Three primer were designed based on the consevered area of the genetic of the ATCC VR-2332 strain and LV strain. And the nest RT-PCR of testing porcine reproductive and respiratory syndrome virus were developed. The nest RT-PCR against ATCC VR-2332 strain, LV strain and B13 strain were done by this method.The DNA fragment were obtained specially from the three strains isolated from different region. The size were 430bp (430bp) , 410bp (413bp) and 410 bp (413bp) separately. But the DNA fragment were not obtained from HCV, PPV and PRV. Its sensitivity was 10-2 TCID50. It's sensitivity increased 10000 times than one step RT-PCR. It should make the method of testing porcine reproductive and respiratory syndrome virus more sensitive, fast and accurate.

OBJECTIVETo explore the effect of total flavonoids of Herba Epimedium (FHE) on BMP-2/RunX2/OSX signaling pathway in promoting osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).

METHODSPassage 3 BMSCs were randomly divided into the control group, the experimental group, and the inhibitor group. BMSCs in the control group were cultured in 0.2% dimethyl sulfoxide + Osteogenuxic Supplement (OS) fluid + DMEM/F12 culture media. BMSCs in the experimental group were intervened by 20 microg/mL FHE. BMSCs in the inhibitor group were intervened by 20 microg/mL FHE and 1 microg/mL NOGGIN recombinant protein. At day 9 alkaline phosphatase (ALP) activity was measured. Calcium nodules were stained by alizarin red staining and the density was observed. The transcription expression of osteogenic differentiation-related proteins (type I collagen, osteocalcin, and osteopontin) and related factors of BMP-2/RunX2/OSX signaling pathway was assayed by RT-PCR.

RESULTSCompared with the control group, ALP activities were enhanced and the density of calcium nodules significantly increased; type I collagen, osteocalcin, and osteopontin expression levels were increased in the experimental group. The expression of osteogenesis-related transcription factor was also increased in the experimental group. Noggin recombinant protein inhibited FHE promoting BMSCs osteogenesis in the inhibitor group. Compared with the experimental group, ALP activity decreased (P < 0.05), the density of calcium nodules was lowered, expression levels of type I collagen, osteocalcin, osteopontin significantly decreased (P < 0.05) in the inhibitor group.

CONCLUSION20 microg/mL FHE promoted osteogenic differentiation process of BMSCs by BMP-2/RunX2/OSX signaling pathway.

Bone Morphogenetic Protein 2 ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Collagen Type I ; metabolism ; Core Binding Factor Alpha 1 Subunit ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Epimedium ; chemistry ; Flavonoids ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Osteocalcin ; metabolism ; Osteogenesis ; drug effects ; Osteopontin ; metabolism ; Signal Transduction ; Sp7 Transcription Factor ; Transcription Factors ; metabolism

A nest RT-PCR/restriction test has been developed in order to distinguish the lapinised vaccine strain from field isolates of classical swine fever virus. The restriction enzyme cut sites mapping of the major coding sequence of E2 gene lapinised vaccine strain and ShiMen strain of classical swine fever virus have been compared. Ten and sixteen unique restriction markers have been found in the lapinised vaccine strain and ShiMen strain. The restriction enzyme cut sites mapping of the twenty six unique restriction marker in the major coding sequence of E2 gene of 17 classical swine fever field isolates have been analyzed. Only 3 sites (HgaI、Hin8I及Hsp92I) are present in the lapinised vaccine strain sequence. Two pans of nested primers and a criteria of analysis have been designed for HgaI restriction marker site. The tests have been conducted first on the lapinised vaccine strain and ShiMen strain of classical swine fever virus resulting in predicted restrection patterns. Finally, the tests have been applied to 5 field isolates of different gene group analyzed by phylogenetic study. The result showed that only HCLV strain gene can be cut to 2 fragment by Hgal , and ShiMen strain and 5 field isolates cant be cut At the same time the sensitivity and specificity of nest RT-PCR have been tested. The sensitivity is 0. 2MLD. The specific fragment of BDV and BVDV were not obtained by the nest RT-PCR. These results showed that the development of the nest RT-PCR/restriction tests is very important for the control and perish of classical swine fever in china.

