AIM: To investigate the apoptosis and Bcl-2/Bax expression in the early follicles of women at reproductive age. METHODS: 12 ovarian specimens were collected from reproductive women (aged 23-38 years) undergoing gynaecological operation. Histopathological examination of these specimens was performed to confirm its' morphological normalities. Using TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling) assay and immunohistochemistry method, cell apoptosis and Bcl-2/Bax expression were examined in the early follicles including mainly primordial, intermediary and primary follicles. RESULTS: 18.75% of the oocytes were found TUNEL positive in the early follicles, but no granulosa cells in these follicles were found TUNEL positive. Bax expression was detected in 76.07% of the oocytes in the early follicles, but Bcl-2 expression was negative in these oocytes. In addition, Bcl-2/Bax expression were not present in the granulosa cells in early follicles. CONCLUSION: The oocyte apoptosis occurs in the early follicles of reproductive woman, and pro-apoptotic protein Bax may play a role in regulating this process. It suggests that Bax mediated oocyte apoptosis may be the molecular mechanism of the early follicle atresia in the ovaries of reproductive woman. [

Objective To investigate the feasibility of aortic and mitral valves replacement under video-assisted thoracoscope.Methods The subjects included 16 dogs(Experimental Group) and a clinical volunteer(Clinical Group).The procedure was performed under extracorporeal circulation with femoral artery and vein intubation.An incision 4~5 cm in length and two 1.5 cm ports were made in the right chest wall.The superior and inferior vena cava were cross-clamped by a self-made clamp and the myocardium was protected by cold cardioplegic coronary perfusion.Artificial mechanical aortic and mitral valves were intermittently sutured.Results In the Experimental Group: the time of extracorporeal circulation was 104~196 min(143.2?46.5 min) and the ascending aorta cross-clamped time was 58~128 min(82.4?26.1 min).Autopsy findings showed satisfactory valve suture fixation,without thread loosing or paravalvular rupture.In the Clinical Group: the time of extracorporeal circulation was 157 min,the ascending aorta cross-clamped time was 112 min,the time of mechanical ventilation was 10 h,and the drainage volume,150 ml.The patient was discharged from hospital on the 10 postoperative day.Follow-up observations for 9 months found no paravalvular leakage or other complications.Echocardiography showed normal prosthetic valve movement.Conclusions Video-assisted thoracoscopic aortic and mitral valves replacement is technically feasible.

This experiment was carried out in young adult albino mice,which were injectedwith Ehrlich ascites tumor cells intraperitoneally to produce Ehrlich ascites tumor.The animals were divided into 2 groups.The first group received cAMP plusaminophylline for 5 to 13 days after inoculation.The second group received saline ascontrol.We found that in the administration of cAMP together with aminophylline,thegrowth of Ehrlich ascites tumor cells was inhibited to the extent of 53% at the 9thdays after inoculation.Early or later than the 9th day,the inhibitory rates were marklylower.By using cAMP immunocytochemical method,it was found that on the9th day of inoculation,there was an increase of the intensity of intracellualr cAMPspecific fluorescence in tumor cells of the cAMP treated mice in comparison withthose of the control.It also inhibited the 3',5'-cAMP-PDE activity on the 5th to 7thday after inoculation.Under the dark field microscope on the surface of the Ehrlich tumor cells therewere numerous“brush-like”microvilli,but they were not visible on the surfaceafter treated with cAMP together with aminophylline.The agglutination by theCon.A,was decreased markly on the 7th to 9th day after the inoculation.The possible relationship between the level of intracellular cAMP and of the3',5'-cAMP-PDE,and especially the inhibition of the formation of microvilli onthe treated tumor cell surface is discussed in this paper.

