1.In vitro and in vivo study on the regulation of microRNA-101 in human gastric cancer proliferation, migration and invasion
Jing DU ; Wei HUANG ; Youwei CHEN ; Hongjun RUAN
Chinese Journal of Digestion 2015;(8):510-515
Objective To investigate the expression of microRNA‐101 (miRNA‐101 ) in human gastric cancer ,and to explore its effects on proliferation ,migration and invasion of gastric cancer cell . Methods The expression of miRNA‐101 in 28 human gastric cancer tissues ,human gastric cell lines BGC‐823 , SGC‐7901 , MKN‐45 , AGS and human normal gastric epithelial cell line GES‐1 were determined by real time polymerase chain reaction (PCR) .Recombinant miRNA‐101 adenovirus vector was constructed . The effects of miRNA‐101 on gastric cancer proliferation was detected with cell proliferation assay .The ability of gastric cancer cell migration and invasion was assessed with Transwell assay .Gastric xenograft cancer model was established in BALB /c nude mice and the tumor size was compared .The t test was used for the statistical analysis .Results The expression of miRNA‐101 in gastric cancer tissues was 0 .661 ± 0 .396 ,which was lower than that of corresponding para carcinoma tissues (1 .128 ± 0 .697) ,and the difference was statistically significant (t = 10 .091 , P < 0 .01) .The expression of miRNA‐101 in normal gastric epithelial cell line GES‐1 was higher than those of gastric cancer cell lines BGC‐823 ,SGC‐7901 , MKN‐45 and AGS . There was significant suppression role of miRNA‐101 on MKN‐45 cells proliferation , and which also had inhibition role on cell migration and invasion of gastric cancer cell lines BGC‐823 ,SGC‐7901 ,MKN‐45 and AGS .At five weeks after MKN‐45 gastric xenograft cancer nude mice model established ,the tumor size of Ad‐miRNA‐101 group ((333 .56 ± 46 .71) mm3 ) was smaller than that of Ad‐enhanced green fluorescent protein (EGFP) group (806 .41 ± 51 .83) mm3 ,and the difference was statistically significant (t = 21 .431 , P < 0 .01 ) .Conclusion In gastric tissues and cells ,miRNA‐101 is a tumor suppressive miRNA and its downregulated expression involved in the genesis and development of gastric cancer ,which may be a new target of biological target therapy in gastric cancer .
2.Study on value of IHA in diagnosis of schistosomiasis japonica
Jie ZHOU ; Wei GUAN ; Furong WEI ; Yao RUAN ; Shengming LI ; Jing XU ; Tiewu JIA ; Xiaonong ZHOU
Chinese Journal of Schistosomiasis Control 2016;28(4):375-380
Objective To evaluate the value of indirect hemagglutination test(IHA)in schistosomiasis diagnosis. Meth?ods The literature concerned schistosomiasis diagnosis with IHA in the databases of Medline,CNKI,VIP and Wanfang Data from 1982 to 2014 was collected and evaluated. Results Totally 21 articles which were satisfied with the research criteria were analyzed with the Meta?analysis method. The IHA method had high value in schistosomiasis diagnosis,the AUCSROC of IHA in laboratory evaluation was 0.990 6,while in filed evaluation was 0.832 9,and the difference between them was significant(Z=4.50,P<0.05). Conclusion The diagnosis value of IHA in field evaluation is less than that in laboratory. In the process of the elimination of schistosomiasis,developing a new and higher sensitive reagent in schistosomiasis diagnosis is needed.
3.Diagnosis value of serum NKX2-1 for primary lung cancer.
Li YANG ; Wei-jing RUAN ; En-guo CHEN ; Ke-jing YING
Journal of Zhejiang University. Medical sciences 2012;41(5):535-539
OBJECTIVETo evaluate serum Nkx2-1 (NKX homeobox-1) levels in diagnosis of primary lung cancer.
METHODSThe serum NKX2-1 and CEA (carcinoma embryonic antigen) levels were measured in 61 patients with primary lung cancer admitted from May 2009 to December 2010 and 49 healthy individuals served as controls. The receiver operating characteristic curve (ROC) of NKX2-1 in diagnosis for primary lung cancer was analyzed. The value of serum NKX2-1 in diagnosing primary lung cancer was compared with that of CEA by X(2) test and Kappa test.
RESULTSThe serum Nkx2-1 levels in lung cancer were significantly higher than those in controls [(1.4206 ±0.1257)ng/ml compared with (0.7646 ±0.0734)ng/ml,P<0.01]. ROC analysis showed the area under the curve of serum NKX2-1 was 0.859. The Kappa value of NKX2-1 was higher than that of CEA (0.586 compared with 0.396,P<0.05). Combination of serum NKX2-1 with CEA improved the Kappa value to 0.704, and also had high sensitivity (83.6%) and specificity (87.0%) for diagnosis of primary lung cancer.
