1.Induction of apoptosis and cell cycle arrest in human endometrial cancer cells by adiponectin
Li CONG ; Wei WANG ; Qiang LI ; Zijian ZHAO ; Jinchao ZHANG
Chinese Journal of Endocrinology and Metabolism 2010;26(2):148-149
Two human endometrial carcinoma cell lines, HEC-1-A and RL95-2, were treated with adiponectin,and then changes of cell count, apoptosis and cell cycle arrest were measured. The expression of phospho-AMPK(Thr172) and AMPK was determined by Western Blot. The results showed that adiponectin may exert direct anti-proliferative effects on HEC-1-A and RL95-2 cells by inducing cell cycle arrest and apoptosis, which may be mediated by AMPK pathway.
2.The effect of mutation of hepatitis B virus core protein dimer interface domain related to nucleocapsid formation on HBV replication
Kai DENG ; Dong JIANG ; Jinchao HAN ; Xiaoben PAN ; Hao WANG ; Lai WEI
Chinese Journal of Microbiology and Immunology 2010;30(8):730-736
Objective To investigate the effect of hepatitis B virus core protein (HBc) dimer interfaces amino acids mutation on nucleocapsid assembly and HBV DNA replication. Methods Based on HBc three dimension structure, four HBc dimer interfaces domain mutation plasmids, pHBc14-18M,pHBc120-135M,pHBc23-39M and pHBc122-139M were constructed in pcDNA3.1 vector by PCR site-directed mutagenesis, there was a flag-tag at the C-terminal of all mutants for easy detection. Wild type core protein plasmid 1-183flag was also constructed as a positive control. The 4 mutants were cotransfected HepG2 cells with pHBV1.2 core negative plasmid (pHBV1.2-core-) ,which contained 1.2 copies of HBV whole genome but the core protein would not express due to a stop codon. The capsid formation, HBV pregenome(pgRNA) and HBV DNA replication mediate were analyzed by native agarose gel electrophoresis and Western blot, Northern blot and Southern blot , respectively. The 4 mutants were also cotransfected HepG2 cells with HBV wild type plasmid pHBV1.2 and examined by Southern blot. Virions in the medium were determined by native agarose gel electrophoresis and Western blot. Results Cotransfecting HepG2 cells with pHBV1.2-core- plasmid, pHBc14-18M,pHBc120-135M and pHBc122-139M mutant groups formed nucleocapsid-like structure but pHBc23-39M could not, Northern and Southern blot displayed no signal in all mutants except 1-183flag conrol group. In pHBV1.2 cotransfection experiment, HBV DNA replication was blocked in pHBc14-18M, pHBc120-135M and pHBc122-139M mutant groups, sharply decreased in pHBc120-135M and pHBc122-139M groups, correspondingly virons production in medium were also inhibited. pHBc23-39M mutant exerted no influence on HBV replication. Conclusion pHBc23-39M mutant can neither form nucleocapsid-like structure nor interact with wild type HBc dimmer to interfere HBV replication.On the contrast, pHBc14-18M, pHBc120-135M and pHBc122-139M mutants can form nucleocapsid-like structure by themselves, but this structure does not support HBV DNA synthesis. Besides, they can effectively inhibit wild type HBV DNA replication by contacting with wild HBc dimmers resulting in nucleocapsid dysfunction.
3.Effects of insulin and β-adrenergic agonist on adiponectin secretion and expression in rats
Gang LI ; Li CONG ; Na YIN ; Qiang LI ; Wei WANG ; Jinchao ZHANG
Chinese Journal of Endocrinology and Metabolism 2011;27(1):62-65
Objective To explore the potential mechanisms of regulating adiponectin secretion and expression in vivo in rats.Methods To observe the regulation of adiponectin by fasing-refeeding and β-adrenergic agonists, male Wistar rats were fasted for 18 h and allowed to refeed or a β3-adrenergic receptor agonist was infused into refeeding rats.The effects of insulin clamp on adiponectin secretion and expression, including euglycemichyperinsulinemic clamp and hyperglycemic-hyperinsulinemic clamp, were also investigated.Plasma adiponectin level was determined by radioimmunoassay.Adiponectin mRNA expression in adipose tissue of rats was detected by realtime PCR.Results (1) Refeeding 18 h fasted rats increased plasma adiponectin concentration (about 2-fold) and adipose tissue adiponectin expression (about 3-fold), which were completely blocked by administration of β-adrenergic agonist.(2) Hyperinsulinemic clamp increased plasma adiponectin concentration and adiponectin gene expression in adipose tissue.Conclusions Adiponectin secretion and expression are acutely regulated in vivo by nutritional status.Insulin and β-adrenergic agonists regulate adiponectin secretion and expression in adipose tissue.
