1.Molecular cloning and sequencing of apoA1 gene
Rong FANG ; Baikun MA ; Yufeng HUANG ;
Journal of Medical Postgraduates 2003;0(08):-
Objectives:Identification, clone and sequence of the chinese mature apoA1 peptide gene.Methods:Total RNA was prepared from Chinese fetal liver tissue, cDNA fragment encoding human apoA1 was amplified by RT PCR using specific primers, and cloned into pGEM T vector. Inserted apoA1 gene was sequenced by ABI 377 DNA Sequencer. Results:Analysis indicates that the DNA fragment is 739 base pairs in length and has 100% nucleotide homology with that reported previously. Conclusions:Human apoA1 gene was successfully cloned from fetal liver tissue.
2.The study of mannose-binding lectin gene polymorphism and serum concentration in patients with systemic lupus erythematosus
Aili WANG ; Qian CHENG ; Baikun MA ; Rong FANG
Chinese Journal of Rheumatology 2000;0(06):-
Objective To investigate the relationship between polymorphism of codon54of the man-nose-binding lectin(MBL)gene and serum concentration in patients with systemic lupus erythematosus(SLE).Methods The MBL gene54(GGC/GAC)polymorphism was analyzed by polymerase chain reaction and restrict fragment length polymorphism(PCR-RFLP)in44patients with SLE and45healthy controls.Serum levels of MBL were also determined by ELISA.Results The frequencies of the wild homozygons(GGC/GGC)and het-erozygotse(GGC/GAC)of MBL codon54did not differ significantly between patients(GGC/GGC71%,GGC/GAC27%,GAC/GAC2%)and healthy controls(GGC/GGC78%,GGC/GAC22%,GAC/GAC2%).The serum MBL level of the three genotypes was(1.8?0.6)mg/L(n=31),(1.5?0.6)mg/L(n=12),1.17mg/L(n=1)respective-ly in SLE group;(3.0?1.0)mg/L(n=35),(2.2?1.0)mg/L(n=10)in control group.The(GGC/GGC and GGC/GAC)genotypes in SLE group were significantly lower than the control group.Conclusion The condon54mutation of the MBL gene may be not associated with low serum level of MBL in patients with SLE.
3.Preliminary study on the treatment of ischemic heart disease with angiopoietin 1 gene transfection by adenovirus vector in rabbits
Shilin CHEN ; Hua JING ; Jieshou LI ; Long YI ; Baikun MA ; Rong FANG
Chinese Journal of Geriatrics 2000;0(06):-
Objective To study the potential effects of angiopoietin 1(Ang1) via adenovirus mediated gene transfer on ischemic heart disease in rabbits. Methods Forty-five male New Zealand white rabbits underwent high positioned double-ligation of the anterior descending left coronary artery, and were divided into three groups: Ang1 group (n=15) received direct myocardium injection of Ang1 recombinant adenovirus; DMEM group (n=15) and LacZ group (n=15) received only DMEM and LacZ recombinant adenovirus as controls respectively. The myocardial infarcted size was evaluated by N-BT macroscopic standing and the degree of angiogenesis was assessed by use of immunohistochemical analysis. The echocardiographic changes were measured on before operation and the 7 th, 14 th, and 28 th postoperative days. Results Animals in three groups had no significant difference in the percentage of infarction size of left ventricle at the 7 th, 14 th day and capillary density at the 7 th day. Animals in Ang1 group showed less infarcted size than DMEM group or LacZ group at the 28 th day and higher capillary density at the 14 th and the 28 th day. The results from echocardiographic measurements showed that animals in all three groups had no significant difference in the left ventricular systolic function before operation and at the 7 th , 14 th day, but the left ventricular systolic function in Ang1 group was better than that in DMEM group or LacZ group at the the 28 th day(P
4.The analysis of the blood fatty in different genotypes of matrix metalloproteinases in the carotid atherosclerosis patients with essential hypertension
Xinjuan XU ; Yanping MA ; Rong REN ; Fang ZHAO ; Junshi ZHANG ; Guangmei HU
Chinese Journal of Postgraduates of Medicine 2014;37(10):46-49
Objective To investigate the blood fatty levels in different genotypes of matrix metalloproteinases (MMP) and the carotid atherosclerosis (CAS) in patients with essential hypertension (EH).Methods Four hundred and eighty-eight cases of Han and Uygur in patients with EH were selected,and MMP-2-735 C/T and MMP-3-1171 5A/6A polymorphism was measured by polymerase chain reactionrestriction fragment length polymorphism.They were divided into two groups:EH with CAS group (CAS group,293 cases,Han 171 cases,Uygur 122 cases) and EH without CAS group (NS group,195 cases,Han 105 cases,Uygur 90 cases).Blood fatty factors were analyzed according to the different MMP genotypes.Results According to MMP-2 genotypes,in Hart CAS group,the low density lipoprotein cholesterol (LDL-C) levels in MMP-2 CT+TT genotypes were significantly higher than those in CC genotype(P < 0.05),and high density lipoprotein cholesterol levels were significantly lower than those in CC genotype(P < 0.05); In Uygur CAS group,the triglyceride levels in MMP-2 CT+TT genotypes were significantly higher than those in CC genotype(P < 0.05).According to MMP-3 genotypes in Han NS group,the total cholesterol and LDL-C levels in 6A/6A genotype were significantly higher than those in 5A/5A+5A/6A genotypes (P < 0.05).Conclusion The blood fatty levels are different in patients with different MMP genotypes,which may probably influence the CAS.
