Objective To investigate the relationships between serum inflammatory mediators and myocardial injury in rats with sepsis.Methods Thirty male Sprague Dawley (SD) rats were randomly divided into normal control group and sepsis (0,24,48,72 h) groups.The rats subjected to sepsis were induced with cecum ligation perforation (CLP).Tumor necrosis factor alpha (TNF-α),interleukin-6 (IL-6),cardiac troponin Ⅰ (cTnI) and B-type natriuretic peptide precursor (NT-proBNP) in the venous blood were measured at 0 h,24 h,48 h,and 72 h after CLP.The lactic acid (Lac) of the arterial blood,cardiac index (CI),acute physiology and chronic health evaluation (APACHE) Ⅱ score were also determined simultaneously.Results The levels of TNF-α,IL-6,cTnI,NT-proBNP,and Lac in sepsis (24 h,48 h,and 72 h) groups were significantly elevated compared to normal control and sepsis (0 h) group.In addition,the number of sepsis (24 h,48 h,and 72 h) groups progressively increased (P ＜0.05).Conclusions It existed a close relationship between serum inflammatory mediators and myocardial injury,and made myocardium damage after CLP.

Objective To explore the mechanism of synthesis metabolism intensified by human growth hormone(GH) on the basis of total parenteral nutrition(TPN) for postoperative gastrointestinal carcinoma patients.Methods Totally 94 cases of postoperative gastrointestinal carcinoma patients were randomly divided into TPN group and TPN+GH group.The levels of TNF-?,IL-1,IL-6,CRP were detected on postoperative day 1,3 and 7.Results The levels of TNF-?,IL-1,IL-6,CRP of TPN+GH group were significantly lower than those of TPN group(P

Objective To investigate the influence of bushenhuoxue drug-containing serum on PI3K and Akt signaling pathway in purified retinal ganglion cell (RGCs) in vitro of Sprague-Dawley (SD) rats,and to explore the protective mechanisms of bushenhuoxue recipe on RGCs.Methods At first,bushenhuoxue drug-containing serum was prepared,and the RGCs of SD rats were purified;after the apoptotic model of pressurized and purified RGCs was established successfully in vitro using open pressure control system,RGCs were dealt with 50 g · L-1,100 g · L-1,200 g · L-1 concentration gradient of bushenhuoxue drug-containing serum.Then the subjected cells were divided into normal culture group (N group),control group (C group),50 g · L-1 bushenhuoxue group (50 g · L-1 BSHX group),100 g · L-1 bushenhuoxue group (100 g · L-1 BSHX group),200 g · L-1 bushenhuoxue group (200 g · L-1 BSHX group).Finally,cell apoptotic rate was detected by Annexin V-FITC/PI staining,while real-time quantitative PCR (qRT-PCR) and Western blot were used to detect the mRNA and protein expression of PI3K and Akt in each group respectively.Results The results of qRT-PCR detection showed that PI3K,Akt mRNA expression level in C group (0.04 ±0.01) was decreased compared with N group (1.00 ± 0.04),and the difference was statistically significant (all P＜0.05),while PI3K,Akt mRNA levels in 50 g · L-1,100 g · L-1 and 200 g · L-1 BXHX group (0.18 ±0.01,0.21 ±0.02,0.22 ±0.01,0.36 ±0.01,0.84 ±0.10,1.07 ± 0.17) were increased compared with the C group,and the difference was statistically significant (all P ＜0.05).The Western blot results of each group showed that PI3K,Akt protein expression level in C group was decreased compared with N group,with statistical difference (all P ＜ 0.05),while PI3 K,Akt protein expression levels in 50 g · L-1,100 g · L-1 and 200 g · L-1 BSHX group were increased compared with C group,with staffstical difference (all P ＜ 0.05).Conclusion Bushenhuoxue drug-containing serum may inhibit the RGCs apoptosis induced by pressure,which may be related to the activation of PBK/Akt signal transduction pathway.

There exist a number of mercury-resistant bacterial in environment, Mer operon is involved in the resistant mechanism, MerRTPA of Mer operon encodes the proteins related to the regulation, transport and reduction of mercury ion, respectively. The toxic mercury ion is transported by MerTP from medium to cytoplasmic mercuric reductase, MerA, and deoxidized to non-toxic and volatile element mercury, Hg(0). Bacterial mercury-resistant system originated from ancient times, and evolved into the Mer operon with diversity by gene integration and insertion. Mercury-resistant bacteria highly specifically absorb mercury ion, and can be used in recovering the mercury-polluted environment as well as the genetic selective marker.

