3.Establishment of a 15 loci multiplex amplification system and the genetic poly- morphism in Xinjiang Uygur population.
Juan GUI ; Hai-Bo LIU ; Qin-Xiang LIAO ; Xu XU ; Di LU ; Li YUAN
Journal of Forensic Medicine 2015;31(1):23-27
OBJECTIVE:
To develop a five fluorescence-labeled multiplex amplification system for 15 loci and study genetic polymorphism in Xinjiang Uygur population.
METHODS:
The STR loci were screened. The alleles were named according to the number of repeats by sequencing. The sensitivity, species specificity, identity and stability of the five fluorescence-labeled multiplex amplification system for the 15 loci were all tested. Then, the genetic polymorphism was analyzed in Xinjiang Uygur population and compared with other ethnic groups including Xizang Tibetan, Xiuyan Manchu, and Guangzhou Han population.
RESULTS:
The 15 loci multiplex amplification system was established. The sensitivity was 0.3 ng with good species specificity, identity and stability. The distributions of genotype for 13 STR loci in Uygur population were in accordance with Hardy-Weinberg equilibrium with no genetic linkage between these loci. Most loci showed statistically significant among different populations.
CONCLUSION
The established system has application value in forensic evidence. The 13 STR loci in Uygur population have
Alleles
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Ethnicity/genetics*
;
Gene Frequency
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Genetic Linkage
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Genotype
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Humans
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Multiplex Polymerase Chain Reaction/methods*
;
Polymorphism, Genetic
4.Preliminary processing, processing and usage of Dendrobii Caulis in history.
Wen-yu YANG ; Sheng TANG ; Dong-jun SHI ; Xiang-gui CHEN ; Ming-yuan LI ; Xian-fu TANG ; Chang-jiang YUAN
China Journal of Chinese Materia Medica 2015;40(14):2893-2897
On account of the dense cuticles of the fresh stem and the light, hard and pliable texture of the dried stem, Dendrobii Caulis is difficult to dry or pulverize. So, it is very important to the ancient doctors that Dendrobii Caulis should be properly treated and applied to keep or evoke its medicinal effects. The current textual research results about the preliminary processing, processing and usage methods of Dendrobii Caulis showed that: (1) In history the clinical use of fresh or processed Dendrobii Caulis as teas and tinctures were very common. (2) Its roots and rhizomes would be removed before using. (3) Some ancillary approaches were applied to shorten drying times, such as rinsing with boiling mulberry-ash soup, washing or soaking with liquor, mixing with rice pulp and then basking, etc. (4) According to the ancients knowledge, the sufficient pulverization, by means of slicing, rasping, hitting or pestling techniques, was necessary for Dendrobii Caulis to take its effects. (5) The heat processing methods for Dendrobii Caulis included stir-baking, stir-frying, steaming, decocting and stewing techniques, usually with liquor as an auxiliary material. Among above mentioned, steaming by pretreating with liquor was most commonly used, and this scheme was colorfully drawn in Bu Yi Lei Gong Pao Zhi Bian Lan (Ming Dynasty, 1591 CE) ; moreover, decocting in advance or long-time simmering so as to prepare paste products were recommended in the Qing Dynasty. (6) Some different processing programs involving stir-baking with grit, air-tightly baking with ondol (Kangs), fumigating with sulfur, which appeared in modern times and brought attractive outward appearance of the drug, went against ancients original intentions of ensuring drug efficacy.
Dendrobium
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History, Ancient
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Medicine, Chinese Traditional
;
history
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Technology, Pharmaceutical
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history
5.Effect of metallothionein on myocyte apoptosis and energy supply of isolated rabbit heart muscle during perfusion with ropivacaine.
Yao-min ZHU ; Zu-yi YUAN ; Xiang LIU ; Gui-xia JING
Journal of Southern Medical University 2011;31(8):1425-1427
OBJECTIVE[corrected] To assess the effects of metallothionein on myocyte apoptosis and energy supply of isolated rabbit heart muscle during perfusion with ropivacaine..
