1.Application of real-time quantitative PCR technique to analysis of DNA in forensic science
Jinping HAO ; Qiaowen WU ; Jiangwei YAN
Chinese Journal of Forensic Medicine 1986;0(02):-
Objective To apply the real-time quantitative PCR assay with the TaqMan Probe to DNA analysis in Forensic science . Methods Using the TaqMan method to quantitate DNA abstracted from variety of biological samples common in Forensic science . Results The quantity of sample DNA can be obtained. Inhibiting factors in the abstracted DNA might be evaluated. And subsequent procedures of STR testing were then directed. Conclusion Real-time quantitative PCR is a necessary technique for DNA analysis in Forensic science.
2.Repurposing old drugs for a new life:metformin shows promising in cancer care
Jinping LI ; Zhifeng NING ; Fuxing LIU ; Hao ZHANG
Chinese Journal of Biochemical Pharmaceutics 2016;36(8):16-21
Development of small molecule drugs is important for targeted and personalized therapy for cancers.However,it is staggering to take new compounds from the research bench to clinical trial pipelines and eventually to the clinical practice,due to the time,effort and costs.Most drugs need five years of development after discovery in the laboratory before they are ready to be tested for efficacy and safety.Compared to development of new drugs from scratch, increasing research efforts have been made to turn existing drugs to new uses to treat cancers, which may bypass years of costly work.Metformin used in cancer care is a good example of giving old drugs a new life.The activity of metformin on anti-cancer has recently drawn significant attention.Epidemiological and experimental studies have shown that metformin can cut down the incidence of cancer in diabetic patients and reduce metabolism-related cancers.Meanwhile, recent study has found metformin can induce autophagy of esophageal cancer cells by inhibiting inflammatory signaling pathway.In recent years, increasing studies have shown that metformin plays a major role in suppressing cancers via multiple mechanisms.In this mini-review,we summarize the updates of the research on metformin in cancer prevention and treatment.
3.Effects of baicalin on mitochondria apoptotic pathway of trophoblast cells in a preeclampsia rat model
Yonghong WANG ; Jiao SONG ; Jinping DONG ; Taotao YANG ; Min HAO
Chinese Journal of Perinatal Medicine 2016;19(12):933-939
Objective To explore the effects of baicalin in the treatment of a preeclampsia (PE) rat model by detecting the expression of X-linked inhibitor of apoptosis protein (XIAP) and cysteine containing aspartate-9 (Caspase-9) and observing the ultrastructure of mitochondria in trophoblast cells.Methods Forty-eight pregnant Wistar rats were randomly divided into two groups:12 in the control group and 36 in the PE model group.The PE model was established with subcutaneous injection of l-nitro arginine methyl ester with 100 mg/kg per day from the 13th day of pregnancy.Beginning from the 16th day of pregnancy,the PE rats were injected with different doses of baicalin till cesarean section,and divided into three groups:non-intervention PE model group treated with saline (NIP group),low-dose baicalin intervention group (IDB group) at 50 mg/kg per day,and high-dose baicalin intervention group (HDB group) at 100 mg/kg per day.The rat tail artery blood pressure and 24-h urine protein level were measured at pregnant day 10,16 and 20.The levels of XIAP and Caspase-9 in placenta were measured by immunohistochemistry.The ultrastructure of mitochondria of trophoblast cells of the rat placenta was observed under electron microscope.T test,F test and LSD-t test were applied for statistical analysis.Results (1) On pregnant day 10,no significant differences were observed in rat tail artery systolic blood pressure,diastolic blood pressure and 24-h urine protein level between the control group and PE model