4.Relationship between serum retinol-binding protein 4 and non-alcoholic fatty liver disease
Binsong LIU ; Qiang LI ; Wei WANG ; Lin GUO ; Jing WANG ; Shu LIU ; Jinchao ZHANG
Chinese Journal of Endocrinology and Metabolism 2013;(1):58-61
Two hundred and sixty-one subjects were recruited from in-patients and subjects for phaysical Check-up,and were divided into normal control group (NC),nonalcoholic fatty liver disease group (NAFLD),type 2 diabetes mellitus group (T2DM),and T2DM accompanied by NAFLD group (DMN).According to the result of ultrasonic examination,the patients with T2DM were further divided into non-NAFLD group,light fatty liver group (NAFLD-L group),moderate fatty liver group(NAFLD-M group),and severe fatty liver(NAFLD-S group).Fasting plasma glucose,blood lipid,liver function,kidney function,and serum retinol-binding protein 4 (RBP4) levels were determined.The risk of various indicators for NAFLD was determined by correlation analysis and logistic regression analysis.The results showed that fasting glucose levels in diabetics with or without NAFLD were significantly higher than those in NC and NAFLD groups(P<0.01).Triglyceride (TG) level in DMN group was significantly higher than those in other three groups(all P<0.01),while high density lipoprotein-cholesterol level was lower than those in other three groups(all P<0.01).Systolic blood pressure and diastolic blood pressure in DMN group were higher than those in NC and T2DM groups (P<0.05 or P<0.01).Serum RBP4 level in patients with NAFLD was significantly higher compared with the subjects without NAFLD [45.00 (38.75,51.00) mg/L vs 51.00 (43.00,62.00) mg/L,P <0.01],and was rising with the progress of NAFLD [NAFLD-L group 44.00 (37.00,51.00) mg/L,NAFLD-M group 52.00(46.00,63.00) mg/L,and NAFLD-S group 78.5 (72.75,83.00) mg/L,all P<0.01].Logistic regression analysis showed that the RBP4 level was an independent factor associated with NAFLD (P =0.029).In addition,serum RBP4 level was correlated with body mass index,waist-to-hip ratio,serum gamma-glutamyl transpeptidase,total cholesterol,TG,aspartate aminotransferase,alanine aminotransferase,prealbumin,creatinine,blood urea nitrogen,and uric acid.These resuhs suggest that serum RBP4 is an independent risk factor of NAFLD.
5.Effects of Ephedra on the body mass and levels of blood lipids, blood glucose and hormones in ovarietomized obese rats
Jinchao SI ; Xiaoyu ZHANG ; Meiling LIN ; Yonghui DING ; Wei LI ; Songyi QU ; Tianzhen ZHENG
Chinese Journal of Tissue Engineering Research 2006;10(47):209-212
BACKGROUND: Ephedra, a Chinese medicine, is often used to treat obesity with relatively satisfying results recently. However, the effects of Ephedra on the perimenopausal and postmenopausal obese women remain unclear.OBJECTIVE: To observe the effects of oral Ephedra decoction on body mass and the levels of blood lipids, blood glucose and hormone in ovarietomized obese rats.DESIGN: A completely randomized and controlled experiment.SETTING: Institute of Physiology and Psychology, School of Basic Medical Sciences, Lanzhou University.MATERIALS: The experiment was performed in the Key Laboratory of Pre-clinical Study for New Drugs of Gansu Province and the Laboratory of Institute of Physiology and Psychology, School of Basic Medical Sciences,Lanzhou University from February 2006 to June 2006. Forty-four healthy female SD rats were randomly divided into four groups with 11 rats in each group, namely sham-operated group, ovariectomized group, estrogen replacement therapy group and Ephedra group.METHODS: ① After having been narcotized by cloraminone (110 mg/kg),rats were underwent a bilateral ovariectomy except those in the sham-operated group, which were also operated, but their ovaries were not cut off. ②Rats in the sham-operated group and ovariectomized group were subcutaneously injected with sesame oil (0.2 mL/each rat) every day postoperatively till the end of the experiment. ③ The rats in the estrogen replacement therapy group were given estradiol (1 mg/kg) by subcutaneous injection every day postoperatively till the end of the experiment. ④ The rats