CONCLUSIONSerum NKX2-1 protein can be used as a marker for diagnosis of lung cancer, the combination of NKX2-1 with CEA may further improve the diagnostic value.
Biomarkers, Tumor ; blood ; Carcinoembryonic Antigen ; blood ; Case-Control Studies ; Humans ; Lung Neoplasms ; blood ; diagnosis ; Nuclear Proteins ; blood ; Sensitivity and Specificity ; Thyroid Nuclear Factor 1 ; Transcription Factors ; blood
4.The Pubmed Bibliometric Analysis of Trend in the Research on Age-related Hearing Loss
Min GUO ; Tao WEI ; Yuping NA ; Chaowu JIANG ; Congjun YE ; Jingyu GAO ; Lizhu YANG ; Jing NA ; Biao RUAN
Journal of Audiology and Speech Pathology 2015;(5):527-533
Objective This study aimed to define research status of age -related hearing loss ,and provide the basis and direction for future research .Methods We have retrieved all relevant literatures on age -related hearing loss from Pubmed ,and conduct an objective analysis of the existing literatures by Bibliometric analytics and co -word analysis method using co -occurrence bibliographic information mining system and SPSS22 .0 software for data analysis .Results There were a large number of articles and journals about presbycusis and age -related hearing loss .Many countries were involved in the research .Literatures and core authors were mainly from developed coun‐tries such as Europe and the United States .The quantity and quality of Chinese literatures were in a leading position in Asia .The researches focused on the common characteristics of patients ,the epidemiology ,characteristics of hear‐ing ,treatment and laboratory studies .There were some new research directions in recent 5 years ,such as factors as‐sociated with the younger ages before developing presbycusis ,standard design and use of questionnaires ,prevention and control .Conclusion Age-related hearing loss will continue to be a hot topic with growing focus on micro and macro development of multi -disciplinary cooperation .The penetration will be the trend for the future research while the prevention will become a new focus of research .
5.Construction, fermentation and purification of high polymer spider dragline silk protein containing RGD peptide.
Chao-Ran RUAN ; Jing-Xing HUANG ; Mei-Hong WEI ; Min LI
Chinese Journal of Biotechnology 2007;23(5):858-861
Spider silk is a natural protein fibroin with excellent character as it is light and tenacious. It has a wild potential applications in the biomedical field due to its good biocompatibility and degradation. Arginine-glycine-aspartic acid (RGD) is a highly conserved amino acid sequence of many adhesion protein. Biological materials binding with RGD peptide in the surface can promote cells adhesion, migration and proliferation. Our lab had constructed the 16 muhimers with the introduced RGD peptide codons which involve cell adhesion for the first time. It was found that the mechanical capability of the 16 mulimer protein was very limited because of the big gap in molecular weight with nature spider proteins when it was used to made biomaterial scaffold.In this paper,based on the 16 multimers of the highly, repetitive sequence of spider dragline silk and with RGD peptide condons which has been constructed by our lab forestall, it was used to construct the 32 and 64 multimers sequence of spider dragline silk by the strategy of "head to tail". The 32 and 64 multimers were ligated into prokaryotic expression vector pET-30a, and then the B121 (DE3) pLysS. The fragments were in agreement with the desired through digestion, agarose gel electrophoresis respectively. By registration into the GenBank data-base, the serial numbers of DQ469929 and DQ837297 were gained respectively. The expression of recombinant protein was introduced by the addition of IPTG. SDS-PAGE analysis shows that the molecular weight of products expressed here are 102 kD and 196.6kD in agreement with the desired respectively. It was the first time for the high polymer spider dragline silk protein expressed in prokaryotic biology. Furthermore, a larger quantity of synthetical proteins with high density fermentation were searched after, and a suit of high efficient purification methods for 32 multimers protein were established.
Animals
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Escherichia coli
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genetics
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metabolism
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Fermentation
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Fibroins
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biosynthesis
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genetics
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Molecular Sequence Data
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Oligopeptides
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biosynthesis
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chemistry
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genetics
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Polymers
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metabolism
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Protein Engineering
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Recombinant Proteins
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biosynthesis
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genetics
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isolation & purification
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Transformation, Bacterial
6.VNTR polymorphism of C6orf37 in Chinese population.
Jing CUI ; Wei WANG ; Min WANG ; Jie LIN ; Yu MA ; Wen-jing RUAN ; Jing XU ; Mao-de LAI
Journal of Zhejiang University. Medical sciences 2006;35(4):354-359
OBJECTIVETo identify a novel VNTR in C6orf37 and to detect the C6orf37 VNTR polymorphism distribution in Chinese population.
