1.Cloning and expression analysis of a key device of HMGR gene involved in ginsenoside biosynthesis of Panax ginseng via synthetic biology approach.
Hongmei LUO ; Jingyuan SONG ; Xueying LI ; Chao SUN ; Chunfang LI ; Xiang LUO ; Ying LI ; Shilin CHEN
Acta Pharmaceutica Sinica 2013;48(2):219-27
3-Hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR), the first enzyme of mavalonic acid pathway, is one of the key devices involved in ginsenoside biosynthesis based on synthetic biology approach. The open reading frame of a novel HMGR gene from Panax ginseng (PgHMGR2) was cloned and analyzed in this study. PgHMGR2-encoding protein showed 71.6% sequence similarity to a P. ginseng HMGR in GenBank. The full-length cDNA sequence of PgHMGR2 containing 1 770 bp, which encodes 589 amino acids, was cloned by RT-PCR strategy from P. ginseng. The bioinformatic analysis showed that PgHMGR2-encoding protein contained two transmembrane regions and the HMG_CoA_reductase domain, without signal peptide. The protein sequence of PgHMGR2 had the highest sequence similarity (99%) with Panax quinquefolius HMGR (GenBank accession No. ACV65036). The expression level of PgHMGR2 was the highest in flower based on a real-time PCR analysis, followed by leaf and root, and the lowest was in stem. The result will provide a foundation for exploring the molecular function of PgHMGR2 involved in ginsenoside biosynthesis based on synthetic biology approach in P. ginseng plants.
2.Treatment and prognostic factors of gastrointestinal stromal tumors
Sunyang FANG ; Xuejun YU ; Yimin DONG ; Xinchuang CHEN ; Henggui LUO ; Chunfang ZHU
Chinese Journal of General Surgery 1993;0(02):-
0.05).The infiltration of tumor to the muscularis mucosa or serosa layers was significantly correlated with the risk of GISTs(?~2=4.87,P
3.Genetic polymorphism of CRYAB gene rs3212227 and rs6894567 in Chinese guangxi populations
Junli WANG ; Huixiong YUAN ; Guijiang WEI ; Chunfang WANG ; Chunying LUO ; Yesheng WEI ; Zhaoquan HUANG
Chongqing Medicine 2013;(31):3744-3746,3749
Objective To study the frequencies of allele and genotype distribution of alpha-B-crystallin (CRYAB ) gene rs3212227 and rs6894567 single nucleotide polymorphism (SNP) in Chinese guangxi populations ,and to Compare the distribution differences among different ethnic .Methods The CRYAB gene rs3212227 and rs6894567 polymorphisms were detected by the pol-ymerase chain reaction-single base extension (PCR-SBE) technique and DNA sequencing methods in 199 Chinese guangxi popula-tions ,frequencies of allele and genotype of CRYAB gene SNP loci ,rs3212227、rs6894567 were analyzed in guangxi populations com-pared with other the four populations (HapMap-CEU ,HapMap-YRI ,HapMap-JPT and HapMap-HCB) from Human Genome Pro-ject group (Hapmap) data .Results There were CRYAB gene polymorphisms in Guangxi populations .The frequencies of allele and genotype distribution of CRYAB gene rs3212227、rs6894567 polymorphisms had significant difference compared with HapMap-CEU and HapMap-YRI populations (P<0 .05) ,and had no significant difference compared with HapMap-JPT and HapMap-HCB (P>0 .05) .Conclusion The frequencies of allele and genotype distribution of CRYAB gene rs3212227、rs6894567 polymorphisms are significantly difference compared with others ethnic populations ,and this variation might account for a variety of clinical mani-festation and morbidity of of some CRYAB related diseases .
