OBJECTIVETo observe the effect of Cangfu Congxian Decoction (CCD) on oxidative stress in granulosa cells of polycystic ovary syndrome (PCOS) patients.

METHODSForty PCOS patients underwent in vitro fertilization-embryo transfer (IVF-ET) were assigned to the treatment group and the control group 1 according to random digit table, 20 in each group. Patients in the treatment group took CCD (200 mL, once in the morning and once in the afternoon) 2 months before IVF-ET, while those in the control group 1 took no Chinese medical decoction. Recruited were another 20 patients undergoing IVF-ET for tubal factors (as the control group 2). The clinical effect of IVF-ET were observed, including oocyte retrieval number, 2 pronuclear (2PN) fertilization rate, good quality embryo rate, clinical pregnancy rate, and ovarian hyperstimulation syndrome (OHSS) induced transplantation cancel rate. The expression of relative oxygen species (ROS) in granulosa cells was detected using cell immunofluorescence combined with confocal microscopy and FCM.

RESULTSCompared with the control group 1, occyte retrieval number, 2PN fertilization rate, and good quality embryo rate increased in the control group 2 and the treatment group (P <0. 05). OHSS induced transplantation cancel rate decreased in the control group 2 (P < 0.05). Fluorescence intensity of ROS decreased in the treatment group and the control group 2, as compared with the control group 1 (P < 0.01).

CONCLUSIONCCD increased good quality embryo rate by down-regulating the expression of ROS protein in ovarian granulosa cells, and correcting in vivo oxidative stress.

Drugs, Chinese Herbal ; therapeutic use ; Embryo Transfer ; Female ; Fertilization in Vitro ; Granulosa Cells ; drug effects ; metabolism ; Humans ; Oocyte Retrieval ; Ovarian Hyperstimulation Syndrome ; prevention & control ; Oxidative Stress ; drug effects ; Polycystic Ovary Syndrome ; drug therapy ; Pregnancy ; Pregnancy Rate ; Reactive Oxygen Species ; metabolism

Objective To study the clinical,cardiopulmonary functional and hemodynamic profiles of systemic sclerosis patients with pulmonary hypertension (SSc-PAH) compared with those of idiopathic pulmonary hypertension (IPAH).Methods Patients diagnosed with SSc-PAH or IPAH by right heart catheterization were consecutively enrolled into the study between 2011 and 2013 in Peking Union Medical College Hospital (PUMCH).Cases with pulmonary hypertension related to other diseases were excluded.Demographic data,laboratory parameters,6 minutes walk distance (6MWD),pulmonary function and hemodynamic variables at the time of diagnosis were collected and compared between the two groups.Results A total of 20 SSc-PAH patients including 19 females and one male with age of (43.1 ± 12.2) years,and 18IPAH patients including 16 females and 2 males with age of (38.4 ± 12.4) years were enrolled in this study.Subjects in both groups had similar mean pulmonary arterial pressure,cardiac index and pulmonary vascular resistance (PVR) when recruited.Compared with IPAH patients,SSc-PAH patients showed significantly decreased all parameters including forced vital capacity (FVC)％ [(77.1 ± 13.2)％ vs (88.6 ± 14.9)％,P =0.026],diffusing capacity of the lung for carbon monoxide (DLCO) ％ [(46.2 ±13.1) ％ vs (66.6± 13.3)％,P ＜0.001],DLCO/alveolar ventilation(VA) [(55.1 ± 14.3)％ vs (75.1 ± 11.5)％,P ＜0.001],and 6MWD [(365.6 ±85.1) m vs (454.3 ± 136.8) m,P =0.034].In subgroup analysis of SSc-PAH patients,elevated PVR (OR 2.122,95％ CI 1.093-4.119,P =0.026) and decreased DLCO％ (OR 0.916,95％ CI 0.842-0.996,P =0.040) were independently associated with reduced 6MWD.Conclusions Under the similar hemodynamic condition,SSc-PAH patients had more severe restrictive ventilation dysfunction and diffusion capacity dysfunction.Decreased 6MWD in SSc-PAHpatients was probably related to the impairment of pulmonary function.