in the Ephedra group freely drank 1% water extracts from Ephedra postoperatively, later the concentration of Ephedra gradually increased to 8% on the sixth day, which lasted until the end of the experiment. ⑤ The food intake was monitored daily, and body mass was measured every ten days. ⑥ At the end of the experiment, all the rats were fasted for 12 hours and collected blood samples for the measurement of serum indexes. The body mass and body length were measured to calculate the Lee's index [(g)×103/body length (cm)] at the same time.MAIN OUTCOME MEASURES: ① Body mass and Lee's index at different time points in each group. ② Food intake at different time points in each group. ③ Levels of blood lipids and blood glucose in each group. ④Levels of estrogen, progesterone and insulin in each group.RESULTS: Forty-four rats all entered the analysis of results. ① Result of body mass and Lee's index at different time points: The body masses on the 20th, 30th, 40th and 50th days in the ovariectomized group were (256.4±14.3),(271.3±16.1), (276.4±12.7), (285.7±24.2) g, which were significantly higher than those in the sham-operated group [(226.5±11.5), (241.8±12.6),(243.1±13.5), (251.1±22.4) g, P < 0.05-0.01], and the Lee's index in the ovariectomized group was greater than that in the sham-operated group(317.2±13.5, 280.4±11.2, P < 0.01). The body masses on the 40th and 50th days in the estrogen replacement therapy group were (243.7±14.8) and(246.2±11.9) g, which were significantly lower than those in the ovariectomized group (P < 0.05-0.01), and the Lee's index (289.9±13.5) was lower than that in the ovariectomized group (P < 0.01). The body masses on the 40th and 50th days in the Ephedra group were (245.4 ±14.1) and(252.4±14.9) g, and the Lee's index was 294.4±11.0, which were all lower than those in the ovariectomized group (P < 0.05). ② Result of Food in take at different time points: The food intakes on the 30th, 40th and 50th days in the Ephedra group were (17.8±2.4), (22.3±3.9), (26.1±3.5) g per day,which were decreased as compared with those in the ovariectomized group[(25.9±4.7), (28.5±5.3), (32.8±5.5) g per day, P < 0.05]. ③ Levels of blood lipids and blood glucose: The levels of triglyceride, cholesterol and low density lipoprotein cholesterol (LDL-C) in the ovariectomized group were (1.73±0.32), (1.45±0.50), (0.78±0.19) mmol/L, which were higher than those in the sham-operated group [(0.94±0.29), (1.05±0.30), (0.08±0.11) mmol/L, P < 0.01]. After the estrogen replacement therapy, the levels of triglyceride, cholesterol, LDL-C and blood glucose were (1.10±0.34),(1.14±0.30), (0.17±0.05), (5.88±1.21) mmol/L, which were lower than those in the ovariectomized group (P < 0.05-0.01), but the level of high density lipoprotein cholesterol (HDL-C) was higher than that in the ovariectomized group [(1.11±0.31), (0.88±0.21) mmol/L, P < 0.05]. The levels of triglyceride, cholesterol, LDL-C and HDL-C in the Ephedra group were (0.97±0.16), (1.11±0.20), (0.59±0.07) and (0.45±0.061) mmol/L, which were lower than those in the ovariectomized group (P < 0.05-0.01). ④ The serum levels of estrogen, progesterone and insulin in each group: The serum levels of estrogen and progesterone in the ovariectomized group were lower than those in the sham-operated group [(17.09±9.00), (28.51 ±7.99) μg/L;(58.69±12.11), (62.73±10.93) μg/L, P < 0.01], the serum level of insulin was higher than that in the sham-operated group [(31.74±6.69),(23.75±6.66) mU/L, P < 0.01]. The serum levels of estrogen in the estro gen replacement therapy and Ephedra group were (36.03±8.83) and (30.18±8.61) ng/L, which were higher than those in the ovariectomized group(P < 0.05-0.01), the level of insulin were (21.34±4.57), (24.86±6.20) mU/L,which were lower than those in the ovariectomized group (P < 0.05-0.01).The serum level of progesterone in the Ephedra group [(17.68±6.19) μg/L]was lower than that in the ovariectomized group (P < 0.01).CONCLUSION: Ephedra can promote loss of body mass, reduce levels of the blood lipids and insulin, and increase the serum levels of hormones in ovariectomized obese rats.
