Object\ To develop a convenient and practical method for the identification of Carapax Trionycis Methods\ Based on the sequence variations of 12S rRNA gene between Pelodiscus sinensis and other softshell turtles, a pair of allele specific primers was designed to distinguish P. sinensis from other species of Trionychidae. DNA were extracted and anplified and Carapax Trionycis could be identified accurately by polymerase chain reaction (PCR) using the primers Results\ Ten samples of turtle shell from different sources were indentified by the allele specific PCR with the primers The result indicated that three samples were substitutes of Carapax Trionycis, consilient with the result from DNA sequence analysis The mitochondrial 12S rRNA gene fragment of P. maculatus and a faked imitation had also been sequenced Conclusion\ The primers could be used as key components in Carapax Trionycis identification kit

AIM　It is difficult to identify the Chinese crude drug snake gallbladder accurately by morphological and microscopical characteristics or chemical components only. In order to solve the problem, the technique based on DNA molecular marker was introduced into the authentication of snake gallbladder. METHODS　DNA templates were extracted from the membrane or the bile of snake gallbladder, and also from the muscle of the original animal Elaphe schrenckii. About 400 bp DNA fragments of 12S rRNA gene were amplified from the templates and sequenced subsequently. RESULTS　Enough amounts of DNA templates could be extracted from a bit of membrane or bile of snake gallbladder. The sequence of amplicons from the membrane, bile and muscle of the same individual were identical completely. CONCLUSION　The technique of DNA molecular marker could be used for the authentication of snake gallbladder and bile. The results indicate that the technique could be used for the identification of crude drugs from other animal secretion. DNA sequence analysis also demonstrated that the origins of commercial snake gallbladder were complicated and more efficient quality control was necessary for supervising the crude drug in the market.

Objective To compare the degree of pain in patients after radical gastrectomy under different anesthetic regimens.Methods One hundred and two ASA Ⅰ or Ⅱ patients of both sexes,aged 50-75 yr,weighing 45-70 kg,undergoing elective radical gastrectomy,were randomly divided into 3 groups ( n =34 each):general anesthesia (GA) group,combined general-subcostal transversus abdominis plane block (CGTA) group and combined general-epidural anesthesia (CGEA) group.The patients were sent to the postanesthesia care unit (PACU) after tracheal extubation,and the VAS score on arrival in the PACU was recorded.The degree of pain was evaluated by VAS score,and when VAS scores ＞ 3,the patients received intravenous morphine titration.When VAS scores ≤ 3,morphine titration was stopped and all the patients were connected to patient-controlled intravenous analgesia and/or epidural analgesia pump.The total amount of morphine consumed was recorded at the end of titration,and the occurrence of adverse reactions was also observed.Results Compared with groups GA and CGTA,the incidence of moderate to severe postoperative pain was significantly decreased in group CGEA (P ＜0.01).The incidence of severe postoperative pain,the VAS score on arrival in the PACU and the total amount of morphine consumed were decreased gradually in groups GA,CGTA and CGEA ( P ＜ 0.01 ).The incidence of sedation was significantly lower in group CGEA than in group GA (P ＜ 0.01 ).There were no significant differences in the other adverse reactions among the three groups ( P ＞ 0.05 ).Conclusion The degree of pain is reduced gradually in patients after radical gastrectomy under GA,CGTA and CGEA.

OBJECTIVETo establish HPLC fingerprint for identification and evaluation of the quality of Trachelospermum jasminoides.

METHODThe analysis was performed on a ZORBAX Eclipse XDB-C18 analytical column (4.6 mm x 150 mm, 5 microm) with H2O (A) and methanol (B) as mobile phases in gradient mode. The elutin conditions were 0-15 min, changed from 10% B to 30% B; 15-40 min, to 40% B; 40-60 min, to 60% B. The column temperature was set at 30 degrees C and the flow rate was 0.8 mL x min(-1) with the detection wavelength of 230 nm.

RESULTThe HPLC chromatographic fingerprint of T. jasminoides, showing 19 characteristic peaks, was established from 14 lots of T. jasminoides. The similarity of every lot of T. jasminoides was calculated as good in general, the similarity of 10 lots was above 0.9 and the other 4 lots had low similarity.

CONCLUSIONThe chromatographic fingerprint of T. jasminoides with high characteristics and specificity can be used to control its quality.

Apocynaceae ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; standards ; Quality Control

Viral hepatitis is a major liver disease caused by virus infection .Viral hepatitis is popular in China , mainly caused by hepatitis B and hepatitis C viruses .Experimental animal model is a necessary platform for the research on mechanism of viral infection and pathogenicity , for treatment and vaccine development .Up to date, a great progress in the development of viral hepatitis animal models has been achieved in spite of the most of findings are limited to hepatitis B and C.Here, we summarize the recent findings of viral hepatitis animal models , focusing on the tree shrew animal model and its modeling strategy .

