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

Recent studies have overturned the previous belief that adult cardiocyte had been thought to permanently withdraw from cell-cycle activity.At present,targeting cardiocyte proliferation is one of current major therapeutic strategies for myocardial injury and repair following injury.Therefore,the author review the research progress in cell-cycle regulation of cardiocyte proliferation by systematically searched the relevant studies.

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.

Objective To observe the intervention effect of vitamin C (Vit C) and adenosine triphosphate (ATP) on myocardial fibrosis in rats.Methods Forty male SD rats were selected,body weight were 125-140 g,and they were divided into 8 groups according to body weight using a random number table method.Four rats for control group,3 rats for model group,6 rats for Vit C early group,6 rats for ATP early group,6 rats for Vit C + ATP early group,5 rats for Vit C late group,5 rats for ATP late group,and 5 rats for Vit C + ATP late group.Rats in model group and these intervention groups were induced with doxorubicin (2 mg/kg each week) for 6 weeks,and control group was given the same amount of normal saline.All early groups were intragastrically administered with Vit C (200 mg·kg-1·d-1),ATP (45 mg·kg-1·d-1) and Vit C + ATP (200 mg·kg-1·d-1 + 45 mg·kg-1 ·d-1) in the fourth week;these late groups were intragastrically administered with the same dose in the sixth week;each group was continuously administered for 21 days.Three days after the last intervention,cardiac ultrasonography was performed in all surviving rats,and left ventricular end diastolic diameter (LVEDD),left ventricular end systolic diameter (LVESD),left ventricular ejection fraction (LVEF),and left ventricular fractional shortening (LVFS) were recorded.The rats were sacrificed and the hearts were taken.HE staining and Masson staining were used to observe the pathological changes of myocardial tissue and the collagen volume fraction (CVF) was calculate.Serum cardiac troponin Ⅰ (CTn-Ⅰ) and type Ⅰ procollagen amino terminal peptide (PINP) levels were determined by enzyme-linked immunosorbent assay (ELISA).Results Compared with control group [(3.65 ± 0.25) mm,(80.63 ± 3.03)％,(43.57 ± 2.54)％],LVESD [(5.07 ± 0.58),(4.06 ± 0.68),(4.71 ± 0.43),(4.87 ± 0.44),(4.79 ± 0.59),(5.07 ± 0.62),(4.97 ± 0.29) mm] of model group and each intervention groups were increased,LVEF [(62.17 ± 4.92)％,(71.28 ± 3.54)％,(65.03 ± 3.35)％,(59.81 ± 2.45)％,(60.42 ± 9.22)％,(60.15 ± 3.06)％,(60.65 ± 2.05)％],and LVFS [(30.05 ± 2.95)％,(36.44 ± 2.90)％,(31.63 ± 2.15)％,(26.95 ± 1.05)％,(28.35 ± 6.84)％,(27.79 ± 2.41)％,(28.38 ± 1.42)％] were decreased (P ＜ 0.05);compared with model group,LVESD was decreased,LVEF and LVFS were increased in Vit C early group (P ＜ 0.05).HE staining showed that the myocardial pathology of each early group improved to different degrees,such as cardiomyocyte degeneration,necrosis and fibrosis,inflammatory cell infiltration,moderate degree of interstitial edema,Vit C early group and Vit C + ATP early group were more pronounced.Masson staining showed significant improvement in fibrosis in the Vit C early group and Vit C + ATP early group,and collagen fibers were significantly reduced.Compared with the control group [(0.52 ± 0.14)％],the CVF [(27.11 ± 5.05)％,(9.80 ± 1.84)％,(16.55 ± 2.21)％,(5.06 ± 1.45)％,(12.11 ± 2.12)％,(15.71 ± 1.56)％,(16.93 ± 2.76)％] of myocardial tissue in model group and each intervention groups were significantly increased (P ＜ 0.05).There were no significant differences in CTn-Ⅰ and PINP levels between the eight groups (P ＞ 0.05).Conclusions Vit C can reduce myocardial fibrosis and improve cardiac function in the early stage.The effect of ATP alone to improve fibrosis is not obvious.