Dengue virus (DENV) nonstructural protein 1 (NS1) is a highly conserved 46-kDa protein that contains 2 glycosylation sites (Asn-130 and Asn-207) and 12 conserved cysteine (Cys) residues. Here, we performed site-directed mutagenesis to generate systematic mutants of viral strain TSV01. The results of the subsequent analysis showed that an alanine substitution at the second N-linked glycan Asn-207 in NS1 delayed viral RNA synthesis, reduced virus plaque size, and weakened the cytopathic effect. Three mutants at Cys sites (Cys-4, Cys-55, Cys-291) and a C-terminal deletion (ΔC) mutant signiifcantly impaired RNA synthesis, and consequently abolished viral growth, whereas alanine mutations at Asn-130 and Glu-173 resulted in phenotypes that were similar to the wild-type (WT) virus. Further analysis showed that the Asn-207 mutation slightly delayed viral replication. These results suggest that the three conserved disulifde bonds and the second N-linked glycan in NS1 are required for DENV-2 replication.

Objective To investigate the diagnostic value of low field MRI for adenomyosis. Methods MRI features of adenomyosis pathologically proved in 18 cases were retrospectively analysed.Results 15 cases were diffusive adenomyosis,the junctional zones of uterus were exteusive thickened to 12.0～32.6 mm,mean 16.2 mm,diffusive high signal intensity distributed over in myometrium which was low signal intensity in 11 cases,it was typical “snowing sign”,lower signal intensity in the myometrium in another 4 cases on T 2WI and fat-suppression imaging. A little high signal intensity was found in 6 cases on T 1WI. 3 cases were focal adenomyosis(adenomyoma), 4 lesions totally. The adenomyoma’s boundaries were not distinct and their shapes were roundish or irregular. The lesions were low signal intensity or diffusive high signal intensity distributed in the low signal intensity fields on T 2WI and fat-suppression imaging. A little high signal intensity was found in 2 lesions on T 1WI. Conclusion T 2WI and fat-suppression imaging of low field MRI are very useful techniques of the diagnosis of adenomyosis.

Objective To observe the effects of comprehensive rehabilitation program on severe traumatic brain injury young patients when the time from injury to admission was delayed. Methods Eleven severe traumatic brain injury young patients received comprehensive rehabilitation program, including the neurodevelopmental techniques. They were evaluated with Functional Independence Measure (FIM) and Berg Balance Scale. One Way ANOVA was used to assess the functional status of patients during the course of treatment. Results After 6 to 12 months' treatment, the motor, cognitive and total scores of FIM and the Berg scores of all patients increased significantly ( P

Objective:To establish a rapid HPLC testing method for chloroquine phosphate,hydroxychloroquine sulfate and amodiaquine hydrochloride.Methods:The chromatographic separation was performed on a GRACE prevail C18(53 mm×7 mm,3 μm)column,and the column temperature was maintained at 30℃.Acetonitrile-0.3% triethylamine acetonitrile solution (adjusting pH to 3.0 with phosphoric acid) (12∶88) was used as the mobile phase,the flow rate was 1.0 ml· min-1 and the UV detection wavelength was 254 nm.The qualitative research was performed using relative retention time and spectral similarity as the double indicators.The relative correction factor in the quantification analysis was used for the content determination.Results:Three anti-malarial drugs showed good behavior in one chromatographic system.The rapid HPLC testing analysis could be achieved.The qualitative research was more accurate by using the double indicators (UV spectral similarity and relative capacity factor).The HPLC qualitative accuracy was increased.The relative correction factor method for the quantification could effectively reduce the use of reference substances and speed up the analysis of HPLC.Conclusion:The method is rapid and simple,and suitable for the rapid determination of drugs.

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.

Objective:To establish a method to determine ferrous succinate in ferrous succinate tablets .Methods:Ion chromatog-raphy was performed on an IonPac AG19(4 mm ×250 mm)anion exchange chromatography column with a guard column of Ion Pac TM AG19( 4 mm ×50 mm);30.00 mmol· L-1 KOH was used as the isocratic eluent at a flow rate of 1.00 ml· min-1 with suppressed conductivity detection ( Curb:75 mA) at the temperature of 30℃.The sample size was 25 μl .Results:The linear range of succinic acid was 2.005 1-25.064 0μg· ml-1(r=0.999 5).The detection limit and quantitation limit was 0.12ng and 0.42ng, respectively. The recovery was 101.45%(RSD=0.43%, n=6).Conclusion:The method is simple, rapid, accurate, reproducible and sensitive , and can be used to determine the ferrous succinate content in ferrous succinate tablets .

