1.Mechanisms of heterogeneous electrophysiology in heart and the relation to electropharmacology and toxicology
Chinese Journal of Pharmacology and Toxicology 2001;15(2):95-104
It is believed that cardiac electrophysiological properties are not homogeneous, and the heterogeneity exists between pacemaker and conduction cells, atrial and ventricular cells, and among endocardium, midmyocardium, and epicardium of the transmural ventricular wall. The heterogeneous electrophysiology is related to the differential distribution and expression of transmembrane ionic channels. The present review involves literatures regarding the intrinsic biological basis of cardiac heterogeneous electrophysiology.
2.Clinical research in influence of cognitive-behavioral method on behavior selection of hospitalized patients with type 2 diabetes
Jinxiu MA ; Guirong RONG ; Yunxiang CHEN ; Guirong TANG
Chinese Journal of Practical Nursing 2010;26(19):4-6
Objective To explore the effect of cognitive-behavioral method on healthy behavior selection of in-patients with type 2 diabetes. Methods All patients with type 2 diabetes involved in this study were divided into the experimental group(99 patients) and the control group(96 patients). The experimental group was given health education with cognitive behavioral method, and the control group received only common health education. The comparison on the knowledge about diabetes mellitus and indicators of glucose metabolism on the first and 6th months between the two groups was performed. Results The knowledge about diabetes mellitus and indicators of glucose metabolism on the first and 6th months in the experimental group were better than those in the control group. Conclusions Cognitive-behavioral method can increase the treatment compliance of in-patients with type 2 diabetes and improve the glucose metabolism indicators, and created favorable condition for prevention of diabetes complications.
3.Expression, purification and bioactivity of a recombinant fusion protein consisting of anti-HBs single chain Fv and interleukin-2
Zhouyao YU ; Wenyin CHEN ; Guirong RAO
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
Objective To study the expression, purification and bioactivity of a recombinant fusion protein consisting of anti-HBs single chain Fv and interleukin-2. Methods The engineering bacterium M15[pQE-ScFv-IL-2] which can express the fusion protein consisting of anti-HBsAg single chain Fv and interleukin-2, was induced by IPTG, then a 43kD recombinant protein was identified by SDS-PAGE and Western-blot analysis. Results The ratio of target protein to total protein of host reached 18%. Further analysis confirmed that the recombinant protein formed inclusion body in the cytoplasm of bacteria. 95% purity could be achieved after two-step purification of ScFv-IL-2, including Ni metal chelating chromatography (the first) and ion-exchange chromatogram (the second). The bioactivity assay of the purified product showed that the antibody-cytokine fusion protein could bind to HBsAg specifically and stimulate the proliferation of CTLL-2. Conclusion These results suggest that the fusion protein retains the bioactivity of its parental molecules, and may be a potential gene-engineering targeting drug for the treatment of chronic hepatitis B and other relevant diseases.
4.Expression of recombinant humanized anti-HBsAg Fab fragment in genetically engineered Pichia pastoris in high-density fermentation
Wenyin CHEN ; Guirong RAO ; Kuanyua SU
Medical Journal of Chinese People's Liberation Army 2001;0(09):-
Objective To study the expression in high density fermentation of anti HBsAg Fab fragment in Pichia pastoris , and the purification and activity detection of expressed target protein. Method The high density fermentation of genetically engineered Pichia pastoris was proceeded in a 5L bioreactor using fed batch fermentation. The fermentation temperature was set at 28-30℃,the pH was 5 0-5 5, and the DO was kept over 20%. When the absorbance (OD 600 ) of the broth reached 400-450 (the first time of fermentation), 200-250 (the second time), and 300-350 (the third time), the induced phase was initiated, and the methanol concentration was 0 5%-1%. The fermentation ended after 96h's induction, the target protein was purified by affinity chromatography, and its activity was assessed by ELISA. Results It showed that the optimum initial cell density during methanol induced phase should be 300-350, which was good for control of the fermentation process and the expression of recombinant Fab. At the end of the fed batch phase, a yield of about 245mg/l of Fab was reached, and 98% purity could be reached as demonstrated by affinity chromatography. The results of ELISA showed that the supernatant of fermentation and the purified recombinant Fab could bind to HBsAg specifically. Conclusion The success of high density fermentation lays a sound foundation of mass production and clinical applications of recombinant humanized anti HBsAg Fab fragment