Objective To review the experience of perioperative management and effect of coronary artery bypass grafting (CABG) for patients of aged＞80 years.Methods We studied 118 cases with CABG for patients of age＞80 years from January 2002 to December 2012.The other 1034 cases with CABG for patients aged 60-80 years were enrolled as control group.Logistic regression analysis was used to assess the effect of age on operative mortality and morbidity.Results The recent mortality was higher in group aged＞80 years [6.8％(8 cases) vs.3.1％(32 cases)].Through multivariate logistic regression,the patients aged＞80 years versus control were concerned about some postoperative adverse events as follows:higher mortality (OR =3.45,95 ％ CI:2.86-4.23),dialysis (OR=3.56,95％CI:3.01-4.32) and re-intubation(OR=3.87,95％CI:3.45-4.87),delayed healing of incision(OR=4.05,95 ％ CI:3.47 5.74),prolonged mechanical ventilation(OR=3.76,95 ％ CI:3.435.01),prolonged ICU stay (OR =2.98,95 ％ CI:2.67 4.12),prolonged hospital stay (OR =2.87,95％CI:2.36-3.96).Conclusions Age＞80 years is an important factor of postoperative mortality and morbidity for CABG.We need pay more attention to perioperative management.

This study was carried out to investigate the effects of ATP on cultured human stomach carcinoma MGC-803 cells. After ATP(0.16—0.23 mg/ml, 24—120 hrs)treatment, inhibition of cell growth rate was observed. The inhibition rate reached 80% after 96 hr exposure to ATP. At the same time, anti-tubulin immunofluorescent staining showed increase of organized microtubules (MT) in ATP-treated MGC-803 cells, in contrast with the poor MT immunofluorescence in control MGC-803 cells. By using rhodamie phalloidin combined staining, increased number of actin bundles, took the place of actin dots and patches which were seen in control MGC-803 cells. Immunofluorescent staining of fibronectin in ATP-treated MGC-803 cells was also increased evidently as compared with the control MGC-803 cells. Concomitant with the improvement of MT and stress fiber organization, the cell Concohas become more flattened. It appeared that the increase of MT and stress fibers was closely related to the recovery of cell morphology in ATP treated tumor cells. It is therefore concluded that ATP not only may inhibit tumor cell proliferation, but also may induce differentiation of cell morphology of tumor cells through its action on cytoskeletal structures.

Objective To observe the effect of Ginkgolide B of various consistency on the differentiation of neuron stem cells (NSCs).MethodsNSCs were cultured in differentiation medium containing Ginkgolide B of various consistency for 3 and 7 days, the neurites length and cell body area were measured by inverted phase-contrast micrograph, then neurofilament-200 (NF-200), glial fibrillary acidic protein (GFAP), adenomatus polyposis coli (CC-1) expression were detected and counted by fluorescence microscope. The suppressor of cytokine signaling-2 (SOCS2), inhibitor of DNA binding-2 (Id2) were alsoimmunostained. The percentage of positive cells were counted respectively.ResultsThe neurites length and cell body area in Ginkgolide B groups were obviously larger than that in the control group. The percentage of NF, GFAP positive cells in Ginkgolide B groups increased with dosage increasing of Ginkgolide B. Compared with the normal control group, the percentage of SOCS2 positive cells increased significantly ( P<0.01) and the percentage of Id2 positive cells decreased significantly ( P<0.01) in Ginkgolide B groups.ConclusionGinkgolide B can promote NSCs to differentiate into neuron and astrocyte, the percentage of astrocyte is increased with a dosage-dependent relationship with Ginkgolide B.

Cocaine is one of the main addiction drugs in the world. Scientists are always trying to discover why the addiction happens and find the methods to cure the cocaine addiction. The classics mechanism is that the cocaine binds with the dopamine transporter (DAT), then the retake of dopamine was blocked, and this resulted in the sustained excitement of the dopaminergic neuron. Now, it is found that the cocaine influence some systems relate to the gene expression of the "reward" circuit. This influence finally leads to the change of neuron dendritic plastisity in that area. All the changes are sustaining and these may be the foundation of some behavior effects about the cocaine addiction. At present, the main therapy orientation is decrease the contact between the cocaine and the neuron by idiosyncratic antibody, vaccine or enzyme. Here, related mechanism and therapies were mainly reviewed.

