2.miRNA-101 inhibits the expression of the enhancer of zeste homolog 2 in androgen-independent prostate cancer LNCaP cell line.
Jian-xin LIU ; Qi-fa ZHANG ; Chang-hai TIAN ; Yong ZHANG ; Xiao-zhou HAN ; Hao GUO
National Journal of Andrology 2015;21(6):500-503
OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.
METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.
RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.
CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.
Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection
3.Cloning and expression analysis of the expansin gene RgEXPA10 in Rehmannia glutinosa.
Feng-Qing WANG ; Yan ZHOU ; Yong HUANG ; Ming-Jie LI ; Yun-He TIAN ; Fa-Jie FENG ; Xin-Jian CHEN ; Zhang ZHONG-YI
Acta Pharmaceutica Sinica 2015;50(2):233-240
Using cDNA from Rehmannia glutinosa leaf as template, a 972 bp fragment of expansin gene which containing a 762 bp ORF that encoded 253 amino acids, was cloned, named RgEXPA10, which GenBank accession number for this gene is KF011918. A 1 207 bp genomic sequence of RgEXPA10 was amplified by PCR with leaf DNA as template, sequencing analysis revealed that three exons and two introns in RgEXPA10 genomic sequence, and which GenBank accession number is KF011919. Molecular and bioinformatic analyses indicated that RgEXPA10 protein have DPBB_1 and Pollen_allerg_1 domain, also including a 26 aa nuclear localization signal and a 19 aa transmembrane region. Phylogenetic analysis revealed that RgEXPA10 showed the highest homology with AtEXPA8 among the 26 α-expansins in Arabidopsis thaliana. However, the RgEXPA10 indicated the highest homology with the expansin from Solanum lycopersicum among 22 plant species. Expression patterns using qRT-PCR analysis showed that RgEXPA10 mainly expressed in unfolded leaf, followed by the tuberous root at stage of expanding period, and rarely expressed in senescing leaf. And RgEXPA10 showed higher expression level in tuberous root at 60 and 90 days after emergence. The transcription level of RgEXPA10 significantly reduced under all the three stresses including continuous cropping conditions, salinity and waterlogging. This study will lay foundations for molecular function in development and regulation of different stresses for R. glutinosa.
Amino Acid Sequence
;
Cloning, Molecular
;
DNA, Complementary
;
Gene Expression Regulation, Plant
;
Genes, Plant
;
Molecular Sequence Data
;
Phylogeny
;
Plant Leaves
;
Plant Proteins
;
genetics
;
Plant Roots
;
Rehmannia
;
genetics
4.Treatment of infantile asthma in remission stage with Chinese medicine and new moxibustion-massage apparatus.
Chinese Acupuncture & Moxibustion 2012;32(2):163-165
OBJECTIVETo compare differences of effects between new moxibustion-massage apparatus and suspended moxibustion for infantile asthma.
METHODSSixty patients were divided into 2 groups randomly, observation group and control group, 30 cases in each. Tiaoyuan Yuping Decoction, a decoction made by the hospital, was applied for both groups. New moxibustion-massage apparatus was applied for the observation group, and suspended moxibustion was adopted for the control group. Moxibustion was applied on acupoints of Fengmen (BL 12), Feishu (BL 13), Geshu (BL 17), Pishu (BL 20), Juque (CV 14) and Zhongwan (CV 12) for 3 months, and therapeutic effect was observed.
RESULTSThe remarkably effective rate of the observe group is 66.7% (20/30) and the total effective rate is 90.0% (27/30), which are better than 40.0% (12/30) and 83.3% (25/30) of the control group. The asthma of attack of both groups after the treatment obviously decreased (both P < 0.05). The asthma of attack of the observation group is obviously lower than that of the control group (P < 0.05). And the effect is approved without side-effects such as dermal infection.
CONCLUSIONIt is held that the combination of new moxibustion-massage apparatus and Chinese medicine for infantile asthma in remission stage has obvious therapeutic effect, which can reduce the attack of asthma and alleviate symptoms. The result is better than suspended moxibustion.
Asthma ; drug therapy ; therapy ; Child ; Child, Preschool ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Massage ; Moxibustion
5.Genetic analysis of the NS genes of H5N1 avian influenza viruses isolated from sewage in poultry markets.
