Objective To produce prokaryotic recombinant protein glyceraldehyde-3-phosphate dehydrogenase of Clonorchis sinensis (CsGAPDH), analyze its enzyme activity and immunological function. Methods The recombinant CsGAPDH was purified according to the protocol of GST?Bind~TM kit and was digested with thrombin proteinase and eluted with wash buffer. The BALB/c mice were inoculated with the purified protein. The antisera collected from the mice were used to detect the titres of IgG antibodies by ELISA, and Western blotting was used to identify the specificity of the antisera with the purified CsGAPDH. S-P immunohistochemistry method was used to confirm the expression and distribution of CsGAPDH in adult Clonorchis sinensis with the polyclonal antibodies from immunized BALB/c mice. The CsGAPDH catalytic activity was evaluated employing the conventional substrate glyceraldehydes-3-phosphate (3-GAP). Results SDS-PAGE showed a single purified protein band. Gel scanning analysis revealed that the protein purity of CsGAPDH was 90%. ELISA analysis showed an increased IgG value. S-P immunohistochemistry analysis demonstrated that the recombinant plasmid pGEX-4T-1-GAPDH expressed and distributed in muscle cell membrane of immune mice. Western blotting result suggested that CsGAPDH protein contained essential epitopes with high antigenic activities. This protein CsGAPDH could catalyzed 3-GAP with enzymatic active unit of 2 872 U min~-1ml~-1. Conclusion The recombinant protein CsGAPDH shows a proper enzymatic activity and immunogenicity.

Objective To construct prokaryotic recombinant plasmids of transcriptional co-activator (TC) gene of Clonorchis sinensis, express and purify the recombinant protein and analyze its biological function. Methods A pair of primers was designed according to the known sequence of TC gene. The TC gene fragment was amplified by PCR. After purification and digestion with BamHⅠ and SalⅠ , the TC gene was connected to the prokaryotic expression vectors, pGEX-4T-1 and pET30a(+). By cloning target gene into these vectors, pGEX-4T-1 and pET30a(+), prokaryotic recombinant plasmids of TC gene were constructed and transferred into E.coli BL21. The positive expressed recombinants were detected by SDS-PAGE and Western blotting. Immobilized metal (Ni 2+ ) chelation affinity chromatography was used to purify His-TC produced by the expression of the recombinant protein pET30a(+)-TC. Results The recombinant plasmids, pGEX-4T-1-TC and pET30a(+)-TC, were constructed successfully. SDS-PAGE testified that the molecular weight of the recombinant protein was correct. Western blot analysis of GST-TC recombinant protein testified that the recombinant protein could be recognized by immunized rabbit serum, which means the protein is GST-immune active and the clone can express recombinant Clonorchis sinensis antigen. After affinity chromatography of the pET-TC protein, there was only one protein band with expected size on the SDS-PAGE gel. Conclusion The TC gene was screened from cDNA library of adult Clonorchis sinensis, cloned, expressed and purified. The purified protein of TC gene will be of importance for further research on the biological function of the gene.

Objective:To detect the contents of hyaluronic acid(HA),Laminin(LN) and probe their role in the pathologic course of chronic renal failure(CRF).Methods:The serum HA and LN from 37 patients with CRF and 40 healthy adults were measured by RIA.T test was used in data(x?s) analysis.Results:The HA and LN in CRF serum evidently increased than those of normal controls,respectively(P

A slight halophilic denitrifying bacteria(designated GYL)was screened from the activated sludge which was used to treat high-salinity wastewater.According to the results of morphological observation,physiological and biochemical test,sequence analysis of the 16S rDNA,strain GYL was identified as Halo-monas sp..This strain could survive at 10% salinity and the optimal salinity range for growth was 2%~7%.The suitable pH value for growth was 7.5~8.5 and sucrose was the most effective carbon source.The nitro-gen removal efficiency exceeded 80% when the temperature ranged from 25?C to 30?C.Meanwhile hetero-trophic nitrification characteristics of this strain were measured.Results showed that this strain was able to realize SND and ammonia removal rate was 98.3%.It showed that this strain could perform the whole proc-ess of bacteria denitrification independently.

Ancient literature of ethnic medicine , as a concentrated expression of the ethnic culture of Chinese medicine , has a distinct regional and national character . In this paper , the questionnaire , interview , literature and other methods , were applied to elaborate the research situation of ethnic medicine ancient literature in Sichuan province from the aspect of the literature resources investigation, excavation arrangement, and etc. Ac-cording to the different national medical status differences , we pointed that the national pharmaceutical develop-ment ignored literature research . And we proposed recommendations on how to protect and use the ethnic medicine ancient literature . This paper provided a scientific basis for the further promotion of ethnic medicine literature excavations work in Sic huan province .

