1.Correlation between bone marrow stromal stem cells and apoptosis in epilepsy
Hao WANG ; Xiangyang REN ; Congmin MA ; Chao HUANG ; Haitao ZHOU
Chinese Journal of Tissue Engineering Research 2016;20(28):4117-4122
BACKGROUND:There is a close relationship between epilepsy and apoptosis. The appearance of epilepsy can lead to the loss of neurons in the hippocampus, triggering a series of programmed cel death. OBJECTIVE:To investigate the effect of bone marrow stromal stem cel transplantation on apoptosis in epilepsy. METHODS:After modeled to be of epilepsy 45, Sprague-Dawley model rats were randomly divided into three groups, fol owed by given no intervention (moldel group), normal saline (normal saline group) or bone marrow stromal stem cel transplantation (transplantation group). At 1, 2 and 4 weeks after modeling, the number of Bax-positive cel s, Bcl-2-positive cel s and Bax/Bcl-2 were detected by immunohistochemistry. RESULTS AND CONCLUSION:The number of Bax-positive cel s, Bcl-2-positive cel s and Bax/Bcl-2 presented no obvious changes in the normal saline group at different time points. However, the number of Bax-positive cel s and Bax/Bcl-2 in the transplantation group was significantly decreased, while the number of Bcl-2-positive cel s significantly increased compared with the other two groups at 1, 2 and 4 weeks after modeling (P<0.05). Moreover, the above indicators varied significantly in the transplantation group at different time points after modeling (P<0.05). These results show that bone marrow stromal stem cel transplantation can affect the apoptosis and effectively reduce the apoptosis in rats with epilepsy by up-regulating the number of Bax-positive cel s and down-regulating the number of Bcl-2-positive cel s.
2.Isolation and Identification of Promoter Sequence of Interferon Regulatory Factor-3 and Detection of Its Promotor Activity in Human Embryonic Kidney-293 Cells
wei, REN ; hua-gao, XU ; chao, LU ; guo-ping, ZHOU
Journal of Applied Clinical Pediatrics 2006;0(17):-
Objective To construct a luciferase reporter plasmid containing interferon regulatory factor 3(IRF-3)human gene promoter and to evaluate promoter activity in human embryonic kidney(HEK)-293 cells.Methods The 1 000 bp fragment was amplified by PCR with human genomic DNA as a template and was directionally cloned into pGL3-basic multiple cloning sites to construct the luciferase repor-ter plasmid pGL3-pIRF-3.Transfection of HEK-293 cells with the promoter-driven lucife-rase construct was performed to induce lucife-rase gene expression and calculate the relative luciferase activity unit(RLU).Promoter sequence of 1 000 bp upstream of transcription initiation site of IRF-3 was analyzed by using Promoter 2.0 Prediction software.Results DNA sequencing and restriction endonuclease analysis verified the successful construction of the plasmid pGL3-pIRF-3.This IRF-3 promoter exhibited a strong promoter activity with an increase of 42.2-fold of RLU in HEK-293 cells when compared with pGL-3 basic vector.The transfection experiment confirmed that the levels of its activation were significantly higher than that in controls in HEK-293 cells.Function analysis of IRF-3 promoter disclosed seve-ral GATA-1 and specific protein 1(Sp1) sites and E2F in minimal promoter region.Conclusion The plasmid pGL3-pIRF-3 promoter is successfully constructed and has a strong basal promoter activity in HEK-293 cells.
3.Interleukin-1 receptor type 1 signaling induces excessive inflammatory responses in H1N1 influenza ;virus infection
Rongrong REN ; Xiaonan REN ; Boyin QIN ; Mengjiao YUAN ; Hua YANG ; Chao WANG ; Shun LI ; Xiaohui ZHOU
Chinese Journal of Microbiology and Immunology 2016;36(12):887-893
Objective To investigate the role of interleukin-1 receptor type 1 (IL-1R1) signaling in H1N1 influenza virus infection. Methods IL-1R1 knockout ( IL-1R1-/-) mice and wild type ( WT) mice were infected intranasally with 2×104 TCID50(50% tissue culture infective dose) of influenza virus H1N1 PR8. Changes in clinical signs, survivals and bodyweights of those mice were monitored daily for 14 consecutive days. Three mice from each group were sacrificed at 3, 7 and 14 days post infection (d. p. i), from which whole lungs were harvested. A part of the lobes was fixed in 4% paraformaldehyde for histopatho-logical assessment and the rest were split and stored at-80 centigrade for further analysis. Real-time quanti-tative PCR and cytometric bead array ( CBA) were performed to detect viral loads in lungs and inflammatory cytokines in supernatants of lung homogenates. Results The mice in both groups showed severe symptoms after the infection of PR8. The maximum bodyweight loss of IL-1R1-/- mice [(24. 22±0. 80) % at 8 d. p. i] was lower than that of WT mice [(28. 03±1. 51)% at 9 d. p. i] (P<0. 05). The IL-1R1-/- mice with PR8 infection showed a higher survival rate (90%) as compared with that of the control group (40%) (P<0. 05). No statistical differences in virus loads were observed between the two groups at 3, 7 and 14 d. p. i. The lung weight to body weight ratio of IL-1R1-/-mice [(1. 42±0. 03) %] was lower than that of WT mice [(1. 79±0. 08) %] at 3 d. p. i (P<0. 05). Pathological changes in IL-1R1-/- mice were less severe than those in WT mice. CBA detection assay revealed that the proinflammatory cytokines in lungs of IL-1R1-/-mice were less than those in WT mice. Conclusion IL-1R1 signaling plays a pathogenic role in mice infec-ted with 2×104 TCID50 of influenza virus PR8 by promoting inflammatory responses.
