2.Association of CA repeats polymorphism in the promoter region of the insulin-like growth factor- Ⅰ gene with type 2 diabetes
Min YANG ; Wei LING ; Jing LIN ; Changlin NI
Chinese Journal of Endocrinology and Metabolism 2010;26(12):1042-1044
Objective To investigate the association of CA repeats polymorphism in the promoter region of human IGF-I gene with type 2 diabetes in narthen Han nationality. Methods Gender-age matched subjects with type 2 diabetes and normal glucose tolerance (NGT) were enrolled for this study, 147 subjects in type 2 diabetes group and 159 subjects in NGT group. Genomic DNA was extracted by standard methods. PCR, Genescan,Genotyper, and direct sequencing were conducted to screen CA repeats polymorphism in the promoter region of the human IGF-I gene. Results No significant association was observed between any ( CA), repeat genotype and type 2 diabetes. A lowered serum uric acid was seen in genotypes that included alleles with larger than (CA) 20 repeats [(4.18 ± 1.25 vs 4.63 ± 1.36) mg/dl, P = 0.03]. Conclusion Alleles with larger than (CA) 20 repeats may be a protective factor for type 2 diabetes.
3.A novel citrinin derivative from the marine-source fungus Penicillium citrinum.
Min NI ; Wei-Ling LIN ; Peng YANG ; Shui-Chun MAO
Acta Pharmaceutica Sinica 2015;50(2):203-206
A novel citrinin derivative, penicitrinol L (1), along with two known analogues, penidicitrinin B (2) and pennicitrinone A (3) were isolated from the marine-source fungus Penicillium citrinum. The structure of the new compound was elucidated by spectroscopic methods including one and two-dimensional NMR as well as high-resolution mass spectrometric analysis. Furthermore, compound 1 showed modest cytotoxic activity against HL-60 cell line and compound 3 showed weak cytotoxic activity against A375 cell line.
Antineoplastic Agents
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chemistry
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isolation & purification
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Citrinin
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analogs & derivatives
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chemistry
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isolation & purification
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HL-60 Cells
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Humans
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Magnetic Resonance Spectroscopy
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Penicillium
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chemistry
4.Up-regulation of calreticulin expression and its significance in patients with rheumatoid arthritis in vivo
Yichao WANG ; Wei WEI ; Chunyou WAN ; Min NI ; Na ZHANG ; Hongyi LIU ; Fang ZHENG
Chinese Journal of Rheumatology 2013;(1):27-30,封3
Objective To investigate the role of calreticulin (CRT) in the pathogenesis of rheumatoid arthritis (RA) by analyzing its expression in the sera,synovial fluid and synovial membrane in patients with RA.Methods Levels of CRT in the sera from patients with RA,osteoarthritis (OA),systemic lupus erythematosus (SLE),other autoimmune diseases and health control (HC) were detected by enzyme linked immunosorbent assay (ELISA) and Western bloting.CRT levels in synovial fluid from RA and OA patients were measured by ELISA.Pathological methods were employed to analyze the expression and localization of CRT in synovial membrane.ANVOA,SNK-q test were used for statistical analysis.Results CRT was found to be significantly up-regulated in sera from RA [(4.8±2.4) ng/ml] than that from in OA [(3.6±0.9)ng/ml],SLE [(4.0±1.5) ng/ml],other autoimmune diseases [(3.9±0.8) ng/ml] and HC [(3.7±0.6) ng/ml].Only the monomer form of CRT in the serum could be detected.Pathological results showed that in RA synovium,CRT was mainly expressed in the lining and sublining layers,endothelial cells and perivascular areas; while in OA,only few CRT staining was seen in perivascular areas and the synovial lines.Conclusion In RA,increased levels of CRT are detected in the sera.In addition,high expression of CRT is observed in synovium and its distribution are different from OA.Our results suggest that CRT may be involved in RA joint inflammation and pannus formation.CRT may become a potential serological marker in the diagnosis of RA.
