ObjectiveTo establish an animal model of intracerebral hemorrhage (ICH) in rat and to observe the changes in behavior, brain edema and tissue structure during absorption of the hematoma.MethodsAn animal model of ICH was established by stereotactically infusing 100 μl (the first time 20 μl, the second time 40 μl, the third time 40 μl) of autologous caudal artery blood into caudate nucleus in the rats. The behavior of rats with ICH was assessed by Bederson score, beam-walking test, bilateral forepaws grasp and measurement of forelimb placing. The brain edema and the change in histology were observed.ResultsThe successful ratio of this model was 75%. There were significant behavioral changes and brain edema at 6 h after the ICH in rats of the experimental group compared to those of the control group. The worse behavioral changes and brain edema was at 48～72 h after hemorrhage. In the earlier period, the nerve cells were swelling. A lot of glial cells appeared around the hematoma in the late period.ConclusionThis ICH model is reproducible, and the obvious behavioral change and histological changes are similar to clinical patient with ICH.

OBJECTIVETo explore the effect of salvia injection (SI) combined with composite amino acid (CAA) in treating fetal growth restriction (FGR).

METHODSA retrospective analysis was conducted on 106 pregnant women with FGR hospitalized from January 2007 to January 2008. Patients were randomized into 2 groups equally, the treated group (53 cases) treated with SI plus CAA, and the control group treated with CAA alone, all for 7 days. The clinical effect and umbilical blood flow (S/D) in patients were observed.

RESULTS(1) The total effective rate in the treated group was 81.13%, it was 88.80% (16/18) for patients in the gestation period of 24+ -28 weeks, 80.00% (12/15) for those of 28+ -32 weeks, and 75.00 (15/20) for 32+ -36 weeks, while in the control group, the corresponding rates were 69.81%, 77.77% (14/18), 66.66% (10/15), and 65.00% (13/20), respectively. The difference between the two groups was significant (P < 0.05). (2) After treatment, S/D significantly lowered in patients of the treated group (P < 0.05, P < 0.01), no matter how long the gestation period was, but it was insignificantly changed in the control group.

CONCLUSIONThe combined treatment with SI and CAA on FGR could improve the condition of the fetus.

Adult ; Amino Acids ; therapeutic use ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Fetal Growth Retardation ; drug therapy ; Gestational Age ; Humans ; Infusions, Intravenous ; Phytotherapy ; Pregnancy ; Retrospective Studies ; Salvia miltiorrhiza ; chemistry ; Young Adult

OBJECTIVETo investigate variations and significance of plasma fibrinogen in patients with severe acute respiratory syndrome (SARS).

METHODSTotally 148 patients with SARS were divided into the following groups: initial stage group (44 cases) and after the initial stage group (104 cases), common type group (87 cases) and severe type group (61 cases), unilobar lung involvement group (49 cases), bilobar lung involvement group (53 cases) and diffuse lung involvement group (46 cases). The values of plasma fibrinogen of the 148 SARS patients were analyzed and compared among the different groups.

RESULTSThe mean value of plasma fibrinogen (x +/- SD, 522.29 +/- 154.87 mg/dl) of the 148 cases was higher than the normal value (p less than 0.01). There were significant differences between the initial stage and after initial stage groups, between the common and severe type groups, and among the unilobar, bilobar and diffuse lung involvement groups (p less than 0.05).

CONCLUSIONtheses results suggested that elevation of peripheral blood fibrinogen in SARS patients may play an important role in development and progress of the disease and its treatment.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Fibrinogen ; metabolism ; Humans ; Male ; Middle Aged ; Prognosis ; Severe Acute Respiratory Syndrome ; blood ; Young Adult

OBJECTIVETo explore the correlation between the expressions of hypoxia inducible factor-1α (HIF-1α) and insulin in pancreatic cancer.

