1.Investigation on the genotyping of Acinetobacter baumannii by DiversiLab system
Yanyan QU ; Wenfei WANG ; Hua ZHOU ; Yiqi FU ; Yunsong YU
Chinese Journal of Laboratory Medicine 2010;33(5):425-429
Objective To evaluate the usefulness of DiversiLab system for genotyping of Acinetobacter baumannii.Methods Fifty-eight non-duplicated clinical Acinetobacter baumannii isolated from 15 cities in China in 2005 were typed by rep-PCR-based DiversiLab system.The results were compared with those of PFGE and multilocus sequence typing. Simpson's index of diversity was used to compare the discriminatory power among DiversiLab system, PFGE and MLST.Results Fifty-eight Acinetobacter baumannii isolates were differentiated into 5 clusters and 25 unique types by DiversiLab system. MLST identified 35 distinct sequence types, which fell into one clone complex of CC22 and 35 singletons, while PFGE resolved 5 pulsotypes and 34 unique types.Simpson's diversity indices for DiversiLab system, MLST and PFGE were O.876, O.944 and 0.961, respectively.Conclusions The discriminatory power of DiversiLab system is lower than that of PFGE and MLST.But as a simple, fast and reproducible typing method, it could be used as a first-line typing tool for the analysis of a large number of isolates.
2.Serial research of traceability and harmonization of thyroid-stimulating hormone detection approved by IFCC
Xiang YU ; Wenfei CHEN ; Xiaoliu YU ; Ming JIANG ; Ke LI ; Zhiguang YU
International Journal of Laboratory Medicine 2016;37(6):751-753
Objective To implement the harmonization of thyroid-stimulating hormone(TSH) of Mindray assay system by par-ticipating in harmonization research program of International Federation of Clinical Chemistry (IFCC)-Standardization of Thyroid Function Tests(C-STFT ) .Methods A total of three combinations of TSH reagents and calibrators were used to measure 20 serum samples of healthy human .Within-run precision and batch-to-batch variation were assessed .The concept of harmonization was dem-onstrated by comparing our test results with All Procedure Trimmed Mean (APTM ) provided by IFCC and recalibrating with Mater Calibrators .Results The within-run precision was 1 .96% ,batch-to-batch variation was 0 .47% - 1 .15% ,depending on the level of TSH analyte .There existed a positive bias compared to APTM values .After recalibration with Mater Calibrators and Passing &Bablok regression ,the slope of method comparison was 1 .0 ,and correlation coefficient was more than 0 .975 .Conclusion By using a panel of real human specimen and recalibration based on APTM ,the test results of Mindray assay system could be harmonized with mainstream manufacturers globally .
3.FGF-21 protects H9c2 cardiomyoblasts against hydrogen peroxide-induced oxidative stress injury.
Miaomiao HAN ; Wenfei WANG ; Mingyao LIU ; Deshan LI ; Bing ZHOU ; Yinhang YU ; Guiping REN
Acta Pharmaceutica Sinica 2014;49(4):470-5
Fibroblast growth factor-21 (FGF-21) is an important metabolism regulator, however, whether FGF-21 has effects on cardiovascular remains unclear. In this study, H2O2-induced injury in H9c2 cells was used as a cell model, the anti-apoptosis potential and mechanism of FGF-21 against oxidative injury were evaluated by MTT assay, flow cytometry assay and real-time PCR. The results showed that FGF-21 could increase the cell survival of H2O2-induced injury in H9c2 cells and prevent H9c2 cells from oxidative stress-induced apoptosis. Furthermore, FGF-21 can elevate SOD activity and regulate Bcl-2/Bax expression in H9c2 cells. The results suggest that FGF-21 have protective effect against the H2O2-induced apoptosis in H9c2 cells.
4.Effect of Pulmonary Rehabilitation on Acute Exacerbation of Chronic Obstructive Pulmonary Disease
Wenfei YAO ; Chunlin TU ; Yuhua FU ; Kaishun ZHAO ; Yanfang YU ; Jianrong HU
Chinese Journal of Rehabilitation Theory and Practice 2017;23(1):101-105
Objective To investigate the availability and safety of pulmonary rehabilitation for hospitalized patients with acute exacerba-tion of chronic obstructive pulmonary disease (COPD). Methods Seventy-two hospitalized patients with acute exacerbation of COPD were randomly included into test group (n=36) and control group (n=36) from June, 2015 to June, 2016. All the patients accepted management of anti-infection, phlegm elimination, antiasthma, etc., as well as the guidance of expectoration and health education; while the test group ac-cepted pulmonary rehabilitation from the third day of admission to discharge. Their strength of hand grip, 1-minute sit-to-stand test (STST), the days of hospitalization, lung function parameters, modified Medical Research Council (mMRC) scores and COPD Assessment Test (CAT) scores were measured before and after treatment. Results Compared with the control group, the strength of hand grip (t=2.985, P<0.01) and number of STST (t=2.024, P<0.05) increased, while the scores of CAT (t=3.222, P<0.01) and mMRC (t=2.212, P<0.05) de-creased in the test group. The hospital stay seemed to be shorter in the test group than in the control group, but there was no significant dif-ference (t=1.433, P>0.05). There was no significant difference in lung function after treatment in both groups (Z<1.031, P>0.05). Conclu-sion Pulmonary rehabilitation is effective on hospitalized patients with acute exacerbation of COPD in muscle strength, capability of activi-ties, and relieve the symptoms.