In gene manipulation, different selectable markers with various linkers are necessary. In order to get selectable markers directly, we constructed from pBlueScript SK(-) a series of particular plasmids, pSKsymKm, pSKsymBle, pSKsymEry, pSKsymHyg and pSKsymGm, each contains Kanamycin, Bleomycin, Erythromycin, Hygromycin or Gentamycin resistance cassette. By restriction enzyme digestion and gel extraction, any of five antibiotic resistance genes with specific ends can be conveniently obtained.

OBJECTIVESTo investigate whether antisense Bmi-1 plasmid could inhibit the proliferation of Jurkat cells.

METHODSThe antisense plasmid was constructed by PCR amplification of a 171 bp segment spanning Bmi-1 start codon and zinc finger structure and the PCR product was subsequently inserted reversely to plasmid pLNCX2. The final construct was confirmed through restriction enzyme digestion. G418 was added into the medium after the plasmid was successfully introduced into Jurkat cells by using lipofectin-mediated DNA transfection. The proliferation of Jurkat cells were determined by MTT and colony formation assays. Cell cycle was determined by flow cytometry. The p16 expression of Jurkat cells was studied by immunofluorescent histochemistry.

RESULTSThe growth rate of antisense Bmi-1 transfected Jurkat cells was significantly lower than that of the controls, and the colony forming capacity of the transfected cells decreased significantly (P < 0.01), the colony numbers being (90.7 +/- 9.07)/10(3) cells, (83.3 +/- 6.11)/10(3) cells and (56.0 +/- 5.56)/10(3) cells for control cells, empty plasmid transfected Jurkat cells and antisense Bmi-1 transfected Jurkat cells, respectively. The percentage of G, phase cells was increased and the p16 expression of antisense Bmi-1 transfected cells was significantly upregulated than that of control cells.

CONCLUSIONAntisense Bmi-1 can inhibit the growth and upregulate the expression of p16 of Jurkat cells in vitro.

Cell Cycle ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Jurkat Cells ; Nuclear Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; Plasmids ; genetics ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; Repressor Proteins ; genetics ; Transfection

Objective To assess the liver histopathological characteristics of chronic hepatitis B (CHB) patients with alanine aminotransferase(ALT)lower than 2 times the upper limit of normal (ULN) through liver biopsy, and try to provide subjective evidence for clinical anti-viral treatment.Methods From October 2005 to August 2010, patients accepted liver biopsy in department of infectious disease, Sichuan provincial people's hospital were enrolled. The criteria for liver biopsy was as follow, (1) HBsAg-positive for more than 6 months, (2) HBeAg-positive patients with HBV DNA ≥103 copies/ml or HBeAg-negative patients with HBV DNA≥ 104copies/ml, (3) ALT was lower than 2 times ULN for more than 6 months,and without any hepatic protectants, (4) never accepted any antiviral treatment before, including IFN or nucleoside analogues, (5) willing to accept liver biopsy. Before liver biopsy, routine blood test, prothrombin time, liver function test, hepatitis B antigen and antibody test, HBV DNA quantification were examined. The biopsy position was located under routine ultrasound, liver biopsy were performed to assess the grading of inflammation and necrosis and the degree of fibrosis. The correlation between all the factors and liver inflammation and fibrosis were analyzed. Results Totally 383 cases (240 males and 143 females) met the diagnostic criteria, aged from 16 to 59 years old and the mean age was 28.0 years old. Cases of liver inflammation in G0, G1, G2, G3andG4 grade was 2 cases (0.5％), 165 cases (43.1％), 191 cases (49.9％), 25 cases (6.5 ％ ) and 0 cases (0 ％ ) respectively, cases≥G2 grade accounted 56.4 ％ of total. Meanwhile,stage of fibrosis in S0, S1, S2, S3 and S4 was 103 cases (26.9％), 265 cases (69. 2％), 13 cases (3.4％), 2 cases (0.5％) and 0 cases (0％) respectively, percentage of liver fibrosis in S2stage and over was only 3.9％. The occurrence of serious liver inflammation was associated with age, ALT levels, HBV DNA levels and HBeAg status (P＜0.05). There was no obvious association between HBV DNA level and liver fibrosis (P＞0.05). Conclusions There were obvious liver inflammation and different degree of liver fibrosis in CHB patients with alanine aminotransferase(ALT)lower than 2 times ULN. The degree of liver injury assessed by liver biopsys is recommended as an evaluation for the necessary of anti-viral therapy.