group (all P>0.05).On pregnant day 16 and 20,the systolic blood pressure,diastolic blood pressure and 24-h urine protein level of NIP,IDB and HDB groups were significantly higher than those of control group [pregnant day 16:systolic blood pressure:(137.74±5.21),(136.15±4.86),(138.28±4.79) and (110.57±3.79) mmHg (1 mmHg=0.133 kPa),diastolic blood pressure:(89.58 ± 5.50),(88.45 ± 8.59),(89.42 ± 6.29) and (80.28 ± 7.36) mmHg,24-h urine protein:(7.78 ± 0.45),(7.53 ± 0.54),(7.86± 0.57) and (6.45 ± 0.56) mg;pregnant day 20:systolic blood pressure:(145.26 ± 4.67),(131.28 ± 4.34),(130.93 ± 5.33) and (110.40 ± 6.92) mmHg,diastolic blood pressure:(89.87±6.55),(85.34±7.33),(84.64±7.36) and (80.19±7.34) mmHg,24-h urine protein:(11.18±0.42),(9.65±0.54),(9.06±0.56) and (6.31 ±0.45) mg] (all P<0.01).On pregnant day 20,the systolic and diastolic blood pressure and 24-h urine protein level in IDB and HDB groups were significantly lower than in NIP group (all P<0.05),but showed no significant differences between IDB and HDB groups (allP>0.05).(2) Compared with NIP group,the expression of XIAP in control group,IDB and HDB groups were significantly increased(210.39±0.78,180.56±0.82,195.36±0.96 and 192.84± 1.06,all P<0.01).There was no significant difference in the expression of XIAP between IDB and HDB groups (P=0.66).The expression of Caspase-9 in control group,IDB and HDB groups were significantly decreased compared with NIP group (210.36±0.55,195.53±0.96,198.42± 1.01 and 185.25±0.64,all P<0.01).There was no significant difference in the expression of Caspase-9 between IDB and HDB groups (P=0.65).Ultrastructure of mitochondria in NIP group showed different degrees of damage,matrix swelling,and mitochondrial cristae bresk or disappearance.In IDB group,mitochondrial matrix swelling was not obvious,and mitochondrial cristae were visible.In HDB group,mitochondrial cristae were neat and clear.Conclusions Baicalin may play an important role in the treatment of preeclampsia by reversing the trophoblast apoptosis and improving the ultrastructure of mitochondria through its regulation of XIAP expression and downregulation of Caspase-9 expression.
4.High glucose stimulates the expression of transforming growth factor-β1 and fibronectin via SREBP-1 in HKC cells
Jun HAO ; Jinping ZHANG ; Jiangbo GU ; Shuxia LIU ; Huijun DUAN
Chinese Journal of Endocrinology and Metabolism 2009;25(4):442-444
significantly increased (all P < 0.01). Following SREBP-1 was down-regulated by siRNA, high glucose-stimulated TGF-β1 and FN protein expressions were decreased by 17.9% and 24.6% ,respectively(all P<0.01).
5.Magnetic Affinity Immunoassay Based Enzyme-Labeled Phage Displayed Antibody
Xihui MU ; Zhaoyang TONG ; Qibin HUANG ; Bing LIU ; Zhiwei LIU ; Lanqun HAO ; Jinping ZHANG
Chinese Journal of Analytical Chemistry 2014;(6):785-790
A new magnetic affinity immunoassay (MAIA) strategy based on enzyme-labeled phage displayed antibody was developed. The assay consisted of a sandwich format in which immobilized polyclonal antibody (pcAb) on magnetic microparticle was used for capture probe, and enzyme-labeled phage displayed antibody for specific detection probe to increase enzyme amount and enhance detection signal. By the proposed method,β-bungarotoxin (β-BGT) was successfully detected. A linear relationship between absorbance value and the concentration of β-BGT in the range of 0. 016-62. 5 μg / L was obtained. The linear regression equation was Y=0. 641X+1. 355 (R =0. 9925, n = 13, p<0. 0001) with a detection limit of 0. 016 μg / L. In comparison with the traditional ELISA, this method gave a 10-fold better sensitivity in β-BGT detection. This strategy also gave a 4-fold better sensitivity comparing with the MAIA based on enzyme labeled monoclonal antibody (mcAb). Due to low detection limit, acceptable reproducibility and high specificity, this method holds great promise in toxin trace detection.
