Objective To isolate endophytic fungi from the rare medicinal plants ephedra, detect the production of ephedrine alkaloids, and explore the optimum fermentation conditions.Methods The endophytic fungi were isolated from the herbaceous stems treated.The ephedrine alkaloids were preliminarily detected by Dragendorff's reagent.The fungi strains producing ephedrine alkaloids were screened by HPLC method with reference substance of ephedrine, pseudoephedrine and methephedrine.After the optimum carbon source and nitrogen source were determined with total ephedrine alkaloids as evaluation indicators by HPLC, the orthogonal test of L16 (45) was designed to investigate five levels of carbon source amount, nitrogen source amount, pH value, temperature, fermentation time.Results The ephedra herbaceous stems endophytic fungus-Es2 mycelium extract after fermentation contained ephedrine, pseudoephedrine and methephedrine.Es3 mycelium extract containing pseudoephedrine after fermentation.Lactose and ammonium sulfate were as the most suitable carbon source and nitrogen source of Es2 fermentation to produce ephedrine alkaloids.Es produced the highest yield of ephedra alkaloids when the carbon source 30 g/L, nitrogen source 4 g/L, pH5.0, temperature 30℃, fermented for 8 days.Conclusion The ephedrine alkaloids of industrial production could be realized by microorganism fermentation method, which has very important significance to alleviate the ephedrine market demand pressure on the environment.

Objective To investigate the effects of berberine on basolateral calcium-activated potassium current I_K(Ca)and cAMP-activated potassium current I_K(cAMP)and its mechanism in treatment of secretory diarrhea.Methods The intact colonic crypt cells were isolated with EDTA solution.The effects of berberine(50,100,500?mol/L)on I_K(Ca)and I_K(cAMP)were detected by patch clamp technique under the conventional whole cell patch clamp mode.The solution of PSS was served as control.Results Berberine could significantly inhibite I_K(Ca)and I_K(cAMP)of rat colonic crypt cells(both P

OBJECTIVE:To evaluate the curative effect and safety of dexamethasone for severe asthma and severe sepsis systematically. METHODS:Randomized controlled trials about dexamethasone for severe asthma and severe sepsis were retrieved from database,such as Cochrane library(1996～2007),PubMed(1966～2007),EMCC(1995～2007),CHKD(1994～2007)and pertinent literatures. The qualities of included studies were evaluated and extracted data were analyzed by Meta-analysis respectively. RESULTS:11 trials containing 1 435 cases were up to included standard. Therapy results of severe asthma:(1)in remission rate of the total symptom,difference between dexamethasone and prednisone was not statistically significant [OR 1.44,95%CI(0.75, 2.75);P=0.27];(2)In the incidence rate of ADR,dexamethasone was lower than prednisone [OR 0.13,95%CI(0.04,0.44);P= 0.001];(3)In the recurrence rate,no statistical significance was noted in difference between dexamethasone and prednisone [OR 0.58,95%CI(0.25,1.31);P=0.19]. Therapy results of severe sepsis:(1)In the mortality,difference between dexamethasone and placebo was not statistically significant [OR 0.85,95%CI(0.41,1.78);P=0.67];(2)In the incidence rate of ADR,no statistical significance was noted in difference between hydrocortisone and placebo [OR 1.05,95%CI(0.51,2.19);P=0.89]. CONCLUSION: Dexamethasone is similar to prednisone in the treatment of severe asthma but better than prednisone in drug safety. The safety of dexamethasone is similar to placebo in the treatment of severe sepsis.