Objective Toevaluatea modified Ziehl-Neelsen(Z-N) stain in the diagnosis of tuberculous meningitis. Methods Cerebrospinal fluid specimens from 35 patients were stained by using the modified Ziehl-Neelsen staining. Re-sults The positive rate was 94.29% in 35 patients with tuberculous meningitisand the intracellular acid-fast bacilli was detected in 53.40%of all specimens. One case was stained positive in 15 patients with non-tuberculous meningitis. Con-clusion The modified Ziehl-Neelsen stain not only significantly improves the detection rates of tuberculous meningitisbut alsois able to identify intracellular M.tuberculosisin cerebrospinal fluidspecimen.Thus, the modified Z-N stain can be a convenient tool for diagnosing tuberculous meningitis.

Objective To investegate the effect of ginsenoside Rg1 on the apoptosis related protein FLICE-inhibitory protein(FLIP),Fas-associated death domain protein (FADD)and Caspase-3 in the subatania nigra(SN)of 1-methyl-4-phenyl-1,2,3,6-tetrahyd-ropyridine (MPTP)-induced mouse models of Parkinson’s disease(PD), and to investigate the role of FADD and FLIP in the pathogenesis of PD and the protective effect of ginsenosides Rg1 on dopaminergic neurons.Methods 45 C57BL/6N mice were randomly divided into control group,model group and ginsenoside Rg1 group (n=15).The mice in model group were injected with MPTP by intraperitoneal,the mice in Rg1 group were injected with ginsenoside Rg1 before injecting MPTP,and the mice in control group were injected with normal saline by intraperitoneal. The behavioral changes of the mice in various groups were observed, and immunohistochemistry and Western blotting methods were used to observe the expressions of tyrosine hydroxylase (TH),FADD,FLIP and Caspase-3 in substantia nigra of the mice.Results Compared with control group,the mice in model group presented with typical symptoms of PD, the TH-positive neurons in the subatania nigra was significantly reduced (P<0.01 ), the number of FADD, FLIP and Caspase-3 positive cells was significantly increased(P<0.01),and the cytoplasm was deeply stained;the protein expression levels of FADD,FLIP and Caspase-3 were significantly increased (P<0.01).Compared with model group,the PD symptoms of the mice in ginsenoside Rg1 group reduced, the number of TH-positive neurons was significantly increased, the number of positive cells of FLIP,FADD and Caspase-3 were significantly reduced(P<0.01),and the cytoplasm was lightly stained;the protein expression levels of FADD, FLIP and Caspase-3 were significantly reduced (P<0.01 ). Nonlinear correlation analysis found that there was a positive relationship between the number of FADD and Caspase-3 positive cells (r=0.791,P<0.05).Conclusion Ginsenoside Rg1 may play a neural protective effect dopaminergic on neurons by modulating the FADD and FLIP expressions in SN of PD model mice.