4.Study of genetic polymorphisms of miR-146a,miR-149 gene in Guangxi popula-tion
Hongcheng LUO ; Chunfang WANG ; Ming LEI ; Ying WEI ; Tan TAN ; Yesheng WEI
Chinese Journal of Immunology 2016;32(9):1282-1285
Objective:To study the frequencies of allele and genotype distribution of miR-146a C>G(rs2910164) and miR-149 T>C( rs2292832) gene, and to analyze the statistical differences between different racial and nationalities.Methods:The Polymerase Chain Reaction-Single Base Extension ( PCR-SBE) technique and DNA sequencing methods were used for the determination of the SNP in miR-146a C>G and miR-149 T>C gene,and compared with the European, African, Japanese and People in Beijing from the Human Genome Project (HapMap).Results:There were no statistical differences of allele and genotype distribution in miR-146a C>G,miR-149 T>C between female and male group (P>0.05).There were significant difference frequencies of allele and genotype distribution of miR-146a C>G and miR-149 T>C gene by compared with the European, African and People in Beijing( P<0.05).Conclusion:There were gene Polymorphisms of miR-146a C>G and miR-149 T>C in Guangxi populations, and there were significant differences by compared with other ethnic populations, which may play an important role in the human inherited disease research.
5.Salvia miltiorrhiza as medicinal model plant.
Jingyuan SONG ; Hongmei LUO ; Chunfang LI ; Chao SUN ; Jiang XU ; Shilin CHEN
Acta Pharmaceutica Sinica 2013;48(7):1099-106
Research on medicinal model organism is one of the core technologies to promote the modernization of traditional Chinese medicine (TCM). The research progress of Salvia miltiorrhiza as medicinal model plant is summarized in this paper. The genome of S. miltiorrhiza is small and its life cycle is short, as well as this plant can be stably genetically transformed. Because S. miltiorrhiza possesses the important medicinal and economic values, recently the transcriptome and genome of S. miltiorrhiza have been significantly recovered. The research prospect of S. miltiorrhiza as medicinal model plant in TCM was discussed, including biosynthesis of active components and their genetic regulation, relationship between quality of TCM and ecological environments, and selective breeding of good quality lines. Furthermore, as medicinal model plant, the construction of mutant library for S. miltiorrhiza, the genome map with high quality, and the functional genome should be investigated. Accompanying modern investigation of life sciences, the platform for medicinal model plant, S. miltiorrhiza, will be promoted to be established. It is important to develop the ethnopharmacology and new drugs around the world.
6.Proliferation inhibition effect by silencing PLCε in the bladder cancer cell lineBIU-87
Yongcan GUO ; Chunli LUO ; Xiaozhong CAI ; Jianhong XIE ; Liping OU ; Yi ZHAO ; Chunfang Lü ; Huiying JI ; Xiaohou WU
Chinese Journal of Urology 2010;31(7):467-470
Objective To study the proliferation inhibition effect by silencing PLCε gene expression with RNA interference in BIU-87 cells. Methods The specific short hairpin RNA recombinant plasmids were constructed by gene clone technology.The expression level of PLCε protein and mRNA were detected by Western blot and RT-PCR respectively after transfected recombinant plasmids into BIU-87 cells.The influence on proliferation was check by MTT.The changes of proliferating cell nuclear antigen(PCNA)were analyzed by immunocytochemical method,and the distribution of cell cycle was analyzed using flow cytometry. Results After transfected with the specific recombinant plasmids,PCNA expression was decreased 33.08%,and the analysis of cell cycle indicated that cells of G0/G1 phase were increased comparision with(40.75±2.30)%and(40.00±1.76)0A,and its G2/M phase cells(8.16±0.51)%were decreased strikingly compared with group control(31.20±1.76)%and group NP(35.94±1.58)%.Cells were blocked at G0/G1 phase,the cell proliferation was inhibited obviously. Conclusion PLCε may play an important role in proliferation of bladder cancer cells,which could be a potential target of biological treatment on bladder cancer in the future.
7.Association of SNP of leukocyte differentiation antigen-CD40 gene and its serum level with ischemic stroke.
Jianming CHEN ; Huatuo HUANG ; Qiuran YUAN ; Hongcheng LUO ; Yang XIANG ; Chunfang WANG ; Lanqing MENG ; Yesheng WEI
Chinese Journal of Preventive Medicine 2016;50(3):239-243
OBJECTIVETo investigate the association of SNP of CD40 gene and its serum levels with ischemic stroke (IS).
