1.Effectiveness of propafenone for paroxysmal supraventricular tachycardia
Jun KE ; Yuanzhong LI ; Fangzhou CHENG
Clinical Medicine of China 2001;0(07):-
Objective To observe the clinical efficacy and safety of propafenone on paroxysmal supraventricular tachycardia (PSVT).Methods 37 patients with PSVT were injected with 70 mg propafenone intravenously in 5 minutes.The unrecovered patients in 20 minutes were injected with 70 mg propafenone intravenously again,who were given 70 mg propafenone intravenously once again if not controlled in late 30 minuted.Blood pressure,heart beat,12 lead electrocardiogram were recorded before and after use of drug.Results The significant effective rate was 77.5% and effective rate was 21.6% with overall effective rate of 97.3% and effective time of 1~55 (7.1?2.8) minutes.The average accumulated dose of propafenone was 105 mg.Some side effects were observed in part of patients,including slight reduction of blood pressure,P R interval.QRS wave and Q T interval prolonged,temporary type sino atrial block,vertigo and nausea,etc whice could vanish gradually if not treated.Conclusion Propafenone by intravenous injection can quickly contro PSVT with saftey and effectiveness.
2.CLONING AND EXPRESSION OF PRES1 GENE OF HEPATITIS B VIRUS IN YEAST
Yinying LU ; Ke LI ; Ju CHENG
Medical Journal of Chinese People's Liberation Army 1981;0(04):-
To investigate the potential role of hepatitis B virus(HBV) preS1 protein in mediating HBV adhesion to liver cell, we prepared recombinant proteins of HBV preS1 in yeast. PCR was performed to amplify the gene of HBV preS1 from the plasmid pCP10/HBV ayw subtype containing the whole fragment of HBV and the PCR product was cloned into pGEM T vector. The gene of HBV preS1 was cut from pGEM T vector and cloned into yeast expression plasmid pGBKT7, the pGBKT7 plamids containing preSl were transformed into yeast cell AH109. The yeast protein was isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting. The results showed that the presence of HBV presl proteins in yeast cells was confirmed by Western slot analysis. the molecular weight of the expressed product was about 30000 Da. The findings indicated that HBV preS1 was successfully expressed in yeast system.
3.SCREENING AND CLONING OF THE GENES OF PROTEIN INTERACTING WITH THE NONSTRUCTURAL PROTEIN 5A OF HEPATITIS C VIRUS
Medical Journal of Chinese People's Liberation Army 1982;0(01):-
Protein protein binding is the basis of virus and host cell interactions. With the application of technology of studying of protein interactions, more knowledge of replication and pathogenesis of hepatitis C virus (HCV) could be acquired. Non structure protein NS5A is one of the important regulatory factors in virus replication , transcription and signal transduction, but there are controversy in effect on HCV pathogenesis and resistance to interferon ?(IFN?). In order to describe the relationship between NS5A and host proteins, we use yeast two hybrid system 3 to screen the gene encoding proteins that could interact with NS5A from human hepatocyte library. Thirty five clones were obtained including apo A1, apo A2, apo B100, haplotype mitochondrion complete genome, phosphatidic acid phosphatase type 2B, albumin similar to tumor endothelial marker 5 precursor, matrix metalloproteinase 14, and three of the Homo sapiens hypothetical proteins. The study paved a way for further studies on the pathogenesis of HCV NS5A
4.SCREENING AND CLONING OF GENE OF HEPATOCYTE PROTEIN INTERACTING WITH HCV NS3 PROTEIN
Medical Journal of Chinese People's Liberation Army 1982;0(01):-
To clone the genes of hepatocyte protein interacting with hepatitis C virus NS3 protein, "bait" plasmids of hepatitis C virus NS3 were constructed. After verifying that hepatitis C virus NS3 protein could be steadily expressed in AH109 yeast strain, yeast two hybrid assay was berformed by mating AH109 with Y187 that pre transformed with liver cDNA library plasmids pACT2, and the diploidy yeast cells were plated on quadruple dropout (QDO) medium and assayed for X ? gal activity. Nineteen yeast colonies that could grow on QDO and had ? gal activity were obtained, then the library plasmids were extracted and sequenced. The gene sequences from the 19 positive colonies were aligned with the genes deposited in GenBank. It was found hepatitis C virus NS3 protein could interact with some proteins which have different functions.