6.Analysis for the Occurrence Rate of Permanent Pacemaker Implantation With Relevant Risk Factors in Patients After Mechanical Heart Valve Replacement
Jinping ZHANG ; Yong JIANG ; Weichun WU ; Jingjin WANG ; Li ZHANG ; Minghui ZHANG ; Hao WANG
Chinese Circulation Journal 2016;31(6):569-572
Objective: To explore the occurrence rate of permanent pacemaker implantation (PPI) with relevant risk factors in patients after mechanical aortic valve replacement. Methods: A total of 1986 consecutive patients with mechanical aortic valve replacement were enrolled in this study. According to PPI conduction caused by severe arrhythmia , the patients were divided into 2 groups: PPI group,n=61 including 27 male with the average age of (53.6 ± 9.03) years and Non-PPI group,n=1925. The median follow-up time was (4.47 ± 4.36) years after valve replacement. Results: The patients in PPI group were with the elder age and higher ratio of pre-existing atrial ifbrillation (AF) than those in Non-PPI group,P<0.05. The overall PPI occurrence rate was 3.07% after valve replacement and the short term incidence rate (within 30 days) was 0.55%, midterm (from 30 day to 1 year) was 0.03%, long term (>1 year) was 2.22%. For PPI indications, there were 70.5% patients with high degree A-V block including 30 of AF combining long intervals, 12 of high degree A-V block, 1 of complete left bundle branch block (LBBB) and 14.8% patients with sick sinus syndrome/sinus arrest/ sinus bradycardia. Conclusion: PPI incidence was at a relative low level, the long term occurrence rate was higher than both short term and midterm; elder age, pre-existing AF could be the high risk factors for PPI requirement, and the major PPI indication was high degree AV block in clinical practice.
7.Effects of acetylcholine on electrical remodeling of human atrial fibers.
Chengkun, XIA ; Jiahong, XIA ; Zhigang, SHUI ; Jinping, LIU ; Wei, SUN ; Yimei, DU ; Guoliang, HAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(2):164-8
Autonomic nervous system activation can result in significant changes of atrial electrophysiology and facilitate induction of atrial fibrillation. By recording influence of different concentrations of acetylcholine (ACh) on atrial fibers (AF), we investigated the role of the increased vagal tone in electrical remodeling in atrial fibrillation. Parameters of action potentials and force contraction (Fc) in atrial fibers were recorded by using standard intracellular microelectrode technique and force transducer. It was found that: (1) ACh at 0.1 μmol/L had no significant influence on spontaneous action potentials (SAPs) and Fc (n=6, P>0.05); ACh at both 1.0 and 10.0 μmol/L shortened action potential duration (APD) and Fc of human AF from right atrium (n=6, P<0.05); there was no significant difference in shortening APD between 10.0 and 1.0 μmol/L of ACh; (2) ACh at 0.1 μmol/L had no significant desensitization (n=6, P>0.05), but ACh at 1.0 and 10.0 μmol/L had desensitization (n=6, P<0.05) to SAPs and Fc. The desensitization of ACh on APD in AF was concentration- and time-dependent. It was shown that APD was longer than the control along with extending time of continuous Tyrode's solution perfusion after desensitization. It is concluded that ACh changes the electrophysiological characteristics of human AF, indicating that increased vagal tone plays a role in the development of a vulnerable substrate for atrial electrical remodeling in atrial fibrillation.
8.Long-term Follow-up Study of Ascending Aortic Diameter Changes in Patients After Mechanical Aortic Valve Replacement
Jinping ZHANG ; Yong JIANG ; Weichun WU ; Jiande WANG ; Nan XU ; Jianrong LI ; Hao WANG
Chinese Circulation Journal 2016;31(3):267-271
Objective: To retrospectively analyze the ascending aortic diameter (AAD) changes in patients after mechanical aortic valve replacement (AVR).
Methods: The medical records and echocardiography reports in patients who received AVR or bivalve valve replacement (BVR) in our hospital from 2000-01 to 2001-12 were retrieved, the retrieval conditions were as aortic valve structure must be mechanical and the follow-up echocardiography examination should be more than 9 years. The clinical information, pre- and post-operative 2-dimentional transthoracic echocardiography reports were collected, the follow-up echocardiography periods were ≤3-year, 3-year<-≤6-year, 6-year<-≤9-year and >9-year. AAD changes at different periods were compared. According to baseline AAD, the patients were divided into AAD<35 mm group and AAD≥35 mm group in order to observe the ascending aortic events.