To study the relation between drug release and the drug status within curcumin-loaded microsphere, SPG (shirasu porous glass) membrane emulsification was used to prepare the curcumin-PLGA (polylactic-co-glycolic acid) microspheres with three levels of drug loading respectively, and the in vitro release was studied with high-performance liquid chromatography (HPLC). The morphology of microspheres was observed with scanning electron microscopy (SEM), and the drug status was studied with X-ray diffraction (XRD), differential scanning calorimetry (DSC) and infrared analysis (IR). The drug loading of microspheres was (5.85 ± 0.21)%, (11.71 ± 0.39)%, (15.41 ± 0.40)%, respectively. No chemical connection was found between curcumin and PLGA. According to the results of XRD, curcumin dispersed in PLGA as amorphous form within the microspheres of the lowest drug loading, while (2.12 ± 0.64)% and (5.66 ± 0.07)% curcumin crystals was detected in the other two kinds of microspheres, respectively, indicating that the drug status was different within three kinds of microspheres. In the data analysis, we found that PLGA had a limited capacity of dissolving curcumin. When the drug loading exceeded the limit, the excess curcumin would exist in the form of crystals in microspheres independently. Meanwhile, this factor contributes to the difference in drug release behavior of the three groups of microspheres.
Calorimetry, Differential Scanning ; Curcumin ; chemistry ; Drug Liberation ; Lactic Acid ; Microscopy, Electron, Scanning ; Microspheres ; Polyglycolic Acid ; X-Ray Diffraction

Objective To investigate the changes and clinical significance of calcitonin gene related peptide (CGRP) and interleu-kin 1α(IL-1α) before and after warm acupuncture therapy in the patients with lumbar disc herniation (LDH ) .Methods Serum CGRP and IL-1αcontents were detected before and after the warm acupuncture therapy in 60 cases of lumbar disc herniation and 15 healthy control cases by ELISA method for conducting the statistical analysis .The changes of the McGill pain scores and serum CGRP and IL-1αconcentrations before and after the warm acupuncture therapy were observed .Results The McGill pain scores af-ter the warm acupuncture therapy in the LDH patients were significantly decreased ,the difference was statistically significant (P<0 .01) ,and serum CGRP and IL-1α concentrations were significantly decreased ,the difference was statistically significant (P<0 .01) .Conclusion CGRP and IL-1αcan be used as the observation index of the effects for the warm acupuncture therapy in trea-ting LDH ,which can provide the basis for the conservative treatment of LDH .

We wished to screen the cell line that stably expresses the HPV18E5 protein, and to ascertain the influence of HPV18E5 protein on cell proliferation and the cell cycle. The HPV18E5 gene was amplified by the polymerase chain reaction. Then, the His-tag pSecTag-HPV18E5 eukaryotic expression vector was constructed by digestion ligation and connection. The recombinant plasmid was transfected into Balb/c3T3 cells with lipofectamine, and positive cell lines were screened by a culture medium containing bleomycin. HPV18E5 expression in cells was confirmed by western blotting and immuno-enzymatic methods. The influence of HPV18E5 on cell proliferation and the cell cycle were detected by Cell Counting Kit-8 and flow cytometry, respectively. The pSecTag-HPV18E5 eukaryotic expression vector was constructed. After 21-day selection in a culture medium containing 400 μg/mL bleomycin, stably expressing HPV18E5 protein cells were harvested. Compared with control groups, cell proliferation in HPV18E5 stably expressed cells was obviously increased, as was the S phase in the cell cycle. Our results suggested that HPV18E5 influences cell proliferation and the cell cycle. Our study has laid the foundation of the biologic properties of HPV18E5 protein, which will aid further studies on the mechanism of action of carcinogenesis.
Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Human papillomavirus 18 ; genetics ; metabolism ; Humans ; Oncogene Proteins, Viral ; genetics ; metabolism ; Papillomavirus Infections ; physiopathology ; virology ; Transfection

Objective:To analyze the morbid risk factors of chronic tubulointerstitial nephropathy(CTN)with an unknown origin in Wenzhhou City.Method:810 residents in several big districts were investigated.652 subjects,which had complete data and were coincided with the inclusion standard,were analyzed by an unmatched case-control study.The case group contained 19 cases coincided with the diagnostic criteria for CTN with unknown causes.The controlled group contained 633 cases.Thirteen probable risk factors were selected to be investigated:age,sex,body height,body weight, occupation,education,allergic history,addiction to smoking or wine,family history,the respective exposure to heavy met- als,radioactive ray and poisons.A history of medications,including caulis aristolochiae manshuriensis,antipyretic analge- sic,antibiotics,etc.was also investigated.The odds ratio(OR)of CTN with an unknown origin caused by risk factors were calculated with Logistic regression analysis.The obtained data were investigated,and the risk factors were screened out. All the data were analyzed with SPSS 12.0.Result:The homogeneity test showed there was no significant difference in age, sex,occupation and education between the case and controlled groups.The exposure to heavy metals or caulis aristolochiae manshuriensis was screened as the valid risk factor of CTN with OR 17.50 and 103.594 respoetively(P