Objective To analysis the immunological characteristics of patients with severe H1N1 influenza A, and to provide theoretical basis for predicting the prognosis of the disease. Methods A retrospective analysis was conducted. The clinical data of 15 patients diagnosed with severe H1N1 influenza A and admitted to Shanghai General Hospital of Nanjing Medical University from October 2015 to December 2016 were collected. All the patients were divided into survival and death groups according to 28-day survival. Clinical characteristics, treatment algorithm, organ function, inflammatory reaction and immune cell status were compared, and Cox regression was used to decide the risk factors of 28-day death in patients with severe H1N1 infection A. Results All 15 patients with severe H1N1 infection A were enrolled, most of who presented with cough (93.3%), fever (86.7%), sputum production (80.0%), shortness of breath (73.3%), myalgia (40.0%) and fatigue (40.0%). All had been received anti-virus, antibiotics, mechanical ventilation and anti-coagulation therapy; some were treated with prone position, neuromuscular blocker and extracorporeal membrane oxygenation (ECMO). The incidences of acute myocardial and kidney injury were high, and the acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score (14.1±6.1) and sequential organ failure assessment (SOFA) score (9.6±4.1) implicated the critical condition. Of 15 patients, 4 patients died in 28 days, while 11 were cured and discharged. Compared with survival group, the patients in death group had higher levels of APACHE Ⅱscore (22.7±3.8 vs. 11.8±3.8), troponin [cTn (μg/L): 0.52 (0.07, 2.02) vs. 0.15 (0.10, 0.45)] and blood urea nitrogen [BUN (mmol/L): 11.9 (6.7, 29.1) vs. 3.9 (2.7, 6.8)] and a lower level of blood platelets count [PLT (×109/L): 76±33 vs. 146±49, all P < 0.05]. The levels of C-reactive protein (CRP) and interleukin-6 (IL-6) within 24 hours of admission in death group were significantly higher than those of survival group [CRP (mg/L): 172.2±88.5 vs. 74.8±33.1, IL-6 (ng/L):283.3 (140.1, 711.0) vs. 18.5 (12.7, 71.4), both P < 0.01]. Compared with survival group, the expressions of CD3+, CD4+, CD8+ T cells and natural killer cell (NK cell) in death group were significantly decreased (CD3+ T cell: 0.348±0.119 vs. 0.573±0.106, CD4+ T cell: 0.135±0.046 vs. 0.344±0.098, CD8+ T cell: 0.089±0.057 vs. 0.208±0.054, NK cell: 0.124±0.057 vs. 0.252±0.182, all P < 0.05), but there were no significant differences in CD4+/CD8+ ratio and human leucocyte antigen-DR positive (HLA-DR+) T cell between death group and survival group (CD4+/CD8+ ratio:1.57±0.26 vs. 1.83±0.54, HLA-DR+ T cell: 0.035±0.022 vs. 0.062±0.036, both P > 0.05). B lymphocyte in death group was significantly higher than that of survival group (0.477±0.136 vs. 0.229±0.121, P < 0.01). Cox regression analysis revealed that APACHE Ⅱ score [risk ratio (RR) = 20.4, 95% confidence interval (95%CI) = 5.3-31.2, P = 0.017], CD4+ T cell (RR = 11.1, 95%CI = 5.1-20.0, P = 0.048) and CD8+ T cell (RR = 9.1, 95%CI = 4.3-16.7, P = 0.049) were independently risk factors of 28-day survival of patients with severe H1N1 influenza A. Conclusion Immunological paralysis and severe inflammatory response were early complicated with severe H1N1 influenza A, and these were significantly associated with prognosis.

OBJECTIVETo observe the effect of Bushen Wenyang Huayu Recipe (BWHR) on hypoxia inducible factor-1α (HIF-1α), proline hydroxylase2 (PHD2), von Hippel Lindau disease (VHL) suppressor gene expressions in endometriosis (EM) rats with Shen yang deficiency blood stasis syndrome (SYDBSS), and to explore the pathogenesis of EM and the mechanism of BWHR for treating EM.

METHODSTotally 50 SD rats were randomly divided into five groups, i.e., the blank control group, the sham-operation group, the model group, the Chinese medicine (CM) group, and the Western medicine (WM) group, 10 in each group. Rats in the blank control group and the sham-operation group were fed routinely. Rats in the rest 3 groups received 30-day "extended refrigerator freezing and ice water immersion" and combined with " autotransplantation" to establish EM rat model with SYDBSS. One Milliliter BWHR at 3.33 g/mL was administered to rats in the CM group by gastrogavage. Gestrinone at the daily dose of 0. 5 mg/kg was administered to rats in the WM group by gastrogavage. Equal volume of normal saline was administered to rats in the model group, the blank control group, and the sham-operation group. The size and morphology of ectopic foci in rats were observed after 4 weeks of medication. Expressions of serum CA125, plasma cyclic adenosine monophosphate (cAMP), and plasma cyclic guanosine monophosphate (cGMP) were detected by radioimmunoassay. Morphological changes of eutopic endometrium and ectopic tissue were observed under the optical microscope by HE staining. Protein expressions and contents of HIF-lα, PHD2, and VHL were detected by immunohistochemical SABC method and Western blot. mRNA expressions of HIF-1α, PHD2, and VHL were detected by RT-PCR.

RESULTSThe ectopic foci grew significantly in the model group. Their volumes were obviously contracted after treated by CM and WM. Compared with the blank control group and the sham-operation group, serum CA125 and plasma cGMP obviously increased, cAMP obviously decreased (P < 0.05); expressions and contents of HIF-1α mRNA and protein all decreased (P < 0.05); mRNA and protein expressions and contents of PHD2 and VHL all decreased in the model group (P < 0.05). Compared with model group, levels of CA125 and cGMP obviously decreased; cAMP levels obviously increased, expressions and contents of HIF-1α mRNA and protein all increased, mRNA and protein expressions and contents of PHD2 and VHL all increased in the WM group and the CM group (P < 0.05). Compared with the CM group, PHD2 protein contents were higher in the WM group (P < 0.05). HIF-1α was negatively correlated with PHD2 (r = -0.799, P = 0.00). HIF-1α was negatively correlated with VHL (r = -0. 625, P = 0.003).