Objective: To observe the clinical effect of Zheng's Jin Gou Diao Yu (gold-hook-fishing) acupuncture method versus ordinary acupuncture on superficial fascia for treating neck type cervical spondylopathy in Kyrgyz. Methods:A total of 64 Kyrgyz patients conforming to the diagnostic criteria of neck type cervical spondylopathy were included. The patients were randomized into a Zheng's Jin Gou Diao Yu (gold-hook-fishing) acupuncture group and a conventional acupuncture group, with 32 cases in each group. Patients in the Zheng's Jin Gou Diao Yu (gold-hook-fishing) acupuncture group were treated with Zheng's Jin Gou Diao Yu (gold-hook-fishing) acupuncture method to stimulate the superficial fascia, the stimulation sites were primarily located at bilateral sides of the cervical vertebra as well as the trigger points in shoulder-neck region; patients in the conventional acupuncture group were punctured at the same acupoints, with twirling reducing method, and the depth of insertion was determined by the treated region. Patients in both groups received treatment every day for a succession of 5 d as a course, with a 2-day interval between 2 courses, and the whole treatment lasted for 3 courses. After 3 courses of treatment, the McGill pain questionnaire (MPQ) and neck disability index (NDI) were measured to compare the clinical effect between the two groups. Results: After treatment, scores of MPQ and NDI scale dropped when compared with those before treatment, and the differences showed statistical significance (allP<0.05); scores of MPQ and NDI in the Jin Gou Diao Yu (gold-hook-fishing) acupuncture group were substantially lower than those in the conventional acupuncture group, and the differences showed statistical significance (allP<0.05). The total effective rate was 96.9％ in the Jin Gou Diao Yu (gold-hook-fishing) acupuncture group and the cured rate was 78.1％, which were higher than 81.3％ and 40.6％ in the conventional acupuncture group, and the differences showed statistical significance (bothP<0.05). Conclusion: Zheng's Jin Gou Diao Yu (gold-hook-fishing) acupuncture method by stimulating superficial fascia to treat cervical spondylopathy in Kyrgyz can produce a better clinical effect than conventional acupuncture treatment, and is effective in improving pain and stiffness in patients, and thus is worth clinical popularization.

Metabonomics is a new method to study on the metabolic network and the relationship between body and environment, which conforms to the way of traditional Chinese medicine (TCM) research. In the study process of modernization of traditional Chinese medicine, effectively conjunction with metabonomics method will facilitate the integration of TCM with modern biological science and technology, and promote the modernization of TCM. This paper introduce the application of metabonomics in the research of toxicity mechanism of TCM, compatibility mechanism of TCM formula, pharmacology effect of TCM and processing mechanism of TCM. This paper summarize the problems in the TCM metabonomics research and prospect its bright future.
Animals ; Drug Therapy ; Drugs, Chinese Herbal ; adverse effects ; analysis ; metabolism ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; methods ; trends ; Metabolomics ; methods ; trends

OBJECTIVE: To establish an HPLC method for the determination of Paraquat in biological samples. METHODS: Paraquat in biological samples was extracted by C18 columns which were pre-treated with cetyl-trimethyl ammonium bromide (CTAB) and soudium dodecyl sulphate (SDS), and analysed by HPLC/DAD. RESULTS: The detection limit of the method was 1 ng x mL(-1), and the average recoveries were 81%-94%. CONCLUSION The method can be used to analysis of paraquat in biological samples.
Animals ; Chromatography, High Pressure Liquid/methods* ; Herbicides/chemistry* ; Liver/chemistry* ; Paraquat/analysis* ; Rabbits ; Sensitivity and Specificity ; Sodium Dodecyl Sulfate ; Solvents ; Swine

The shape and structure of granules are controlled by the granulation process, which is one of the main factors to determine the nature of the solid dosage forms. In this article, three kinds of granules of a traditional Chinese medicine for improving appetite and promoting digestion, namely, Jianwei Granules, were prepared using granulation technologies as pendular granulation, high speed stirring granulation, and fluidized bed granulation and the powder properties of them were investigated. Meanwhile, synchrotron radiation X-ray computed micro tomography (SR-µCT) was applied to quantitatively determine the irregular internal structures of the granules. The three-dimensional (3D) structure models were obtained by 3D reconstruction, which were more accurately to characterize the three-dimensional structures of the particles through the quantitative data. The models were also used to quantitatively compare the structural differences of granules prepared by different granulation processes with the same formula, so as to characterize how the production process plays a role in the pharmaceutical behaviors of the granules. To focus on the irregularity of the particle structure, the box counting method was used to calculate the fractal dimensions of the granules. The results showed that the fractal dimension is more sensitive to reflect the minor differences in the structure features than the conventional parameters, and capable to specifically distinct granules in structure. It is proved that the fractal dimension could quantitatively characterize the structural information of irregular granules. It is the first time suggested by our research that the fractal dimension difference (Df,c) between two fractal dimension parameters, namely, the volume matrix fractal dimension and the surface matrix fractal dimension, is a new index to characterize granules with irregular structures and evaluate the effects of production processes on the structures of granules as a new indicator for the granulating process control and optimization.
Drugs, Chinese Herbal ; analysis ; Fractals ; Medicine, Chinese Traditional ; Powders ; Quantitative Structure-Activity Relationship ; Synchrotrons ; Technology, Pharmaceutical ; Tomography, X-Ray Computed

OBJECTIVETo provide evidence for intervention measures on safe injection through a survey on the safety of injection in the health facilities in Wulong county, Chongqing city, China.