METHODSSixty New Zealand white male rabbits were randomized into 3 equal groups. In group I, the rabbits received a intreaperitioneal injection of distilled water 24 h before isolation of the heart with perfusion by Langendoff model; in group II, distilled water was injected intreaperitioneally, and 24 h later the heart was isolated and perfused with Langendoff model and ropivacaine; in group III, 3.6% ZnSO(4) was injected intreaperitioneally and the isolated heart was perfused with Langendoff model and ropivacaine. The myocardial metallothionein content, myocyte apoptosis, and myocardial ATP, ADP and AMP content were detected.
RESULTSThe myocardial metallothionein content was significantly higher in group III than in the other two groups; the percent of myocyte apoptosis was the highest in group II, and was significantly higher in group III than in group I. The myocardial content of ATP was the highest in group I, and was significantly higher in group III than in group II.
CONCLUSIONMetallothionein can significantly inhibit myocyte apoptosis and alleviate energy supply disorder induced by ropivacaine.
Amides ; pharmacology ; Animals ; Apoptosis ; drug effects ; Energy Metabolism ; drug effects ; In Vitro Techniques ; Male ; Metallothionein ; pharmacology ; Myocardium ; cytology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Perfusion ; Rabbits
6.Impact of 5-fluorouracil on glucose metabolism and pancreatic pathology in rats.
Jue-ping FENG ; Ji-gui CHEN ; Xiang-lin YUAN ; Ya-ping WANG ; Jing FANG ; Can LIU
Chinese Journal of Gastrointestinal Surgery 2010;13(12):935-938
OBJECTIVETo explore the impact of 5-fluorouracil (5-FU) on glucose metabolism and pancreatic pathology.
METHODSTwenty Wistar rats were divided into 5-FU group(n=10, chemotherapy was administered intraperitoneally to animals at a dose of 20 mg/kg daily for continuous 5 days) and control group (n=10, sodium chloride was administered intraperitoneally to animals with the same dose at the same time ). Glucose tolerance was evaluated 2 and 7 days following 5-FU treatment by serial measurement of blood glucose before and after an oral glucose load. Plasma insulin concentration was determined by radioimmunoassay. Pancreatic pathology was examined with morphological method and the ultrastructural changes of β cells were observed by transmission electron microscope.
RESULTSFasting blood glucose level was significantly higher in the 5-FU group than that in the control group [(7.6±0.9) mmol/L vs. (4.6±0.6) mmol/L at day 2; (8.9±1.0) mmol/L vs. (4.7±0.6) mmol/L at day 7, P<0.01]. Insulin releasing test indicated that the early phase insulin response to glucose load was significantly diminished in animals treated with 5-FU at day 2. Insulin level was significantly lower in the 5-FU group than that in the control group at 30 min (P<0.01). The peak secretion time of plasma insulin in 5-FU group was at 60 min, similar to the control group; and plasma insulin level decreased more slowly. Plasma insulin level was higher in 5-FU groups than in control groups on 120 min and 180 min. At day 7, Insulin level was lower in the 5-FU group than that in the control group on 60 min, and the peak secretion time of plasma insulin was delayed to 120 min. Plasma insulin level was significantly increased in 5-FU group than that in control group on 180 min(P<0.01). No gross histopathological damage to the pancreas was observed at day 2 and 7 following administration of 5-FU. The structural changes of mitochondria were mainly the quantities of secretory granule diminished at day 7 under transmission electron microscope. Dilated rough endoplasmic reticula, swollen mitochondria, and the presence of adipose drops in lysosomes were found in few cells.
CONCLUSIONS5-FU-induced hyperglycemia appears to be mediated in part by a relatively deficient insulin secretion to glucose stimulation. A relative deficiency in insulin secretion following 5-FU treatment appears to be related to β cells function impairs with islet cell ultrastructural changes induced by 5-FU.