METHODSRT-PCR and sequencing were conducted to identify VNTR alleles in the variable region of C6orf37.SSLP and DHPLC were applied in detecting the VNTR genotypes in 166 Chinese individuals.
RESULTA novel VNTR sequence was found in the second exon of C6orf37, which was composed of 15 base pairs encoding 5-amino-acid (G-G-D-F-G). The repeat times ranged from 3 to 5. There were three common alleles containing three repeats (a), four repeats (b) and five repeats (c), respectively, which produced three homozygotes (a/a, b/b and c/c) and three heterozygotes (a/b, a/c and b/c). The frequency of a, b, c alleles were 0.145, 0.304, 0.551, respectively in Chinese population. Heterozygosity (H) was 0.583. Polymorphism information content (PIC) was 0.510. The screened result of DHPLC was consistent with that of SSLP.
CONCLUSIONA novel highly polymorphic VNTR in C6orf37 exists in Chinese population. DHPLC is the most efficient technique for screening VNTR polymorphism.
Asian Continental Ancestry Group ; genetics ; Base Sequence ; Chromatography, High Pressure Liquid ; methods ; Colorectal Neoplasms ; genetics ; pathology ; Exons ; genetics ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Minisatellite Repeats ; genetics ; Molecular Sequence Data ; Polymorphism, Genetic ; genetics ; Proteins ; genetics
7.Genetic Diversity and Natural Selection in 42 kDa Region of Plasmodium vivax Merozoite Surface Protein-1 from China-Myanmar Endemic Border.
Xia ZHOU ; Ernest TAMBO ; Jing SU ; Qiang FANG ; Wei RUAN ; Jun Hu CHEN ; Ming Bo YIN ; Xiao Nong ZHOU
The Korean Journal of Parasitology 2017;55(5):473-480
Plasmodium vivax merozoite surface protein-1 (PvMSP1) gene codes for a major malaria vaccine candidate antigen. However, its polymorphic nature represents an obstacle to the design of a protective vaccine. In this study, we analyzed the genetic polymorphism and natural selection of the C-terminal 42 kDa fragment within PvMSP1 gene (Pv MSP142) from 77 P. vivax isolates, collected from imported cases of China-Myanmar border (CMB) areas in Yunnan province and the inland cases from Anhui, Yunnan, and Zhejiang province in China during 2009–2012. Totally, 41 haplotypes were identified and 30 of them were new haplotypes. The differences between the rates of non-synonymous and synonymous mutations suggest that PvMSP142 has evolved under natural selection, and a high selective pressure preferentially acted on regions identified of PvMSP133. Our results also demonstrated that PvMSP142 of P. vivax isolates collected on China-Myanmar border areas display higher genetic polymorphisms than those collected from inland of China. Such results have significant implications for understanding the dynamic of the P. vivax population and may be useful information towards China malaria elimination campaign strategies.
China
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Genetic Variation*
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Haplotypes
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Malaria
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Merozoite Surface Protein 1*
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Merozoites*
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Myanmar
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Plasmodium vivax*
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Plasmodium*
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Polymorphism, Genetic
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Selection, Genetic*
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Silent Mutation
8.Effects of chronic administration of melatonin on spatial learning ability and long-term potentiation in lead-exposed and control rats.
Xiu-Jing CAO ; Ming WANG ; Wei-Heng CHEN ; Da-Miao ZHU ; Jia-Qi SHE ; Di-Yun RUAN
Biomedical and Environmental Sciences 2009;22(1):70-75
OBJECTIVETo explore the changes in spatial learning performance and long-term potentiation (LTP) which is recognized as a component of the cellular basis of learning and memory in normal and lead-exposed rats after administration of melatonin (MT) for two months.
METHODSExperiment was performed in adult male Wistar rats (12 controls, 12 exposed to melatonin treatment, 10 exposed to lead and 10 exposed to lead and melatonin treatment). The lead-exposed rats received 0.2% lead acetate solution from their birth day while the control rats drank tap water. Melatonin (3 mg/kg) or vehicle was administered to the control and lead-exposed rats from the time of their weaning by gastric gavage each day for 60 days, depending on their groups. At the age of 81-90 days, all the animals were subjected to Morris water maze test and then used for extracellular recording of LTP in the dentate gyrus (DG) area of the hippocampus in vivo.
RESULTSLow dose of melatonin given from weaning for two months impaired LTP in the DG area of hippocampus and induced learning and memory deficit in the control rats. When melatonin was administered over a prolonged period to the lead-exposed rats, it exacerbated LTP impairment, learning and memory deficit induced by lead.
CONCLUSIONMelatonin is not suitable for normal and lead-exposed children.