5.CLONING OF THE HEPATITIS C VIRUS NONSTRUCTURAL PROTEIN 5A BINDING PROTEIN 37 CODING GENE
Medical Journal of Chinese People's Liberation Army 2001;0(09):-
The nonstructural protein 5A (NS5A) of the hepatitis C virus (HCV) has been shown to interact with a variety of cellular proteins and implicated in the regulation of cell growth, interferon resistance, and other cellular signaling pathways. Using the yeast-two hybrid method, we have isolated a clone that encodes a novel NS5A--associated binding protein: NS5ABP37. Reverse transcription polymerase chain reaction (RT-PCR) method was employed to amplify the full fragment,and the plasmid pGADT7-NS5ABP37 with the Saccharomyces cerevisiae vector pGADT7 was constructed. To prove the interaction, yeast cell Y187 transformed with pGADT7-NS5ABP37 was mated with yeast cell AH109 containing pGBKT7-NS5A to verify the interaction between the novel protein coded by the new gene NS5ABP37 and NS5A.
6.Cloning and expression of NS3 gene of hepatitis C virus in yeast
Chinese Journal of Infectious Diseases 1999;0(01):-
Objective HCV NS3 protein plays an important role in disease caused by HCV. We investigate the gene expression of HCV NS3 in yeast for future study of the function of the protein. Methods PCR was performed to amplify the gene of HCV NS3 from the plasmid pBRTM/HCV containing the whole fragment of HCV and the gene was cloned into pGEM T vector. Thereafter, HCV NS3 gene was cut from pGEM T vector and cloned into yeast expression plasmid pGBKT7, and recombinant pGBKT7∶NS3 was transformed into yeast AH109. The yeast protein was isolated and analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting. Results HCV NS3 gene was successfully cloned into pGBKT7. The results of SDS PAGE and Western blotting assay showed that the molecular weight of the expressed product was about 22000 Da and HCV NS3 protein was existed within yeast cells.Conclusions HCV NS3 was successfully expressed in yeast expression system.
7.Application of new barrel theory in improving evaluation of hospitalized patients
Xiaojin LI ; Caixia KE ; Shouzhen CHENG
Modern Clinical Nursing 2014;(8):53-57
Objective To study the effect of new barrel theory in improving evaluation of hospitalized patients. Methods Eight hundred hospitalized patients from September 2012 to August 2013 were randomized equally into the control group and the observation group.The last one or two items affecting patient satisfaction from the control group were used as objectives to be improved.Causal effect analysis was done pertinent to the items and the worksheet was bettered and improved and then enforced.The two groups were compared after intervention with new barrel theory in terms of satisfaction of patients during admission and discharge. Result The satisfaction of patients in the observation group during admission and discharge was significantly better than that in the control group(P<0.05).Conclusion The new barrel theory used to detect the flaws in nursing service and improve the workflow can improve assessment from the patients so that the management quality can be enhanced.
8.Experimental study on the transplantation of neural stem cells derived from bone marrow of crab-eating macaque
Yiquan KE ; Gang LI ; Ruxiang XU ; Xiaodan JIANG ; Wenping CHENG
Chinese Journal of Tissue Engineering Research 2005;9(5):200-201
BACKGROUND: Researches indicated that bone marrow stem cells (BMSCs) could differentiated into neural stem cells in vitro, but what was the role of neural stem cells(NSCs) in the recovery of cortical injury,whether the NSC is capable of growing and migration in injured still remained unknown.OBJECTIVE: To explore the growing state of autograft NSC derived from crab-eating macaque BMSC transplanted in brain.DESIGN: Prospective case control study based on experimental animals.SETTING: Department of neurosurgery in a hospital of a military medical university.MATERIALS: This study was carried out at Center Laboratory of Neurological Research Institute, Zhujiang Hospital affiliated to the First Military Medical University of Chinese PLA. Six healthy adult crab-eating macaques were purchased from the South China Primate Animal Center.INTERVENTIONS: BMSCs harvested from six crab-eating macaques were cultured in vitro and induced to differentiate into neural stem cells, which then labeled by bromodeoxyuridine(BrdU) and autografted into brains.MAIN OUTCOME MEASURES: Brain tissues underwent hematoxylin and eosin(HE) staining and immunohistochemical staining before observed under optical microscope.RESULTS: The results of HE staining showed that the cell number in injured brain vas obviously higher in both instant and delayed transplanting groups than sham-transplanting group; moreover cells were proved reacting to BrdU by immunohistochemical staining in cortical injuries of both groups at 1-6 months following stem cells autograft, as well as at neighboring white matters at half year later, but no BrdU positive cells could be found in traumatic controls, sham-transplanting group and normal brains.CONCLUSION: NSCs derived from in vitro cultured BMSCs were proved capable of surviving, proliferating, differentiating and migrating in cortex after autograft, so that BMSCs is considered as replacing cells or the source of NSCs; moreover autograft stem cells could survive, proliferate and migrate in old cortical traumatic focus.