5.Study on the expression of serum amyloid protein A in vivo in patients with rheumatoid arthritis
Yun MU ; Wei WEI ; Fujiang ZHANG ; Xuguo SUN ; Hongyi LIU ; Min NI ; Fang ZHENG
Chinese Journal of Rheumatology 2011;15(11):783-785
ObjectiveTo investigate the role of SAA in rheumatoid arthritis(RA) pathogenesis by analyzing the expression of serum amyloid protein A(SAA) in serum,synovial fluid and synovial membrane in patients with RA.MethodsSAA levels in the serum and synovial fluid in each group were detected.Sera SAA was tested by Western blotting,while the expression of SAA in RA and osteoarthritis(OA) synovium was detected by immunohistochemistry.Comparisons between groups were performed by t-tests or KruskaWallis test.ResultsThe serum levels of SAA were significantly higher in RA [(318±132) μg/L] than those in OA [(127±47) μg/L] and healey controls [(127±41) μg/L,P<0.01].In RA,the SAA levels in the synovial fluid [ (571±473) μg/L ] were significantly higher when compared to those in O A [ (129±33) μg/L](t=2.46,P=0.04).Western blotting results showed that SAA bands were found in the serum samples of each group,and higher expression of SAA were seen in RA.Pathology study had showed that SAA was observed mainly in endothelial cells,synovial fibroblasts,macrophages and perivascular areas in RA synovium.In OA,SAA was observed in perivascular areas and synovial fibroblasts.ConclusionIn RA,SAA levels in both serum and synovial fluid are significantly higher than those in the controls.High expression of SAA in RA synovium can be observed.Our results suggest that SAA may play a role in inflammation reaction and joint destruction of RA.
6.The attenuating effect of ursodeoxycholic acid on endoplasmic reticulum stress-mediated pancreatic β-cell apoptosis in streptozotocin-induced diabetic rats
Min ZHU ; Xing SHI ; Shining NI ; Wei GU ; Mei GUO ; Li FEI ; Xiaoqin PAN ; Qianqi LIU
Chinese Journal of Endocrinology and Metabolism 2011;27(11):935-940
Objective To clarify the protective effect of nrsodeoxycholic acid ( UDCA ) on endoplasmic reticulum stress-mediated apoptosis in pancreatic β-cell of streptozotocin ( STZ )-induced diabetic rats.Methods Rats( n =40) received a single injection STZ( 50 mg/kg) intra-peritoneally and formed a β-cell injury model.Weight-matched normal rats( the control group,n =10 ) were injected with the buffer alone.STZ-treated rats with persistent random blood glucose higher than 16.7 mmol/L for 1 week were considered as diabetic status( n=14 ),then divided randomly into STZ-induced diabetes mellitus ( DM ) group ( n =7 ) and UDCA-treated DM group ( n =7 ).UDCA (40 mg· kg- 1,d-1 ) was administered daily by intragastric intubations throughout the experimental period (30 d).During the entire experiment,blood glucose in all rats was assessed.By the end of the experiment,all rats were sacrificed with the pancreas removed and the blood sample collected immediately.Fasting insulin levels were assayed by radioimmunoassay.The morphological changes of pancreatic β-cells apoptosis were determined by TUNEL assay.RNA in pancreas was abstracted and microarray containing 89 pieces of apoptosis related genes was applied.The related gene expressions were detected by RT-PCR and Western blot.Results The concentration of blood glucose in diabetic rats was gradually decreased after UDCA treatment,but at the end of the experiment it was still higher than that in the normal control group.The treatment with UDCA raised the fasting insulin level in diabetic rats,but this concentration was significantly lower as compared to the control group.Based on TUNEL stained tissue sections,the percentage of β-cell apoptosis of UDCA-treated DM group was significantly lower than that of STZ-induced diabetic group(P<0.05 ).Among 89 genes,42 genes up-regulated and 46 genes down-regulated in diabetic rats,some of which were ameliorated by UDCA treatment.The expressions of Caspase-3,Bax,Bip,and CHOP mRNA in pancreas of DM group were significantly up-regulated as compared with those in the control group ( P < 0.05 ) ; while the expression of Bcl-2 mRNA was markedly down-regulated (P<0.05 ).However,these parameters in the U DCA-treated animals showed a marked improvement.Conclusion Ursodeoxycholic acid seems to protect pancreatic β-cell from apoptosis in STZ-induced diabetes by attenuating the severity of endoplasmic reticulum stress.
