2.Effect of Buyang Huanwu decoction and its simple prescription (Naojian tablet) on CDK4/Cyclin D1 expression of rats with cerebral ischemia.
Fang LIU ; Yu-hong WANG ; Guang-xian CAI ; Yan SHE ; Le SHAO ; Xiang-yi XIA
China Journal of Chinese Materia Medica 2015;40(20):4058-4062
To evaluate the regulating effect of Buyang Huanwu decoction and its simple prescription (Naojian tablet) on CDK4/Cyclin D1 expression in hippocampus tissues of rats with cerebral ischemia, SD rats were divided into the sham-operation group, the model group, the Buyang Huanwu decoction group (ig, 3.15 g · kg⁻¹) and the simple prescription group (ig, 2.41 g · kg⁻¹). Each group was further divided into five subgroups based on time points after the administration, i. e. 1 d, 3 d, 7 d, 14 d and 28 d, respectively. CDK4/Cyclin D1 expressions of the group at different time points were examined by using immunohistochemistry and real-time qPCR. According to the results, the cerebral ischemia model group showed higher CDK4/Cyclin D1 expression than the sham-operation groups (P < 0.05), suggesting that the cell cycle signal pathway would be activated by the cerebral ischemic injury. Both Buyang Huanwu decoction and simple prescription groups showed significantly lower cyclin expression than the model group at 3 d, 7 d, 14 d, 28 d (P < 0.05), indicating both Buyang Huanwu decoction and its simple prescription could play the neuroprotective effect through the cell cycle signal pathway.
Animals
;
Brain Ischemia
;
drug therapy
;
genetics
;
metabolism
;
physiopathology
;
Cell Cycle
;
drug effects
;
Cyclin D1
;
genetics
;
metabolism
;
Cyclin-Dependent Kinase 4
;
genetics
;
metabolism
;
Drugs, Chinese Herbal
;
administration & dosage
;
Humans
;
Male
;
Rats
;
Rats, Sprague-Dawley
3.Prediction of CTL epitopes of Tat exon 1 region in HIV-1 CRF07_BC strains in China
Zhiming FANG ; Hui XING ; Zhefeng MENG ; Kunxue HONG ; Lingjie LIAO ; Xiang HE ; Jianxin Lü ; Yiming SHAO
Chinese Journal of Microbiology and Immunology 2010;30(2):135-139
Objective To predict the CTL epitopes of Tat exon 1 region in HIV-1 CRF07_BC strains, which were prevailing in China. Methods Total of 236 plasma samples were from the 3rd National HIV Molecular Epidemic Survey (NMES3). All the subjects were infected with HIV-1 CRF07_BC viruses. The tat exon 1 region was amplified by reverse transcription reaction and nested polymerase chain reaction (nested-PCR), then the PCR products were sequenced. The distribution of CTL epitopes of this region were predicted by on-line software BIMAS HLA Peptide Binding Predictions and statistics software. Results To-tal of 236 CRF07_BC strains were from 16 provinces, mainly in intravenous drug asers(58.9%)and then sex(25.0%). It was showed that there were 12 CTL epitopes of 236 Tat exon 1 region of CRF07_BC strains mainly located in proline-rich region, cysteine-rich region and core-region. Those epitopes were banded by 5 HLA presenting molecules in genotype(A * 2501 ,A * 2902, B * 15,B * 5301 and Cw * 1203) and 6 HLA presenting molecules in serotype (B53, B58 ,B57 ,A3 ,A68 and Cw12). The frequency of single amino acid substitution was more than 50% in 7 CTL epitopes. Conclusion The CTL epitopes in Tat exon 1 of CRF07 _BC strains were located in different functional regions, and there were some amino acid variations in them.
4.Cloning,Sequence Analysis and Expression in E.coli of the EP0 Gene of Pseudorabies Virus Ea Strain
Liu-Rong, FANG ; Huan-chun, CHEN ; Shao-bo, XIAO ; Xiang-Ru, MA ; Ge-fei, WANG
Virologica Sinica 2001;16(2):183-187
The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea,and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.