Animals ; Blood Glucose ; drug effects ; metabolism ; Female ; Fluorouracil ; pharmacology ; Insulin ; blood ; Male ; Pancreas ; drug effects ; pathology ; Rats ; Rats, Wistar
7.Regulatory Mechanism of Electroacupuncture Therapy for Diaphragm Function of Chronic Obstructive Pulmonary Disease Model Rats with Muscular Dystrophy
Jian-Xiong CHEN ; Chang-Sheng LIU ; Ying HE ; Gui-Yuan LI ; Xiang-Ao KONG ; Juan TONG
Journal of Guangzhou University of Traditional Chinese Medicine 2018;35(2):265-271
Objective To investigate the effect of electroacupuncture on diaphragm function of chronic obstructive pulmonary disease (COPD) rats with muscular dystrophy, and to explore the regulatory mechanism. Methods Forty male rats were randomly divided into blank group, model group, electroacupuncture group, exercise group, electroacupuncture plus exercise group, 8 rats in each group. After successful establishment of COPD rat model with muscular dystrophy, the modeled rats in various intervention groups were given electroacupuncture and/or exercise treatment. After the last treatment, the changes of rat body mass were observed, the rat lung function was detected, and the mRNA expression levels of myosin heavy chains (MHC) of MHC-1, MHC-2 and diaphragmatic related signal proteins of Atrogin-1, muscle ring-finger protein-1(MuRF-1), MyoD were detected by real-time quantitative polymerase chain reaction (qPCR). Results (1) Compared with the blank group, inspiratory resistance (IR) and functional residual mass (FRC) in the model group were increased (P < 0.05) , and the dynamic lung compliance(Cydn) was decreased(P<0.05). Compared with the model group, IR and FRC in the intervention groups were decreased (P < 0.05), but the differences among the three intervention groups were insignificant(P>0.05). (2) Compared with the blank group, the mRNA expression levels of MHC-1, Atrogin-1, MuRF-1, MyoD in the model group were increased (P<0.05), and the mRNA expression level of MHC-2 was decreased (P < 0.05). Compared with the model group, the mRNA expression levels of MHC-1, Atrogin-1, MuRF-1, MyoD in the intervention groups were decreased (P < 0.05) , and the mRNA expression level of MHC-2 was increased(P<0.05). Compared with the exercise group, the mRNA expression levels of Atrogin-1, MuRF-1, MyoD in the electroacupuncture group were decreased (P<0.05), and the mRNA expression level of MHC-2 was increased (P<0.05) , but the above indexes in electroacupuncture plus exercise group showed no obvious changes(P>0.05). Conclusion Electroacupuncture can improve respiratory function of COPD rats with muscular dystrophy, and the possible mechanism is related with the increase of MHC-2 mRNA expression and with the decrease of Atrogin-1, MuRF-1, MyoD mRNA expression, which result into the regulation of ubiquitin proteasome pathway(UPP), reduction of myosin loss, and the relief of diaphragmatic atrophy.
8.Recombinant fibronectin polypeptide CH50 improves positive immune regulation in tumor microenvironment
Jin-Yi XIANG ; Gui-Mei ZHANG ; Hui GENG ; Ye YUAN ; Yi LIU ; Dong LI ; Han XIAO ; Feng-Hua WU ; Zuo-Hua FENG
Chinese Journal of Cancer Biotherapy 1995;0(02):-
Objective:To investigate the inhibitory effect of in vivo non-targeting transfection of recombinant fibronectin polypeptide CH50 against tumors and to study the related mechanisms.Methods:After inoculated with tumor cells, BALB/c mice were injected with CH50 plasmids,control plasmids,and normal saline separately.The growth of the tumor was observed;the expression of genes (such as B7-1,B7-H1 etc.) in tumor tissues was detected by RT-PCR;and the count of T lymphocytes in local tumor tissues was analyzed by flow cytometry.Results:The tumor growth was obviously suppressed by in vivo CH50 expression.The expression of genes (B7-1 and B7-H1) was up-regulated along with the growth of tumor.CH50 increased the ratios of B7-1/B7-H1 and B7-1/B7-DC and suppressed the up-regulation of IL-10 and TGF-?genes.The direct action of CH50 on H22 cells resulted in the down-regulatoin of TGF-?gene.The count of T lymphoeytes in tumor tissues of CH50 treatment group was significantly higher than that in other groups.Conclusion:Ex- pression of CH50 by non-targeting transfection can effectively inhibit the growth of tumor;the regulation of the immuno- regulatory genes in tumor mieroenvironment is an important part of the treatment mechanism of CH50.
9.Comparative analysis of variable region of white spot syndrome virus genome in Penaeus vannamei in Guangxi, China.