Results: A total of 141 patients were enrolled form 595 echocardiography reports which included 75 male, the patients were at the mean age of (45.5 ± 11.2) years with mean follow-up time of (7.59 ± 3.38) years. Compared with baseline level, the follow-up AAD was similar between ≤3-year and 3-year<-≤6-year patients,P>0.05; while the follow-up AAD was different between 6-year<-≤9-year and >9-year patients,P<0.05. The patients with second operation in AAD≥35 mm group were much higher than those in AAD<35 mm group (24.0% vs 12.9%). There were 5 (20%) patients suffered from ascending aortic events in AAD≥35 mm group.
Conclusion: AAD dilatation were gradually occurring after mechanical AVR, the patients with AAD≥35 mm had the higher risk for ascending aortic events, therefore special attention should be taken in patients with aortic valve disease combining AAD dilatation during surgical treatment.
9.Cardiomyocyte Differentiation from P19 Cells Induced by NKX2-5
Jinping ZHANG ; Jun HAO ; Huijuan WANG ; Suxia SHAO ; Duo WANG ; Yanhong GUO ; Chunfang ZHAO ; Lei ZHANG
Journal of China Medical University 2010;(3):197-200
Objective To identify the rale of NKX2-5 gene in cardiomyocyte differentiation and its mechanism.Methods P19 cells were divided into transfected and non-transfected groups.In the transfected group,P19 cells were with stable expression of NKX2-5 gene.The P19 cells were cultured in suspension for 4 days,and the formed aggregates were transferred to Petri dish for adherent culture.On days 4,8,12,and 16 of the adherent culture,the expressions of ct-saicomeric actin(α-SA)and cardiac troponin T(cTnT)were detected with double-labeling immunofluorescence and Western blot.The ultrastruetural changes were observed on day 16.Results In the transfected group,no expression of α-SA and cTnT was found on day 4,and the expression of these 2 proteins or co-expression existed on days 8,12,and 16.There were early cell junction and myofilament-like structure in the cytoplasm of some cells in the transfected group.In the non-transfected group,these 2 proteins were negative,and no differentiated cell was found.Conclusion Stable expression of NKX2-5 gene can induce cardiomyocyte differentiation from P19 cells,but the P19 cells with stable expression of JVKX2-5 gene is not suitable to be an in vitro model of cardiac development.
10.Aptamer-Based Microcantilever Sensor for O-ethyl S-[2-(diisopropylamino) ethyl] methylphosphonothiolate, Sarin Detection and Kinetic Analysis
Zhiwei LIU ; Zhaoyang TONG ; Lanqun HAO ; Bing LIU ; Xihui MU ; Jinping ZHANG ; Chuan GAO
Chinese Journal of Analytical Chemistry 2014;(8):1144-1149
A new method for O-ethyl S-[2-( diisopropylamino) ethyl] methylphosphonothiolate ( VX) , sarin detection and its kinetic analysis based on piezoresistive microcantilever aptasensor was developed, where VX, sarin aptamers were immobilized on the microcantilever surface by biotin-avidin binding system. A linear relationship between the response voltage and the concentration of VX in the range of 2-60μg/L was obtained. The linear regression equation was △Ue=0. 886C-1. 039 (n=5, R=0. 984, p<0. 001) and the detection limit was 2μg/L ( S/N≥3 ) . A linear relationship between the response voltage and the concentration of sarin in the range of 10-60 μg/L was obtained, the linear regression equation was △Ue=0. 716C-2. 304 ( n=5, R=0 . 996 , p<0 . 001 ) and the detection limit was 10 μg/L ( S/N≥3 ) . The sensor showed no response for O-butyl methylphosphonochloridate, a structural analog of VX and sarin, which indicated high specificity and good anti-interference ability. On this basis, a reaction kinetic model based on receptor-ligand binding and the relationship with output voltage change was established. Response voltage (△Ue ) and response time( t0 ) were obtained from the fitting equation on different concentrations of VX, sarin fitted well with the measured values.