Xin-Hua OU ; Ru-Sheng ZHANG ; Ke-Yun SONG ; Tian-Mu CHEN ; Ru-Chun LIU ; Bian-Cheng SUN ; Fa-Ming CHEN
Chinese Journal of Virology 2012;28(3):265-271
In order to investigate the transmission risk of H5N1 avian influenza viruses (AIV) from sewage in Changsha poultry markets, the evolution relationship and molecular characteristics of non-structural (NS) genes of H5N1 AIV from sewage were analyzed. Nine H5N1 AIV environmental sewage specimens were collected from Changsha poultry markets. The NS genes were amplifyed by PCR and then sequenced with TA cloning. Amino acid(aa) sequence alignment and phylogenetic tree analysis were conducted by Lasergene and Mega5 software. Eight NS genes TA cloning were constructed successfully. Phylogenetic tree indicated that they were belonged to the allele A subgroup. Aa homology analysis showed 90.1% 92.5% identity in NS1 proteins and 91.0% - 92.6% identity in NS2 proteins compared with reference viruses of the allele A (A/chicken/ Hubei/ w h/ 1999). The homologies of the amino sequences of NS1 and NS2 in this study were 93.8%-100.0% and 98.4%-100.0%, respectively. The C terminal of all eight H5N1 NS1 proteins from sewage in poultry markets carried a ESEV of PL motif and the 92 amino acids were E, furthermore, the 80 to 84aa were missed which were the characteristics of highly pathogenic AIV. The NS genes of H5N1 AIV from sewage in poultry markets have molecular characteristics of highly pathogenic and have the potential risk of H5N1 virus spreading.
Amino Acid Sequence
;
Animals
;
Influenza A Virus, H5N1 Subtype
;
chemistry
;
classification
;
genetics
;
isolation & purification
;
Influenza in Birds
;
transmission
;
virology
;
Molecular Sequence Data
;
Phylogeny
;
Poultry
;
Sequence Homology, Amino Acid
;
Sewage
;
virology
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Viral Nonstructural Proteins
;
chemistry
;
genetics
6.Video-assisted thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in thoracic cavity: analysis of 60 cases.
Xiang-xiang SUN ; Mei-qing XU ; Ming-fa GUO ; Chang-qing LIU ; Shi-bin XU ; Xin-yu MEI ; Jie-yong TIAN ; Zheng-hua ZHANG ; Da-zhong WEI
Chinese Journal of Surgery 2013;51(4):354-357
OBJECTIVETo investigate the feasibility and safety of video-assisted thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity.
METHODSThe clinical data of 120 patients who underwent esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity from March to December 2011 was analyzed retrospectively. In the video-assisted thoracoscopic surgery group, there were 60 patients [41 male and 19 female patients with aver age of (62 ± 7) years old] who underwent video-assisted thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity. In the routine thoracotomy group, there were 60 patients [39 male and 21 female patients with aver age of (62 ± 9) years old] who underwent routine thoracotomy esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity. Operation time, intra-operative blood loss, postoperative total thoracic drainage in 3 days, total number of harvested lymph nodes, hospitalization, cost of hospitalization and complications were compared between the two groups.
RESULTThe operations were carried out successfully in two groups. There was no perioperative death in all patients. There was no statistical difference in intra-operative blood loss, postoperative total thoracic drainage and cost of hospitalization between the two groups. Operation time of rideo-assisted thoracoscopic surgery group was significantly longer than that of thoracotomy group ((188 ± 38) minutes vs. (138 ± 50) minutes, t = 6.171, P = 0.000), but postoperative hospitalization was significantly lower ((14 ± 3) d vs. (18 ± 6) d, t = -4.093, P = 0.000) and total number of harvested lymph nodes was lower (17 ± 9 vs. 21 ± 11, t = -2.058, P = 0.042). There was significantly statistical difference in total postoperative main complication (25.0% vs. 48.3%, χ(2) = 7.033, P = 0.008). And postoperative incisional infection of VATE group patients was significantly lower than that of thoracotomy group patients (6.7% vs. 25.0%, χ(2) = 7.566, P = 0.006).
CONCLUSIONSVideo-assisted thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity is technically feasible and safe, with minimized trauma and quick recovery. The recent result is satisfactory.
Aged ; Aged, 80 and over ; Anastomosis, Surgical ; methods ; Esophageal Neoplasms ; surgery ; Esophagectomy ; methods ; Female ; Humans ; Lymph Node Excision ; Male ; Middle Aged ; Retrospective Studies ; Thoracic Surgery, Video-Assisted ; Thoracotomy
7.m6A Regulates Liver Metabolic Disorders and Hepatogenous Diabetes
Li YUHUAN ; Zhang QINGYANG ; Cui GUANSHEN ; Zhao FANG ; Tian XIN ; Sun BAO-FA ; Yang YING ; Li WEI
Genomics, Proteomics & Bioinformatics 2020;18(4):371-383
N6-methyladenosine (m6A) is one of the most abundant modifications on mRNAs and plays important roles in various biological processes. The formation of m6A is catalyzed by a methyltransferase complex (MTC) containing a key factor methyltransferase-like 3 (Mettl3). How-ever, the functions of Mettl3 and m6A modification in hepatic lipid and glucose metabolism remain unclear. Here, we showed that both Mettl3 expression and m6A level increased in the livers of mice with high fat diet (HFD)-induced metabolic disorders. Overexpression of Mettl3 aggravated HFD-induced liver metabolic disorders and insulin resistance. In contrast, hepatocyte-specific knockout of Mettl3 significantly alleviated HFD-induced metabolic disorders by slowing weight gain, reducing lipid accumulation, and improving insulin sensitivity. Mechanistically, Mettl3 depletion-mediated m6A loss caused extended RNA half-lives of metabolism-related genes, which consequently pro-tected mice against HFD-induced metabolic syndrome. Our findings reveal a critical role of Mettl3-mediated m6A in HFD-induced metabolic disorders and hepatogenous diabetes.