4.Diversity and tissue distribution of fungal endophytes in Alpinia officinarum: an important south-China medicinal plant.
Ren-Chao ZHOU ; Juan HUANG ; Ze-En LI ; Shu-Bin LI
China Journal of Chinese Materia Medica 2014;39(16):3023-3029
In the present study, terminal-restriction fragment length polymorphism (T-RFLP) technique was applied to assess the diversity and tissue distribution of the fungal endophyte communities of Alpinia officinarum collected from Longtang town in Xuwen county, Guangdong province, China, at which the pharmacological effect of the medicine plant is traditional considered to be the significantly higher than that in any other growth areas in China. A total of 28 distinct Terminal-Restriction Fragment (T-RFs) were detected with HhaI Mono-digestion targeted amplified fungal nuclear ribosomal internal transcribed spacer region sequences (rDNA ITS) from the root, rhizome, stem, and leaf internal tissues of A. officinarum plant, indicating that at least 28 distinct fungal species were able to colonize the internal tissue of the host plant. The rDNA ITS-T-RFLP profiles obtained from different tissues of the host plant were obvious distinct. And the numbers of total T-RFs, and the dominant T-RFs detected from various tissues were significantly different. Based on the obtained T-RFLP profiles, Shannon's diversity index and the Shannon's evenness index were calculated, which were significantly different among tissues (P < 0.05). Furthermore, two types of active chemicals, total volatile oils by water vapor distillation method and galangin by methanol extraction-HPLC method, were examined in the each tissue of the tested plant. Both of tested components were detected in all of the four tissues of the medicine plant with varying contents. And the highest was in rhizome tissue. Correlation analysis revealed there were significant negative correlations between both of the tested active components contents and calculated Shannon's diversity index, as well as the Shannon's evenness index of the fungal endophyte communities of the host plant (P = 0, Pearson correlation coefficient ≤ -0.962), and significant positive correlations between both of the tested active components contents and 325 bp dominant T-RF linkage to Pestalotiopsis (P = 0, Pearson correlation coefficient ≥ 0.975). In conclusion, A. officinarum is colonized by diverse fungal endophytes communities. The diversity of the fungal endophytes was found in the A. officinarum varied with differences of the tissue types of the host plants and was closely correlated with the accumulation of main active components, total volatile oils and galangin contents in the host plant tissue.
Alpinia
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chemistry
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microbiology
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Biodiversity
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China
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DNA, Fungal
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genetics
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DNA, Ribosomal
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genetics
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Drugs, Chinese Herbal
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analysis
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Endophytes
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classification
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genetics
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growth & development
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isolation & purification
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Fungi
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classification
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genetics
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growth & development
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isolation & purification
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Phylogeny
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Plants, Medicinal
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chemistry
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microbiology
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Polymorphism, Restriction Fragment Length
5.Functional analysis of conserved sequences in the area of the promoter of CD2 associated protein
Xinming SU ; Guoping ZHOU ; Wei REN ; Chao LU ; Jiqing CHEN ; Shenghua WU ; Yuanjun WU
Chinese Journal of Microbiology and Immunology 2008;28(9):805-809
Objective To identify the important regulatory elements in the promoter of human CD2 associated protein(CD2AP) by conserved sequence analysis among different species and luciferase functional detection. Methods The promoter sequences of CD2AP from different species were analyzed by BLAST. Plasmids containing different length of deletion mutations of human CD2AP promoter were constructed. Pro-moter activities were tested in 3 kinds of cells from different species by luciferase analysis and were tested in HEK-293 cells treated with all-trans-retinoic acid. Results Homologous sequence comparison in CD2AP promoter area among human, cattle and pig showed that putative specific protein 1 (Sp1) sites and down-stream promoter element (DPE) were highly evolutional]y conserved. Progressive deletion luciferase analysis of DNA fragments revealed similar promoter activity style among 3 different cell lines from 3 different spe-cies, HEK-293, BHK-21 and Vero cells. One basic promoter activity located within 500 bp upstream of ATG. Fragments of further upstream 100 bp or more had drastically 10 times increased promoter activity. Two putative Sp1 sites were in this 100 bp region. All-trans-retinoic acid decreased the luciferase activity of CD2AP promoter. Conclusion Putative Spl sites and DPE have important functions in the promoter activity of CD2AP.