5.Study on the purification of recombinant human anti-HBsAg Fab fragment
Guirong RAO ; Kuanyuan SU ; Wenyin CHEN
Medical Journal of Chinese People's Liberation Army 1981;0(04):-
Objective The aim of the study was to develop a purification procedure on Pichia pastoris GS115/Fab expressing human anti-HBsAg Fab fragment. Methods Purity and yield ratio and conjugated activity of purified Fab fragment were analyzed with three purified ways of goat anti human Fab affinity chromatography and 14F7 monoclonal antibody affinity column, as well as Ion exchange Size exclusion column. Results 98% purity was reached through 14F7 monoclonal antibody column, and 95% purity was gained after goat anti human Fab fragment column. But yield ratio of the two affinity columns was low, being 35% and 55%, respectively. ForIon exchange Size exclusion column, purity and yield ratio of Fab fragment were very good, being 93.8% and 80% or more, respectively. Results of ELISA analysis showed that purified Fab fragment through three columns could bind to HBsAg specifically. Conclusion The purification process of recombinant anti-HBsAg Fab fragment was established. It lays a foundation for industrialization and clinical research of human anti-HBsAg Fab fragment.
6.Screening and identification of potential targets of carthamin against sepsis
Song GUO ; Yubin XU ; Guirong CHEN
Chinese Critical Care Medicine 2021;33(1):23-27
Objective:To screen and identify the potential targets of carthamin against sepsis by studying the characteristics of carthamin.Methods:The pharmacological parameters and molecular characteristics of carthamin were analyzed with the aid of Traditional Chinese Medicine Systems Pharmacology (TCMSP). The targets of carthamin were screened by SwissTargetprediction (a website providing compound target prediction) and Drug Repositioning and Adverse drug Reaction via Chemical-Protein Interactome (DRAR-CPI). The anti-sepsis targets were selected from the three databases of Online Mendelian Inheritance in Man (OMIM), Comparative Toxicogenomics Database (CTD) and Therapeutic Targets Database (TTD). The targets of carthamin screened by the two websites and disease targets selected from the three databases were matched to screen the targets of carthamin against sepsis. The anti-sepsis potential targets of carthamin were identified by molecular docking software.Results:The oral bioavailability of carthamin was 41.15%, the drug-likeness was 0.24, and the rotational bond number was 1, which indicated that carthamin was well absorbed by oral administration and showed good drug formation. A total of 115 potential targets of carthamin were screened by SwissTargetprediction and DRAR-CPI; 149 disease targets were found from OMIM, CTD and TTD databases; 115 target proteins of carthamin screened by the two websites were matched with the disease targets , and 10 target proteins were found to be both molecular targets and disease targets. The 10 target proteins were coagulation factor Ⅸ (F9), adenosine A1 receptor (ADORA1), nitric oxide synthase 2 (NOS2), mitogen activity protein kinase 1 (MAPK1), cathepsin G (CTSG), neutrophil elastase (ELANE), protein C (PROC), lipocalin 2 (LCN2), glucose-6-phosphate dehydrogenase (G6PD) and prostaglandin endoperoxidase 2 (PTGS2). Molecular docking software analysis showed that carthamin had the ability to bind to the above 10 target proteins, which were potential targets of carthamin against sepsis. Carthamin could interact with the key amino acid residues of the targeted proteins, so as to play the corresponding efficacy.Conclusion:Carthamin combines with the targets could reduce the tissues and organs damage of sepsis by regulating CTSG, ELANE and LCN2, reduce inflammatory response of sepsis by regulating ADORA1, PTGS2, NOS2, MAPK1 and mediating PROC and F9 to inhibit clotting, and improve oxidative stress, reduce the incidence of sepsis by regulating G6PD, finally, prevented and treated sepsis.