CONCLUSIONSBWHR could effectively treat EM. Its mechanism might be associated with reducing contents of HIF-1α, serum CA125, and plasma cGMP, and up-regulating expressions of PHD2, VHL, and cAMP.

Animals ; Cyclic AMP ; Drugs, Chinese Herbal ; therapeutic use ; Endometriosis ; drug therapy ; metabolism ; Female ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Proline ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Up-Regulation ; Yang Deficiency ; drug therapy ; metabolism

Objective By means of animal study,investigated the gut barrier function in severe acute pancreatitis ( SAP),and role of inflammatory factors releasing,gut mucosa oxidative stress,cell apoptosis in it.Methods The animal experiment was done in the animal center of first people' s hospital,shanghai jiaotong university.Twenty four BALB/c mice were randomized ( random number) divided into two groups with twelve mice each group.The SAP group,mice received six intraperitoneal injections of cerulein at 1-hour intervals, the dose was 50μg/kg, then given one intraperitoneal injection of 10 mg/kg lipopolysaccharide ( LPS from E.Coli) for the induction of severe acute pancreatitis.The control ( sham operation) group,the mice received intraperitoneal injection of 2 ml normal saline for six times at 1-hour intervals.All the animals of each group were averaged to two batches,4 h and 8h after being operated respectively,to be anesthetized and adopted blood and tissue specimen.Then we observed the pathological change of pancreas and gut,scored it.We measured the blood value of diamine oxidase ( DAO),amylase and tumor necrosis factor-α (TNF-α).We detected content of malondialdehyde (MDA),superoxide dismutase (SOD),glutathione (GSH) and activity of xanthine oxidase (XO) in gut mucosa.We detected the casepase-3 activity and cell apopotosis by means of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) in gut mucosa,and conculated the apopotosis index (AI).Then using the PASW 18.0 software,we analyzed the data by anova and t-test,to make sure if the values were statistically different between the two groups and the mechanism of gut barrier dysfunction in panreatitis.Results At 4 h and 8 h after operation,the SAP-group-mice had significantly higher pancreas pathological score (P ＜0.01 ),blood amylase value ( P ＜ 0.05 ),gut pathological score and blood DAO and TNF-α value ( P ＜0.01 ),compared with the contral-group-mice.The gut mucosa MDA content and XO activity of mice in SAP group were significantly higher than which in control group ( P ＜ 0.01 ). The SAP-group-mice had significantly lower gut mucosa SOD content ( P ＜ 0.01 ) and GSH content ( P ＜ 0.05 ),compared with the contral-group-mice.The gut mucosa cells of mice in SAP group had significantly higher caspase-3 activity and apoptosis index than which in control group ( P ＜ 0.01 ).Conclusions In severe acute pancreatitis,inflammatory factors such as TNF-αwere waterfall-style released,induced gut mucosa suffer from ischemia-reperfusion injury,then serious oxidative stress developed in mucosa and activated caspase-3 pathway,inducing gut mucosa cells apoptose seriously,which was an important mechanism of gut barrier dysfunction.

Objective To study the effects of inhibing AKT2 by siRNA on SMMC7721 liver cancer cells proliferation,apoptosis,migration and invasion.Methods The siRNA targeting AKT2 was designedandthe SMMC7721AKT2- siRNAplasmidwasconstructed andtransfected into SMMC7721 cells.The stable cell lines were screened by G418.The effects of AKT2 by siRNA on SMMC7721 liver cancer cells,growth inhibition was evaluated by MTT assay.Cell cycle was analyzed by flowcytometry (FCM).Protein of P27 and CyclinD1 was evaluated by Western-blot.The ability of migration and invasion was evaluated by wound healing and Transwell assay.ResultsThe growth of SMMC7721 cells was significantly inhibited by siRNA (P＜0.05).Flow cytometry display that AKT2 by siRNA can induce G1 phase arrest,the ratio of G1 phase increased homologously and S phase declined homologously.The protein of CyclinD1 was declined and the protein of P27 was increased by Western-blot.Wound healing and Transwell assay show that the ability of cells,migration and invasion was inhibited by AKT2 by siRNA.Conclusion AKT2 by siRNA can significantly inhibit the growth of SMMC7721 cells,arrest cell cycle.AKT2 by siRNA can inhibit the ability of invasion and migration of SMMC7721 cells.

