Objective: To study the effect of GuiFu capsule on sexual hormone and other relative factors in rats with endometriosis.Methods: Endometriosis was induced in rats by surgery of self-endometrium transplant.Rats with successful models were randomized into GuiFu capsule group and Danazol group.Expressions of ER,PR,VEGF and NK cells were examined by immunohistochemical stainin;the level of E2 and P was detected by radioimmunoassay.Results: GuiFu capsule can reduce the level of E2 and P in the blood of rats(P

Based on the gene sequence of AIV matrix protein 2(M2) and two cytotoxic T-lymphocyte epitopes derived from nucleoprotein,the prokaryotic expression vector pET-3M2e-NP1-NP2 was constructed.The target gene was expressed in the solvable form in BL21(DE3) when induced with 1.0 mmol/L IPTG and Western blot analysis showed that the expression product had good immunogencity.Purified fusion protein was mixed with various amount of adjuvant,including Freund's,Vash oil and chitosan,and then immunized 20-day-old chicken by intramuscular injection and boosted 3weeks later.Blood samples were collected weekly following the primary vaccination.The anti-M2e antibody was detected with ELISA method with the synthesized peptide as coated antigen;the neutralizing ability of anti-serum was evaluated on MDCK cell line and chick embryo,the CD4+ and CD8+ T lymphocyte amounts in peripheral blood of immunized chicken was measured by flow cytometry.Results showed that the fusion protein can induce immunological reaction,and the antibody can bind with the viral M2 protein that expressed on the surface of MDCK cells.Flow cytometry result showed that CD4+ and CD8+ T lymphocyte in peripheral blood increased obviously following immunization(P

M2e gene of three copies for H5N1 subtype AIV was synthesized and fused with human mycobacterium tuberculosis hsp70 gene.The fused gene was cloned into the prokaryotic expression vector to get pET-3M2e and pET-3M2e-hsp70.Recombinant protein r3M2e and r3M2e-hsp70 were successfully expressed induced with IPTG and purified with Ni2+-NTA collumn.Following that,the immunity of the recombinant protein was analysized with Western blot.20-day-old AIV non-immunized chickens were vaccined with r3M2e and r3M2e hsp70,at the same time,Trx and KLH-M2e inoculated chickens were served as vector and positive controls.Two weeks after the primary vaccination,every group was boosted with the same vaccine as in the primary vaccination.The humoral immunity of the vaccined chickens was evaluated with antibody detection against M2e,cytopathic suppression test,and indirect fluorescence assay.The cellular immunity was estimated according to lymphocyte subtype analysis with flow cytometry and M2e specific cytokine detection.Four weeks after the boost vaccine,all groups were challenged with 100EID50 AIV of H9N2 subtype,and the virus from swabs was detected with Real-time PCR.Results indicated that r3M2e hsp70 vaccined chicken developed the better humoral and cellular immune response,also,made a better performance compared with r3M2e vaccined group in virus challenge.

In this research,Microtox technology was used to evaluate the comprehensive toxicity of Shen-Fu injection,which was an exclusive product of the pharmaceutical company.Firstly,vibrio fischeri was used as test strain.The best test system and methodology were identified.Under the best test conditions,vibrio fischeri was firstly used in the luminescent bacteria comprehensive toxicity of the exclusive product Shen-Fu injection.The results showed that in the 2 mL reaction system,the best recovery liquid volume was 0.9 mL/ freeze-dried vial,50 μL bacterial suspensions/ sample,the detection time was 10 min after adding bacterial suspensions,the pH was 5-10,the luminous intensity was 800 000-12 000 000 for 10 rin.The RSD of replication experiment and intermediate precision test was ? and ?,respectively,which fitted to the requirements.There was no significant difference on EC50 values among 9 batches of tested Shen-Fu injection (41.07％,RSD ＜ 5％).It was concluded that there was significant concentration-effect relationship between Shen-Fu injection and the toxicity of vibrio fischeri.There was no significant difference on EC50 values among different batches.It indicated that there was small quality volatility among different batches of Shen-Fu injection.The control on product manufacturing was strong.