6.Evaluation on the intervention of Nitroglycerin on ~(99m)TC-MIBI myocardial tomography imaging for myocardial viability
Yumei XIAO ; Jinning YANG ; Fan LI ; Mingjian XIAN ; Xuejing JIN ; Rongguang GUAN ; Jinchao CHEN ; Wei HUANG ; Yu KE
Journal of Chinese Physician 2001;0(05):-
Objective To study the value of the intervention of Nitroglycerin(NTG) on ~(99m)Tc-MIBI myocardial tomography imaging in the estimation of myocardial viability.Method According to the analysis of autologous electrocardiogram(ECG),a total of 66 patients with unstable angina(UA) was divided into group A and group B.The patients in group A were without old myocardial infarction and those of group B were with old myocardial infarction.The patients in the two groups were respectively underwent the resting ~(99m)Tc-MIBI myocardial tomography imaging and the NTG administration intervened the next day.The imaging was collected and tested by the computer.Results Of 594 myocardial segments in the 66 cases,242 segments(40.7%) in testing myocardial imaging were proved to be perfusion abnormal,while after the intervention of NTG administration,the perfusion of 114 segments(47.1%) had been improved according to the imaging.Conclusion The intervention of NTG administration on ~(99m)Tc-MIBI myocardial tomography imaging for myocardial viability is simple,safe,objective and accurate,which provides a forceful means for the post-operative evaluation and the selection of indication before the rebuilding of coronary artery of the UA patients.
7.Selective neurotomy of anterior ethmoid nerve and squeezing operation of inferior turbinate for the treatment of perennial allergic rhinitis.
Yongxin WEI ; Jinchao LIN ; Qiaoling CHEN ; Zhizhong SHEN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;23(11):499-500
OBJECTIVE:
To explore the treatment effect of selective neurotomy of anterior ethmoid nerve and squeezing operation of inferior turbinate in the treatment of perennial allergic rhinitis (PAR).
METHOD:
Seventy cases of perennial allergic rhinitis were selected and subjected to selective neurotomy of anterior ethmoid nerve and squeezing operation of inferior turbinate,and the treatment effect was observed by analysis of the the symptoms and signs score of all cases preoperatively and postoperatively.
RESULT:
The total effective rate were 90.0% at 1 year follow-up.
CONCLUSION
Selective neurotomy of anterior ethmoid nerve and squeezing operation of inferior turbinate were effective for the patients with PAR. It is worthy to be popularized for its convent and rare complications.
Adolescent
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Adult
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Denervation
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methods
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Ethmoid Bone
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innervation
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Female
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Humans
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Male
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Middle Aged
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Rhinitis, Allergic, Perennial
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surgery
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Turbinates
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surgery
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Young Adult
8.The accuracy of real-time continuous monitoring system at different stages and its association with glucose excursion
Yingying WANG ; Qiang LI ; Ping YU ; Ping GAO ; Yanhui WU ; Can CUI ; Wei WANG ; Lili CHEN ; Hong QIAO ; Lihong WANG ; Ying XIANG ; Zhong ZHANG ; Jinchao ZHANG
Chinese Journal of Endocrinology and Metabolism 2011;27(3):224-228
Objective To study the accuracy of real-time continuous monitoring system (RT-CGMS) at different stages and its association with glucose excursion. Methods Totally 33 patients with type 1 diabetes or type 2diabetes were under surveillance of RT-CGMS for 5 d. Capillary glucose values were measured 7 times daily.Correlation coefficient, error grid analysis (EGA), and Bland-Altman analysis methods were used to assess the correlation, accuracy and agreement of RT-CGMS at different stages and in general level; The mean amplitude of glucose excursion (MAGE) and the frequency of glucose excursion ( FGE ) were also calculated. Results ( 1 ) The correlation coefficient of RT-CGMS with capillary glucose values at fasting, postprandial stages, and in general level were 0.94,0.92, and 0.93 respectively( P<0.01 ). (2) EGA showed that 98.82%, 98.39%, and 98.64% of the results fell in the A and B zones and 1. 18%, 1.61%, and 1.36% fell in the D zone respectively at fasting,postprandial stages, and in general level. There is no result fell in C and E zones. ( 3 ) The agreement analysis showed that RT-CGMS readings were in close agreement with capillary glucose values at fasting, postprandial periods, and in general level. (4)The MAGE at fasting, postprandial periods, and in general level were (3.57±2.66), (4.07±3.09), and (4. 02 ±3.04) mmol/L (P>0. 05), (0±0. 5), (3± 1), and( 1 ±3) d for FGE (P<0. 01 ).Conclusion RT-CGMS at fasting stage has higher accuracy than postprandial stage and general level, FGE at fasting stage is higher than postprandial stage and general level.
