Streptococcus gordonii is a nonpathogenic gram-positive commensal bacterium and component of the normal microbial flora of the human oral cavity. It is suitable to be as a mucosa vaccine vector due to its special bionomics. knowing the bionomics of Streptococcus gordonii,the general expressing system,and the application of it in mucosa vaccine,will provid important reference for the further development of its mucosa vaccine.

To construct expressing vector carrying esat6 gene and express this protein in Streptococcus gor-donii GP251. esat6 gene was amplified by PCR with specific primer from genome of Mycobacterium tuber-culosis (MTB)H37Rv. Inserted esat6 into the pMD18-T vector by T/A clone to get recombinant vector pMD18-esat6. Then digested pMD18-esat6 with restriction enzyme, esat6 was cloned to vector PSMB104 and expressed in Streptococcus gordonii GP251. The expression of esat6 protein was detected by Tricine-SDS-PAGE and Western-blot, ELISA technique was also used to detect its secretory volume. Re-striction endonuclease, PCR, Tric ine-SDS-PAGE and Western-blot confirmed that esat6 gene was cloned into expressing vector successfully, and a 10 kD protein secreted in Streptococcus gordonii GP251, this pro-tein has a good immunogenicity. The expression vector of esat6 gene was constructed, and esat6 protein ex-pressed in Streptococcus gordonii1 successfully, it will be benefit for future study.

Myopia is the most important cause of visual impairment in adolescents. However, its etiology is complex. In recent years, a large number of epidemiological studies have been done on risk factors of myopia. Most of these studies is cross- sectional study, not longitudinal cohort study. Overall, the incidence of myopia is the result of the interaction between genetic susceptibility and environmental exposure. This review is about the risk factors for myopia.

OBJECTIVETo investigate the triterpenoids from root of Achyranthes bidentata in Henan.

METHODSephadex, normal-and reversed-phase column chromatographies were applied for the isolation and purification. The structure determinations were performed by means of physiochemical properties, MS and NMR data analyses.

RESULTSeven compounds were isolated from the water soluble fraction in root of A. bidentata, and determined as achyranthoside A (1), achyranthoside E (2), momordin Ib (3), chikusetsusaponin IVa (4), chikusetsusaponin IVa methyl ester (5), chikusetsusaponin V (6), chikusetsusaponin V methyl ester (7).

CONCLUSIONCompounds 1 and 2 were isolated from the natural resources for the first time.

Achyranthes ; chemistry ; Molecular Conformation ; Molecular Structure ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Saponins ; chemistry ; isolation & purification

OBJECTIVEAn accurate and reliable analytical method for-simultaneous determination of six active components (scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C) in plants of Erycibe was developed.

METHODScopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C in the samples were well separated in analytical HPLC by gradual elution with methanol-0.1% formic acid solution. The chromatographic condictions: Agilent Poroshell 120 EC-C18 column, flowing rate being 1 mL x min(-1), detecting wavelength at 345 nm.

RESULTGood linearities of scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C were in the range of 0.026 8-2.68, 0.027 0-2.70, 0.008 1-0.81, 0.018 8-1.88, 0.017 6-1.76, 0.019 6-1.96 μg, respectively (r > 0.999 6). The average recoveries of the six components were 98.1%, 98.7%, 100.8%, 100.4%, 99.7%, 101.1%; the relative standard deviations were 2.67%, 2.86%, 2.62%, 1.98%, 2.76%, 2.19%.

CONCLUSIONThe method is simple, feasible and reproducible and can be used for the quality control of plants of Erycibe.

China ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Convolvulaceae ; chemistry ; Coumarins ; analysis ; Drugs, Chinese Herbal ; analysis ; Glucosides ; analysis ; Scopoletin ; analysis

Objective To in ve stigate the effects of melatonin on oxidative stress and renal function in kidne ys of diabetic rats. Methods Diabetic model of rats was induced by streptozocin. Male diabetic SD rats were assigned to 3 group s: (1) Untreated (DMC); (2) DM+Mel 1, melatonin 0.2 mg?kg -1 ?d -1 b y gavage, and (3) DM+Mel 2, melatonin 5 mg?kg -1 ?d -1 by gavage, se ven rats were in each group. Seven normal rats were assigned to normal control ( NC) group. Eight weeks later, systemic and renal intrinsic anti-oxidant enzyme activities, lipid peroxide levels and urinary protein excretion, creatinine clea rance rate and kidney weight were evaluated. Results Glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities were lower and lipid peroxide levels were higher in renal cortex of diabetic ra ts than those in NC group. SOD and GSH-Px activities in renal cortex increased and plasma triglycerides level and urinary protein excretion decreased in DM+Mel 1 group. DM+Mel 2 group showed changes similar to DM+Mel 1 group. These dose s also reduced lipid peroxide levels in renal cortex and kidney weight/body weig ht ratio of diabetic rats. Moreover, larger dose melatonin reversed the lowered SODactivitiesine rythrocytesand the raised lipid peroxide levels in plasma of dia betic rats. Conclusion Melatonin may inhibit oxi dative stress in kidneys and improve renal function in diabetic rats. Overproduc tion of reactive oxygen free radicals and insufficieny in intrinsic anti-oxidan t enzymes may contribute to the development of diabetic nephropathy.