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.
Cloning, Molecular ; DNA, Complementary ; Datura ; enzymology ; genetics ; Escherichia coli ; Hyoscyamine ; chemistry ; Mixed Function Oxygenases ; genetics ; Plant Leaves ; enzymology ; Plant Roots ; enzymology ; Recombinant Proteins ; genetics ; Scopolamine Hydrobromide ; chemistry

RT-PCR amplification of ginseng ?-amyrin synthase gene was successfully performed based on the total RNAs extracted from ginseng hairy roots induced by Agrobacterium rhizogenes A4 using a modified guanidine isothiocyanate-method. Sequence analysis of this gene revealed that its sequence was consistent with the sequence of a previously reported ginseng ?-amyrin synthase gene (GenBank No. AB009030). This gene was inserted into pMD-119T simple vector and transformed into E.coli DH5?. Furthermore antisense plant expression vector of this gene was constructed using the pBI121 vector, laying foundation for studies on antisense regulation of ginseng ?-amyrin synthase gene.

Genetic engineering is a powerful tool in Panax ginseng breeding.Genetic transformation and plant regeneration are the premise and foundation involved in genetic engineering of Panax ginseng.Ginseng can be regenerated through organogenesis or somatic embryogenesis and indirect somatic embryogenesis is mainly used for its regeneration.Summurized the factors influencing plant regeneration such as different explants,different carbohydrates,somatic embryo optimization and hormone-free approach.Ginseng transformation has been achieved by Agrobacterium tumefaciens and Agrobacterium rhizogenes and transgenic ginseng with good characters was obtained by introducing genes associated with biosynthesis of ginsenosides or herbicide gene.Hairy root culture system can supply large scale of ginsenosides,thus effect of rolC genes on ginseng hairy root induction,regeneration and bioreactor culture of hairy root were discussed.Additionally,problems that are present in genetic engineering of Panax ginseng were also discussed in this review.

Objectives To investigate the clinical significance of dual energy spectral CT (DESCT) in quantitatively differentiating peripheral lung cancers from pulmonary inflammatory masses.Methods Sixty patients with 35 lung cancers and 25 inflammatory masses underwent DESCT to get arterial phase (AP) images and venous phase (VP) images.Iodine concentrations in the central and peripheral zone of the masses were measured and normalized to the aorta as normalised iodine concentration (NIC).The difference of NIC between central and peripheral zone of the masses (dNIC) was calculated.The spectral attenuation curve was obtained automatically and the slope of curve (λHU) was also calculated in the two groups.The quantitative parameters was presented as M (Q1,Q3),and Wilcoxon signed rank test was used to compare above two independent samples.Receiver operating characteristic (ROC) curves were generated to calculate the sensitivity and specificity.Results NICs in the central zone of peripheral lung cancers were significantly lower than that of inflammatory masses:mean NICs were 0.03 (0,0.05) versus 0.12 (0.07,0.20) in AP,and 0.14 (0.12,0.19) versus 0.30 (0.21,0.57) in VP (Z=-4.14,-3.70,respectively,P＜0.01).While the dNIC values of lung cancers were significantly higher than that of inflammatory masses:dNIC values were 0.08 (0.05,0.11) versus 0.04 (-0.02,0.08) in AP,and 0.23 (0.17,0.34)versus 0.07 (-0.04,0.08) in VP(Z=-2.56,-4.00,respectively,P＜0.05).Mean λHU values of lung cancers were also lower than inflammatory masses:1.03 (0.67,1.67)versus 2.75 (1.61,3.19) in AP,and 1.58 (1.30,2.17) versus 3.25 (2.37,4.54) in VP (Z=-3.90,-4.42 respectively,P＜0.01).According to ROC curves,cutoff value of λHU =2.11 in VP had the highest sensitivity (89％) and specificity (91％) in differentiating peripheral lung cancers from inflammatory masses.Conclusions Contrast-enhanced dual energy spectral CT imaging with some quantitative parameters such as normalised iodine concentration,dNIC,and the slope of spectral attenuation curves may be a promising new method for differentiating peripheral lung cancers from inflammatory masses.