9.Study on norovirus GⅡ.4/Sydney 2012 variant in China
Huan MAI ; Yan GAO ; Xiaoben PAN ; Jinchao HAN ; Xu CONG ; Lai WEI
Chinese Journal of Epidemiology 2014;35(2):157-162
Objective To study the nucleotide and amino acid sequences of norovirus G Ⅱ.4/Sydney 2012 variants,in China.Methods Twenty-two stool specimens,confirmed as G Ⅱ.4/Sydney 2012-positive were collected from Beijing in the winter of 2012-2013.RT-PCR was performed to target the complete capsid gene.G Ⅱ.4/Sydney 2012 strains from other regions in China were searched and obtained from the GenBank.Nucleotide and amino acid sequences of G Ⅱ.4/Sydney 2012 strains were analyzed,using the CLUSTAL X (Version 1.83)and followed by phylogenetic analysis using Mega version 5.1.Results The complete major capsid nucleotide sequences of thirty-eight G Ⅱ.4/Sydney 2012 strains from seven regions in China were obtained.The VP1 nucleotide and amino acid sequences diversity were 0.1%-3.3% and 0-3.1%,respectively.Result from phylogenetic analysis demonstrated that the G Ⅱ.4/Sydney 2012 variant shared a common ancestor with both the dominant norovirus G Ⅱ.4 variants Apeldoom 2008 and the New Orleans 2009.G Ⅱ.4/Sydney 2012 variants appeared to have had two A/D/E site combinations at the amino acid level,TSRN-GTT-SNT and TSRN-STT-SNT.Conclusion G Ⅱ.4/Sydney 2012 variant had been circulating in many regions in China.There seemed a high nucleotide and amino acid identity among G Ⅱ.4/Sydney 2012 strains collected from China.G Ⅱ.4/Sydney 2012 variants showed different A/D/E site combination from other pandemic G Ⅱ.4 variants.
10.LncRNA PART1 Attenuates Myocardial Ischemia-Reperfusion Injury by Regulating TFAP2C/DUSP5 Axis via miR-302a-3p
Min ZENG ; Xin WEI ; Jinchao ZHOU ; Siqi LUO
Korean Circulation Journal 2024;54(5):233-252
Background and Objectives:
Myocardial ischemia-reperfusion injury (MIRI) refers to the damage of cardiac function caused by restoration of blood flow perfusion in ischemic myocardium. However, long non-coding RNA prostate androgen regulated transcript 1 (PART1)’s role in MIRI remain unclear.
Methods:
Immunofluorescence detected LC3 expression. Intermolecular relationships were verified by dual luciferase reporter assay. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, flow cytometry and transferase-mediated dUTP nick-end labeling (TUNEL) assays analyzed cell viability and apoptosis. The release of lactate dehydrogenase was tested via enzyme-linked immunosorbent assay (ELISA). Left anterior descending coronary artery surgery induced a MIRI mouse model. Infarct area was detected by 2,3,5-triphenyltetrazolium chloride staining. Hematoxylin and eosin staining examined myocardial injury. ELISA evaluated myocardial marker (creatine kinase MB) level.
Results:
PART1 was decreased in hypoxia/reoxygenation (H/R) induced AC16 cells and MIRI mice. PART1 upregulation attenuated the increased levels of Bax, beclin-1 and the ratio of LC3II/I, and enhanced the decrease of Bcl-2 and p62 expression in H/R-treated cells.PART1 upregulation alleviated H/R-triggered autophagy and apoptosis via miR-302a-3p. Mechanically, PART1 targeted miR-302a-3p to upregulate transcription factor activating enhancer-binding protein 2C (TFAP2C). TFAP2C silencing reversed the protected effects of miR-302a-3p inhibitor on H/R treated AC16 cells. We further established TFAP2C combined to dual-specificity phosphatase 5 (DUSP5) promoter and activated DUSP5. TFAP2C upregulation suppressed H/R-stimulated autophagy and apoptosis through upregulating DUSP5.Overexpressed PART1 reduced myocardial infarction area and attenuated MIRI in mice.
Conclusion
PART1 improved the autophagy and apoptosis in H/R-exposed AC16 cells through miR-302a-3p/TFAP2C/DUSP5 axis, which might provide novel targets for MIRI treatment.