1.Transcriptional activation function of intracellular domain of EPOR and G-CSFR protein in yeast: a comparative study
Bo CHEN ; Xiaoling YUAN ; Zhenyu YAO
Medical Journal of Chinese People's Liberation Army 2001;0(08):-
Objective To investigate the feasibility of searching for proteins which interact with intracellular domain of hematopoietic growth factor receptors using yeast two-hybrid system. Methods RT-PCR method was performed to amplify the genes of intracellular domains of G-CSF receptor and EPO receptor in NFS-60 and BET-2 cells of mice. The genes were cloned into yeast expression plasmid pGBKT7 vector,and then transformed into yeast AH109. The yeast proteins were isolated and analyzed with Western blotting. Transcriptional activation was analyzed by the ?-galactosidase colony-lift filter assay. Results The intracellular domains of G-CSF receptor and EPO receptor genes were successfully cloned into pGBKT7 vector. The results of Western blotting assay showed that both proteins were expressed in the yeast cells. The ?-galactosidase colony-lift filter assay demonstrated that G-CSF receptor alone had no activity of transcriptional activation,while the EPO receptor alone could activate transcription. Conclusion The findings suggested that intracellular domain of G-CSF receptor could be used as a bait to find interacting proteins using yeast two-hybrid system,while that of the EPO receptor could not. Therefore the system could not be applied to all hematopoietic factor receptor.
2.Observation on Therapeutic Effects of Compound Glycyrrhizin Combined with Halometasone Cream in Treating Vitiligo
Xinghai YUAN ; Xinmin YAO ; Yuping WANG ; Bo ZHU
China Pharmacy 2005;0(14):-
OBJECTIVE:To observe the therapeutic effects of compound glycyrrhizin combined with halometasone cream on vitiligo.METHODS:95patients with vitiligo were randomly divided into treatment group and control group.64patients in treatment group had taken compound glycyrrhizin continuously for60days at a dose of1~3tablets with3times per day,meanwhile,0.05%halometasone cream had been used locally with2times per day.While31other patients in control group had only been treated with0.05%halometasone cream with2times per day for60days.RESULTS:The efficacy rate in treatment group was62.5%as compared with32.3%in control group,there existed significant difference between the two groups(P
3.The mechanism of alteronol inhibiting the proliferation of human promyelocytic leukemia HL-60 cells.
Liangliang LIU ; Na CHEN ; Xuan YUAN ; Ying YAO ; Bo ZHANG ; Qiusheng ZHENG
Acta Pharmaceutica Sinica 2012;47(11):1477-82
This study is to investigate the mechanism of human promyelocytic leukemia HL-60 cells proliferation induced by alteronol in vitro. Human promyelocytic leukemia HL-60 cells cultured in vitro were treated with different concentrations of alteronol. Inhibition rate was detected by SRB assay. Cellular morphological changes were observed by Hoechst and AO/EB (acridine orange/ethidium bromide dye) staining. The apoptosis rate was determined by Annexin V-FITC/PI assay. Cell cycle distribution was determined by flow cytometry. Western blotting analysis was carried out to determine the cell cycle related proteins. The proliferation of HL-60 cells treated with alteronol was inhibited in a concentration-dependent manner. Based on cell viability assay, observation on cell morphology and apoptosis rate, it confirmed that alteronol played an obvious role in proliferation inhibition of human promyelocytic leukemia HL-60 cells, but it did not induce apoptosis in human promyelocytic leukemia HL-60 cells in different concentrations groups. Alteronol could effectively inhibit the proliferation of human promyelocytic leukemia HL-60 cells inducing cell cycle arrest at G1 phase, as well as, alteration expression of cell cycle proteins level of CyclinD1 and pRb.
4.Regular expression of discoidin domain receptor 2 in the improved adjuvant-induced animal model for rheumatoid arthritis.
Wei LI ; Yuan-qiang ZHANG ; Xin-ping LIU ; Li-bo YAO ; Lan SUN
Chinese Medical Sciences Journal 2005;20(2):133-137
OBJECTIVETo investigate the expression of discoidin domain receptor 2 (DDR2) of fibroblast-like synovial cells in improved adjuvant-induced animal (AIA) model for rheumatoid arthritis (RA) and to provide evidence for DDR2's antagonist use clinically.
METHODSAIA was modified by administrating 0.1 mL of complete Freund's adjuvant (CFA, mixed with 5 mg Bacillus Calmette-Guerin vaccine/mL) into rats' right hind paws and 0.125 mL tumor necrosis factor-alpha (2 U/mL) into right ankles and subpatellar fatty tissue. The expression of DDR2 in fibroblast-like synovial cells was assessed using immunohistochemistry, immunofluorescence histochemistry, and in situ hybridization methods. Levels of anti-collagen II antibody were measured using enzyme-linked immunosorbent assay.
