OBJECTIVE To investigate the incidence of integrons in Pseudomonas aeruginosa strains isolated from blood samples in Peking University People′s Hospital and to analyze the correlation between integrons and drug resistance of P.aeruginosa.METHODS Forty-two strains of clinically isolated P.aeruginosa were collected.The antibiotics susceptibility was tested by K-B methods.Integrase gene of integron was amplified by PCR using degenerate primers.The integrons were classified by restriction fragment length polymorphism(RFLP) analysis of positive PCR products with Hinf Ⅰ restriction enzyme.RESULTS The drug-resistance rates of 42 strains of P.aeruginosa against 20 kinds of antibiotics ranged from 9.5% to 100%.Twenty-three strains were resistant to 12 kinds of antibiotics.Nineteen of the 42 isolates(45.2%) contained integrons,all of which were revealed as class Ⅰ of integrons by RFLP analysis.Neither class Ⅱ nor class Ⅲ of integron was detected.The positive percentage of integrons was increased by years.CONCLUSIONS Class Ⅰ integrons are widespread in isolates from blood samples in our hospital.The presence of integrons is closely associated with multi-drug resistance of P.aeruginosa.

Objective To compare the clinical efficacy of moxibustion Zhiyin with knee chest decubitus,ear acupoint magnetic bead press combined with knee chest decubitus and knee chest decubitus in correction of breech position,thus to provide more safe and effective correction method for the clinical treatment.Methods 150 pregnant women with breech were selected.They were randomly divided into observation group Ⅰ,observation group Ⅱ and control group according to the digital table,50 cases in each group.The control group was treated with knee chest decubitus to correct breech position,the observation group Ⅰ was given moxibustion Zhiyin combined with knee chest position,the observation group Ⅱ was given ear acupoint magnetic bead press combined with knee chest decubitus.Results The effective rates of the observation group Ⅰ and observation group Ⅱ were 96％,92％,respectively,which were higher than 76％ of the control group,the difference were statistically significant (χ2=5.09,4.18,all P＜0.05).The effective rate between the observation group Ⅰ and observation group Ⅱ showed no significant difference (P＞0.05).Conclusion Moxibustion Zhiyin combined with knee chest decubitus and ear acupoint magnetic bead press combined with knee chest decubitus in the correction of breech position have short course of treatment,high success rate,and are more economical and convenient,with high safety and good curative effect,which are worthy of promotion.

OBJECTIVETo observe the effect of Zhenqing Recipe (ZQR) on non-alcoholic fatty liver (NAFL), and the expression of hepatic salt-inducible kinase 1 (SIK1) and sterol-regulatory element binding protein-ic (SREBP-lc) in type 2 diabetes rats.

METHODSA rat model of type 2 diabetes was established by high fat/sucrose diet combined with intraperitoneal injection of small dose streptozotocin (STZ) . Modeled rats were randomly divided into the model group, the ZQR group, and the metformin group, 8 in each group. Eight rats were recruited as a normal control group. ZQR at the daily dose of 12 g crude drugs/kg was administered to rats in the ZQR group by gastrogavage. Metformin suspension at the daily dose of 150 mg/kg was administered to rats in the metformin group by gastrogavage. Equal volume of distilled water was administered to rats in the normal control group and the model group. All medication lasted for 12 weeks. The levels of fasting blood glucose (FBG), free fatty acid (FFA), serum triglyceride (TG), serum total cholesterol (TC), serum alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were detected. The body weight and wet liver weight were weighed, and the liver weight index calculated. The liver TG content was measured. The pathological changes of liver and the expression of SIK1 were observed by HE staining and immunohistochemistry. The mRNA and protein expression of SIK1 and SREBP-1c were detected using RT-PCR and Western blot.

RESULTSCompared with the normal control group, FBG, FFA, TG, TC, ALT, AST, liver weight index, and liver TG contents significantly increased (P < 0.01); liver steatosis was severe, the mRNA and protein expression of SIK1 obviously decreased (P < 0.01); mRNA and protein expression of SREBP-1c increased (P < 0.01). After drug therapy, compared with the model group, FBG, FFA, TG, TC, ALT, AST, and liver weight index significantly decreased, liver TG contents significantly decreased, the mRNA and protein expression of SIK1 obviously increased, while mRNA and protein expression of SREBP-1c obviously decreased (P < 0.05, P < 0.01) in the ZQR group and the metformin group (P < 0.05, P < 0.01); and the pathological changes were also improved. All the indices were improved more in the ZQR group (all P < 0.05).

