1.Experimental research of GuiFu capsule in treating endometriosis
Yan WANG ; Shuang-Hou CHEN ; Rui-Hua LIU ;
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(11):-
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
2.Conjunctive Use of Various Adjuvant and Fusion Protein Which Composed of M2e and N P Genes of Avian Influenza Virus,and the Influence on Immunogenicity
Hai XU ; Hong-Yan HOU ; Bi-Hua DENG ; Qi-Sheng ZHENG ; Ji-Bo HOU ;
China Biotechnology 2006;0(02):-
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
3.Study on the Immune Efficiency for General Vaccine Against Avian Influenza Virus Using Human Mycobacterium Tuberculosis hsp70 as the Carrier for Peptide Epitopes
Qi-Sheng ZHENG ; Gong-Bao XU ; Hong-Yan HOU ; Xue-Hua ZHANG ; Ji-Bo HOU ;
China Biotechnology 2006;0(12):-
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.
4.Microtox-based Fast Testing Method of Comprehensive Toxicity of Shen-Fu Injection
Limin LUO ; Liangchun YAN ; Xinlian HOU ; Hua HUA ; Jianwei GAN ; Junning ZHAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(3):491-496
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.
5.Changes of neuronal activities after gut electrical stimulation with different parameters and locations in lateral hypothalamus area of obese rats.
Yun, YAN ; Xue-Lian, XIANG ; Wei, QIAN ; Jun-Ying, XU ; Xiao-Hua, HOU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):510-5
This study tested the effects of the gastrointestinal pulse train electrical stimulation with different parameters and at different locations on the neuronal activities of the lateral hypothalamus area (LHA) in obese rats in order to find the optimal stimulation parameter and location. Eight gastric electrical stimulations (GES) with different parameters were performed and the neuronal activities of gastric-distension responsive (GD-R) neurons in LHA were observed. The effects of stimulations with 8 parameters were compared to find the optimal parameter. Then the optimal parameter was used to perform electrical stimulation at duodenum and ileum, and the effects of the duodenal and ileac stimulation on the GD-R neurons in LHA were compared with the gastric stimulation of optimal parameter. The results showed that GES with the lowest energy parameter (0.3 ms, 3 mA, 20 Hz, 2 s on, 3 s off) activated the least neurons. The effects of GES with other parameters whose pulse width was 0.3 ms were not significantly different from those of the lowest energy parameter. Most gastric stimulations whose pulse width was 3 ms activated more LHA neurons than the smallest energy parameter stimulation, and the effects of those 3 ms gastric stimulations were similar. Accordingly, the lowest energy parameter was recognized as the optimal parameter. The effects of stimulations with the optimal parameter at stomach, duodenum and ileum on the LHA neuronal activities were not different. Collectively, gastrointestinal electrical stimulation (GIES) with relatively large pulse width might have stronger effects to the neuronal activities of GD-R neurons in LHA of obese rats. The effects of the GIES at different locations (stomach, duodenum and ileum) on those neurons are similar, and GES is preferential because of its easy clinical performance and safety.
6.Expression and implication of toll-like receptors TLR2, TLR4 and TLR9 in colonic mucosa of patients with ulcerative colitis.
Yan, TAN ; Kai-Fang, ZOU ; Wei, QIAN ; Sheng, CHEN ; Xiao-Hua, HOU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(5):785-90
Toll-like receptors (TLRs) family may play important roles in inflammatory bowel disease. This study examined the expression of TLR2, TLR4 and TLR9 in the colonic tissues of patients with ulcerative colitis (UC) and explored their roles in the pathogenesis of UC. Colonic biopsies were taken from the colon of 30 patients with mild or moderate UC (at active phase) and 10 healthy controls during colonoscopy. TLR2, TLR4 and TLR9 protein expression levels were immunohistochemically detected. The mRNA expression levels of TLR2, TLR4 and TLR9 were assessed by reverse transcription polymerase chain reaction (RT-PCR). The disease activity index (DAI), colonoscopic and histologic grades and fecal microbial flora were determined. Histological examination showed that the intestinal mucous membrane of UC patients underwent acute inflammation changes. Immunohistochemistry exhibited that the expression levels of TLR2, TLR4 and TLR9 in colon epithelia and inflammatory cells were higher in UC patients than in control group (P<0.01). The mRNA expression levels of TLR2, TLR4 and TLR9 were increased in UC patients but were not detected in the normal controls. Expression levels of TLR2, TLR4 and TLR9 were positively correlated, and bore close correlation with DAI, colonoscopic and histologic grades and fecal microbial flora. An important mechanism of UC might be that abnormal activation of mucosal immunity by intestinal dysbacteriosis caused dysregulation of TLRS that mediates innate immunity.
7.Cloning of Ginseng ? AS Gene and the Construction of Its Antisense Plant Expression Vector
Shou-Jing ZHAO ; Chun-Xi HOU ; Yan-Long LIANG ; Jian XUE ; Jian-Hua WANG ;
China Biotechnology 2006;0(04):-
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.
8.Advances in Genetic Transformation of Panax Ginseng
Chun-Xi HOU ; Shou-Jing ZHAO ; Yan-Long LIANG ; Jian-Hua WANG ;
China Biotechnology 2006;0(09):-
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.
9.Differentiating peripheral lung cancers from inflammatory masses using dual energy spectral CT imaging
Weishu HOU ; Yan YIN ; Jiejun CHENG ; Jianrong XU ; Xiaolan HUA ; Huawei WU
Chinese Journal of Radiology 2014;48(10):832-835
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.
10.Clinical characteristics of 3090 caes of hemorrhagic fever with renal syndrome and risk factor analysis
Yuan, XING ; Zheng-hua, CAI ; Tie-jun, HOU ; Hong-yan, ZHANG
Chinese Journal of Endemiology 2013;(1):91-94
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.