Objective To evaluate diagnostic value of X-ray computed tomography(CT) neck masses in children.Methods Clinic,pathologic diagnosis and CT scans of 26 patients from Jan.2004 to Jun.2006 with neck masses which location,density,edge and near construction were reviewed retrospectively.Results Seven cases in neck anterior area,13 cases in neck lateral area,6 cases in other areas or unconfirmed.Eight cases with lower density than cervical muscles,2 cases with same density,2 cases with higher density,14 cases with mixed density.Inflammatory masses were observed in 13 cases,congenital malformation in 11 cases,thyroiditis in 1 case,pharyngeal tumor-like proliferation in 1 case.Conclusion CT scan is the best choice of diagnosis of neck masses in children,and is valuable to diagnosis associated with history and physical examination.

High altitude polycythemia (HAPC) had become one of the main common chronic diseases, which had seriously threatened the health of Plateau people. In the Tibetan medicine classic bookSi-Bu Yi-Dian, there were recordings on HAPC treatment methods and medications, which had the unique advantages of identified therapeutic effect with little side effect. This article analyzed Tibetan medicine in the prevention and treatment of HAPC from aspects such as etiology and pathogenesis, clinical treatment advantages and modern innovation study. Questions were also raised on lacking of standardization on HAPC clinical effectiveness, as well as Tibetan medicine compound material basis and action mechanisms were unclear. It was proposed that based on the inheritance of Tibetan medicine theoretical basis and clinical therapeutic effect, the Tibetan medicine original thinking should be combined with modern science and technology, in order to strengthen the analysis of ancient literature collection in HAPC treatment and data mining in medication experiences. The clinical treatment standards and medication plan should be standardized. Methods of systems biology, such as metabolomics, can be used in the further study of the scientific connotation of HAPC treatment by Tibetan medicine.

Objective To systematically evaluate the clinical efficacy of relieving fever with sweet and warm drugs (Ganwen Chure Therapy) for noninfectious fever caused by vital-energy deficiency based on Meta-analysis. Methods Literature about RCTs of Ganwen Chure Therapy for noninfectious fever caused by vital-energy deficiency in CNKI, Wanfang Database, CBM, VIP, Pub Med, Cochrane Library, and Embase was retrieved by computers from establishment of database to April 2017. After two researchers independently conducted literature screening, cross-checking, data extraction, and literature quality evaluation, cumulative Meta-analysis was performed on the outcome indicators in order of publication time and sample size, and the trend of the results was tested. Then the quality of the literature based on GRADE was under an overall evaluation. Results Totally 27 articles were included in this study, involving 2599 patients. The results of cumulative Meta-analysis showed that the total effective rates of using Ganwen Chure Therapy only [OR=3.875, 95％CI (2.87, 5.24), Z=8.82, P=0.000 1]and Ganwen Chure Therapy combined with routine therapy [OR=5.791, 95％CI (3.55, 9.45), Z=7.03, P=0.000 1]were better than the routine therapy, with statistical significance. Trend test showed that there was variability in the timing of drug combination study, showing that the cumulative Meta-analysis results were not stable. Conclusion Ganwen Chure Therapy has certain efficacy for noninfectious fever caused by vital-energy deficiency. However, the overall quality of the included studies was low, with relatively high homogeneity. There are biases in publication, yet more high-quality clinical research is needed for further verification.

Objective To investigate the expression of immunohistochemistry of gastrin(GAS),somatostatin(SS),proliferating cell nuclear antigen(PCNA) and Fas-ligand(Fas-L) in the sinus ventriculi of children with pediatric gastritis and to explore the significance of their expression in the pathogenesis of pediatric chronic gastritis.Methods Fifty cases of the sinus ventriculi mucosa samples were enrolled in 3 groups:chronic gastritis,helicobacter pylori(Hp) positive(group A,n=20);chronic gastritis,Hp negative(group B,n=19);control group,normal sinus ventriculi mucosa,Hp negative(group C,n=11).Immunohistochemistry En Vision were carried out including GAS,SS,PCNA and Fas-L.Results In the expression of GAS and SS,the values of group A and B were comparatively higher than those of group C,but there was no significant difference among them in statistics.In the expression of PCNA,the value of group A was comparatively higher and that of group B.The value difference between 2 groups was significant(P=0.019);in the expression of Fas-L,no significant difference was found among these 3 groups.Conclusions Expressions of GAS and SS both increase in children with chronic gastritis and maybe the increase of GAS and SS play a role in the pathogenesis of pediatric chronic gastritis;Hp infection promotes the multiplication of the sinus ventriculi membrana mucosa epithelium cell in pediatric chronic gastritis.