CONCLUSIONIn this experiment, we found that the expression of SIK1 decreased in NAFL rats with type 2 diabetes. ZQR could alleviate lesion of NAFL type 2 diabetes rats possibly by up-regulating hepatic SIK1 expression at mRNA and protein levels.

Animals ; Diabetes Mellitus, Type 2 ; complications ; drug therapy ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Fatty Liver ; complications ; drug therapy ; metabolism ; Liver ; metabolism ; Male ; Protein-Serine-Threonine Kinases ; metabolism ; Rats ; Rats, Wistar ; Sterol Regulatory Element Binding Protein 1 ; metabolism

Background:Tryptophan(TRP)is an essential amino acid,and can produce 5-hydroxytryptamine(5-HT)via 5-HT signal pathway and kynurenine( KYN)metabolic pathway under the catalysis of enzyme,thereby participating in the pathogenesis of visceral hypersensitivity in diarrhea-predominant irritable bowel syndrome(IBS-D). Aims:To investigate the relationship between visceral sensitivity and TRP metabolic pathway in patients with IBS-D. Methods:Thirty patients with IBS-D and 30 healthy controls from June 2012 to January 2014 at Guangdong Provincial TCM Hospital were enrolled. Score of gastrointestinal symptom rating scale( GSRS)was evaluated. Visceral sensitivity was measured by anorectal manometry. RT-PCR and Western blotting were used to detect mRNA and protein expressions of colon mucosal IDo, respectively. Serum 5-HT,5-HIAA,TRP,IDo,KYN,KYNA concentrations and IDo activity,KAT activity were determined by high performance liquid chromatography assay. Results:Compared with control group,GSRS score was significantly increased(P < 0. 05),initial perception threshold,defecation threshold,pain threshold were significantly decreased(P < 0. 05),anorectal constriction pressure was significantly increased( P < 0. 05),serum 5-HT,5-HIAA concentrations were significantly increased(P < 0. 05),mRNA and protein expressions of IDo were significantly increased (P < 0. 05),serum KYN was significantly increased(P < 0. 05),KYNA was significantly decreased(P < 0. 01),IDo activity was significantly incseased(P < 0. 01),and KAT activity was significantly decreased in IBS-D group(P < 0. 01). Correlation analysis showed that initial perception threshold,defecation threshold,pain threshold and anorectal constriction pressure were correlated with 5-HT,5-HIAA,TRP,KYN,KYNA,IDo activity and KAT activity in patients with IBS-D (P < 0. 05). Conclusions:TRP metabolic pathway is correlated with visceral hypersensitivity in patients with IBS-D.

OBJECTIVETo investigate the molecular mechanisms underlying in the treatment of inflammatory bowel disease by Pulsatilla Decoction.

METHODSForty Wistar male rats were randomly divided into 5 groups( n = 8)control group, model group, model + positive control group (mesalazine), Pulsatilla Decoction treatment group, in addition, the Pulsatilla Decoction treatment group was divided into middle and high dose group. Intragastric administration was used in the positive control group and Pulsatilla Decoction treatment group. The expression of interleukin-1beta (IL-1beta), interleukin-6(IL-6) and tumor necrosis factor-alpha (TNF-alpha) were detected by real time PCR after extraction of RNA from colons.

RESULTSCompared with the model group, positive medicine and Pulsatilla Decoction group, especially high-dose group, could effectively inhibit the expression of IL-1beta, IL-6 and TNF-alpha.

CONCLUSIONPulsatilla Decoction could exert its effect in the treatment of inflammatory bowel disease by inhibiting the expression of proinflammatory cytokines.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Inflammatory Bowel Diseases ; drug therapy ; metabolism ; Interleukin-1beta ; genetics ; metabolism ; Interleukin-6 ; genetics ; metabolism ; Male ; Phytotherapy ; Pulsatilla ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