RESULTSGiven the terms mentioned above, we found a more practical rat model, apparently decreasing immunization time (average 3-5 days). DDR2 can be detected upon the 15th day of immunization; expression gradually increased with time going on, and reaching a peak 35 days after immunization before gradually decreasing. Serum anti-collagen II antibody showed similar expression patterns as DDR2, but reached peak later than DDR2, about 40 days after immunization.
CONCLUSIONRegular expression of DDR2 in animal models infers its important role in the pathological process of RA.
Animals ; Antibodies ; blood ; Arthritis, Experimental ; etiology ; metabolism ; Arthritis, Rheumatoid ; chemically induced ; metabolism ; BCG Vaccine ; Collagen Type II ; immunology ; Discoidin Domain Receptors ; Female ; Fibroblasts ; drug effects ; metabolism ; pathology ; Freund's Adjuvant ; Rats ; Rats, Sprague-Dawley ; Receptor Protein-Tyrosine Kinases ; metabolism ; Receptors, Mitogen ; metabolism ; Synovial Fluid ; cytology ; metabolism
5.Evaluation of the application of laparoscopic sleeve gastrectomy in bariatric and metabolic surgery.
Chinese Journal of Gastrointestinal Surgery 2013;16(10):1017-1020
Bariatric surgery is one of the most effective treatment options for obesity and type 2 diabetes mellitus. In recent years, laparoscopic sleeve gastrectomy has become increasingly popular due to simple operation, high safety and significant efficacy. In this article, the progress and the application evaluation of laparoscopic sleeve gastrectomy for the treatment of obesity and type 2 diabetes mellitus in the field of bariatric and metabolic surgery are reviewed.
Bariatric Surgery
;
Diabetes Mellitus, Type 2
;
complications
;
Gastrectomy
;
Humans
;
Laparoscopy
;
Obesity, Morbid
;
complications
;
Treatment Outcome
6.Prenatal sonographic diagnosis on agenesis of fetal corpus callosum
Bo GONG ; Shengli LI ; Congying CHEN ; Juling LIU ; Yuan YAO ; Huaxuan WEN ; Jingru BI ; Xiaodong YANG ; Yumei LIAO ; Xiulan CHEN
Chinese Journal of Ultrasonography 2008;17(3):224-226
Objective To assess the value of prenatal sonographic diagnosis on fetal agenesis of corpus callosum(ACC). Methods A retrospective study was on prenatal sonographic findings of 10 cases with ACC malformation and their abnormalities in central nervous system (CNS) or extra-CNS.Results The special sonographic findings established the diagnosis of ACC malformation in all 10 cases,with 7cases diagnosed absence agenesis. Among all patients with ACC,6 cases were accompanied with abnormalities in extra-CNS,8 in CNS and 5 in both. Conclusions Prenatal ultrasonography plays a vital role in accurate diagnosis on fetal ACC. Attentions should be paid to the indirect encephalic features and complicated abnormalities so as to make accurate and prompt diagnosis.
7.Research advances on relationship between gut microbiota and anorexia nervosa
Runxue YUAN ; Shining BO ; Gaiqi YAO
Chinese Journal of General Practitioners 2021;20(7):799-802
Anorexia nervosa is a mental disorder characterized by an intense fear of gaining weight, a distortion of body image, longstanding dietary restriction and low body weight, which has an impact on the cognition, behavior and emotion of patients to varying degrees. Recently, thanks to the advances of microbiota-gut-brain axis research, the role of gut microbiota in the mechanisms of anorexia nervosa has been elucidated. The application of probiotics has been proved to alleviate anxiety, depression and improve appetite, which is expected to be an important measure for the treatment of anorexia nervosa in the future.
8.Changes of interleukin-1 beta, tumor necrosis factor alpha and interleukin-6 in brain and plasma after brain injury in rats.
Tao ZHU ; Zhi YAO ; Han-na YUAN ; Bo-gang LU ; Shu-yuan YANG
Chinese Journal of Traumatology 2004;7(1):32-35
OBJECTIVETo study the changes of interleukin-1 beta (IL-1beta), tumor necrosis factor alpha (TNFalpha) and interleukin-6 (IL-6) levels in brain and plasma after brain injury and to assess the relationship between the cytokine levels and injury severity in rats.