METHODSHIF-1α and insulin expression was detected by immunohistochemistry in the center and the edge of pancreatic adenocarcinoma specimens of 65 cases. Western blot was used to detect HIF-1α expression and insulin level in the center and the edge of pancreatic adenocarcinoma specimens of 28 cases. The relationship between HIF-1α expression and insulin level in the pancreatic cancer was analyzed.

RESULTSThe results of immunohistochemistry and Western blot showed that HIF-1α protein expression was high in both the center and the edge of pancreatic cancers (P > 0.05), and insulin level was significantly higher at the edge of specimen than that in the center (P < 0.05). HIF-1α protein and insulin levels were positively correlated at the edge of cancer tissue (r = 0.374, P < 0.05), but no significant correlation between them in the center of cancerous tissue (r = -0.145, P > 0.05).

CONCLUSIONInsulin may promote the local invasion and metastasis of pancreatic cancer by up-regulating HIF-1α.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Blotting, Western ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Immunohistochemistry ; Insulin ; metabolism ; Male ; Middle Aged ; Pancreatic Neoplasms ; metabolism ; pathology

OBJECTIVESTo determine whether -238G/A and -857C/T polymorphisms of tumor necrosis factor-alpha (TNF-alpha) gene promoter were associated with outcomes of hepatitis B virus infection.

METHODSA total of 246 HBV self-limited infected subjects and 443 chronic hepatitis B (HB) patients were recruited in this case-control study. TNF-alpha-238G/A and -857C/T gene promoter polymorphisms were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).

RESULTSThe frequency of TNF-alpha-238 GG (90.7%) in chronic HB group was significantly lower than that (95.1%) in self-limited group (P = 0.041). The frequency of TNF-alpha-857 CC (79.7%) in chronic HB patients was significantly higher than that (70.9%) in self-limited infected subjects (P = 0.021). Multiple logistic regression analysis revealed that both TNF-alpha-238GA and -857CC were independently associated with chronic HB.

CONCLUSIONSTNF-alpha promoter variants are likely to play a substantial role in influencing the outcomes of HBV infection.

Adult ; Case-Control Studies ; Female ; Haplotypes ; Hepatitis B ; genetics ; physiopathology ; Hepatitis B virus ; pathogenicity ; Hepatitis B, Chronic ; genetics ; physiopathology ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Promoter Regions, Genetic ; genetics ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo investigate the infection of Cryptosporidium and its epidemiological characteristics in AIDS patients of Southern China.

METHODSStool samples colleted from AIDS confirmed patients. The samples were detected for oocyst of Cryptosporidium by acid fast bacteria stain and indirect fluorescent antibody stain respectively, CD4 count was detected by Flow Cytometry.

RESULTS212 samples of fresh stool obtained from the AIDS patients who live in Guangdong and Yunnan province. The total infection rate of Cryptosporidium in AIDS patients was 4.25% (9/212), the infectious rate of oocyst in the group of 50- 59-years-old was significantly higher than those in 30-39 (P < 0.01); the infectious rate of oocyst in patients with antiretroviral therapy (ART) was also significantly lower (P = 0.0000); we found the patients coinfected with Cryptosporidium with CD4 count all below 100 cells/microl. However, there were no any difference between the infectious rate to the patient's gender, areas and stool shape.

CONCLUSIONAIDS patients infected by Cryptosporidium are not rare in southern China, and the infectious rate was lower than western country. Patients received ART could decrease the infectious rate of Cryptosporidium, Cryptosporidium always happen in patient whose CD4 count was very low (< 100 cells/microl).

AIDS-Related Opportunistic Infections ; parasitology ; Acquired Immunodeficiency Syndrome ; complications ; parasitology ; Animals ; Antigens, Protozoan ; CD4 Lymphocyte Count ; China ; Cryptosporidiosis ; diagnosis ; etiology ; immunology ; parasitology ; Cryptosporidium ; chemistry ; isolation & purification ; Feces ; parasitology ; Flow Cytometry ; HIV Infections ; parasitology ; Humans ; Oocysts ; Staining and Labeling

Animals ; Cattle ; Fatty Liver ; Inflammation ; Liver ; Liver Extracts ; Non-alcoholic Fatty Liver Disease ; Rats

OBJECTIVETo clone the FimA gene of fimbriae from Porphyromonas gingivalis (P. gingivalis) and to construct prokaryotic expression vector which was induced in E.coli BL21(DE3)pLyS in the form of fusion protein expression and to identify, purify the product of its expression.