5.FGF-21 Improves Glucose Uptake and Glycogen Synthesis of Insulin-resistant Liver Cells
Mingyao LIU ; Wenfei WANG ; Yixue YU ; Yuting HOU ; Guiping REN ; Deshan LI
Progress in Biochemistry and Biophysics 2006;0(10):-
Insulin resistance in insulin sensitive organ results in metabolic disorder such as hyperglycemia, hyperinsulinemia and hyper triglyceridemia which are common features of type 2 diabetes.Insulin resistance in liver cells mainly causes impaired glycogen synthesis, failed to suppress glucose production which is the major contribution to hyperglycemia.FGF-21 as a new metabolic regulator can control fasting blood glucose.The mechanism of FGF-21 effects on regulating plasma glucose has little to known.In order to establish an in vitro insulin resistant model of liver cells and evaluate the effects and mechanism of FGF-21 on glucose metabolism in the cell model, HepG2 cells were incubated with 10-7 mol/L insulin for 24 h to build insulin-resistant cell model.To evaluate the cells for insulin resistance, the cells were stimulated with fresh insulin for 24 h and the glucose uptake by these cells was carried out.The insulin-resistant cells were treated with different concentrations of FGF-21 for 24 h and insulin-treated cells were used as a control.The glucose uptake by the cells was detected by the method of glucose oxidizes/peroxides(GOD-POD);the synergy between insulin and FGF-21 was evaluated.The mRNA expression of GLUT1 in the insulin-resistant cells was detected by the real-time PCR.Glycogen synthesis of the cells was examined by the anthrone method.The results showed that HepG2 cells treated with 10-7 mol/L insulin for 24 h became resistant to insulin and the insulin resistance status was maintained for 48 h without change of cell morphology.FGF-21 could stimulate glucose consumption of the insulin-resistant model in a dose-dependent manner.The glucose consumption and glycogen synthesis of the insulin-resistant model were significantly improved by FGF-21 treatment.FGF-21 showed strong synergy with insulin in glucose uptake and glycogen synthesis of the model cells.While the cells became resistant to insulin, FGF-21 could increase the mRNA expression of GLUT1.Thus, It is concluded that FGF-21 stimulates glucose uptake in insulin resistant HepG2 cells through GLUT1 expression, stimulates glycogen synthesis and improves the glucose metabolism in the insulin resistant liver cell model.
6.Effect of FGF-21 on learning and memory ability and antioxidant capacity in brain tissue of D-galactose-induced aging mice.
Yinhang YU ; Guiping REN ; Yaonan LIU ; Susu QU ; Fuliang BAI ; Tong ZHANG ; Wenfei WANG ; Guiyou TIAN ; Xianlong YE ; Deshan LI
Acta Pharmaceutica Sinica 2014;49(7):1000-6
This study aims to investigate the effects of fibroblast growth factor 21 (FGF-21) on learning and memory abilities and antioxidant capacity of D-galactose-induced aging mice. Kunming mice (37.1 +/- 0.62) g were randomly divided into normal control group, model group and FGF-21 high, medium and low dose groups (n = 8). Each group was injected in cervical part subcutaneously with D-galactose 180 mg x kg(-1) x d(-1) once a day for 8 weeks. At the same time, FGF-21-treated mice were administered with FGF-21 by giving subcutaneous injection in cervical part at the daily doses of 5, 2 and 1 mg x kg(-1) x d(-1). The normal control group was given with normal saline by subcutaneous injection in cervical part. At seventh week of the experiment, the learning and memory abilities of mice were determined by water maze and jumping stand tests. At the end of the experiment, the mice were sacrificed and the cells damage of hippocampus was observed by HE staining in each group. Reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and total antioxidant capacity (T-AOC) in the brain of mice were determined. The results showed that different doses of FGF-21 could reduce the time reaching the end (P < 0.01 or P < 0.05) and the number of touching blind side (P < 0.01 or P < 0.05) in the water maze comparing with the model group. It could also prolong the latency time (P < 0.05) and decrease the number of errors (P < 0.01 or P < 0.05) in the step down test. The result of HE staining showed that FGF-21 could significantly reduce brain cell damage in the hippocampus. The ROS and MDA levels of three different doses FGF-21 treatment group reduced significantly than that of the model group [(5.58 +/- 1.07), (7.78 +/- 1.92), (9.03 +/- 1.77) vs (12.75 +/- 2.02) pmol (DCF) x min(-1) x mg(-1), P < 0.01 or P < 0.05], [(2.92 +/- 0.71), (4.21 +/- 0.81), (4.41 +/- 0.97) vs (5.62 +/- 0.63) nmol x mg(-1) (protein), P < 0.01]. Comparing with the model group, the activities of SOD, GPx, CAT and T-AOC of the three different doses FGF-21 treatment groups were also improved in a dose-dependent manner. This study demonstrates that FGF-21 can ameliorate learning and memory abilities of D-galactose induced aging mice, improve the antioxidant abilities in brain tissue and delay brain aging. This finding provides a theoretical support for clinical application of FGF-21 as a novel therapeutics for preventing aging.