5.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.
6.SCREENING OF MONASCUS PRODUCING ONLY YE LLOW PIGMENTS
Mei-Rong MA ; Hui-Ying FANG ; Zheng-Xiang WANG ; Yan LI ; Ge-Jian ZHU ;
Microbiology 1992;0(04):-
mutants which didnt produce red pigmen ts on malt extract agar plate were obtained.The 8 stable mutants were cultured on solid medium.Two samples wer e yellow,the others were white.The extracted samples were scanned in visible len gth.2 yellow samples showed only one absorptive peak at 370nm,the 6 white sample s showed no absorptive peak.The mutants producing only yellow pigments on solid medium were tested in liquid culture.The results indicated their ability to pro duce only yellow pigments were stable.
7.Expression of Fascin-1 and β-catenin protein in colorectal adenocarcinoma and its relevance with K-ras gene mutations
Gui MA ; Lan WANG ; Shu-Ping MA ; Fang BIAN ; Yan-Ni REN ; Qing-Rong HU ; Rong YANG
Chinese Journal of Clinical and Experimental Pathology 2019;35(1):27-32
Purpose To examine the expression of Fascin-1 and β-catenin protein and K-ras gene mutation in colorectal adenocarcinoma, and to explore their role in progression of colorectal neoplasm and their relevance. Methods Fascin-1 and β-catenin were analyzed by use of immunohistochemistry En Vision two-step. K-ras gene mutation was detected by ARMS method.Relationship between overexpression of Fascin-1, the nuclear expression of β-catenin, and the mutations of K-ras gene and clinicopathologic parameters was analyzed, the correlation between them was also analyzed. Results In 112 colorectal adenocarcinoma samples, the overexpression rate of Fascin-1 protein was 27.7% (31/112), significantly higher than non-neoplastic mucosa (P < 0.01). The high nuclear expression rate of β-catenin was 29.5% (33/112) in adenocarcinoma and non-neoplastic mucosa respectively with a significant difference between two groups (P < 0.01). High expression rate of Fascin-1 protein and β-catenin were correlated significantly with lymph node metastasis (P = 0.022, P = 0.027), and TNM staging (P =0.042, P = 0.019) in colorectal adenocarcinoma. The overexpression of Fascin-1 protein was correlated with tumor location (P = 0.004). The mutation rate of K-ras gene was 34.8% (39/112), which showed no correlation with age, gender, tumor size, grade of differentiation, lymph node metastasis and TNM staging (P> 0.05). There was a correlation between the overexpressison of Fascin-1 protein, the nuclear expression of β-catenin and the mutation of K-ras gene (rs= 0.252, rs= 0.258, P < 0.05). The overexpression of Fascin-1 protein positively correlated with the nuclear expression of β-catenin (rs= 0.213, P < 0.05). Conclusion Fascin-1 protein and β-catenin protein are involved in invasion and metastasis of colorectal cancer and are associated with K-ras gene mutation. K-ras may promote the overexpression of Fascin-1 by virtue of nuclear expression ofβ-catenin, which provided a new research direction on the treatment of K-ras gene mutated colorectal adenocarcinoma.
8.Design method of constitution regulating and healthcare foods based on medicinal property combination mode.