6.Updates on Budd-Chiari syndrome associated with hepatocelluar carcinoma
Chao LIU ; Qinghui ZHANG ; Gang WU ; Pengli ZHOU ; Xinwei HAN ; Jianzhuang REN ; Miao XU
Chinese Journal of Hepatobiliary Surgery 2015;21(11):786-789
The onset of hepatocelluar carcinoma, one of the serious complications of primary Budd-Chiari syndrome, is associated with poor prognosis.Although so, the diagnosis and treatment of such disease has still not been standardized at recent.In this paper, we overviewed the recent advances on Budd-Chiari syndrome associated with hepatocelluar carcinoma.
7.Study on the thickness-change of different thickness thermoplastic materials after thermoforming and saliva immersion.
Ning ZHANG ; Yu-xing BAI ; Kun-ya ZHANG ; Chao-chao REN ; Jie-min ZHOU ; Peng QI
West China Journal of Stomatology 2010;28(1):99-101
OBJECTIVETo survey and compare the thickness-change of different thickness thermoplastic materials under different test condition and make sure the relationship between the thickness-change and the material initial thickness in order to provide a guide in selecting the suitable thickness thermoplastic in practice.
METHODSTo choose Biolon, the thickness include 1.0 mm, 0.75 mm, 0.5 mm. Used Electron Vernier caliper to measure the thickness-change of different thickness thermoplastic materials under different processing mode. The data was analyzed by SPSS 10.0.
RESULTSAfter thermoforming the thickness of thermoplastic became thinner, the thickness of Biolon 0.75 mm decreased by 0.14 mm, Biolon 1.0 mm decreased by 0.22 mm and Biolon 0.5 mm decreased by 0.14 mm. After saliva immersion the thickness became thicker. The thickness of Biolon 0.75 mm increased by 0.02 mm, Biolon 1.0 mm increased by 0.03 mm and Biolon 0.5 mm increased by 0.02 mm.
CONCLUSION1)The influence of different processing mode to the thickness-change had relation to the material initial thickness. 2)The Biolon 0.75 mm had certain superiority in thickness stability compared to the homogeneous brand through the above research.
Dental Materials ; Humans ; Materials Testing ; Saliva
8.Expression and its clinical significance of gastric and intestinal phenotypic markers in 47 Siewert type Ⅱ and type Ⅲ early gastroesophageal junctional cancers
Yuhan JIANG ; Zhiguo ZHAO ; Zhibo CAO ; Yingfa ZHOU ; Chao LIU ; Jingli REN ; Guiming HU ; Yanling HUANG ; Yanan WANG
Chinese Journal of Digestion 2017;37(3):183-189
Objective To investigate the expression of gastric and intestinal phenotypic markers in Siewert typeⅡand Ⅲ early gastroesophageal junction(GEJ) cancer, and to explore its correlation with clinic-pathological features.Methods From April 2010 to July 2015, 53 cases diagnosed as early GEJ cancer were enrolled.The gastric and intestinal phenotypic markers such as mucin5AC(MUC5AC),mucin6(MUC6),mucin2(MUC2),caudal related homeodomain transcription 2(CDX2) and cluster of differentiation 10(CD10) were detected, and then the patients were divided into gastric type, gastrointestinal type, intestinal type and non-classified type according to the results of immunohistochemical staining.Combined with Siewert classification the clinicopathological features were analyzed.Chi square test or Fisher′s exact test was performed for statistical analysis.Results In the cancer tissues of 47 patients with Siewert type Ⅱand Ⅲ early GEJ cancer, the case numbers of positive expression of MUC5AC,MUC6,MUC2, CDX2 and CD10 were 21(44.7%),19(40.4%),31(66.0%),27(57.4%) and 17(36.2%),respectively;the case numbers of gastric type, gastrointestinal type, intestinal type and non-classified type were 11(23.4%),14(29.8%),21(44.7%) and one(2.1%), respectively.The positive expression rates of MUC5AC and MUC6 in Siewert typeⅡwere 55.9%(19/34) and 50.0%(17/34),which were higher than those of Siewert typeⅢ(2/13), and the positive expression rate of MUC2 was 55.9%(19/34), which was lower than that of Siewert typeⅢ(12/13), and the differences were statistically significant (x2=6.240,4.679 and 4.053;all P<0.05).In Siewert typeⅡ, the proportion of intestinal type was 32.4%(11/34), which was lower than that of Siewert typeⅢ(10/13), and the differences were statistically significant (x2=7.142,P=0.010).In patients with Siewert typeⅡand Ⅲ early cancer, males predominated in intestinal type which were mostly well differentiated type with less submucosal carcinoma.The maximum diameter of tumor was less than those of gastric type and gastrointestinal type.In paracancerous mucosal tissues, the incidences of intestinal metaplasia in gastrointestinal type and intestinal type were 11/14 and 81.0%(17/21), which were higher than that of gastric type (3/11);the incidences of atrophy in gastrointestinal type and intestinal type were 12/14 and 85.7%(18/21),which were higher than that of gastric type (4/11),and the differences were statistically significant (Fisher′s exact test,all P<0.05).Conclusions Siewert typeⅡand Ⅲ early GEJ cancer can directly originated not only from gastric mucosa, but also from gastrointestinal and intestinal mucosa.Atrophy and intestinal metaplasia could exist before cancer genesis.