9.Inflammatory response and peroxisome proliferator-activated receptor isotypes expression in the brain of spontaneously hypertensive rats
Li SUN ; Yan CHENG ; Jianpu ZHENG ; Yan KE ; Ka BIAN
Chinese Journal of Neurology 2008;41(2):118-122
Objective To evaluate the inflammatory response and the expressions of peroxisome proliferator-activated receptor(PPAR)isoforms(PPARα,PPARβ/δ,and PPARγ)in the brain(cortex,striatum,cerebellum)of spontaneously hypertensive rats(SHR).Methods Brain tissues(cortex,striatum,and cerebellum)were dissected from SHR and age-matched control Wistar-Kyoto rats.Myeloperoxidase(MPO)activity was measured in brain tissues as an index of neutrophil accumulation and the carbonyl protein content was analyzed by spectrophotometry to evaluate the protein oxidation.RT-PCR and Western blotting were performed to examine the expressions of inflammatory mediators(IL-1β,TNFα,ICAM-1,and iNOS)and nuclear factor PPARs(PPARα,PPARβ/δ,and PPARγ),respectively.Results(1)Systolic blood pressure of SHR was significantly higher than that of Wistar-Kyoto rats,(205.4±9.4)mm Hg versus(130.4±7.9)mm Hg(t=14.96,P<0.01).(2)MPO activity of cortex,striatum,and cerebellum were markedly higher in SHR than in Wistar-Kyoto rats.Carbonyl protein levels of cortex,striatum,and cerebellum in Wistar-Kyoto rats and SHR were(3.27±0.43)nmol/mg versus(11.87±1.11)nmol/mg,(4.02±1.04)nmol/mg versus(14.06±1.36)nmol/mg,(5.94±0.71)nmol/mg versus(14.95±1.82)nmol/mg,indicating significantly higher levels of protein oxidation in SHR than Wistar-Kyoto rats(t=17.70,14.36,11.30,P<0.05).Consistently,the expression of pro-inflammatory mediators(IL-1β,TNFα,ICAM-1,and iNOS)was upregulated when compared with Wistar-Kyoto rats.The difference between SHR and control Wistar-Kyoto rats was statistically significant except the mRNA expression of IL-1β in striatum,cerebellum and TNFα in cerebellum of SHR.All the above experimental data indicated the occurrence of inflammatory status in the brain tissue of hypertension.(3)mRNA and protein levels of brain PPAR isoforms(PPARα,PPARβ/δ,and PPARγ)of SHR increased significantly when compared with Wistar-Kyoto rats.Specifically.protein levels of PPARα in cortex.striatum,and cerebellum of SHR increased by 644.78%,791.95%,and 42.85%;PPARβ/δ increased by 106.72%,94.12%,and 161.44%;PPARγ was up-regulated by 2700.16%,790.81%,and 875.00%compared with that of Wistar-Kyoto rats,respectively.Conclusions The brain(cortex,striatum,and cerebellum)from SHR shows marked inflammatory status and increased expression of all PPAR isoforms.Increases in PPARs expression may play a compensatory role in the inflammatory response of the brain in SHR.
10.Multivariate analysis for risk factors of bile leakage after hepatectomy
Xiang CHENG ; Ke LI ; Yuzhe WU ; Qichang ZHENG
Chinese Journal of General Surgery 2013;28(11):850-853
Objective To analyse risk factors for bile leakage after liver resection.Methods Between January 2011 and December 2012,469 patients underwent elective hepatectomy.We prospectively collected and retrospectively analyzed demographic data,pathological variables,and perioperative variables.Univariate analysis screened the related factors of bile leakage after liver resection.Multivariate analysis identified the independent risk factors of postoperative bile leakage.Results 469 patients were included in the analysis.The prevalence of bile leakage was 22.6% (n =106).Univariate analysis identified the following risk factors as male gender,portal hypertension,steatosis,cirrhosis,ChildPugh grade,ascites,operative time,intraoperative transfusion,intraoperative blood loss,portal triad clamping,microwave solidification,lymphadenectomy,number of tumor,tumor margin,tumor capsular,diameter of tumor,portal vein invasion or portal branch thrombosis,number of abdominal drains.Multivariate analysis identified 4 independent risk factors for postoperative bile leakage:Cirrhosis [OR =13.2 (2.3,76.9),P =0.004],steatosis [OR =73.1 (17.7,301.5),P < 0.001],infusion volume of the surgery day [OR=1.0 (1.0,1.0),P=0.019] and diameter of tumor [OR=1.2 (1.1,1.3),P=0.003].Conclusions Cirrhosis,steatosis,transfusion volume of the surgery day,and tumor size were risk factors for bile leakage after major liver resection.