Fang DONG ; Su-rong YAN ; Li MA ; Wen-qiang ZU ; Li DU ; Yan-ling ZHANG ; Yun WANG
China Journal of Chinese Materia Medica 2015;40(18):3660-3663
Currently, the herbal prescription therapy for corresponding constitutional diseases is a common constitution regulating method. This method has an obvious effect in treating and regulating constitution-related diseases. However, for people who do not have disease, they prefer to regulate constitution with dietary therapy. In this paper, the researchers came up with a design method of constitution regulating and healthcare foods based on medicinal property combination mode of clinical empirical formulas. With "Yupinfeng San", a common formula for Qi-insufficiency constitution and specific endowment constitution, as the example for constitution regulating and healthcare foods, the researchers proved the effectiveness and rationality of healthcare food schemes in terms of the efficacy of single herb and the modern pharmacological study.
Diet
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Diet Therapy
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Humans
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Plants, Medicinal
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chemistry
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metabolism
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Prescriptions
9.Investigation on association of activating transcription factor 6 Ala145Pro variant with glucose and lipid metabolism in Chinese
Cheng HU ; Wei-Ping JIA ; Hui WAN ; Rong ZHANG ; Congrong WANG ; Xiaojing MA ; Qichen FANG ; Kunsan XIANG ;
Chinese Journal of Endocrinology and Metabolism 1986;0(04):-
Objective To investigate the association of activating transcription factor 6(ATF6)gene Ala145Pro(GCG→CCG)variant with glucose and lipid metabolism in Chinese.Methods The genotypes were determined by PCR-RFLP in 689 Chinese in Shanghai.Among them,361 subjects showed normal glucose regulation,250 cases were newly-diagnosed diabetic patients without taking any drug and 78 cases were probands of early-onset type 2 diabetes pedigrees.The following phenotypes were measured:body height and weight to calculate BMI;waist,hip and femoral circumference to calculate waist-to-hip ratio and waist-to-femoral ratio;blood pressure;plasma glucose levels obtained at 0 and 120 minute during 75 g oral glucose tolerance test;fasting serum lipid profile including total cholesterol,triglyceride,high density lipoprotein-cholesterol and low density lipoprotein-cholesterol;body fat percentage and distribution.Results(1)The frequency of C allele is significantly lower in probands from early-onset type 2 diabetes patients compared with subjects with normal glucose regulation(P=0.035).(2)In subjects with normal glucose regulation,the CC+CG genotype had a significantly lower level of high density lipoprotein cholesterol as compared with GG genotype(P=0.014).(3)In type 2 diabetic patients,the CC+CG genotype had a significantly higher level of low density lipoprotein-cholesterol as compared with GG genotype(P=0.041).Conclusion These findings suggest that variant of ATF6 plays a role in glucose and lipid metabolism in Chinese.
10.Risk factors of cerebral microbleeds in patients with ischemic stroke
Yangkun CHEN ; Yonglin LIU ; Zhuoxin NI ; Weimin XIAO ; Genpei LUO ; Runxiong LI ; Jianfeng QU ; Rong MA ; Xuewen FANG
Chinese Journal of Nervous and Mental Diseases 2016;42(4):234-239
Objective To investigate the clinical, neuroimaging and serum risk factors of cerebral microbleeds (CMBs) in patients with ischemic stroke and find the associations between these risk factors and the location and num?bers of CMBs were also analyzed. Methods One hundred and fifty-three patients with acute ischemic stroke were re?cruited in this study and their data werewas retrospectively analyzed. All of the patients underwent MRI- susceptibility weighted imaging (SWI). The location and numbers of CMBs were recordedexamined. The severity of WMLs was assessed using the Fazekas scale. Logistic regressions were performed to find the predictors of the presence of CMBs. The relation?ships between these risk factors and the location and numbers of CMBs were also analyzed. Results Fifty-nine(38.6%) cases had at least one CMB. The frequency of cortical-subcortical, deep and infratentorial CMBs were 34.0%, 24.8%and 27.5%, respectively. Multivariate logistic regression showed that male sex, hypertension and moderate-to-severe deep white matter hyperintensities (DWMH) were independent risk factors of the presence of CMBs. Adjusted with age and sex, partial correlation showed that hypertension only correlated with the numbers of deep CMBs significantly (r=0.174, P=0.032). Moderate-to-severe DWMH significantly correlated with the numbers of cortical-subcortical and deep CMBs (r=0.285, P<0.001 and r=0.258, P=0.001, respectively). Conclusion Male sex, hypertension and moderate-to-severe DWMH were are independent risk factors of CMBs in patients with ischemic stroke. Hypertension correlates with Deep deep CMBs, while Moderatemoderate-to-severe DWMH correlates with cortical-subcortical and deep CMBs.