7.The Value of Ultrasonography in the Diagnosis of Idiopathic Retroperitoneal Fibrosis
Chunguang WANG ; Yantao CHEN ; Guirong WANG ; Ying NA
Chinese Journal of Medical Imaging 2010;(1):72-74
Purpose To explore the ultrasonic characteristics of idiopathic retroperitoneal fibrosis (IRPF) and assess the diagnostic value of ultrasound. Materials and Methods Ultrasonic images of 11 cases of IRPF were retrospectively analyzed.Results The hypoechogenic masses encasing the abdominal aorta were detected in all cases, among which the encasement of inferior vena cava was found in 4 cases, the involvement of iliac artery in 3 cases and hydronephrosis in 9 cases.Conclusion IRPF demonstrated ultrasonic characteristics that would facilitate its detection and diagnosis.
8.Predictive value of umbilical cord blood bilirubin level for neonatal pathological jaundice
Shanxia WU ; Siguo FENG ; Zhengshan CHEN ; Guirong WU ; Guanghui FANG
Chinese Journal of Postgraduates of Medicine 2011;34(15):21-22
Objective To investigate the predictive value of umbilical cord blood bilirubin for pathological jaundice in healthy term newborns. Methods Two ml navel string vein blood of baby were collected after giving birth in the normal newborn, and the hemobilirubin was detected by accidentally oxidation method. After birth, the infant's bilirubin level was tested on the forehead by the transcutaneous bilirubinometer at 8:00 -9:00 every morning until discharging from hospital. The ration of pathological jaundice of newborn and its treatment were analyzed in different levels of cord blood hemobilirubin. Results Fifty-nine cases ( 22.96% ) with pathological jaundice were diagnosed in 257 newboms.The concentration of cord blood hemobilirubin in baby with pathological jaundice [(39.68 ±8.10) μmol/L] was significantly higher than that of the normal newborn [(30.05 ±5.51) μmol/L](P<0.01). As the concentration of cord blood hemobilirubin was increased, the incidence of pathological jaundice was raised (P< 0.01), and the cases that needed to intervention treatment was increased(P< 0.01). Conclusion The detection of the level of cord blood hemobilirubin is not only very worthy to estimate the occurrence of pathological jaundice of newborn, but also offer reliable evidence for clinical early diagnosis and treatment.
9.Construction,identification and pharmikinetic research on a therapeutic double plasmid of anti-HBV DNA vaccine
Guirong RAO ; Fuqiang YANG ; Guoyu MO ; Guangming CHEN
Chinese Journal of Immunology 1999;0(12):-
Objective:To develop an effective therapeutic double HBV DNA vaccine with two eukaryotic expressing motif,namely pS2.S and pIIF,a fusion ORF of hIL-2/hIFN-?.Methods:Two genes were amplified,which separately encoded HBV preS2.S middle envelope protein as a vaccine and human IL-2/IFN-? fusion protein as an adjuvant by PCR technique from plasmids pcDNA3.1/preS2.S and pcDNA3.1/hIL-2/IFN-?.Then the genes subcloned into eukaryotic vector pVAX1.The new plasmid was analysed by restriction endonuclease and DNA sequencing.The constructed plasmids were transfected into COS-7 cells in vitro with LipofectamineTM 2000.The expressing products in supernatants were quantified by ELISA.Space structure of fusion hIL-2/IFN-? expressed by the adjuvant plasmid was simulated by CE software.The double plasmid association were injected into BALB/c mice by in situ electroporation method,and specific humoral and cellular immune responses were examined.Results:The gene segments and inserting direction of pVAX1-S2.S(pS2.S)and pVAX1-IL-2IFN-?(pIIF)were both correct by genetic analysises.At 48 h after transfection,HBsAg and the cytokines of IL-2 and IFN-? were respectively 45.1 ng/ml,10.03 ng/ml and 11.5 ng/ml by ELISA.The activity domains of the two cycokines in fusion protein were entirely exposed on surface by CE software.Experiments were pefrormed in mice by injection of plasmid pS2.S.Protective antibodies of anti-HBs were produced in terms of dose-dependment pattern for the efficacy.Co-injection of plasmid pS2.S together with adjuvant pIIF resulted in more evident immune responses and dose of the plasmid pS2.S needed was diminished.There was strongly specific humoral and cell immunity by pS2.S and pIIF co-injection with EP technique.Conclusion:The double plasmids pS2.S and pIIF of anti-HBV are constructed succeedly and induce strongly specific immune activity in vitro and vivo.