5.Changes of end-tidal carbon dioxide during cardiopulmonary resuscitation from ventricular fibrillation versus asphyxial cardiac arrest
Qing-Ming LIN ; Xiang-Shao FANG ; Li-Li ZHOU ; Yue FU ; Jun ZHU ; Zi-Tong HUANG
World Journal of Emergency Medicine 2014;5(2):116-121
BACKGROUND:Partial pressure of end-tidal carbon dioxide (PETCO2) has been used to monitor the effectiveness of precordial compression (PC) and regarded as a prognostic value of outcomes in cardiopulmonary resuscitation (CPR). This study was to investigate changes of PETCO2 during CPR in rats with ventricular fibrillation (VF) versus asphyxial cardiac arrest. METHODS:Sixty-two male Sprague-Dawley (SD) rats were randomly divided into an asphyxial group (n=32) and a VF group (n=30). PETCO2 was measured during CPR from a 6-minute period of VF or asphyxial cardiac arrest. RESULTS:The initial values of PETCO2 immediately after PC in the VF group were significantly lower than those in the asphyxial group (12.8±4.87 mmHg vs. 49.2±8.13 mmHg,P=0.000). In the VF group, the values of PETCO2 after 6 minutes of PC were significantly higher in rats with return of spontaneous circulation (ROSC), compared with those in rats without ROSC (16.5±3.07 mmHg vs. 13.2±2.62 mmHg,P=0.004). In the asphyxial group, the values of PETCO2 after 2 minutes of PC in rats with ROSC were significantly higher than those in rats without ROSC (20.8±3.24 mmHg vs. 13.9±1.50 mmHg,P=0.000). Receiver operator characteristic (ROC) curves of PETCO2 showed significant sensitivity and specificity for predicting ROSC in VF versus asphyxial cardiac arrest. CONCLUSIONS:The initial values of PETCO2 immediately after CPR may be helpful in differentiating the causes of cardiac arrest. Changes of PETCO2 during CPR can predict outcomes of CPR.
6.Co-occlusion of foreign protein into polyhedra with BmNPV polyhedrin.
Xing-Wei XIANG ; Rui YANG ; Lin CHEN ; Xiao-Long HU ; Shao-Fang YU ; Xiao-Feng WU
Chinese Journal of Virology 2011;27(4):366-371
In order to make clear the packing mechanism of the BmNPV polyhedra, a polyhedrin gene negative recombinant baculovirus, vBmBac(polh-)-5B-EGFP, expressing EGFP was constructed, and used to infect BmN cells jointly with wild-type BmNPV. Fluorescent microscopic observation demonstrated that EGFP and polyhedrin were expressed simultaneously, and the EGFP expression and polyhedra formation occurred in most of the jointly infected cells. Analysis of the purified polyhedra from jointly infected BmN cells showed that the foreign proteins were present in the polyhedra. The results indicated that BmNPV polyhedrin could incorporate proteins other than viral proteins into the polyhedra. It implies that a nonspecific recognition mechanism exists in the embedment of BmNPV polyhedra.
Animals
;
Bombyx
;
Gene Expression
;
Green Fluorescent Proteins
;
genetics
;
metabolism
;
Nucleopolyhedrovirus
;
genetics
;
physiology
;
Viral Structural Proteins
;
genetics
;
metabolism
;
Virus Assembly
7.Determination of gentiopicroside in different morphological types of cultivated Gentiana manshurica population.
Chen WANG ; Xiu-fang XING ; Li-xiang MA ; Yang GUAN ; Shao-jun DAI ; Ming-yuan LIU
China Journal of Chinese Materia Medica 2004;29(9):841-844
OBJECTIVETo study the correlation between morphological variation and gentiopicroside content in cultivated Gentiana manshurica roots and to investigate the feasibility of breeding new varieties of high effective constituent content.
METHODGentiopicroside was determined in 5 morphological types of cultivated G. manshurica roots by HPLC, which are normal (or wild) type, white-flowered type, thick-rooted type, broad-leaved type I and broad-leaved type II.
RESULTAmong different types gentiopicroside content is the highest in the roots of thick-rooted type, the contents decrease as following order: normal type, broad-leaved type I white-flowered type and broad-leaved type II, and the gentiopicroside contents in the same type root system are a positive correlation with root ages, as 3-years-age roots > 2-years-age roots > 1-year-age varied with roots.
CONCLUSIONThe contents of effective constituent vary with the morphological variation in cultivated G. manshurica. It is feasible to breed a new variety with high effective constituent with the morphological character as a selecting index.
Genetic Variation ; Gentiana ; anatomy & histology ; chemistry ; classification ; Glucosides ; analysis ; Iridoid Glucosides ; Iridoids ; analysis ; Plant Roots ; anatomy & histology ; chemistry ; Plants, Medicinal ; anatomy & histology ; chemistry ; classification ; Pyrans ; analysis ; Species Specificity
8.Effect of curcumin on pulmonary hypertension and wall collagen of pulmonary arterioles of chronic hypoxic hypercapnic rats.
Quan LIN ; Liang-xing WANG ; Shao-xian CHEN ; Xiang-feng ZHOU ; Xiao-ying HUANG ; Xiao-fang FAN
Chinese Journal of Applied Physiology 2006;22(3):257-261
AIMTo study the effect of curcumin on pulmonary arterial pressure and type I collagen of pulmonary arterioles in pulmonary hypertensive rats induced by chronic hypoxia and hypercapnia.
METHODSThirty six rats were randomly divided into three groups: normal control group (NC), hypoxic hypercapnic group (HH) and hypoxic hypercapnia + curcumin group (HC). Collagen I in pulmonary arterioles was observed by the technique of immunohistochemistry.