METHODSA total of 202 IS patients from a hospital of Baise city were enrolled in case group from May 2013 to November 2014. At the same time, 109 healthy people who had physical check-ups in the outpatient department at the same hospital were enrolled in the control group. All participants were from Guangxi Zhuang Autonomous Region and unrelated to each other. 3 ml venous blood were collected on the premise of informed consent. The single nucleotide polymorphisms of CD40 gene rs1883832 C/T, rs13040307 C/T, rs752118 C/T and rs3765459 G/A were analyzed using a Snapshot SNP genotyping assays, and the serum levels of CD40 were tested by ELISA. t-test was used to compare the serum levels of CD40 between the case and control group, and the genotypes at different locuses in case group; χ(2) test was used to compare the distribution differences of the CD40 gene locuses in different genotypes and allele between the case group and the control group; alleles was established as independent variables, the occurrence of the IS as dependent variable, and expressed relative risk with OR (95%CI) value.
RESULTSIn the case group, the frequency of CC, CT and TT genotypes in CD40 gene rs1883832 C/T were 21.78% (44/202), 49.51% (100/202) and 28.71% (58/202), respectively, and 33.17% (66/199), 48.74% (97/199), 18.09% (36/199) in the control group, respectively, the differences between the two groups was significant (χ(2)=9.57, P=0.008). The CD40 serum levels were (62.7 ± 24.5) pg/ml in the case group, which was higher than that in the control group (45.3 ± 17.2) pg/ml (t=8.97, P<0.001). The serum levels of TT and CT genotypes in CD40 gene were (65.9 ± 26.3) and (64.3 ± 25.9) pg/ml, respectively, and the differences were significant when comparing with CC genotype (t equaled 5.34 and 5.03, respectively, P<0.001). The risk of developing IS was 1.56 times higher in 1883832 T allele carriers than that in rs1883832 C allele carriers (OR=1.56, 95% CI: 1.18-2.06); Combined genotype analysis displayed that CD40 gene rs1883832 C/T, rs13040307 C/T, rs752118 C/T and rs3765459 G/A polymorphisms showed strong linkage disequilibrium, the case group TCCA haplotype was tested to be associated with a significantly increased risk of IS as compared with that in the control group(OR=2.49; 95%CI: 1.13-5.48).
CONCLUSIONCD40 gene rs1883832 C/T polymorphism and its TCCA haplotype were possibly associated with ischemic stroke, and the susceptibility gene for ischemic stroke may be rs1883832 T allele.
Alleles ; CD40 Antigens ; blood ; genetics ; Case-Control Studies ; Cell Differentiation ; China ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Haplotypes ; Humans ; Polymorphism, Single Nucleotide ; Stroke ; blood ; genetics
8.Application study of rSO2-BIS monitoring in children with supracondylar fracture of humerus
Chunfang LUO ; Yinghui FENG ; Zhengbing ZHANG ; Kai XU ; Ming LI ; Weidong OUYANG
China Modern Doctor 2024;62(27):22-25
Objective To explore the application of regional cerebral oxygen saturation(rSO2)-bispectral index(BIS)monitoring anesthesia in children undergoing surgery for supracondylar fractures of the humerus.Methods A total of 60 children with supracondylar fractures of the humerus undergoing surgery in Department of Orthopedi,Jiangxi Provincial Children's Hospital were chosen and randomly number table method segmented into control group and observation group from January 2020 to December 2022,30 cases in each group.After anesthesia,children in observation group were carried out with rSO2-BIS monitoring,while children in control group were given routine anesthesia management.Results The dosage of propofol and long chain fat emulsion/remifentanil injection in observation group were significantly lower than those in control group(P<0.05),while there was no difference in heart rate,blood oxygen saturation and mean arterial pressure at various time points during awakening(P>0.05);The recovery rate of postoperative analgesics of observation group was lower than that of control group,and the extubation time of observation group was shorter than that of control group(P<0.05);The incidence of postoperative adverse reactions and behaviors in observation group were lower than those in control group(P<0.001).The dosage of propofol and long chain fat emulsion/remifentanil injection,as the independent risk factors inducing the occurrence of postoperative adverse reactions and adverse behaviors were proved by multivariate analysis.rSO2-BIS monitoring is a key factor in reducing postoperative adverse behaviors and reactions in pediatric patients(P<0.05).Conclusion The use of rSO2-BIS monitoring during surgery for supracondylar fractures of the humerus in children can reduce the incidence rate of postoperative adverse behaviors and adverse reactions,thereby improving postoperative efficacy.