Objective:To evaluate the changes of neurocognitive function in early stage in patients with first-episode schizophrenia.Methods:In this study,73 cases of patients with first-episode schizophrenia (research group) and 75 cases of health person (control group) were selected from January 2015 to January 2016 in our hospital.The neurocognitive function was evaluated by neuro-psychological testing tool and the data between two group were compared.Results:Scores of delayed recall,total recall,3 trial,2 trial and 1 trial of research group were lower than those of control group in BVMT-R test,and the difference was statistically significant (P＜0.05).In HVLT-R test,the scores of total delay,3 trial and 2 trial of research group were significantly lower than those of control group (P＜0.05).The consuming time of dominant hands and subdominant hands in pegboard tasks were significantly higher in research group than in control group (P＜0.05).Conpletion time of connection test A,color connection 1 and 2 in connection test of research group were significantly higher than those of control group (P＜0.05).Attempt number and correct number in research group in PASAT test were significantly lower than in control group (P＜0.05).Number of search errors in research group was higher than in control group,while number of search correct,search total score and digital sign score were significantly lower than in control group (P＜0.05).Total number of words,color and color / word count in research group were lower than in control group in Stroop color word test,and the difference was statistically significant (P＜0.05).WMS-Ⅲ test results between two group had no significant difference (P＞0.05).Conclusion:The neurocognitive function in early stage in patients with first-episode schizophrenia has been extensively damaged.

OBJECTIVETo investigate clinical characteristics and pathological changes of tissue surrounding prosthesis after hip and knee arthroplasty.

METHODSA total of 67 patients receiving hip and knee arthroplasty were included in the study and pathological changes of the revision specimens were evaluated by microscopic examination.

RESULTSOf 67 patients, there were 25 males and 42 females (ratio of 0.6) with a mean age of 64 years. There were 42 cases of revision hip prosthesis and 25 cases of knee prosthesis. The primary causes for the revision varied, including 20 cases of infection (29.9%, within 3 months in 9 cases,3 to 24 months in 3 cases and over 24 months in 8 cases), 14 cases of pain (20.9%), 13 cases of loosening of the prosthesis (19:4%), 9 cases of joint stiffness (13.4%), 8 cases of prosthetic dislocation (11.9%), and 3 cases of prosthesis fracture (4.5%). Pathological findings in the tissue surrounding the prostheses included debris reaction, histiocytes, acute inflammatory, chronic non-specific inflammation, pigmented villonodular synovitis (PVNS), "pseudomembranous", calcification, necrosis, sequestrum, etc. These histological changes were frequently admixed.

CONCLUSIONSVarious reasons may lead to hip and knee revision arthroplasty. The main pathological findings include infection, debris granulomas, chronic non-specific inflammatory changes, PVNS. The surgical pathology of the prosthesis provids guidances for clinical treatment and basic research.

Arthroplasty, Replacement, Hip ; Arthroplasty, Replacement, Knee ; Female ; Hip Joint ; pathology ; Humans ; Knee Joint ; pathology ; Knee Prosthesis ; Male ; Middle Aged ; Reoperation ; Synovitis, Pigmented Villonodular ; pathology

OBJECTIVE: To investigate the correlation between the polymorphism of genotype of site-1296 in alpha2A-AR receptor gene and the susceptibility of vestibular function. METHOD: Ninety-four blood samples were collected from pilot cadets, consisting of susceptible and tolerance groups to vestibular function. Genomic DNA was isolated, and the coding region of alpha2A-AR receptor gene was amplified by polymerase chain reaction (PCR). The PCR products were analyzed by gene sequencing. Gene frequency was calculated, and, the coincidence between the polymorphism of alpha2A-AR receptor gene in the groups and Hardy-Weinberg balance was evaluated. The allele frequency of the two groups was compared by Chi square test. RESULT: G/C polymorphism was existed in Site-1296 of alpha2A-AR gene regulation zone, including GG, GC, CC. The express of GG Genotype in susceptible group exceeded that of the other group. There were significance differences in both genotype constituent ratio and alleles frequency of the two groups. CONCLUSION The polymorphism of genotype of site-1296 in alpha2A-AR receptor gene is possibly correlated with the susceptibility to vestibular function.
Adult ; Alleles ; Gene Frequency ; Genotype ; Humans ; Male ; Polymorphism, Single Nucleotide ; Receptors, Adrenergic, alpha-2 ; genetics ; Vestibular Function Tests ; Vestibule, Labyrinth ; physiology ; Young Adult