METHODSTo clone the FimA gene of fimbriae from P. gingivalis and to construct prokaryotic expression vector pET15b-FimA vector which was transformed into the competent cells of BL21(DE3)pLyS. The expression of fusion protein was induced by isopropyl beta-D-1-thiogalactopyranoside (IPTG). With anti-6xHis Tag monoclonal antibody as the first antibody, the expressed fusion protein was characterized by Western blot and purified by Co(2+)-NTA affinity chromatography.

RESULTSCloned FimA gene sequences and inserted into expression vector of the FimA sequences were related to the sequence in GenBank database showed 100% homology. IPTG induced and then identified by Western blot showed a fragment of 4.1 x 10(4) has been expressed. Co(2+)-NTA affinity chromatography column was used to obtain high concentrations of FimA purified protein.

CONCLUSIONThe recombinant prokaryotic expression vector of pET15b-FimA was constructed and was expressed and purified successfully in E. coli BL21 (DE3)pLyS. This study laid the experimental foundation to further prepare for monoclonal antibodies of fimbriae of P. gingivalis and to develop the subunit protein vaccine of prevention of periodontitis.

Cloning, Molecular ; Escherichia coli ; Porphyromonas gingivalis ; Recombinant Fusion Proteins ; Recombinant Proteins

OBJECTIVETo clone the glyceraldehydes 3-phosphate dehydrogenase (GAPDH) gene of Porphyromonas gingivalis (P. gingivalis) and to induce its fusion expression in E. coli.

METHODSGAPDH was obtained by PCR and was inserted into cloning vector pMD-18-T to construct clone recon. Double enzymes digest the recon pMD18-T-GAPDH and the prokaryotic expression vector pET-32a and then connect to get the expressing recon pET-32a-GAPDH. The recombinant expression plasmid which had been confirmed by enzymes digestion was transformed to E. coli competent cells BL21 and induced the expression of GAPDH with isopropyl beta-D-1-thiogalactopyranoside (IPTG) of different density.

RESULTSDNA sequencing showed that the fragment was 99.802% the same to the sequence published in NCBI. Under the best density, IPTG could be highly expressed.

CONCLUSIONThe GAPDH had been successfully cloned and expressed in E. coli which gets ready for the following experiment to study the immunity of GAPDH and the homologues antibody preparation.

Cells, Cultured ; Cloning, Molecular ; Cloning, Organism ; Escherichia coli ; Genetic Vectors ; Glyceraldehyde ; Oxidoreductases ; Phosphates ; Polymerase Chain Reaction ; Porphyromonas gingivalis

OBJECTIVETo observe the effect of tetrandrine on activity of collagenase derived from human hypertrophic scar for the sake of clarifying the mechanism as tetrandrine acting on scar.

METHODSThe experimental concentration was controlled below that of cell proliferation inhibited, SDS-PAGE electrophoresis was adopted to separate collagenase from extracellular matrix, and then activated by trypsin analyzed the activity of collagenase with density scanning apparatus. At the same time quantity of extracellular collagen was measured using improved chloraseptine T oxidizing assay, moreover analyzed correlation between activity of collagenase and quantity of extracellular collagen.

RESULTSIn the concentration below the lever of inhibiting fibroblast proliferation, the total activity of collagenase could be significantly increased by tetrandrine with dosage-dependence associated with quantity of extracellular collagen reduced, which was much greater than that of triamcinolone.

CONCLUSIONIncreasing activity of collagenase on degradation of collagen even in a lower concentration was one of the mechanisms of tetrandrine treating hypertrophic scar.

Benzylisoquinolines ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Collagenases ; metabolism ; Fibroblasts ; cytology ; Humans