7.Expression,Purification and Antibody Preparation of Recombinant C8orf32 Protein
Lei ZHU ; Zheng-Xi ZHANG ; Guo-Xin NI ; Xue-Min XU ; Biao-Yang LIN ; Wei LI ;
China Biotechnology 2006;0(04):-
C8orf32 is a gene which has not been functionally characterized,the mRNA level of this gene is significantly higher in breast cancer tissues than that in normal breast tissues.The amplified cDNA fragment was inserted into the pGEX-6P1 vector fused with the upstream GST gene.The expression vector was transformed into the E.coli BL21(DE3) strain and expression of GST-C8orf32 fusion protein was induced by IPTG..After removal of GST tag by site-specific protease,the C8orf32 protein fused with an eight amino acid peptide tag was obtained.The purity of recombinant C8orf32 protein was about 95%.The identity of the purified protein was confirmed by N-terminal sequencing and tandem mass spectrometry.The polyclonal antibody was prepared by immunizing the New Zealand white rabbits with C8orf32 protein.The polyclonal antibody was proved to recognize the C8orf32 protein correctly.The purified C8orf32 protein can be used for structural and functional studies and the polyclonal antibody can be used for tissue specific protein expression profiling.
8.Environmental factors affecting the succinic acid production by Actinobacillus succinogenes CGMCC 1593.
Pu ZHENG ; Wei ZHOU ; Ye NI ; Min JIANG ; Ping WEI ; Zhihao SUN
Chinese Journal of Biotechnology 2008;24(6):1051-1055
Actinobacillus succinogenes is a promising candidate for the production of bio-based succinic acid. Previously, we isolated a succinic acid-producing strain Actinobacillus succinogenes CGMCC 1593 from bovine rumen. In this paper, the influence of the environmental factors such as gas phase, pH, ORP, on succinic acid production by A. succinogenes CGMCC 1593 was studied. The results showed that CO2 was the optimum gas phase for anaerobic fermentation ofA. succinogenes CGMCC 1593 as well as one of the substrate for the succinic acid synthesis. Using MgCO3 as a pH regulator, the pH was maintained within 7.1-6.2 during the anaerobic fermentation for the cell growth and acid production of A. succinogenes CGMCC 1593. Our results showed that low initial ORP was disadvantageous for the growth of A. succinogenes CGMCC 1593 and an ORP of -270 mV was demonstrated to be beneficial to the succinic acid production. By adding Na2S.9H2O to decrease ORP to -270 mV at the end of exponential growth phase in batch culture of A. succinogenes CGMCC 1593, the succinic acid concentration reached 37 g/L and the yield of succinic acid was 129% at 48 h. This work might provide valuable information for further optimization of succinic acid fermentation by A. succinogenes CGMCC 1593.