8.Prevalence of overactive bladder in a community-based male aging population.
Xiang-Hua ZHANG ; Xin LI ; Zheng ZHANG ; Shu-Qing LI ; Ye TIAN ; Yan-Qun NA ; Yi WANG ; Shan CHEN ; Bao-Fa HONG ; Wen-Xue SUN ; Ying-Zhi DIAO
Chinese Journal of Surgery 2010;48(23):1763-1766
OBJECTIVETo investigate the situation of overactive bladder (OAB) in a community-based male population.
METHODSMale participants over 50 years old were randomly selected from multiple communities in Beijing. The evaluation of lower urinary tract symptoms (LUTS) including the International Prostate Symptom Score (IPSS), quality of life (QOL) score, prostate volume and post voiding residue (PVR) by abdominal ultrasonography, and maximum flow rate (Qmax). Definition of OAB was determined as the score of item number 4 in IPSS ≥ 2.
RESULTSOf 1656 male participants enrolled, a total of 1639 men met our study criteria. The mean age was (64 ± 10) years. The prevalence of OAB was 26.3% (431/1639), and was significantly related to age, IPSS, QOL score, prostate volume, PVR and Qmax (P < 0.01). The prevalence of OAB was closely associated with aging (P < 0.01) and the degree of LUTS (P < 0.01).
CONCLUSIONSThe prevalence of OAB increased with aging of the community-based male population. OAB would obviously affect the quality of life of the aging men.
Aged ; Aged, 80 and over ; Aging ; China ; epidemiology ; Humans ; Male ; Middle Aged ; Prevalence ; Quality of Life ; Urinary Bladder, Overactive ; epidemiology
9.Advances on molecular mechanisms of Rehmannia glutinosa consecutive monoculture problem formation in multi-omics era.
Ming-Jie LI ; Fa-Jie FENG ; Bao ZHANG ; Li GU ; Feng-Qing WANG ; Yan-Hui YANG ; Yun-He TIAN ; Xin-Jian CHEN ; Zhong-Yi ZHANG
China Journal of Chinese Materia Medica 2017;42(3):413-419
Although consecutive monoculture problems have been studied for many years, no effective treatments are currently available. The complexity of systems triggered the formation of consecutive monoculture problems was one major cause. This paper elaborated the physiological and ecological mechanisms of consecutive monoculture problem formation based on the interaction relationship among multiple factors presented in the rhizosphere soil of consecutive monoculture plants. At same time, in this paper the multiple interactions among cultivated medicinal plants, autotoxic allelochemicals and rhizosphere microbial were proposed to be most important causes that derived the formation of consecutive monoculture problem. The paper also highlighted the advantage of 'omics' technologies integrating plant functional genomics and metabolomics as well as microbial macro-omics in understanding the multiple factor interaction under a particular ecological environment. Additionally, taking R. glutinosa as an example, the paper reviewed the molecular mechanism for the formation of R. glutinosa consecutive monoculture problem from the perspective of the accumulation of allelopathic autotoxins, the rhizosphere microecology catastrophe and theresponding of consecutive monoculture plants. Simultaneously, the roles of mutilple 'omics' technologies in comprehending these formation mechanism were described in detail. This paper provides finally a new insight to solve systematically the mechanism of consecutive monoculture problem formation on molecular level.
10. Effects of high glucose on expression of OPG and RANKL in rat aortic vascular smooth muscle cells
Hong-Juan CHANG ; Xin MENG ; Tian-Fa LI ; Jun-Li GUO ; You-Ling LAN ; Yue-Qiong KONG ; Xian-Ji MA ; Xiao-Ling LU ; Wei-Ying LU ; Shao-Jiang ZHENG
Asian Pacific Journal of Tropical Medicine 2015;8(3):209-213
Objective: To explore effect of high glucose on expression of osteoprotegerin (OPG) and receptor activator of NF- κ B ligand (RANKE) in rat aortic vascular smooth muscle cells. Methods: SD rats were intraperitoneally injected with streptozotocin, OPG and RANKL expression in rat thoracic aortas were detected by immunohistochemical staining. In cultured vascular smooth muscle cells (VSMCs) (A7r5), qRT-PCR and Western blot analysis were used to examine the mRNA and protein levels of OPG and RANKL. Results: Our results demonstrated that OPG expression was increased in hyperglycemic rat aortic VSMCs, while RANKL expression was decreased. Besides, in vitro experiments high glucose induced OPG expression, but depressed RANKL expression by dose- and time-dependent manner in cultured A7r5. Conclusions: Our findings suggested that high glucose could promote the expression of OPG, and inhibit the expression of RANKL in VSMCs, which may be partly be the molecular mechanism of diabetic vascular calcification.