Objective To study the intervention effect of the fluorosis with the high aluminum plus soybean, selenium powder, spiral algae. Methods Two week-long SD rats were randomly allocated into 5 groups bases on body weight with 8 in each group, respectively being control, the high fluoride aluminum,the high fluoride aluminum plus soybean, the high fluoride aluminum plus selenium and the high fluoride aluminum plus spiral algae group. The control group was fed with corn produced in non-endemic area containing 5.20 mg/L of fluoride and 6.80 mg/L of aluminum and tap water of fluoride 0.70 mg/L and aluminum 0.20 mg/L. Other groups were fed primarily with corn produced in the endemic area with the content of fluoride of 110.00 mg/L and aluminum of 19.70 mg/L and tap water with high aluminum also with fluoride of 0.70 mg/L and aluminum of 90.00 mg/L. The high fluoride aluminum plus soybean group was added soybean in the beginning of the experiment, approximately 30% of the total amount. Selenium (3.00 mg/kg) was given to the high fluoride aluminum plus selenium groups and spiral algae (1000.00 mg/kg) to the high fluoride aluminum plus spiral algae groups after dental fluorosis was formed. The experiment lasted for 22 weeks. In the end of the experiment 24-hour urine and bones of the limbs were collected to test the content of fluorine and aluminum. The ultrastructure of femur spongiosa was observed under electron microscope. Results ① Bone fluoride in high fluoride aluminum group [(204.07 ± 63.78)mg/kg] was higher than that in the control group, high fluoride aluminum plus soybean group, high fluoride aluminum plus selenium group and high fluoride aluminum plus spiral algae group[(30.06 ± 6.11), (54.12 ± 14.56), (30.44 ± 5.02), (105.08 ± 21.07)mg/kg, t = 0.62,0.53,0.62,0.35, all P < 0.01], indicating that it was lower in high fluoride aluminum plus selenium group than high fluoride aluminum plus spiral algae group(t = 0.27, P < 0.01). ② The urinary fluoride levels in high fluoride aluminum group [(4.52 ± 3.09)mg/L] was higher than that in the control group [(0.89± 0.37)mg/L, t = 0.23, P < 0.01] and lower than that in the fluoride aluminum plus selenium group[(10.38 ± 1.58) mg/L, t = 0.34, P < 0.01], it was higher in high fluoride sluminum with selenium group than that in the high fluoride aluminum plus soybean group and high fluoride aluminum plus spiral algae group[(5.56 ± 1.69), (4.38 ± 3.36)mg/L, t = 0.28,0.35, all P < 0.05]. ③ The bone aluminum level in the control group[(3.32 ± 3.02)mg/kg] was lower than that in the high fluoride aluminum group, high fluoride aluminum plus soybean group, high fluoride aluminum plus selenium group and high fluoride aluminum plus spiral algae group [(374.21 ± 56.11), (118.20 ± 23.59), (114.01 ± 22.84), (67.11 ± 11.53)mg/kg, t = 1.42,0.44,0.42,0.24, all P < 0.05], it was higher in the high fluoride aluminum group than high fluoride aluminum plus the soybean and selenium and spiral algae groups (t = 0.56,0.57,0.68, all P < 0.01)lower in high fluoride aluminum plus spiral algae group than the soybean plus selenium group(t = 0.11,0.10, all P < 0.05). ④The urine aluminum level in high fluoride aluminum group [(1.14 ± 0.32)mg/L] was higher than that in the control group and the high fluoride aluminum plus the soybean group [(0.66 ± 0.10) mg/L, t = 1.92,2.24, all P< 0.05] and that in high fluoride aluminum plus the soybean group was lower than aluminum plus the selenium[(1.03 ± 0.22)mg/L] and aluminum plus spiral algae group[(1.10 ± 0.28) mg/L, t = 1.73,2.06, all P < 0.05]. ⑤ Under electron microscope, the collagenous fiber was arranged in disorder in high aluminum fluorine group, the trabeculum of bone merged into patches or mostly vanished, This phenomenon was alleviated in the intervened group compared to the high aluminum fluorine group, in an attenuating order of the selenium group, the spiral algae and the soybean group. Conclusion In this experiment the intervention measure alleviates fluorosis, the selenium is the best, spiral algae the second and soybean the last.