Actinobacillus
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classification
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growth & development
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metabolism
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Anaerobiosis
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Carbon Dioxide
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pharmacology
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Fermentation
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Hydrogen-Ion Concentration
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Oxidation-Reduction
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Succinic Acid
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metabolism
9.Investigation of iodine nutritional status and thyroid function among pregnant and lactating women from rural areas of Jinlin province in 2009
Li-fen, YANG ; Hui-xin, CHEN ; Wei, LI ; Min, WANG ; Dan-ni, YANG ; Hong-yan, L(U) ; Yan-hua, LUO ; Jing-shen, ZHAO ; Li-xia, ZHU
Chinese Journal of Endemiology 2011;30(2):162-164
Objective To investigate the iodine nutritional level and thyroid function of pregnant and lactating women in rural areas of Jilin province. Methods The investigation sites were selected from rural areas of three towns (Baoshan, Mingcheng and Yantongshan of Panshi county, Jilin province) in 2009. The pregnant and lactating women were selected as subjects in these three towns. The blood samples were collected and the thyroid function (including serum TT3, TT4, FT3, FT4) were measured with chemiluminescence, and serum thyroglobulin antibodies(TgAb), thyromicrosome antibody(TMAb), and thyroglobulin(Tg) were measured with radioimmunoassay (RIA). The urine samples were collected three times within one month and were measured for iodine concentration by As-Ce catalytic spectrophotometry method. Results In the pregnant women, serum TT3 was higher than that of healthy pregnant women, accounted for 14.3%(8/56), while serum TT4, TT3, FT4 were lower than those of healthy pregnant women, accounted for 3.6%(2/56),5.4% (3/56), and 1.8%(1/56), respectively. In the lactating women,serum TT3 was higher than that of healthy lactating women, accounted for 3.6%(2/56), while serum TT4, FT4 were lower than those of healthy lactating women, accounted for 1.8%(1/56), respectively. Five per cent to 20% of the pregnant and lactating women had higher TgAb and TMAb. Conclusions Existing salt iodine level is appropriate for pregnant women and lactating women, but there was a tendency towards hypothyroid in some women. Routine monitoring of urinary iodine and thyroid function should be carried out among pregnant and lactating women.
10.Clinicopathological features and prognostic analysis of esophageal sarcomatoid carcinoma.
Peng-cheng CHEN ; Qi-xun CHEN ; Xing-hao NI ; Xin-ming ZHOU ; Wei-min MAO
Chinese Journal of Oncology 2012;34(4):287-290
OBJECTIVETo analyze the clinicopathological characteristics and prognosis of a rare histological type of esophageal cancer-sarcomatoid carcinoma.
METHODSClinicopathological data of 31 patients with esophageal sarcomatoid carcinoma who underwent surgery in the Department of Thoracic Surgery of Zhejiang Cancer Hospital from Jan 2000 to Dec 2009 were collected and analyzed. The survival analysis was performed using Kaplan-Meier method.
RESULTSAll the patients underwent surgery. Of the 31 patients, one received preoperative chemoradiotherapy and postoperative chemotherapy, and 8 received postoperative chemotherapy. All the tumors were located in the middle or lower esophagus. Microscopically, the tumors were composed of both carcinomatous and sarcomatous components, and there was a transition between the two components, but no obvious heterogenous elements such as osteosarcoma, chondrosarcoma or rhabdomyosarcoma were found. In the carcinomatous components, positive expression of CK and EMA was found in all the 31 cases, and positive expression of vimentin in 5 of the 31 cases. In the sarcomatous components, positive expression of CK, EMA and vimentin was found in 29, 28 and 23 cases, respectively. The 1-, 3-, and 5-year survival rates were 80.6%, 55.9% and 33.4%, respectively, and the median survival time was 40 months.
CONCLUSIONSEsophageal sarcomatoid carcinoma is a particular type of esophageal malignancy with unique clinicopathological features. The diversity and complexity of the carcinomatous and sarcomatous components and their potential of transformation and differentiation lead to different prognosis from each other.
Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinosarcoma ; metabolism ; pathology ; surgery ; therapy ; Chemoradiotherapy, Adjuvant ; Chemotherapy, Adjuvant ; Esophageal Neoplasms ; metabolism ; pathology ; surgery ; therapy ; Esophagectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Keratins ; metabolism ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Prognosis ; Survival Rate ; Vimentin ; metabolism