Objective To investigate CT features on increased cerebral vascular density and its pathological mechanism in patients with cyanotic congenital heart disease(CCHD).MethodsPreoperative brain CT scan and clinical data in 82 patients suffering from CCHD were analyzed. According to the increased levels of vascular density,patients were divided into 4 groups:normal,mild,moderate and severe.Relationships between the increased levels of vascular density and Hb,RBC,HCT,as well as the degree of cyanosis,were studied.AVONA was carried out to test blood CT value of cerebral sinuses,Hb,RBC and HCT in different groups. Descriptive analysis and linear regression were adopted to study the correlation between blood CT value and Hb concentration.The relationship of increased vascular density todegrees of cyanosis was analysed by Spearman.Results Among 82 patients,12 patients ( 14.6％ ) werefound in the group of normal vascular density and 70 patients ( 85.4％ ) in the increased vascular density group.Among 70 patients with increased vascular density,22 patients (26.8％ ) with (55.4 ± 2.6) HU,(169 ±6)g/L of Hb,(5.8 ±0.3) × 1012/L of RBC and 0.51 ±0.03 of HCT,29 patients (35.4％) with (61.3 ± 2.9) HU,(209 ± 15 ) g/L,(7.1 ± 0.4) × 1012/L,0.66 ± 0.06 and 19 patients ( 23.2％ ) with ( 68.8 ± 4.2) HU,( 242 ± 23 ) g/L,( 8.3 ± 0.9 ) × 1012/L,0.78 ± 0.08 were observed in the mild,moderateand severe group,respectively.There were significant differences in distribution of blood CT value ( HU),Hb,RBC and HCT in different groups ( F =163.263,134.703,120.974,136.541 ;P ＜ 0.01 ).Blood CT value was positively correlated with Hb concentration ( r =0.98,P ＜ 0.01 ). Vascular density was also positively correlated with the degree of cyanosis ( r =0.86,P ＜ 0.01 ). Conclusions Cerebral vascular density of patients suffering from CCHD presented different levels of increases based on CT scan results due to rise of RBC stimulated by anoxia.The increased level of vascular density was positively correlated with blood Hb concentration,and also associated with RBC accunulation caused by abnormal blood circulation.Moreover,it was positively correlated with the degree of cyanosis.

Objective To investigate the role of intestinal fatty acid binding protein (IFABP) in early diagnosis of acute traumatic intestinal rupture. Methods The patients with suspected acute traumatic intestinal rupture admitted in our emergency department from July 2010 to June 2011 were involved in the study.Their blood samples were taken on admission,1,2,3,4,6,8,12,16,24 and 48 hours after admission.All the patients were given closely medical observation and therapy,and were followed up in aspects of their clinical signs and imageology according to the present diagnosis and treatment routine.Surgical procedures would be carried out as soon as the diagnosis of intestinal rupture was confirmed and the duration between the admission and the final diagnosis was recorded.All the blood samples were determined for the IFABP concentration by means of ELISA.According to the final diagnosis results,the patients were divided into the intestinal rupture group and non-intestinal rupture group.The changes of IFABP concentration and its concentration difference between the two groups at different time points were compared. Results The study involved 33 patients,including 11 patients with confirmed intestinal rupture (intestinal rapture group) and 22 without intestinal rupture (non-intestinal rupture group).The average duration from hospitalization to the final diagnosis in the intestinal rapture group was (7.0 ±2.0) hours.At all the given time points,the IFABP concentration in the intestinal rupture group was significantly higher than that in the non-intestinal rupture group (P ＜ 0.05 ).The IFABP concentration in the intestinal rupture group was ascended on admission,reached the peak one hour later and maintained the level till the surgery,while the IFABP concentration was relatively stable in the non-intestinal rupture group within 24 hours after admission. Conclusion IFABP is the index for early diagnosis of acute traumatic intestinal rupture.