Objective To investigate the role of regulatory B cells ( Breg ) in the immunological pathogenesis of Kawasaki disease ( KD) .Methods Twenty-eight children with acute KD and twenty-eight age-matched healthy subjects were enrolled in this study .Blood samples were collected before and after the treatment of intravenous immunoglobulin (IVIG).The proportions of CD19+CD24hiCD38hi Breg and CD4+CD25+Foxp3+regulatory T cells (Treg), and the expressions of co-stimulatory molecules (CD80, CD86 and PD-L1) were analyzed by flow cytometry .The concentration of IL-10 protein was measured by enzyme-linked immunosorbent assay .Real-time PCR was performed to evaluate the expressions of Foxp 3, CTLA4 and GITR in CD4+T cells and IL-10 in CD19+B cells at mRNA level.Results (1) The proportions of CD19+CD24hi CD38hi Breg in peripheral blood and the expressions of IL-10 at mRNA level in CD19+B cells from patients with KD were lower than those from healthy controls (P<0.05), but significantly increased after treated with IVIG (P<0.05).Upon stimulation of lipopolysaccharide for 48 hours, the concentrations of IL-10 protein in culture supernatant of B cells from patients with KD were still lower than those from healthy controls [(31.06±8.26) pg/ml vs.(46.32±13.91) pg/ml, P<0.05].(2) Compared with healthy controls , the expressions of CD80, CD86 and PD-L1 were remarkably down-regulated in patients with acute KD ( P<0.05).With the treatment of IVIG, the expressions of CD80 and CD86 were significantly up-regulated though lower than those of the control group (P<0.05).There was no significant difference in the expression of PD-L1 before and after treatment (P>0.05).(3) The proportions of CD4+CD25+Foxp3+Treg and the expressions of Foxp3, CTLA-4 and GITR at mRNA level were significantly decreased in KD group compared with those in control group (P<0.05), but were increased to some extent after IVIG treatment (P<0.05). In addition, a positive correlation between the proportion of CD 19+CD24hiCD38hi Breg and the proportion of CD4+CD25+Foxp3+Treg was found in patient with acute KD (r=0.62, P<0.05).Conclusion Breg cell deficiency and its impaired function might be one of the important factors causing immune dysfunction in pa -tients with acute KD .

Objective To investigate the effects of baicalein (Bai) on acute lung injury (ALI) induced by intestinal ischemia/reperfusion (I/R) and its mechanism in mice.Methods Twenty-four male C57BL/6J mice were divided into three groups by random number table:namely sham group,I/R group and Bai+I/R group,with 8 mice in each group.Intestinal I/R induced lung injury model was reproduced by clamping superior mesenteric artery for 90 minutes,followed by reperfusion.Bai (100 mg/kg) was intraperitoneally injected 1 hour before ischemic challenge in the Bai+I/Rgroup.The mice in sham group underwent the similar procedure with I/R group but without vascular occlusion.All mice were sacrificed at 4 hours of reperfusion,and blood was collected from inferior vena cava and lung tissues were harvested.Lung tissues were stained with hematoxylin-eosin (HE),and histological changes were examined under light microscope for pathological score.Lung wet/dry (W/D) ratio was calculated.Lung cell apoptosis was determined by TdT-mediated dUTP nick end labeling (TUNEL) technique.Serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6(IL-6) were determined by enzyme-linked immunosorbent assay (ELISA).The mRNA expressions of TNF-α and IL-6 in lung tissues were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR).The protein expression levels of cytoplasmic inhibitory factor-α of nuclear factor-κB (IκB-α) and nucleus NF-κB were determined by Western Blot.Results Under light microscope,a normal lung tissue structure was shown in the sham group and no evidence of obvious lung injury was found.In the I/R group,the alveolar structure was seriously damaged.The alveolar wall was widened and there was significant interstitial edema and leukocytes infiltration.In the Bai+I/R group,pathological damage was significantly decreased as indicated by reduced lung tissue edema and leukocytes infiltration.Compared with the sham group,the lung pathological scores,W/D ratio and cellular apoptosis in the I/R group were significantly increased.Bothserum TNF-α and IL-6 contents and lung TNF-α and IL-6 mRNA expressions were significantly increased.Furthermore,I/R significantly resulted in a decrease of IκB-α in the cytoplasm and an increase of NF-κB in the nucleus.Notably,Bai treatment significantly attenuated ALI induced by intestinal I/R injury.Compared with the I/R group,the lung pathological scores and W/D ratio in the Bai+I/R group were significantly decreased (lung pathological score:4.59±1.17 vs.6.27±1.34,W/D ratio:3.79±0.28 vs.4.32±0.57),cellular apoptosis was significantly decreased [(4.85 ± 2.47)％ vs.(8.15 ± 2.33)％],both serum TNF-α and IL-6 contents and lung TNF-α and IL-6 mRNA expressions were significantly decreased [serum TNF-α (pg/L):124.18±30.49 vs.167.72 ± 38.65,IL-6 (ng/L):1.65 ± 0.69 vs.2.43 ± 0.57;lung TNF-α mRNA (2-△△Ct:4.75 ± 2.38 vs.7.69 ± 2.32,IL-6 mRNA (2-△△ Ct):16.45 ±4.39 vs.27.69 ± 6.82],additionally,Bai pretreatment significantly increased cytoplasmic IκB-α protein expression (gray value:0.47 ± 0.11 vs.0.27 ± 0.09),while decreased nuclear NF-κB protein expression (gray value:0.57 ± 0.13 vs.1.07 ± 0.14,all P ＜ 0.05).Conclusion Bai could attenuate intestinal I/R injury induced ALI via the inhibition of inflammation and apoptosis.

