Aim To observe the protective effects of ginkgolide B(GB)on cerebral ischemia-reperfusion injury in rats and explore its mechanism.Methods Cerebral ischemia-reperfusion injury in rats was induced by middle cerebral artery occlusion.The neurological out-come and infarct size were evaluated.SOD and LDH activity,LD and MDA levels of ischemia cerebral tissue were measured.The pathomorphology examination was done.Results GB did not improve neurologic defects in 3 h ischemia rats,but it improved neurologic defects in 21 h reperfusion rats.GB(10,5 mg?kg-1)markedly decreased infarct size in rats.SOD activity in GB(10,5 mg?kg-1)groups was higher,and LDH activity and MDA level in GB(10,5,2.5 mg?kg-1)groups were significantly lower than those in vehicle group.LD level did not chang.The cellular and intercellular edema was reduced markedly in the pathomorphology examination.Conclusions GB has protective effects on cerebral ischemia-reperfusion injury in rats.Its role is probably related to its effect on the activity of SOD and LDH,and the level of MDA.

BACKGROUND: Pancreatic stem cells can be differentiated into endocrine functioning cells under a suitable cultivation condition, suggesting that pancreatic stem cells can be used as a new source of islet in treating diabetes mellitus.OBJECTIVE: To discuss the expression of stem cell in different purities of pancreatic islets.DESING: A controlled observation based on cells.SETTING: Third People's Hospital of Mianyang City.MATERIALS: The experiment was conducted in the Center Laboratory (Key Laboratory of Chongqing City), the First Affiliated Hospital of Chongqing Medical University from March 2005 to March 2006. Thirty healthy male SD rats, of clean grade, aged 50 days were used in this study. During the experiment, the disposal of animals corresponded to Animal Ethical Standard. Cytokeratin (CK)-19 and β-actin were designed by Shanghai Invitrogen Biological Co., Ltd.Brdu reagent, mouse anti-rat Brdu monoclonal antibody, rabbit anti-mouse (RAM) CK-19 polyclonal antibody and Brdu immunohistochemical kit were from Boster Co., Ltd. (Wuhan); HistostainTM -DS double immunohistochemical staining kit was purchased from Beijing Zhongshan Company.METHODS: Rats were intraperitoneally injected with Brdu for labeling, and subjected to perfusion with Ⅴ-type collagenase via pancreatic ducts, with pancreas being excised, ripped, beaten up, digested and centrifuged to obtain pancreatic islet sediments. After being precipitated, the sediments were divided into 3 groups. Group A: The isolated sediments of islets were not purified; Group B: The sediments were added slowly with 25% Ficoll-400 and Hanks solution for purification; Group C: The sediments were slowly added with 25% Ficoll-400, 11% Ficoll-400 for purification in order.MAIN OUTCOME MEASURES: Expressions of Brdu and CK-19-positive cells in the islet samples were detected by immunohistochemical method, and mRNA expression of CK-19 in the different purities of islets was detected by reverse transcription-polymerase chain reaction.RESULTS: ① The purity of islets in the group A was the lowest. The purity of islets obtained by different purification techniques showed significances among the three groups (F =89.42, P ＜ 0.05). ②Positive expressions of Brdu and CK-19 in the group A were significantly higher than those in the group B and group C (F =18.64, 22.12, 38.61. P ＜0.01). ③Expression of CK-19 mRNA in the group A was significantly higher than that in other groups, with group C showing the lowest.CONCLUSION: Expressions of Brdu and CK-19-positive cells exist in different purities of islets, suggesting the existence of more stem cells in the low purify of islets.

Objective To analyze the risk factors in the operative mortality in neonates with congenital heart disease.Methods The surgical outcomes of eongenital heart defects in 231 neonates at Shanghai Children's Medical Center Affiliated to Shanghai Jiaotong University of Medicine,during Jan.2011 and Dec.2013 were detected for retrospectively.Patients were analyzed according to Risk Adjustment for Congenital Heart Surgery-1 (RACHS-1).The age,weight at operation,emergency operation,cardiopulmonary bypass,single ventricle surgery,and RACHS-1 score were detected for risk assessments.Results Overall mortality of congenital heart defect in neonates was 9.96％ (23/231 cases).The top three procedures for high mortality were single ventricle palliative surgery.(25.00％,7/28 cases),corrective operation of interrupted aortic arch (21.43％,3/14 cases),and arterial switch operation with ventricular septum defect repair (13.95％,6/43 cases).There was only 1 case of pulmonary atresia with intact ventricular septum repair,and the case was not classified into high mortality category.With the elevation of RACHS-1,the mortality increased.The mortality rate was 6.67％ (2/30 cases) in RACHS-1 as category 2,6.84％ (8/117 cases) in 3,13.75％ (11/80 cases) in 4,and 50.00％ (2/14 cases) in 6.Multivariable analvsis of risk factors for death showed that single ventricle palliative operation,body weight less than 3 000 g,and RACHS-1 were independent risk factors for mortality.Conclusions This study shows the surgical mortality in neonates with congenital heart defect is still high.Further prospective analysis of specific treatment strategies for high risk patients above was needed and the strategy for single ventricle operation in neonates should be optimized.