RESULTS(1) The findings from hemodynamics showed that the mPAP in group HH was significantly higher than that in group NC and HC. Differences of mCAP among groups were not significant (P > 0.05). (2) Light microscopy showed the value of WA/TA (vessel wall area/total area), SMC (the density of medial smooth muscle cells) and thickness of pulmonary arterial media smooth cell layer(PAMT) were significantly higher in group HH than group NC (P < 0.01) and group HC (P < 0.01). (3) Electron microscopy showed that structure of the endothelial cells in pulmonary arterioles in group HC was near to normal, and the proliferation of medial smooth muscle cells and collagen fibers in adventitia was much lighter than those of group HH. (4) Expression of collagen I in pulmonary arterioles was significantly higher in group HH than group NC (P < 0.01) and group HC (P < 0.01).
CONCLUSIONCurcumin can decrease pulmonary arterial pressure, improve pulmonary vessel remodeling and inhibit the deposition of collagen I in pulmonary arterioles.
Animals ; Arterioles ; drug effects ; metabolism ; Collagen Type I ; metabolism ; Curcumin ; pharmacology ; Extracellular Matrix ; metabolism ; Hypercapnia ; metabolism ; physiopathology ; Hypertension, Pulmonary ; metabolism ; physiopathology ; Hypoxia ; metabolism ; physiopathology ; Male ; Pulmonary Artery ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley
9.Studies on the establishment of a method to detect HPV 6b E7-specific antibodies of serum and cervical secretion from patients with condyloma acuminatum and its epidemiological significance
Xiao-Zhi ZHONG ; Shao CHEN ; Shan-Li ZHU ; Xiang-Yang XUE ; Wen-Shu LI ; Le-Dan WANG ; Li-Fang ZHANG
Chinese Journal of Epidemiology 2010;31(2):199-203
Objective To establish a method for detection of the human papi 11 omavirus(HPV)6b E7-specific antibodies in serum and cervical secretion from patients with condyloma acuminatum(CA).Methods A full-length HPV 6b E7 gene was amplified by PCR from the CA tissue to construct the recombinant plasmid pET32a(+)/HPV 6b E7.The expression of prokaryotic protein was analyzed by SDS-PAGE and Western blot,then purified with Ni-NTA Agarose affinity column and used as an diagnostic antigen for establishing indirect ELISA method,to detect specific serum IgG and specific cervical secretion slgA from 56 CA patients,81 healthy control.Sera from 43 cervical cancer was served as control.HPV 6b DNA from 56 CA patients was identified by PCR.Results Data showed that the nucleotide homology of cloned sequence was 99.5%,compared to the standard sequences of HPV 6b E7 gene(GenBank accession number:NC001355).A high level expression of E7 fusion protein was obtained in prokaryotic expression system(40 μg/ml).Based on HPV 6b E7 fusion protein being used as coating antigen,results from ELISA showed that the absorbance rates(A)of serum IgG from CA,cervix cancer and healthy control groups were 1.82±0.48,1.36 ± 0.39 and 1.39 ± 0.27,respectively.The level of IgG antibody in the serum of CA group was significantly higher than that in cervix cancer group and healthy control(P<0.05).The A values of cervical secretion sIgA in CA and healthy control groups were 0.63 ± 0.26 and 0.53 ± 0.06,respectively,while the level of sIgA antibody in the cervical secretion of CA group was also significantly higher than that in healthy controls(P<0.05).The positive rate of HPV 6b E7 DNA in CA tissue was 78.6%(44/56)by PCR method.When compared the results measured by PCR,the HPV 6b E7-specific IgG and sIgA antibodies by ELISA used to detect the patients infected with HPV 6b infection,showed that the sensitivity rates were 68.2%(30/44)and 54.6%(24/44)respectively,and the specificity were all 100%(12/12).Conclusion Based on the serum and cervical secretion specific HPV 6b E7 antibodies from patients with CA to diagnose HPV 6b infection,results showed medium sensitivity and high specificity,and could further be used to investigate the epidemiological characteristics of HPV 6b infection.
10.Effect of Hypertension on Life Quality in Elderly
Li-Na MA ; Zhe TANG ; Shao-Chen GUAN ; Xiang-Hua FANG ; Xiao-Guang WU ; Li-Jun DIAO ; Hong-Jun LIU ; Fei SUN ;
Chinese Journal of Hypertension 2007;0(07):-
Objective To explore the effect of hypertension on the life quality in elderly.Methods A cohort of 1847 elderly in community in Beijing was recruited.Blood pressur,body structural and mental heath,cognition, activity and life satisfaction were evaluated by physical examination,CES-D scale,MMSE,and ADL,IADL scale.Results The life quality of the elderly hypertension was worse than those of normotensive;the morbidi- ties of chronic disease related to hypertension were higher and the cognitive ability worse with the duration of hy- pertension.The prevalence of depression was higher and the vitality was decreased as the rising of blood pres- sure.Conclusion Prevention and control hypertension improve life quality in elderly hypertension.