METHODSA total of 51 male Wistar rats, weighing 280-340 g, were anesthetized with chloral hydrate (400 mg/kg body weight) through intraperitoneal injection and fixed on a stereotaxic instrument. Severe brain injury was created in 16 rats (severe injury group) and moderate brain injury in 18 rats (moderate injury group) by a fluid percussion model, and cytokine levels of IL-1beta, TNFalpha and IL-6 were measured with biological assay. And sham operation was made on the other 17 rats (control group).
RESULTSIn the control group, the levels of IL-1beta, TNFalpha and IL-6 were hardly detected in the cortex of the rats, but in the ipsilateral cortex of the rats in both injury groups, they increased obviously at 8 hours after injury. The increasing degree of these cytokines had no significant difference between the two injury groups. The levels of IL-6 in the plasma of all the rats increased slightly, whereas the levels of IL-1beta and TNFalpha were undetectable.
CONCLUSIONSThe increase of IL-1beta, TNFalpha and IL-6 levels is closely related to brain injury. The increased cytokine levels in the central nervous system are not parallel to those in the peripheral blood. It suggests that inflammatory cytokines play important roles in the secondary neural damage after brain injury.
Animals ; Biomarkers ; analysis ; Blood Chemical Analysis ; Brain ; metabolism ; Brain Injuries ; diagnosis ; metabolism ; Disease Models, Animal ; Injury Severity Score ; Interleukin-1 ; analysis ; metabolism ; Interleukin-6 ; analysis ; metabolism ; Male ; Probability ; Prognosis ; Rats ; Rats, Wistar ; Sensitivity and Specificity ; Tumor Necrosis Factor-alpha ; analysis ; metabolism
9.Effect of coking oven emissions on level of serum oxidation-reduction and peripheral white blood cellular ras P21 and P53 in coke oven workers.
Qiao ZHANG ; Fang ZHOU ; Wu YAO ; Yi-bo ZHAO ; Zhi-yuan LI ; Yu-bao XU ; Yi-ming WU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2008;26(4):212-214
OBJECTIVETo explore the adverse effects of coking oven emissions (COE) on the serum oxidation-reduction and relevant genes in the exposed workers.
METHODSFifty-six coke oven workers and forty controls were investigated. Serum Malondialdehyde (MDA) levels and the activities of total superoxide dismutases (T-SOD) were measured by spectrophotometrical method. Immunohistochemical method was used to assess the P21 and P53 levels in peripheral white blood cells.
RESULTSCompared with controls, the individuals exposed to COE had significantly increased levels of serum MDA [(5.30 +/- 2.29) nmol/mL, P < 0.01] and markedly decreased levels of T-SOD [(100.04 +/- 10.75) NU/mL]. Additionally, the median levels of P53 and P21 were markedly increased in the exposed individuals compared with the controls (21.4% and 23.2%, respectively, all P < 0.05).
CONCLUSIONThe findings indicate that occupational exposure to COE causes the rise of serum oxidation-reduction MDA and the fall of T-SOD, and increasing expression levels of P21 and P53 proteins before the occurrence of apparent clinical symptoms.
Adult ; Air Pollutants, Occupational ; adverse effects ; Case-Control Studies ; Coke ; adverse effects ; Female ; Humans ; Lymphocytes ; metabolism ; Male ; Malondialdehyde ; blood ; Middle Aged ; Occupational Exposure ; adverse effects ; Oxidation-Reduction ; Proto-Oncogene Proteins p21(ras) ; metabolism ; Superoxide Dismutase ; blood ; Tumor Suppressor Protein p53 ; metabolism ; Young Adult
10.Effect of Tibetan medicine zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2.
Xiang-Yang LI ; Yong-Nian LIU ; Yong-Ping LI ; Jun-Bo ZHU ; Xing-Chen YAO ; Yong-Fang LI ; Mei YANG ; Ming YUAN ; Xue-Ru FAN ; Yue-Miao YIN
Acta Pharmaceutica Sinica 2014;49(2):267-272
To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.
Administration, Oral
;
Animals
;
Arylamine N-Acetyltransferase
;
genetics
;
metabolism
;
Caffeine
;
metabolism
;
urine
;
Cytochrome P-450 CYP1A2
;
genetics
;
metabolism
;
Dose-Response Relationship, Drug
;
Drugs, Chinese Herbal
;
administration & dosage
;
pharmacology
;
Female
;
Male
;
Medicine, Tibetan Traditional
;
RNA, Messenger
;
metabolism
;
Rats
;
Rats, Sprague-Dawley
;
Theophylline
;
urine
;
Uracil
;
analogs & derivatives
;
urine
;
Uric Acid
;
analogs & derivatives
;
urine
;
Xanthines
;
urine