In order to study the analgesic effect of Shaoyao Gancao Decoction, this paper discussed material basis and mechanism from the perspective of macromolecules in traditional Chinese medicine. Inspired by the phenomenon of turbidity after boiling Chinese medicine, this experiment took Shaoyao Gancao Decoction as the research object to study the formation process of precipitation during boiling. The results showed that aggregates with a certain shape were formed in the solvent during the boiling process, and the precipitate was obtained by standing and centrifuging. Analysis found that the precipitation was mainly composed of small molecules such as paeoniflorin, albiflorin, liquiritin, glycyrrhizic acid, isoliquiritin and gallic acid, and macromolecules such as protein and polysaccharide. The composition of precipitate was consistent with that of Shaoyao Gancao Decoction, but the analgesic effect of Shaoyao Gancao Decoction after removing the precipitate was significantly reduced. Based on these results, we isolated small molecular compounds, polysaccharides and protein from Shaoyao Gancao Decoction and their contents are 60.4, 700.7 and 207.2 mg·g^-1 respectively. We get the ratio, polysaccharide: small molecule = 11.6∶1, protein: small molecule = 3.4∶1, the precipitate is prepared in the state of boiling. The characterization results showed that the particle size of the precipitate will change significantly after co-heating, and the content determination results showed that the content of the six small molecular compounds which was free in solvent was significantly reduced after the formation of the precipitate. The acetic acid writhing experiment proved that the precipitate has a good analgesic effect, and effectively reduced the levels of inflammatory factors prostaglandin E2 and nitric oxide, and increased the level of anti-inflammatory factor interleukin-10. These results proved that the precipitate in Shaoyao Gancao Decoction is an important material basis for analgesic effect, and macromolecules such as protein and polysaccharide are the main components of the precipitate. The study of macromolecules in the precipitate of Shaoyao Gancao Decoction not only provides new ideas and methods for elucidating the pharmacodynamic material basis of Shaoyao Gancao Decoction, but also provides a reference for analyzing the scientificity of traditional decoction.

This paper, on top of its introduction of the concept of medical ethics counseling, analyzed the necessity of establishing a medical ethics consultation system in China.By reviewing the development of the medical ethics counseling system in other countries, the authors, in view of China′s medical practice, put forward specific recommendations on establishing a medical ethics counseling system in China.Issues on the construction of organ donation ethics counseling system in China were also discussed in focus.

The paper comprehensively understands the current construction situation of 30 provincial disease control websites through scientific evaluation methods and professional evaluation indexes,and finds and analyzes the major problems.The result shows that the website construction of disease control system is in the development phase generally,information publication and online service is provided with certain foundation,but the mutual communication and website design are relatively weak links which can be greatly improved.

The mainly antigenic sites for the adenovirus neutraliation are present on Loop1 and Loop2 of hexon.Majority research were focus in the human adenovirus.Little was known on infectious canine hepatitis virus (ICHV), which was also called canine adenovirus typeⅠ.Here,ICHV (the isolated strain) DNA was isolated and purified from the cultured MDCK cells.The Loop1 and Loop2 fragments were amplified by polymerase chain reaction(PCR) method,and then was connected by ligase T4.The target fragment was then connected with vector pET28a.The nucleotide sequence ecoding Loop1 and Loop2 was determined.The nucleotide sequence identity of Loop1 region between the isolated strain and CLL, RI261 and Toronto A26/61 strains is 100%, 100% and 83.8%, and the nucleotide sequence identity of Loop2 region between the isolated strain and CLL, RI261 and Toronto A26/61 strains is 88.1% , 88.1% and 99.3%, and amino acid identity is 93.6%, 93.6% and 98.6%.The recombinant Loop protein was expressed in E.coli and was approximately 36kDa in size,and then was purified. Then BALB/c mice were injected subcutaneously in the back and armpit with the recombinant Loop protein.The anti-ICHV antibody titers of immunized serum was tested by indirect ELISA and the titers were up to 1:320.Western blot demonstrated that immunized sera could specifically combine with ICHV. The research laid a foundation for creating new genetic engineering products of infectious canine hepatitis virus.

Microbial lipases are important industrial biocatalysts with the character of stereoselectivity,site selectivity and high catalytic activity with few side effects.They have been used widely in many industrial and agricultural fields.The technology of protein engineering has been successfully applied to improve the activity and stability of microbial lipases,which will raise the competitive capacity of microbial lipase preparations and enlarge theirs application fields.The strategies,the problems and the prospects of protein engineering technology which have been applied to modify the microbial lipases was surveied.