METHODSAn investigation on injection safety was conducted among health care providers with a uniform questionnaire.

RESULTSGlass syringes which were not appropriately sterilized was found in 52.2% (12/23) of the health facilities. Injection practices were not correct in 31.4% (11/35) of the health facilities. Used disposable syringes were discarded directly without any disposal in 63.6% (21/33) of the health facilities. At least one of the unsafe injection practices mentioned above existed in 77.1% (27/35) of the health facilities.

CONCLUSIONSThe main unsafe factors in the practice of injection are included: the practice to change needle but not syringe in penicillin allergic test and the injuries caused by used disposable syringes in medical staff. The important measures to improve injection safety were to train the medical staff on the knowledge about safe injection and the risks caused by unsafe injection, and to equip the health facilities with safe tools to destroy the used disposable syringes.

Adult ; China ; epidemiology ; Cross-Sectional Studies ; Disinfection ; statistics & numerical data ; Disposable Equipment ; statistics & numerical data ; Equipment Reuse ; statistics & numerical data ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Injections ; adverse effects ; statistics & numerical data ; Male ; Risk Factors ; Rural Health Services ; Safety ; Sampling Studies ; Surveys and Questionnaires ; Syringes

Objectives To investigate the clinical features,prognosis and risk factors of patients with cryptococcal meningitis.Methods Totally 146 patients with cryptococcal meningitis who were hospitalized in Huashan Hospital from January 2000 to December 2006 were enrolled in this study.The clinical data including diagnosis and misdiagnosis,experimental and etiology tests,treat- ments and prognosis from all the patients were analyzed retrospectively.Results Among the 146 patients enrolled in the study,78 patients(53.4%)had concomitant diseases.The misdiagnosis rate of all patients was 72.6%(106/146).The positive rate of cerebrospinal fluid(CSF)India ink smear was 59.6%(87/106),while 43.2%(63/146)cases of cryptococcus neoformans culture in CSF was positive.The positive rate of Latex agglutination test(LAT)was 91.7%(134/146)in CSF among all patients.The treatments were as follows:combination of Amphotericin B(AmpB)or its lipid formula- tions with flucytosine(5-FC)(98 cases),including combination with Fluconazole initally(62 cases), single therapy of Fluconazole(13 cases).Ommaya implanted for lateral cerebral ventricle drainage(53 cases)and AmpB intrathecal injection(53 cases).The average dose of AmpB is 3.06 g.The course of treatment lasted from 12 weeks to 20 months.There were 104 patients(71.2%)cured,27(18.5%) improved,15(10.3%)died and 34(23.3%)relapsed.Conclusions High misdiagnosis rate is common in patients with cryptococcal meningitis.Immunodeficiency is the major risk factor for cryp- tococcal meningitis.CSF LAT is the most sensitive diagnostic test.Early diagnosis,combination of AmpB with 5-FC antifungal therapy and control of acute intracranial hypertension are the keys to im- prove prognosis of cryptococcal meningitis.

Objective To investigate the expression of Toll-like receptor 3 (TLR3) on dendritic cells(DCs) derived from peripheral blood mononuclear cells(PBMCs) of chronic hepatitis B(CHB) patients and to explore the mechanism of sustained infection of hepatitis B virus (HBV). Methods Twenty CHB patients were randomly screened in the study,and ten healthy persons were recruited as controls.The monocytes isolated from peripheral blood of candidates were incubated with recombinant human granuloeyte macrophage colony-stimulating factor (rhGM-CSF) and rhIL-4 to induce the DCs generation and proliferation.Then the phenotype of DCs was identified by micro- scope.The expressions of the phenotypes[histocompatibility leukocyte antigen(HLA)-DR,CD80, CD86,CD83]of immature and mature DCs were measured by flow cytometer.Furthermore,the ex- pression of TLR3 on mature DCs(mDC) and immature DCs(imDC) was determined by flow cytometry and Western blot analysis respectively.Results As for healthy volunteers,the expressions of CD80, CD86,HLA-DR and CD83 on DCs at the 7th day,which were(82.35?8.67)%,(79.61?10.08)%, (92.79?8.48)% and (83.76?5.47)% respectively,were significantly higher than those at the 5th day which were(28.31?8.79) %,(31.17?11.23)%,(27.61?10.28)% and (23.46?11.53)% respec- tively(P0.05).The expression of TLR3 on imDC was significantly higher than that on mDC at control group (P0.05).And the expression of TLR3 on imDC in CHB patients group was significantly lower than that of control group(P