Animals ; Female ; Lead ; toxicity ; Learning ; drug effects ; Long-Term Potentiation ; drug effects ; Male ; Maze Learning ; drug effects ; Melatonin ; administration & dosage ; toxicity ; Rats ; Spatial Behavior ; drug effects
9.Protective effects of omega-3 fish oil on lead-induced impairment of long-term potentiation in rat dentate gyrus in vivo.
Xiu-Jing CAO ; Jie-Jie CAO ; Ting-Ting CHEN ; Wei-Heng CHEN ; Di-Yun RUAN
Acta Physiologica Sinica 2010;62(3):225-230
In order to evaluate the effect of omega-3 fish oil supplement by gavage (0.4 mL/100 g body weight) on the chronic lead-induced (0.2% lead acetate) impairments of long-term potentiation (LTP) in rat dentate gyrus (DG) in vivo, we designed the experiments which were carried out in four groups of newborn Wistar rats (the control, the lead-exposed, the control with fish oil treatment and the lead-exposed with fish oil treatment, respectively). The excitatory postsynaptic potential (EPSP) and population spike (PS) amplitude were measured in the DG of rats with above different treatments at the age of 80-90 d in response to stimulation applied to the lateral perforant path. The results showed (1) postnatal chronic lead-exposure impaired LTP measured on both EPSP slope and PS amplitude in DG area of the hippocampus; (2) in the control rats, omega-3 fish oil had no effect on LTP while in the lead-exposed rats, omega-3 fish oil had a protective effect on LTP. These results suggest that omega-3 fish oil supplement could protect rats from the lead-induced impairment of LTP. Omega-3 fish oil might be a preventive substance in reducing LTP deficits induced by lead.
Animals
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Animals, Newborn
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Dentate Gyrus
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drug effects
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Excitatory Postsynaptic Potentials
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Fatty Acids, Omega-3
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pharmacology
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Fish Oils
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chemistry
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Lead Poisoning
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physiopathology
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Long-Term Potentiation
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drug effects
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Perforant Pathway
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Rats
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Rats, Wistar
10.Pathological study of lung cancer induced by Yunnan tin mine dusts in F344 rats.
Yong-hua RUAN ; Hai-rong HUA ; Qian GAO ; Jing-ling SONG ; Rui LIANG ; Ke-wei JIN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2007;25(6):331-335
OBJECTIVETo set up animal models of the lung cancer induced by Yunnan tin mineral dusts (no radon) in F344 rats and to explore the process of carcinogenesis and pathologic alterations in various stages of malignant transformation in the animal models.
METHODSOne hundred and ninety F344 rats were randomly divided into Yunnan tin mineral dust group (100 rats), furfural physiological saline group (30 rats), physiological saline group (30 rats) and normal control group (30 rats). The intratracheal instillation with mass fraction of 6% suspension liquid mixture Yunnan tin mineral dusts, volume fraction of 2% furfural physiological saline and physiological saline 0.2 ml was performed in the rates once per week respectively except normal control group. Then the rats were sacrificed in batch periodically after one week. The last rat was exposed to the tin mine dusts for 100 weeks. The morphological process and tumor formation were dynamically observed under LM and TEM. Immunohistochemistry detection of cytokeratin of High MW and low MW was used for tumor classification. Pollak stein was used to evaluate the development of fibrosis of lung in the rats.
RESULTSBronchoalveolar inflammation occurred in the early stage after the intratracheal instillation of Yunnan tin mineral dust was performed in F344 rates. Along with reduction of inflammation, collagen fibrils increased at alveolar interstices. Simple hyperplasia, papillary hyperplasia and metaplasia of the epithelial cells in alveolar and bronchi were observed, followed by atypical adenomatous hyperplasia and squamous dysplasia. Lung cancer was induced in the end. Among the 14 cases of lung cancer, 9 cases were adenocarcinoma, 2 squamous cell carcinoma and 3 mixed carcinoma. No lung cancer occurred in other three control groups. There was a significant difference in the malignancy rate between the experimental group and the three control groups (P < 0.01). The squamous metaplasia and squamous carcinoma were found in alveoli that expressed cytokeratin of High MW. Lung fibrosis was found in 31 cases of in the tin mineral dust group. The greater the mineral dust deposit was, the more serious the alveolar fibrosis was.
CONCLUSIONYunnan tin mineral dusts without radon induce lung cancer in rates. The adenocarcinoma and squamous carcinomas induced in F344 rat lung can occur in the alveoli. The further study on whether type II alveolar epithelial cells are the origin cells of adenocarcinoma and some peripheral squamous lung carcinomas is worthwhile.
Animals ; Disease Models, Animal ; Dust ; Female ; Lung ; drug effects ; pathology ; Lung Neoplasms ; chemically induced ; pathology ; Male ; Rats ; Rats, Inbred F344 ; Tin ; adverse effects