Objective To observe the therapeutic effect of methylphenidate hydrochloride controlled-release tablets (OROS-MPH) on attention-deficit hyperactivity disorder (ADHD). Methods Seventy-two cases of children with ADHD were randomly divided into treatment group (40) and control group (32). Cases of treatment group were given 0.8-1.0 mg??kg-1 of OROS-MPH for three months. Cases of control group were given 1.2-1.4 mg??kg-1 of atomoxetine hydrochloride for three months. After 12 weeks treatment, children were evaluated by Wechsler intelligence test, Integrated visual and auditory continuous performance test (IVA-CPT), the SNAP-Ⅳ effect assessment scale and TESS scale. Results The treatment efficiency was similar in both groups. Attention deficit and hyperactivity in both groups were improved obviously. Wechsler intelligence score was significantly elevated ( P<0. 05), SNAP-Ⅳ score was significantly decreased ( P<0. 05), and IVA-CPT score was increased significantly after treatment ( P<0.05) . The changes of scores on hyperactivity, auditory attention and visual attention were more in OROS-MPH group than those in atomoxetine group(P<0.05). There was loss of appetite in 10 children of OROS-MPH group and in 14 children of atomoxetine group. There was drowsiness in 1 child of OROS-MPH group and in 5 children of atomoxetine group, as well as difficulty to fall asleep in 6 children of OROS-MPH group and 1 child of atomoxetine group (P<0.05). One child developed a transient spasm after 4-month treatment. Conclusion Both of OROS-MPH and atomoxetine hydrochloride can improve learning ability and the symptom of attention deficit and hyperactivity, and they are similarly effective and safe in children with ADHD, but OROS-MPH can work faster.

Objective In this paper we analyzed the major risk factors of hemorrhagic fever with renal syndrome(HFRS) as well as its clinical manifestation,so as to provide a basis for clinical diagnosis and effective control of HFRS in Xi'an.Methods The method of retrospective study was used to collect clinical data of hospitalized patients with HFRS between 2005 and 2010 in the Hospitals for Infectious Diseases in Xi'an city,Zhouzhi and Huxian counties and then clinical manifestations of the patients with HFRS were analyzed and classified according to the national standards for clinical symptoms and test indicators of HFRS.HFRS patients matched by the ratio of 1 ∶ 2 healthy human controls were retrospectively investigated in order to obtain risk factors relevant to HFRS incidence using casecontrol study.Odds ratio(OR) method was used for single factor study.While for the multifactor study,we took the conditional Logistic regression approach.We also built models for both studies.Factor with OR ＞ 1 and P ＜ 0.05 was judged to be a risk factor.Results A total of 3090 cases information of patients with HFRS and 6018 healthy controls were collected.Of the 3090 cases of HFRS patients,sixty vaccinated patients showed atypical clinical manifestations and they were mild or moderate cases and no deaths.A total of 3030 nonvaccinated patients had obvious clinical symptoms.Severe or critical cases accounted for 39.07％(1184/3030)and 60 patients died and the fatality rate was 1.98％ (60/3030).The results of single factor Logistic regression analysis showed that of the 16 factors analyzed,the difference of 11 risk factors between the case group and the control group was statistically significant.Multivariate Logistic regression analysis showed that of the 11 factors,the difference of six factors between the case group and the control group was statistically significant.The main risk factors of suffering HFRS in the order were:exposure to rat pollutants,living in the affected areas,sitting or lying on grass fields in the affected areas,working in the affected areas,house rat infestation,and domesticated cats or dogs(OR =6.826,5.764,4.882,4.857,3.126 and 2.875; P ＜ 0.01 or P ＜ 0.05).Conclusions Vaccines are very useful in the sense that vaccinated HFRS patients tend to have mild symptoms and good prognosis.Health education in this area should be focused on the six risk factors in order to prevent the spreading of HFRS.