Gui-Xiang TONG ; Xiao-Zheng LI ; Xin-Xian WEI ; Xin-Yu YE ; Ming-Yuan WU ; Zhen-Fa QIN ; Liu-Chun LAN ; Jing-Jing ZHOU
Chinese Journal of Virology 2014;30(1):51-56
Comparative analysis of variable region ORF14/15 genes of white spot syndrome virus (WSSV) genome in Guangxi Penaeus vannamei (P. vannamei) could provide useful information for the evaluation of genetic diversity and genetic evolutionary relationship among WSSV isolates from Guangxi, China and other places. Based on geographical and temporal considerations, 40 WSSV-positive P. vannamei samples were collected during the period between May 2010 and July 2013 from Beihai, Qinzhou, and Fangchenggang, which were the main P. vannamei production areas in Guangxi, and the variable region ORF14/15 genes of the WSSV genome from all infected samples were amplified by PCR and then subjected to cloning and sequence analysis. Pairwise and multiple alignment analysis was then conducted to evaluate the degree of genetic divergence between different strains. The variable region ORF14/15 genes from 25 of 40 WSSV positive samples were successfully cloned and sequenced; among the ORF14/15 genes of 25 WSSV-positive strains, 22 was 619 bp in length and 3 was 620 bp. All the 25 Guangxi strains carried a 5949-bp deletion in the ORF14/15 region relative to TH-96-II, which has the longest nucleotide sequence in this region; the deletion of Guangxi strains occurred in the middle region of ORF14/15 gene, with only 190 bp and 429 bp/ 430 bp at 5' and 3' ends, respectively, which were coincident with WSSV-IN-05-I in deletion length and position. Sixteen of 25 Guangxi strains had completely identical nucleotide sequences in the variable re gion, and the homology between other strains also exceeded 97.9%. There were single nucleotide substi tution, deletion, and insertion in the ORF14/15 region of Guangxi strains compared with other strains in GenBank. In the phylogenetic tree based on WSSV variable region ORF14/15, the Guangxi strains were closely related and formed a separate branch with Indian strain IN-05-I, but far from other strains in GenBank. The ORF14/15 gene of WSSV isolates in cultured P. vannamei in Guangxi has a large deletion in the middle of the variable region, and the Guangxi WSSV strains show no significant spatio-temporal differences; the Guangxi strains are closer in genetics to Indian strain IN-05-I than other strains in GenBank.
Animals
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China
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Cloning, Molecular
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Evolution, Molecular
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Genome, Viral
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genetics
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Genomics
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Penaeidae
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virology
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Phylogeny
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White spot syndrome virus 1
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genetics
10.The study of hBMP-4 gene modified tissue-engineered bone.
Xin-quan JIANG ; Zhi-yuan ZHANG ; Jian-guo CHEN ; Gui-xiang MA ; Chuan-jun CHEN
Chinese Journal of Stomatology 2003;38(5):390-392
OBJECTIVEBone marrow stromal cells (bMSCs) of rabbits transferred with mammalian hBMP-4 expression plasmid were used to construct tissue-engineered bone. Gene therapy combined with tissue-engineering technique was explored to further improve osteogenesis.
METHODSpEGFP-hBMP-4 plasmid was constructed by subcloning technique. bMSCs were then transferred with either pEGFP-hBMP-4, pEGFP plasmid by lipofectamine or left uninfected in vitro. The cells from the 3 groups were combined with natural non-organic bone (NNB) to construct tissue-engineered bones, which were subcutaneously implanted into nude mice (6 implants per group) for 4 weeks. Specimens were evaluated through histological and computerized new bone formation analysis.
RESULTSpEGFP-hBMP-4 plasmid was successfully constructed. bMSCs could attach and proliferate on the surface on NNB. In vivo experiment showed that new bone formation in pEGFP-hBMP-4 group was higher than those of the control groups.
CONCLUSIONSTissue-engineered bone using hBMP-4 gene modified bMSCs might be an ideal alternative for the repair of bone.
Animals ; Bone Morphogenetic Protein 4 ; Bone Morphogenetic Proteins ; genetics ; Genetic Therapy ; Humans ; Mice ; Mice, Nude ; Osteogenesis ; Rabbits ; Tissue Engineering