Objective To explore the effect of inferior hypothermy treatment on serum TNF and IL-6 in patients with severe cerebral trauma.Methods 46 patients were randomly divided into two groups:inferior hypothermy group(24 C88e8)and normal group(22 cases).There are the same basic treatments within the two groups,in the inferior hypothermy group we also sive them hypothermy treatment rectal temperature:32～34℃ which need to last for nearly 4～5 days,at the same time we give patients the lyric cocktail.the TNF,IL-6 and GCS grades on the lst and 14th day were tested.Results TNF and IL-6 as compared with normal group are higher than the inferior hypothermy group,the differences between the two groups are of statistical significance(P＜0.01).The difference of GCS grades between the two groups are of stafictical significance(P＜0.05).Conclusion The inferior hypothermy tbempy which inhibits TNF and IL-6 releasing after severe cerebral trauma and the following damages plays a very important role in the cerebral trauma therapy.

OBJECTIVE To provide references for clinical diagnosis of catheter-related bloodstream infections(CRBSIs) and focus on studying the spectras of pathogenic bacteria and the drug sensitiveness. METHODS A total of 137 patients enrolled from Jan 2005 to Dec 2007 in our hospital with positive catheter cultures were admitted to our retrospective analysis.Pathogenic bacteria,contaminant bacteria and the drug sensitiveness of main pathogenic bacteria were analyzed. RESULTS From them 80 patients were diagnosed CRBSIs.Among 92 strains of pathogenic bacteria,43(46.7%) strains were Gram-positives,31(33.7%) coagulase negative staphylococci(CNS) strains,31(33.7%) Gram-negative bacilli strains and 18(19.6%) were Candida strains. CONCLUSIONS CNS are the most common bacteria of CRBSIs,and the second are Candida.The common pathogen show multi-drug resistance.

Objective To describe the clinical feature ,therapeutic approach and prognosis of histoplasmosis for improving clinicians’ awareness of this disease .Methods The clinical data of 7 cases of histoplasmosis treated in Shanghai Huashan Hospital from 2001 to 2014 were reviewed retrospectively .Relevant reports about histoplasmosis from 2001 to 2014 in Chinese mainland were comprehensively reviewed .Results The major clinical manifestations of progressive disseminated histoplasmosis included fever ,hepatosplenomegaly ,lymphadenopathy ,and pancytopenia .Skin lesions and pancytopenia were more common in the patients complicated with HIV/AIDS .The patients with local infection were lack of systemic symptoms or signs . Histological examination found Histoplasmacapsulatum in macrophages in bone marrow or biopsy tissues .Amphotericin B was used most frequently to treat histoplasmosis .Itraconazole was appropriate in mild patients .Conclusions Histoplasmosis is caused by H .capsulatum .The golden standard of diagnosis is any culture positive for H .capsulatum .Antifungal treatments such as amphotericin B and itraconazole are very important .

Objective To examine the impact of case management on hospitalizations of the chronically and severely mentally ill patients in Zhongshan. Methods Patients with severe and chronic mental illness,aged ≥ 15 years and living in pilot area were divided into two groups naturally since the program of case management launched, 65 cases in the group of case management and 112 patients in the group of standard management. Hospitalizations of the two groups before and after case management were compared. Results Data were analysised with MIXED procedure. Length of stay in days per admission (LOS) of both groups decreased with time in years (F=11.02, P=0.001), and the decline in LOS of case management group was greater than that of standard management group (F=9.02, P=0.003). The average admissions of case management group was more than that of standard management group (F=4.98,P=0.03). There was no significant differences in average hospitalization incidents before and after case management in both groups(case management group 5.13%vs. 5.38%, standard management group 7.14%vs. 8.92%, P>0.05). Conclusions Case management was effective in reducing hospitalizations for a group of Chinese with chronic and severe mental illness, and may contribute to the balance on mental health resources between community and hospital.