9.Prevalence of trachoma trichiasis and corneal opacity in the Shaanxi Province of China
Chen-Jing, ZHOU ; Yu-Hong, CHENG ; Qiang, MA ; Jun, JIA ; Yuan, HE ; Li-Ling, ZHANG ; Bai-Chao, REN
International Eye Science 2016;16(8):1534-1536
Abstract?AIM: To assess the current situation of trachoma in Shaanxi Province and analyze its epidemiology and clinical features.?METHODS: The World Health Organization ( WHO ) simplified trachoma grading system was used for the recognition and registration of cases of trachoma. Trachoma rapid assessment ( TRA ) was conducted and 30.3687 million people from Shaanxi province were screened. Eyelids, eyelashes, conjunctiva and cornea were examined.The prevalence of trachoma trichiasis ( TT) in Shaanxi Province was estimated.?RESULTS: Totally 987 cases with TT were collected in Shaanxi province, in which 395 cases were male and 592 cases were female. The overall TT prevalence was 0.0325‰.The age of TT cases ranged from 25-86 years old, and concentrated in the 60-80 years old, only 58 cases were <50 years old.There were 12 cases of TT combined corneal opacity (CO) and the ratio was 1.2%. Sixty-four patients were cured by electrolysis trichiasis, the remaining 923 patients corrected by surgery interventions.?CONCLUSION: Based on the results of this study, trachoma blind is no longer estimated as a public health problem in Shaanxi province, as the detection rate of TT was less than 1‰ which is the goal of “elimination of trachoma” worldwide.
10.Relationship of angiotensin-converting enzyme 2 gene polymorphisms and vulnerability to coronary heart disease in patients with type 2 diabetes mellitus.
Quan-nang YAN ; Chun-sheng XU ; Zhi-liang LI ; Peng ZHOU ; Ren-qing YE ; Ming LI ; Chao-xin JIANG
Journal of Southern Medical University 2008;28(8):1365-1368
OBJECTIVETo investigate the association of angiotensin-converting enzyme 2 (ACE2) gene polymorphisms and coronary artery disease (CAD) in patients with type 2 diabetes mellitus (T2DM).
METHODSThis study involved 121 patients with T2DM and 94 with diabetic macroangiopathy. The polymorphisms of G8790A in ACE2 gene was analyzed using PCR-restriction fragment length polymorphism analysis in these patients, and the clinical, biochemical and echocardiographic data were also analyzed.
RESULTSNo obvious difference was found in the genotyping data between the two groups. Among the male patients with diabetic macroangiopathy, the interventricular septal end-diastolic thickness (IVSTd) were significantly greater in patients of GG genotypes of ACE2 gene G8790A than in those of AA genotypes (P<0.01), and the left ventricular mass (LVMI) and urine protein were also significantly higher in GG genotypes (P<0.05). No similar results were found the uncomplicated diabetic group or the female diabetic patients with CAD.
CONCLUSIONThe ACE2 gene G8790A polymorphism plays a role in the pathogenesis of CAD in patients with type 2 diabetes, suggesting that ACE2 genotyping is helpful to screen the susceptible patients.
Adult ; Aged ; Alleles ; Base Sequence ; Case-Control Studies ; Coronary Disease ; complications ; genetics ; Diabetes Mellitus, Type 2 ; complications ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Peptidyl-Dipeptidase A ; genetics ; Polymorphism, Genetic ; Sequence Analysis, DNA