured for 2 weeks,and fungal DNA was extracted.The DNA samples were sent to Beijing Auwegene Technology Co Ltd for quality inspection,and sequencing analysis of fungal internal transcribed spacer 1 (ITS1) region was done using modern high-throughput DNA sequencing technique and MiSeq PE3000 platform,the rationality of sequencing data was analyzed through the rarefaction curve and the Shannon-Wiener curve trend to be gentle or not,and the true condition of the fungal community in the sample was tested by species composition Coverage ＞ 99％ or not,the difference in fungal community was studied between Keshan disease group and non-Keshan disease group based on the relative abundance of the fungal species in each group.Results A total of 513 869 high quality sequences were obtained,and 528 operational taxonomic unit (OTU) classifications were produced.The trend of rarefaction curve and Shannon-Wiener curve were gradually flattening,they showed that the sequences of the data were reasonable;and coverage ＞ 99％ showed that data could reflect the true situation of the fungal community.Analysis of species composition showed that there was a great overlap of fungal species between Keshan disease group and non-Keshan disease group,the relative abundance of the genus Wickerhamomyces,Candida and Aspergillus in Keshan disease group were higher than that of non-Keshan disease group.Conclusion Genus Wickerhamomyces,Candida and Aspergillus may be the dominant fungi associated with Keshan disease,it can be considered for secondary toxins studies.

Objective To explore expression level of circulating microRNA (miR)-133a and Galectin-3 and their potential clinical application in differential diagnosis between patients with chronic Keshan disease and dilated cardiomyopathy.Methods Twenty-eight patients with chronic Keshan disease and 28 cases of age-and sex-matched healthy people as control from the same severe historical endemic areas of Keshan disease in Heilongjiang Province,and another 28 patients with dilated cardiomyopathy from non-affected areas were chosen for the study.All the subjects were asked for disease history and did physical examination,examined by Doppler echocardiography for left ventricular ejection fraction (LVEF) and left ventricular end-diastolic diameter (LVEDD),and collected fasting venous blood specimen (elbow vein).The plasma miR-133a and the serum Galectin-3 were determined by Real-time PCR and enzyme-linked immunosorbent method,respectively.Meanwhile,the correlation was analyzed between miR-133a,galectin-3,LVEF and LVEDD.Results The miR-133a and Galectin-3 levels in different groups were statistically different (F =48.789,9.485,P ＜ 0.01).The plasma miR-133a level in chronic Keshan disease group and dilated cardiomyopathy group [median (quartile):0.394 (0.271,0.770),1.665 (0.943,2.713)] were both significantly lower than those in control group [2.382 (1.502,3.302],P ＜ 0.01 or ＜ 0.05],and the plasma miR-133a level in chronic Keshan disease group was lower than that in dilated cardiomyopathy group (P ＜ 0.01).There was no significant difference of serum Galectin-3 level between chronic Keshan disease group and dilated cardiomyopathy group [17.710 (9.624,27.799),12.692 (9.376,26.290) μg/L,P ＞ 0.05],but both were significantly higher than those in control group [8.070 (7.135,9.308) μg/L,P ＜ 0.01].The miR-133a was positively correlated with LVEF (rs =0.297,P ＜ 0.01),while negatively correlated with LVEDD,and Galectin-3 (rs =-0.271,-0.318,P ＜ 0.05 or ＜ 0.01);the serum Galectin-3 was negatively correlated with LVEF (rs =-0.392,P ＜ 0.01),and positively correlated with LVEDD (rs =0.385,P ＜ 0.01).Conclusion The combined application of miR-133a,Galectin-3,LVEF and LVEDD may provide assistance in clinical differential diagnosis of chronic Keshan disease and dilated cardiomyopathy.

Heart failure is the leading cause of death in cardiovascular diseases.Despite effectiveness of current clinical treatment,it is not satisfactory in general,so more effective and optimal therapies are under seeking.All available evidences show that cardiac muscles have limited regenerative capacity in adult mammals,while some vertebrates,such as zebrafish and salamander,can completely recover through perfect regeneration following myocardial injury.In-depth investigation into underlying mechanism may facilitate the development of human heart's potential of scarless healing.In this review paper,we summarized recent progresses in distinct cardiac regenerative capacity and their main influencing factors of several model animals through comparative analysis.