In order to confirm the role that the 49th amino acid residue plays in enzymatic inactivity of Glutamine 49 phospholipase A2(Gln49-PLA2),site-directed mutagenesis of its 49th amino acid gene codon was conducted using PCR.Aspartic acid 49 phospholipase A2(Asp49-PLA2-Q49D-PLA2),the mutant of Gln49-PLA2 was expressed in E.coli with pET32a+ vector.The fusion protein,expressed as inclusion body,after being denatured,was on-column refolded and purified by immobilized metal affinity chromatography(IMAC),and then cleaved by Factor Xa.The mature Q49D-PLA2 mutant was obtained by Hitrap SP cation exchange and Superdex 75 gel filtration chromatography,with the recovery rate of 1.3%,and the specific activity of the mature Q49D-PLA2 mutant was 72 U/mg.It has been demonstrated that the 49th glutamine amino acid residue is the main reason in enzymatic inactivity of Gln49-PLA2 and the results are helpful for denatured protein refolding,especially in rich disulfide bonds conditions.

Objective To investigate the advantages of detection for EGFR gene mutations by denaturing high performance liquid chromatography (DHPLC) technology. Methods DHPLC was used to detect EGFR gene mutations at exon 19 and 21 in 49 cases of non-small cell lung cancer (NSCLC) patients,and the direct DNA sequencing was used to verify the accuracy of DHPLC detection. Results EGFR gene mutation was identified from 13 of 49 cases by DHPLC,including deletion mutation at exon 19 in 10 cases (76.92 %) and alternative mutations at exon 21 in 3 cases (23.08 %). Mutation results of DHPLC was consistent with DNA direct sequencing. The results of the direct DNA sequencing were the same as those of DHPLC. The sensitivity of mutation test by DHPLC was 100 %. Conclusion DHPLC technology can be used for large scale screening of EGFR gene mutation with rapid and accuracy.

How to identify active constituents of traditional Chinese medicines (TCMs) and study their interactions are key problems in the development of TCMs. The inhibitory effect of six alkaloids from Rhizoma Coptidis (RC) on Shigella dysenteriae (S. dysenteria) growth had been investigated by microcalorimetry in this study. Main active constituents of RC were confirmed by comparing their contributions to the bacteriostatic effect, and the interactions among active constituents were further researched. According to the result, in 0.8 mg-mL-1 extract of RC, the contributions of six active alkaloids including berberine, coptisine, epiberberine, palmatine and the combination of jatrorrhizine and columbamine were 52.83%, 36.31%, 2.49%, 4.27% and 3.21%, respectively. Therefore, berberine and coptisine were the main active constituents of RC that inhibited the growth of S. dysenteria. The study of interactions among the six alkaloids indicated that, 1 there were some contstituents antagonizing the inhibitory effect of RC, 2 there was a synergy effect between berberine and coptisine, 3 there were additive effects between other four alkaloids and the main active constituents. These results may provide some useful references for the establishment of the quality standard for RC and the development of multi-component TCMs.
Alkaloids ; analysis ; pharmacology ; Berberine ; analogs & derivatives ; analysis ; pharmacology ; Berberine Alkaloids ; analysis ; pharmacology ; Coptis ; chemistry ; Drug Interactions ; Drug Synergism ; Plants, Medicinal ; chemistry ; Quality Control ; Rhizome ; chemistry ; Shigella dysenteriae ; drug effects ; growth & development

The contents of schisandrol A, schisandrol B, schisantherin A, schisandrin A , schisandrin B, schisandrin C in Schisandrae Chinensis Fructus (SCF) were determined simultaneously by HPLC. Collect 100-seed weight, color, pulp content, longitude and latitude of SCF of different batches were collected. SIMCA-P and SPSS were applied to make PLS-DA analysis of 24 batches of SCF and correlation analysis of relevant parameters. According to the 13 parameters, SCF from three different places of origin could be distinguished effectively. It was found that the content of chemical component of SCF increased with latitude and longitude first, and then decrease. The results provide some theoretical basis for study of SCF genuineness and traditional method of identifying just from experience.
China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Quality Control ; Schisandra ; chemistry ; classification