9.Association of single nucleotide polymorphisms of rs3819024 and rs8193037 loci of IL-17A gene with the risk of ischemic stroke.
Huatuo HUANG ; Rong WANG ; Yulan LU ; Chunfang WANG ; Haimei QIN ; Yang XIANG ; Hongcheng LUO ; Junli WANG ; Lanqing MENG ; Yesheng WEI
Chinese Journal of Medical Genetics 2018;35(6):883-886
OBJECTIVE:
To assess the association of polymorphisms of rs3819024 and rs8193037 loci in the promoter region of IL-17A gene with the risk of ischemic stroke (IS) among ethnic Han Chinese from Guangxi.
METHODS:
The polymorphisms of rs3819024 and rs8193037 loci were detected by a SNaPshot assay and DNA sequencing among 392 IS patients and 443 healthy controls with matched age and gender.
RESULTS:
The genotypes, dominant model, recessive model, and alleles of rs3819024 polymorphisms showed no significant difference between the two groups, with the P values calculated as 0.150, 0.227, 0.125, 0.594 and 0.202, respectively, and OR (95% CI) as 1.27(0.92-1.74), 1.28(0.86-1.91), 1.27(0.94-1.72), 1.10(0.78-1.54), and 1.13(0.94-1.38), respectively. The genotypes, dominant model, recessive model, and alleles of rs8193037 polymorphisms also showed no significant difference between the two groups, with the P values calculated as 0.722, 0.352, 0.863, 0.345 and 0.969, respectively, and OR (95% CI) as 0.94(0.65-1.35), 2.25(0.41-12.35), 0.97(0.68-1.38), 2.27(0.41-12.48), and 1.01(0.72-1.40), respectively.
CONCLUSION
Polymorphisms of the rs3819024 and rs8193037 loci of the IL-17A gene are not associated with the susceptibility to IS among ethnic Han Chinese from Guangxi.
Alleles
;
Asian Continental Ancestry Group
;
Brain Ischemia
;
genetics
;
Case-Control Studies
;
China
;
Gene Frequency
;
Genetic Predisposition to Disease
;
Genotype
;
Humans
;
Interleukin-17
;
genetics
;
Polymorphism, Single Nucleotide
;
Stroke
;
genetics
10.Application of BACs-on-Beads in prenatal diagnosis.
Yan ZENG ; Ting WANG ; Tao ZHANG ; Jiaming FAN ; Lifang ZHANG ; Feiyan QIAN ; Tingting LUO ; Weiping CHEN ; Ming CHE ; Chunfang QIAN
Chinese Journal of Medical Genetics 2020;37(8):891-894
OBJECTIVE:
To explore the value of BACs-on-Beads (BoBs) for the practice of prenatal diagnosis.
METHODS:
The results of chromosomal karyotyping and BoBs of 1773 prenatal samples were compared. Microdeletions and microduplications detected by BoBs were subjected to chromosome microarray analysis (CMA) with informed consent from patients.
RESULTS:
BoBs has detected 46 cases of common aneuploidies involving chromosomes 13, 18, and 21, and 16 cases involving X and Y chromosomes. For 4 fetuses with normal results by BoBs, karyotyping analysis of amniotic fluid sample suggested low percentage mosaicisms (< 20%). BoBs has detected none of the 9 common microdeletions, but 14 male fetuses with Xp22 microdeletions and 5 with other microdeletions/microduplications. In 10 cases, the couples had chosen CMA verification, and the results were all consistent.
CONCLUSION
As a rapid diagnostic technique, BoBs has a high accuracy for common aneuploidies, and is capable of discovering certain chromosome microdeletions and microduplications. The difficulty lies in the inability to detect low proportion mosaicisms and the consultation following detection for male fetuses carrying Xp22 microdeletions.