Objective To investigate the transcriptional autoregulation of PhoP/PhoQ under different growth conditions in Yersinia pestis.Methods The entire promoter region of YPO1635 was amplified and cloned into the pRW50 vector containing a promoterless lacZ reporter gene.The recombinant LacZ reporter plasmid was transformed into the wild-type strain (WT) and the phoP mutant strain (ΔphoP),respectively,to measure the promoter activity (the β-galactosidase activity) of the target gene in WT and ΔphoP by using the β-galactosidase enzyme assay system.Total RNAs were extracted from WT and ΔphoP strains,and primer extension assay was employed to detect the promoter activity by examining the amount of primer extension products of YPO1635 in WT and ΔphoP.Results The LacZ fusion results showed that the transcription of YPO1635 was positively regulated by PhoP under L-TMH and brain-heart infusion(BHI) conditions,but it was not regulated in H-TMH medium.The primer extension assay detected two transcriptional start sites located at 90 and 118 bp upstream of the translation initiation site of phoP,named P1 and P2,respectively.Under low Mg2+ TMH conditions,the promoter activity of P1 rather than P2 was positively regulated by PhoP.Under high Mg2+ TMH conditions,the promoter activities of both P1 and P2 showed no obvious difference in the WT and ΔphoP strains.Under rich BHI conditions,both promoters were under negative control of PhoP.Conclusion Different autoregualtion patterns of PhoP/PhoQ under different growth conditions would help Y.pestis to quickly adapt to the changing living environment.

Objective Tamm-Horsfall protein (THP) may play a role in kidney stone formation.The article aimed to conduct a preliminary study on the role of THP in kidney stone formation by investigating the changes of THP in rat urine, pathological changes of renal tissue and the formation of calcium salt crystals after establishing CNPs rat model of kidney stones.Methods Stone samples of 40 patients from February to June 2015 in our department were collected to establish the model of CNPs-induced kidney stone in rats and prepare CNPs suspension.48 SD rats were randomly divided into experimental group (group A) and blank control group (group B).Group A were injected with CNPs and the same amount of sterile saline injection in the group B.The urine of rats was collected after injection at 3h, 6h, 12h, 24h, 1w, 2w, 4w and 8w.ELISA were applied to detect THP levels in the urine.Then the rats were killed to take the kidney tissue.HE staining was used to investigate the pathological changes of the cells and evaluate the formation of the calcium salt crystals.Results THP levels in group A at 24h, 1w, 2w, 4w and 8w ([166.03±3.02], [173.50±1.78], [174.55±2.05], [176.54±2.45], [177.11±1.76]pg/mL) were significantly higher than that at 3h(165.89±2.23pg/mL)(P<0.05), which was the same case in comparison with those of group B ([157.65±2.22], [156.54±1.43], [159.45±3.21], [158.63±2.98], [157.33±2.05]pg/mL).Compared with the calcium salt crystal score at 6h (1 point), the scores at 3,6,12,24h (average score 2 points) increased.At 2w the score increased significantly to 3 points and reached the top score(6.7points) at 8w, which was of significant difference.The score of calcium salt crystals was in positive correlation with THP content (r=0.843,P<0.05).Conclusion THP in urine may contribute to the aggregation of calcium salt crystals and the formation of kidney stones.

Objectives To investigate the prevalence of non-alcoholic fatty liver (NAFLD) disease and its correlation with chronic metabolic diseases in two primary school students in Shanghai. Methods One thousand ifve hundred and thirty-two 7-11 year-old students from two primary schools were enrolled in Septamber-October 2011. The anthropometric indices, blood pressure, screening for pseudoacanthosis nigricans and liver ultrasonography of all subjects were recorded. Results The overall prevalence of NAFLD, obesity, abdominal obesity, pseudoacanthosis nigricans, high systolic blood pressure and high diastolic blood pressure was 6.5%, 26.7%, 16.3%, 5.1%, 1.7%and 1.9%, respectively. The prevalence of NAFLD and abdomi-nal obesity in students of central urban area was signiifcantly higher than that in suburban area (P<0.01). The binary regression analysis revealed a signiifcant association between NAFLD in students with sex, age, obesity, abdominal obesity, pseudoacan-thosis nigricans and high systolic blood pressure (P<0.05). Conclusions NAFLD has close correlation with chronic metabolic disease in children. It is time to adopt prevention, detection and treatment in NAFLD children with symptoms of chronic meta-bolic diseases.

Animals ; Bacterial Proteins ; genetics ; metabolism ; Biofilms ; growth & development ; Gene Expression Regulation, Bacterial ; Lipopolysaccharides ; chemistry ; metabolism ; Operon ; Promoter Regions, Genetic ; Protein Binding ; Siphonaptera ; microbiology ; Species Specificity ; Transcription, Genetic ; Transferases ; genetics ; metabolism ; Virulence ; Yersinia pestis ; genetics ; metabolism ; pathogenicity ; Yersinia pseudotuberculosis ; genetics ; metabolism

OBJECTIVETo establish a method for gene complementation in Vibrio parahaemolyticus using the plasmid pBAD33.

METHODSThe entire coding region of opaR or aphA was amplified by PCR and cloned into pBAD33. The recombinant plasmid was transformed into δopaR and δaphA (the opaR or aphA null mutant strain, respectively) separately to construct the complemented mutant strain C-δaphA and C-δopaR, respectively. RT-PCR was used to verify the transcription of opaR and aphA in the corresponding complemented mutant strains. Primer extension experiments were performed to determine the relative mRNA levels of mfpA (a gene previously characterized to be negatively regulated by AphA and positively by OpaR) in the wild-type strain, δopaR, δaphA, C-δaphA, and C-δopaR.

RESULTSopaR and aphA were transcribed in the corresponding complemented mutant strains, and their mRNA levels were comparable to those detected in the wild-type strains.

CONCLUSIONA method has been established for gene complementation in Vibrio parahaemolyticus using the plasmid pBAD33.

Bacterial Proteins ; genetics ; Gene Expression ; Genetic Complementation Test ; methods ; Plasmids ; genetics ; Promoter Regions, Genetic ; Vibrio parahaemolyticus ; genetics