Objective To investigate the changes and significance of ubiquitination of Foxp3 pro-tein during the acute phage of Kawasaki disease ( KD) .Methods Forty-eight children with KD and twenty-eight age-matched healthy children were recruited in this study.Co-immunoprecipitation and Western blot assays were performed to determine the poly-ubiquitination status of Foxp3 in CD4+T cells.The percentages of CD4+CD25high Foxp3+regulatory T cells ( Treg) and the levels of IL-10, transforming growth factor-β( TGF-β) , cytotoxic T lymphocyte-associated protein-4 ( CTLA4 ) , STIP1 homology and U-Box containing protein 1 (STUB1), heat shock protein 70 (HSP70), ubiquitin-specific-processing protease 7 (USP7) and pSTAT3 were analyzed by flow cytometry analysis.Quantitative real-time PCR was used to evaluate the tran-scription levels of TLR1-10, IL-1R1, IL-1RAP, IL-6Rα, gp130, TNFR1, MyD88 and RIP1 in CD4+T cells.Plasma concentrations of IL-1β, IL-6 and TNF-αwere measured by enzyme-linked immunosorbent as-say.Results (1) The percentages of Treg cells and the levels of IL-10, TGF-β, CTLA4 and forkhead box P3 (Foxp3) in patients with acute KD were lower than those of healthy subjects (P<0.05), while the poly-ubiquitination of Foxp3 protein was significantly enhanced in patients with acute KD [(0.52 ±0.19) vs (0.08±0.02),P<0.05].Meanwhile, the four former items in KD patients with coronary artery lesions (KD-CAL+) were lower than those of KD patients without coronary artery lesions (KD-CAL-) (P<0.05), while the polyubiquitination level of Foxp3 protein in KD-CAL+group was much higher than that of KD-CAL-group [(0.70±0.28) vs (0.43±0.17), P<0.05].The levels of Treg cells, IL-10, TGF-βand CTLA4 in patients with KD were increased and the ubiquitination of Foxp3 protein was inhibited [(0.24±0.10) vs (0.52±0.19), P<0.05] upon the treatment with IVIG.(2) The levels of STUB1 and HSP70 in CD4+T cells were significantly elevated during acute KD, while the levels of USP7 were decreased (P<0.05).The ratios of STUB1/USP7 in patients with acute KD were much higher than those of the control group [(2.65± 0.92) vs (1.09±0.37), P<0.05], but were significantly decreased after IVIG therapy [(1.46±0.53) vs (2.65±0.92), P<0.05].A negative correlation was found between STUB1/USP7 ratio and Foxp3 level during acute KD (r=-0.56, P<0.05).Moreover, KD patients with CAL+showed higher levels of STUB1 and HSP70 and higher ratios of STUB1/USP7 (P<0.05), but lower levels of USP7 as compared with those of KD-CAL-group (P<0.05).(3) The plasma concentrations of inflammatory cytokines (IL-1β, IL-6 and TNF-α), the levels of surface receptors ( IL-1R1/IL-1RAP/TLR4, IL-6Rα/gp130 and TNFR1) and its downstream molecules ( MyD88, pSTAT3 and RIP1) in CD4+T cells were up-regulated during acute KD ( P<0.05), especially in patients with CAL+, but were down-regulated upon the IVIG therapy (P<0.05).No significant differences with other TLRs were found among the groups (P>0.05).Conclusion Hyper-ubiq-uitination of Foxp3 protein might be involved in the immune dysfunction during Kawasaki disease.