7.Optimization and characterization of a novel FGF21 mutant.
Xianlong YE ; Huashan GAO ; Wenfei WANG ; Guiping REN ; Mingyao LIU ; Kun HE ; Yakun ZHANG ; Jingzhuang ZHAO ; Dan YU ; Deshan LI
Acta Pharmaceutica Sinica 2012;47(7):897-903
Fibroblast growth factor 21 (FGF21) is a member of FGF family. It has been demonstrated that FGF21 is an independent, safe and effective regulator of blood glucose levels in vivo. In order to improve the activity of FGF21, we exchanged the beta10-beta12 domain of the human FGF21 with that of the mouse FGF21 to construct a novel FGF21 gene (named hmFGF21), and then subcloned hmFGF21 gene into the SUMO expression vector to create pSUMO-hmFGF21 and transformed it into E. coli Rosetta for expression of the fusion protein SUMO-hmFGF21. Both in vitro and in vivo glucose regulation activity of hmFGF21 was evaluated. The SDS-PAGE result showed that compared with wild-type hFGF21, the soluble expression of hmFGF21 increased about 2-fold. HmFGF21 was more potent in stimulation of glucose uptake in HepG2 cells in vitro. The results of anti-diabetic effect on db/db mice demonstrated that hmFGF21 had better efficacy on controlling the blood glucose of the db/db diabetic animals than wild-type hFGF21. These results suggest that the biological properties of FGF21 are significantly improved by optimization.
8.Immunogenicity of Poria cocos polysaccharides PCP-Ⅰand PCP-Ⅱas vaccine adjuvants
Haixia LI ; Kunlu LIU ; Wenfei LI ; Peiyuan JIA ; Weili YU ; Junhua WU ; Tao HU ; Yuxia WANG ; Junjie SHAN ; Guohui SUN
Chinese Journal of Pharmacology and Toxicology 2017;31(3):255-261
OBJECTIVE To investigate the immunogenicities of Poria cocos polysaccharides, PCP-Ⅰand PCP-Ⅱ, as a vaccine adjuvant. METHODS ①Keyhole limpet hemocyanin (KLH) was linked to PCP-Ⅰor PCP-Ⅱrespectively to prepare immuno-antigen KLH-PCP-Ⅰor KLH-PCP-Ⅱ. Bovine serum albumin (BSA) was also linked to PCP-Ⅰor PCP-Ⅱrespectively to prepare screening-antigen. Rabbits were immunized with KLH-PCP-Ⅰor KLH-PCP-Ⅱplus Freund adjuvant by intradermal injection twice, and serum specific antibody titers were determined by ELISA. ②BALB/c mice were immunized with PCP-Ⅰ or PCP-Ⅱ alone intramuscularly twice, and serum polysaccharide antibody titers were determined by ELISA.③BALB/c mice were co-immunized intramuscularly or subcutaneously with PCP-Ⅰor PCP-Ⅱplus hepatitis B surface antigen (HBsAg) or porcine reproductive and respiratory syndrome virus inactivated vaccine (PRRSV) twice, and serum polysaccharide-antibody titers were determined by ELISA. RESULTS ①Serum anti-KLH and anti-polysaccharides (PCP-Ⅰor PCP-Ⅱ) antibodies were pro?duced after rabbits were immunized with KLH-PCP-Ⅰor KLH-PCP-Ⅱplus Freund adjuvant twice.②Serum anti-PCP-Ⅰor anti-PCP-Ⅱantibodies were not found after mice were immunized with PCP-Ⅰand PCP-Ⅱalone twice.③After mice were immunized with HBsAg or PRRSV plus PCP-Ⅰor PCP-Ⅱtwice, serum anti-PCP-Ⅰor anti-PCP-Ⅱantibodies were not found. CONCLUSION PCP-Ⅰand PCP-Ⅱshow weak immunogenicity, which may be quite safe as a vaccine adjuvant.