Objective To do screening acute lymphoblastic leukemia patients scFv antibody single chain variable region to cre-ate conditions for the expression and obtain further specificity of antibody fragments.Methods In this study,patients with newly diagnosed acute lymphoblastic leukemia serum as coating antigen using phage display technology,screening phage an-tibody specificity from the semi-synthetic human phage antibody libraries,the first to target the immune antigen-coated tab-let,phage library was added,so that with the target antigen-specific binding phage antibody was immobilized on plates immu-nization,could not be specifically bound phages were rinsed.The eluted specific binding phage,E.coli infection.Could get the specific antibody gene containing phagemid.Results After three “adsorption-elution-amplification”screening process,got stronger leukemia patient antigen-specific phage antibody variable region fragment and identification.Conclusion Got better strain affinity antibody fragments,to create the conditions for the next fragment expression,identification and clinical appli-cation.

To study the pharmacokinetics characteristic of loganin, ferulic acid and stilbene glucoside in rat plasma after oral administration of Bushen Tongluo formula. The plasma samples were treated by using liquid-liquid extraction technique, the concentrations were determined by HPLC-UV. Johnson spherigel C18 column (4.6 mm x 250 mm, 5 μm) was adopted and eluted with the of mobile phase of methanol-water containing 0.01% glacial acetic acid in a gradient mode, with the flow rate at 1.0 mL x min(-1), column temperature at 30 degrees C and injection volume of 10 μL. According to the findings, loganin was determined at 235 nm, ferulic acid and stilbene glucoside were determined at 320 nm, with the sample size of 10 μL. The pharmacokinetic parameters of loganin, ferulic acid and stilbene glucoside were calculated by DAS 2. 0 software as follows: C(max) was (0.369 ± 0.042), (0.387 ± 0.071), (0.233 ± 0.044) mg x L(-1); t(max) was (0.226 ± 0.022), (0.282 ± 0.031), (0.233 ± 0.044) h; t(½β) was (6.89 ± 0.20), (10.73 ± 0.11), (6.93 ± 0.09) h; AUC(0-∞) was (1.91 ± 0.36), (3.22 ± 0.52), (1.52 ± 0.33) mg x h x L(-1); AUCO(0-t) was (1.62 ± 0.33), (2.58 ± 0.43), (1.30 ± 0.30) mg x h x L(-1); CL was (20.2 ± 4.0), (1.39 ± 0.23), (31.7 ± 6.9) L x h(-1) x kg(-1), respectively. The results showed that after the oral administration with Bushen Tongluo formula, loganin, ferulic acid and stilbene glucoside showed concentration-time curves in conformity with the two compartment model, with a rapid absorption, loganin and stilbene glucoside was excreted at a moderate speed, and ferulic acid was excreted slowly (but with the highest bioavailability). Bushen Tongluo formula can main maintain plasma concentration with three administrations everyday and so is suitable to be made into common oral preparation.
Administration, Oral ; Animals ; Biological Availability ; Coumaric Acids ; administration & dosage ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Iridoids ; administration & dosage ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; administration & dosage ; blood ; pharmacokinetics

BACKGROUND: Brain-derived neurotrophic factor(BDNF) is a potent dopaminergic neurotrophin. The major pathological change in Parkinson disease(PD) is the degeneration and death of dopaminergic neurons in the substantia nigra pars compacta. There is a possibility that the onset of PD is associated with BDNF genetic polymorphisms.OBJECTIVE: To investigate the relationship between BDNF genetic polymorphisms and sporadic Parkinson disease(SPD) in Chinese population with the expectation of offering some genetic data for the primary rehabilitation and prevention of the disease.DESIGN: Explorative study based on DNA samples of SPD patients as study group and DNA samples of healthy population as control group.SETTING: Neurological Department of Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, and Human Genome Center of Huazhong University of Science and Technology.PARTICIPANTS: Subjects included DNA samples of 85 SPD patients(study group, Han population, living in Huazhong area for a long term) offered by Department of Neurology of Wuhan Union Hospital and DNA samples of health persons(control group, Han population, living in Huazhong area for a long term) offered by Human Genome Center of Huazhong University of Science and Technology.METHODS: The genotype of healthy controls and SPD patients was analyzed with the polymerase chain reaction-restriction fragment length polymorphism technique (PCR-RFLP).MAIN OUTCOME MEASURES: Genotypes and alleles of the two polymorphisms: G196A and C270T of the two groups.RESULTS: G/A genotype was dominant in both SPD patients and control group with frequencies of 50.6% and 52.0% respectively. The C/C genotype occurred with the frequency of 100% in both groups. There were no significant differences in genotype and allele frequencies of G196A and C270T between SPD and control group( P ＞ 0. 05).CONCLUSION: No association existed between BDNF genetic polymorphisms and the onset of SPD in Chinese Han population of Huazhong area.