Objective To investigate the clinical features, diagnosis, treatment and prognosis of muhisystemic invasive fungal diseases. Methods Twenty-one patients with multisystemic invasive fungal diseases who were hospitalized in department of infectious diseases from January 2001 to June 2008 were retrospectively reviewed. The pathogenic bacteria, involved organs, underlying diseases, clinical manifestations, treatments and prognoses of muhisystemic invasive fungal diseases were analyzed. Results Among 21 recruited cases, 17 had underlying diseases and 11 were treated with long-term immunosuppressive agents. The main pathogenic bacteria were Cryptococcus neoformans, Aspergillus and Candida parapsilosis. Lung and brain were involved in 16 cases (skin involve in 2 cases and lymph node involved in 1 case simultaneously), lung and lumbar involved in 2 cases, heart valves involved in 2 cases, and liver, spleen and bone marrow involved in 1 case. Eight cases were cured, 6 were improved and 7 died. Conclusions In this study, most of the 21 cases with multisystemic invasive fungal diseases are immunocompromised. The main pathogenic bacterium is Cryptococcus neoformans. The lung and brain are common organs involved. Prognosis is associated with early diagnosis and active anti-fungal treatment.

Objective To investigate the effects of andrographolide on the expressions of Th1 cytokine [interferon (IFN)-γ] mRNA and Th2 cytokines [interleukin (ID-4, IL-10] mRNA of peripheral blood mononuclear cells (PBMCs) from patients with chronic hepatitis B (CHB) and its anti-hepatitis B virus (HBV) activity. Methods PBMCs from CHB patients were cultured with 10 mg/L andrographolide (experimental group) or 0.1% DMSO (control group). HepG2. 2. 15 cells were stimulated with andrographolide of different concentrations (experimental group) or adefovir (control group). The expressions of IFN-γ mRNA, IL-4 mRNA and IL-10 mRNA of PBMCs and the replication of HBV DNA in HepG2. 2. 15 cell line were detected by fluorescence quantitative PCR. Results The expression of IFN-γ mRNA in PBMCs cultured with 10 mg/L andrographolide for 16 h was higher than that in control group (Z=-2. 78, P=0. 05), and the expressions of IL-4 and IL-10mRNA in experimental group were lower than control group (Z= -3. 82, P<0. 01), while the ratio of IFN-γ/IL-4 mRNA was higher than control group (Z= - 3. 82, P<0. 01). Andrographolide with different concentrations had no effect on the replication of HBV DNA in HepG2. 2.15 cells ((=11. 88, P>0.05). However, adefovir had inhibitory effect on the replication of HBV DNA (t =15. 95,P< 0. 05). Conclusion Andrographolide can regulate the expressions of IFN-γ mRNA, IL-4 mRNA and IL-10 mRNA in PBMCs from CHB patients and improve Thl/Th2 balance, while it has no effect on the replication of HBV DNA.

Objective To establish a novel and convenient method to study the phenotype of drug resistant hepatitis B virus (HBV) isolates,and to analyze the drug susceptibility by replacing the reverse transcriptase (RT) domain of wild-type HBV with that of the drug resistant HBV isolates.Methods Full length of HBV isolates was amplified and cloned from the sera of patients prior to nucleoside/nucleotide analogues (NA) treatment.Wild-type full-length HBV genomes was used to construct the recombinant expression plasmids PHY536207 (genotype B) and PHY97 (genotype C).The restriction enzyme sites were introduced in the upstream and downstream region of reverse transeription (RT) domain to construct plasmid,which were named as mPHY536207 and mPHY97,respectively.Lamivudine (LAM) resistant mutant and adefovir (ADV) resistant mutant were isolated and cloned to construct recombinant expression plasmids PHY634 and PHY6923,respectively.Subsequently,the RT domain of mPHY536207 was replaced by that of drug resistant mutant to construct the plasmids RT634 (LAM-resistant) and RT6923 (ADVresistant).The HBV constructs were transfected into Huh7 cells.The HBsAg levels in supernatant were determined by enzyme-linked immunosobent assay (ELISA),and the amount of intracellular HBV DNA was assayed by real-time polymerase chain reaction and Southern blot analysis.Results The plasmids PHY536207 and PHY97 containing genotype B and genotype C wild-type fulllength HBV genomes were constructed successfully,both of which could replicate in Huh7 cells.Intracellular HBV DNA extracted from cells in each of six-well culture plates was more than 1 × 107 copy/ mL,and the introduction of Pst Ⅰ restriction enzyme site did not affect the viral replication and HBsAg secretion.PHY634 and RT634,in which mutant RT domain was replaced into a wild type HBV expressing vector,exhibited the same HBV DNA replication under the treatment with different doses of LAM,the value of 50％ inhibitory concentration (IC50) was ＞100 μmol/L,while the IC50 of mPHY536207 was 0.18μmol/L.Moreover,wild-type isolate was sensitive to ADV (IC50 =1.2 μmol/L),while PHY6923 and RT6923 were resistant to ADV treatment (IC50 ＞100 μmol/L).Conclusion The phenotypic assay is successfully developed in this study based on replacing RT domain of wild-type HBV strains with that of clinical isolated drug resistant strain.