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.

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 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.

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 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.

Objective To investigate the changes and significances of inducible IL-35-producing regulatory T cells(iTR35) in immunological pathogcnesis of Kawasaki disease (KD).Methods Forty-eight children with KD and 32 age-matched healthy children (healthy control group) consented to participate in this study.Flow cytometry was performed to evaluate the proportions of CD4+ FOXP3-IL-12p35+IL-27EBI3+iTR35 and CD4+CD25high FOXP3+regulatory T cells (Treg),and expression levels of associated molecules such as programmed death-ligand 1 (PD-L1),CD169,programmed death 1 (PD-1),CD43,IL-12p35,Epstein-Barr virus induced 3 (IL-27EBI3),glycoprotein 130(gp130),IL-12 receptor beta 2 (IL-12Rβ2),phosphated signal transducer and activator of transcription 1 (pSTAT1) and phosphated signal transducer and activator of transcription 4 (pSTAT4).Transcription levels of the Sre homology 2 domain-containing protein tyrosine phosphatase 2 (SHP-2),phosphatase and tensin homolog (PTEN),Vavl guanine nucleotide exchange factor(Vav) in CD4+T cells were determined by quantitative real-time PCR.Plasma concentrations of IL-35,IL-10,TNF-α and IL-12 were measured by enzyme-linked immunosorbent assay.Results (1) The proportions of iTR35 and its expressions of IL-12p35 and IL-27EBI3 in patients with acute KD dccreased remarkably[iTR35:(0.72±0.26) ‰ vs (1.65±0.43) ‰,P＜0.05],and restored after treatment [iTR35:(1.58±0.63) ‰ vs (0.72±0.26) ‰,P＜0.05].(2) The proportions of Treg and transcriptional levels of IL-12p35 and IL-27EBI3 were down-regulated during acute phase of KD [Treg:(3.26±1.21) ％ vs (7.26±2.86) ％,P＜0.05],and increased to some extent after therapy [Treg:(5.89±2.60)％ vs (3.26±1.21)％,P＜0.05].Meanwhile,plasma concentrations of IL-35 and IL-10,and expressions of gp130,IL-12Rβ2,pSTAT1 and pSTAT4 in iTR35 of patients with acute KD were found lower than those of the healthy control group (all P＜0.05),and increased after treatment (P＜0.05).Additionally,positive correlations were found between plasma concentrations of IL-35 and the proportion of iTR35 or its expressions of IL-12p35 and IL-27EBI3,respectively.(3) Expressions of PD-L1 and CD169 on CD14 + cells and plasma concentrations of TNF-α and IL-12 were elevated significantly during acute KD(all P＜0.05),as well as expression levels of the ligands (PD-1 and CD43) and its downstream molecules (SHP-2,PTEN,Vav) in CD4 + T cells were found to be lower in patients with acute KD (P＜0.05),and restored remarkably after therapy.Conclusion Insufficiency of iTR35 and its expression of IL-35 might be one of the important factors contributing to immunological dysfunction in KD.

Objective To investigate the role of IL-35-producing regulatory B cells(IL-35 + Breg)in immunological pathogenesis of Kawasaki disease (KD).Methods Thirty-two children with KD and 28 age-matched healthy children were allowed to participate in this study.Flow cytometry was performed to evaluate the proportions of IL-35 + Breg as well as requlatory T cells (Treg)and expression levels of associated molecules such as programmed death-ligand 1 (PD-LI),CD169,programmed death 1 (PD-1),CD43,IL-12p35,epstein-Barr virus induced 3 (IL-27 EBI3).IL-12 receptor beta 2 (IL-12 Rβ2),IL-27 receptor alpha (IL-27 Rα),phosphated signal transducer and activator of transcription 1 (pSTAT1) and phosphated signal transducer and activator of transcription 3 (pSTAT3).Transcription levels of the Src homology 2 domain-containing protein tyrosine phosphatase 2 (SHP-2),phosphatase and tensin homolog (PTEN),vav1 guanine nucleotide exchange factor(Vav) in CD19 + B cells were determined by using quantitative real-time PCR.Plasma concentrations of IL-35,tumor necrosis factor α(TNF-α) and IL-12 were measured by adopting enzyme-linked immunosorbent assay.Results (1) The proportions of IL-35 +Breg and its expressions of IL-12p35,IL-27EBI3 and IL-10 in patients with acute KD were lower than those of healthy controls [IL-35 + Breg:(5.79 ± 2.60) ％ vs (12.65 ± 5.34) ％;F =19.23,9.70,14.30.7.08;all P ＜ 0.05],but they were significantly increased after intravenous immune globulin (IVIG) treatment [IL-35 + Breg:(10.52 ± 4.95) ％;all P ＜ 0.05].(2) The proportions of Treg and its transcriptional levels of IL-12p35 and IL-27 EBI3 were down-regulated during acute KD [Treg:(4.12 ± 1.51) ％ vs (8.06 ± 3.32) ％;F =19.70,17.69,38.22;all P ＜ 0.05],but were increased after therapy [Treg:(7.39 ± 2.85) ％;P ＜ 0.05].A positive correlation was found between the proportions of Treg and IL-35 + Breg during acute KD (r =0.69,P ＜ 0.05).Meanwhile,plasma concentrations of IL-35 and expression levels of IL-12Rβ2,IL-27Rα,pSTAT1 and pSTAT3 in CD19 + B cells were significantly down-regulated in children with acute KD,but they were increased after treatment(F =8.09,7.54,7.69,5.89,12.59,all P ＜ 0.05).(3) Compared with healthy controls,expressions of PD-L1 and CD169 on CD14 + cells and plasma concentrations of TNF-α and IL-12 were elevated during acute KD (F =24.94,16.53,34.71,19.51;all P ＜ 0.05).Expression levels of PD-1,CD43 and its downstream molecules (SHP-2,PTEN,Vav) in CD19 + B cells were down-regulated during acute KD (F =6.43,5.57,19.52,10.37,11.37;all P ＜ 0.05),and restored remarkably after therapy (all P ＜ 0.05).Conclusion Insufficiency of IL-35 + Breg and its expression of IL-35 may be the important factors contributing to immunological dysfunction in KD.

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 .