9.The relationship among the aggressive behavior, hostile attribution bias and childhood trauma in schizophrenic patients
Xuequan CHEN ; Kai WANG ; Yi DONG ; Wenfei LI ; Haitao XIA ; Yifu JI ; Shengchun JIN ; Wei BAI ; Xiaomei CAO ; Jiakuai YU ; Ruibin WANG
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(10):893-896
ObjectiveTo explore the relationship among the aggressive behavior,hostile attribution bias and childhood trauma in schizophrenic patients.Methods 135 schizophrenic patients were tested with Modified Overt Aggression Scale (MOAS),the Chinese-version of the Ambiguous Intentions Hostility Questionnaire (AIHQ-C) and Childhood Trauma Questionnaire (CTQ).According to the score of the MOAS,the patients were divided into the aggressive group ( n =58 ) and the non-aggressive group ( n =77 ).The hostile attribution bias and the childhood trauma were compared between the two groups,and correlation and hierarchical regression analysis were used to investigate the relationships of the variables.ResultsCompared with the non-aggressive patients,the aggressive patients had significantly higher AIHQ-C total hostility bias score (6.27 ± 1.20 vs 5.90 ± 0.97,P <0.05 ),total blame bias score (8.04 ± 1.97 vs 6.91 ± 2.10,P < 0.01 ) and total aggression bias score ( 6.17 ±1.02 vs 5.59 ± 1.04,P < 0.01 ).Correlation analysis showed that the MOAS score,AIHQ scores and the total score of CTQ were significantly positively correlated with each other ( r =0.171 ~ 0.350,P < 0.05 ~0.01 ).Regression analysis indicated the hostile attribution bias directly predicted the aggressive behavior( β =0.342,P <0.05) and completely mediated the relationship between the childhood trauma and the aggressive behavior.ConclusionThe aggressive behavior in schizophrenic patients is associated with the experience of childhood trauma and the attribution style.The childhood trauma indirectly influences the aggressive behavior by the mediating of the hostile attribution bias.
10.The synergistic effect of FGF-21 and insulin on regulating glucose metabolism and its mechanism.
Dan YU ; Cuiyu SUN ; Guopeng SUN ; Guiping REN ; Xianlong YE ; Shenglong ZHU ; Wenfei WANG ; Pengfei XU ; Shujie LI ; Qiang WU ; Zeshan NIU ; Tian SUN ; Mingyao LIU ; Deshan LI
Acta Pharmaceutica Sinica 2014;49(7):977-84
Previous studies proposed that the synergistic effect of fibroblast growth factor-21 (FGF-21) and insulin may be due to the improvement of insulin sensitivity by FGF-21. However, there is no experimental evidence to support this. This study was designed to elucidate the mechanism of synergistic effect of FGF-21 and insulin in the regulation of glucose metabolism. The synergistic effect of FGF-21 and insulin on regulating glucose metabolism was demonstrated by investigating the glucose absorption rate by insulin resistance HepG2 cell model and the blood glucose chances in type 2 diabetic db/db mice after treatments with different concentrations of FGF-21 or/and insulin; The synergistic metabolism was revealed through detecting GLUT1 and GLUT4 transcription levels in the liver by real-time PCR method. The experimental results showed that FGF-21 and insulin have a synergistic effect on the regulation of glucose metabolism. The results of real-time PCR showed that the effective dose of FGF-21 could up-regulate the transcription level of GLUT1 in a dose-dependent manner, but had no effect on the transcription level of GLUT4. Insulin (4 u) alone could up-regulate the transcription level of GLUT4, yet had no effect on that of GLUT1. Ineffective dose 0.1 mg kg(-1) FGF-21 alone could not change the transcription level of GLUT1 or GLUT4. However, when the ineffective dose 0.1 mg x kg(-1) FGF-21 was used in combination with insulin (4 u) significantly increased the transcription levels of both GLUT1 and GLUT4, the transcription level of GLUT1 was similar to that treated with 5 time concentration of FGF-21 alone; the transcription level of GLUT4 is higher than that treated with insulin (4 u) alone. In summary, in the presence of FGF-21, insulin increases the sensitivity of FGF-21 through enhancing GLUT1 transcription. Vice versa, FGF-21 increases the sensitivity of insulin by stimulating GLUT4 transcription in the presence of insulin. FGF-21 and insulin exert a synergistic effect on glucose metabolism through mutual sensitization.