BACKGROUND:As the multipotent differentiation potential, bone marrow mesenchymal stem cells exert an important role in bone metabolism disorders, which is regulated by a variety of hormones and cytokines. Currently, the epigenetic regulatory mechanisms underlying osteogenic differentiation of bone marrow mesenchymal stem cells are unclear, and association between histone deacetylase (Hdac) and osteoporosis needs to be further explored. OBJECTIVE:To investigate the epigenetic mechanisms of bone formation by analyzing the expression of Hdac1, 3, 4 mRNA profile in bone marrow mesenchymal stem cells isolated from ovariectomized mice. METHODS:A total 30 female healthy Kunming mice were randomly divided into sham group and ovariectomy group. After 7 days of adaptive feeding, mice in the ovariectomized group (n=15) were subject to bilateral ovariectomy;mice in sham group (n=15) were sham-operated. RESULTS AND CONCLUSION:In the ovariectomy group, the trabecular bone of the femur was sparse or broken, the width of trabecular bone was narrowed, the trabecular separation was widened, and the trabecular bone volume was reduced. The level of Hdac3 mRNA was lower in bone marrow mesenchymal stem cells from ovariectomized mice compared with controls, but there was no significance between Hdac1, Hdac4 mRNA expression of the two groups. These findings demonstrate that Hdac might play an important role in bone remodeling in the model of estrogen deficiency-induced osteoporosis.

OBJECTIVETo develop an HPLC method for fingerprint determination of the astragalosides injection.

METHODAnalyses were carried out at 25 degrees C on a Zorbax SB-C18 column (4.6 mm x 250 mm,5 microm). Mobile phase A was water, and B was acetonitrile. The analysis followed a linear gradient program. Initial conditions were 15% B; 0 to approximately 65 min, changed to 60% B. The flow rate was 0.8 mL x min(-1). The drift tube temparature of the ELSD was 105 degrees C, and gas flowrate was 2.7 L x min(-1).

RESULTThe RSD of relative retention time of the common peaks in the plant material, the extract and the injection fingerprints was less than 0.1%, and there was good similarity among 10 batches of samples.

CONCLUSIONThe method was reliable and simple which could be used for quality control of the injection.

Astragalus membranaceus ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Injections ; Light ; Plants, Medicinal ; chemistry ; Quality Control ; Saponins ; administration & dosage ; analysis ; chemistry ; isolation & purification ; Scattering, Radiation ; Triterpenes ; analysis

AIM: To investigate the biocolonization of poly 2-hydroxyethy methacrylate (PHEMA) sponge with cornea tissue and evaluate the therapeutic effects of modified porous poly 2-hydroxyethy methacrylate-Polymethyl met-hacrylate (PHEMA-PMMA) Keratoprostheses (KPro) on rabbit and monkey corneas. METHODS:The KPro were made using two-stage polymerization combined with mechanical cutting. The experiment was divided into two groups. In the control group, ten normal rabbit eyes received lamellar implanta-tion of PHEMA sponges. The sponges were obtained 2 weeks, 1,2,3 and 4 months after operation. The cell proliferation and neovascularization inside the sponges were observed using light and transmission electron microscopy (TEM) and immunohistochemistry. In the experimental group, the porous PHEMA-PMMA KPros were inserted into the lamellar pockets of eight rabbit corneas and two monkey corneas (stage I operation). The healing process was investigated by slit-lamp microscopy. The anterior lamellar cornea tissues were removed 3 months after surgery, exposing the under-neath transparent core (stage II operation). The operated eyes were then followed up for 3-6 months.light microscope, fibroblasts started to grow into the cornea 2 weeks after operation; lots of cells, accompanied with new blood vessels, invaded into the cornea 2-3 months after surgery. Invading cells of sponge, as well as keratocytes, were positive for vimentin. Under the electron microscope, the invading cells looked healthy and were surrounded by extracellular matrix and collagen. In 8 rabbit eyes which received KPro implantation, anterior lamellar cornea melting happened in two eyes after the stage II operation. The remaining 6 corneas retained their central cores during observation after the stage II operation.Two monkey operated eyes were found no complication thoughout the whole follow-up.cornea. The modified PHEMA-PMMA KPros have obtained a relatively stable results after implantation into animal corneas.

Objective To investigate the antimicrobial resistance of clinical isolates from a hospital in Chongqing during one year according to CHINET project.Methods Disc diffusion test (K-B method) was employed to study the antimicrobial resistance. WHONET5 was used for data analysis.Results In one year period from 2004 to 2005,690 non-duplicate isolates were collect- ed.Enterobacter isolates showed the lowest resistance rate to imipenem and meropenem.About 37.5% of E.coli and 31.4% of K.pneumoniae isolates produced ESBLs,respectively.All ESBLs-producing strains were susceptible to imipenem and mer- openem.About 37.2%,39.4% and 48.9% of P.aeruginosa isolates were resistant to imipenem,meropenem and ceftazidime, respectively.Pandrug-resistant (PDR) P.aeruginosa was isolated from our hospital.All strains of A.baumannii were sus- ceptible to imipenem and meropenem.About 37.7% of A.baumannii were resistant to cefoperazone-sulbactam.Twenty-nine strains showed the same resistant pattern among non-susceptible strains of A.baumannii,mainly derived from 2 clones by PFGE analysis.Conclusions The surveillance results suggest that prevalent strain resistant to cefoperazone-sulbactam may pres- ent in some ICUs.Resistance rate to cefoperazone-sulbactam increased significantly.