2.Expression, characterization and application of thermostable beta-glucuronidase from Thermotoga maritima.
Zhuo WANG ; Jianjun PEI ; Huazhong LI ; Weilan SHAO
Chinese Journal of Biotechnology 2008;24(8):1407-1412
The gene of beta-glucuronidase from Thermotoga maritima was cloned into the plasmid pHsh, and expressed in Escherichia coli JM109. The recombinant protein was purified to homogeneity by a simple step, heat treatment. The recombinant enzyme had a molecular mass of 65.9 kD. The optimal activity of beta-glucuronidase was found at pH 5.0 and 80 degrees C. The purified enzyme was stable over a pH range from 5.8 to 8.2 and had a half life of 2 h at 80 degrees C. The kinetic experiments at 80 degrees C with p-nitrophenyl-beta- glucuronide as substrate gave a K(m) and V(max) of 0.18 mmol/L and 312 u per mg of protein. The purified enzyme could transform glycyrrhizin to glycyrrhetinic acid.
Cloning, Molecular
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Enzyme Stability
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Escherichia coli
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enzymology
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genetics
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Glucuronidase
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biosynthesis
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genetics
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metabolism
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Glycyrrhetinic Acid
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metabolism
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Glycyrrhizic Acid
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metabolism
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Hot Temperature
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Kinetics
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Recombinant Proteins
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biosynthesis
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genetics
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metabolism
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Thermotoga maritima
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enzymology
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genetics
3.Effect of cilostazol and aspirin on function of platelet assembly rate and change of protein kinase B activity in elderly patients with acute coronary syndrome
Peiliang LIU ; Tao JING ; Zhuo ZHOU ; Yao CHEN ; Xuan LI ; Chunlai SHI ; Lifeng PEI
Chinese Journal of Postgraduates of Medicine 2006;0(25):-
Objective To examine the effect function of platelet(Pt)assemble rate(PLTAR) and the change of protein kinase B(PKB) active by cilostazol (CS)and aspirin (AS)on elderly patients with acute coronary sydrome(ACS). Methods Forty-eight elderly patients with ACS were divided randomly into two groups:CS group (100 mg,n=26),AS group (300 mg,n=22).Twenty-six healthy elderly were into the group of normal control(NC group) . The CS group and AS group were treated by routine anticoagulation and antiplatelet.PLTAR and PKB activity were measured at 10 minutes before treatment and at 7 days after treatment 3.5,6.0,24.0 hours. Results The maximum PLTAR in elderly CS group and AS group was elevated significantly compared with NC group(P
4.Case Report of Mixed Connective Tissue Disease Complicating Pulmonary Hypertension and Its Literature Review
zhi-hong, ZHUO ; pei-chao, TIAN ; huai-li, WANG ; tie-zheng, GAO
Journal of Applied Clinical Pediatrics 2006;0(21):-
Objective To investigate the diagnosis and treatment of mixed connective tissue disease(MCTD)complicating pulmonary hypertension(PAH) in childhood in order to improve the recognition of this disease.Method According to the symptoms,signs,past history,labratory examinations,the child′s disease was diagnosed and treated,and the relative literature was reviewed.Results The main symptom of the child was interruptable apsychia.Ultrasound showed severe PAH,positive of anti-RNP antibody.After given immunosuppressant and decreased PAH,the patient′s condition was more improved.Conclusions MCTD complicating PAH in childhood onstes delitescently,and it′s difficult to diagnose.Recognition should be elevated to diagnose and treat it earlier.The prognosis can be improved.
5.Differential diagnosis of hyperdensities on computed tomography immediately after intra-arteriai thrombolysis in patients with acute ischemic stroke
Jun-Gong ZHAO ; Ming-Hua LI ; Chun FANG ; Ju WANG ; Pei-Lei ZHANG ; Zhuo-Ying DU ; Min LI ;
Journal of Interventional Radiology 1994;0(03):-
Objective The present study was to differentiate the hyperdensities on CT immediately after intra-arterial thrombolysis in patients with acute isehemic stroke.Methods Twenty two patients with acute ischemic stroke were treated with intra-arterial combining with intravenous thrombolysis within 6 hours after onset.All patients underwent nonenhaneed CT scans before,immediately and 24 hours after thrombolytic therapy.The hyperdensities on CT after intra-arterial thrombolysis were analyzed retrospectively.Results Five hyperdense areas were seen in 22 patients immediately after thrombolytie therapy.According to their locations, CT values and follow-up CT scans,the hyperdensities on CT imaging were classified into two groups:contrast enhancement and hemorrhagic transformation.The former was characterized by rapid clearance of the hyperdensity lesion with maximum Hounsifild Unit<90,on the contrary,the latter was noted by persistence of hyperdensity lesion after 24 hours CT scan with maximum Hounsifild Unit>90.Two of the five hyperdense lesion patients were confirmed to be contrast enhancement with location in cerebral cortex,the other were hemorrhagic transformation,mostly located in basal ganglia.Hyperdensity in patients with contrast enhancement showed neurological improvement although no further medical cares were offered.Conclusions Different kind of hyperdensity on CT immediately after intra-arterial thrombolysis in patients with acute ischemic stroke can be differentiated according to its location,CT value and follow-up CT scan.When contrast enhancement occurred,no further medical care is needed.
6.Total hip replacement after failed internal fixation in the elderly patients with displaced femoral neck fractures
Yunpeng LI ; Zhenpeng GUAN ; Zhuo ZHANG ; Zheng PEI ; Bolong KOU ; Jianhao LIN ; Yanlin YUAN ; Diange ZHOU ; Houshan Lü
Chinese Journal of Trauma 2010;26(5):438-441
Objective To investigate the clinical results of the primary total hip replacement (THR) and the secondary THR after failed internal fixation in the elderly patients with displaced femoral neck fracture so as to find the optimal treatment for displaced femoral neck fractures in the elderly patients. Methods From April 2001 to April 2007,16 patients (Study Group) treated with a secondary THR after failed internal fixation and 20 patients (Control Group) treated with a primary THR were enrolled in the study and followed up. There were seven males and nine females, at average age of 66. 5 years (50-85 years) and with mean follow-up period of 58. 25 months (24-96 months) in the Study Group. There were six males and 14 females, at average age of 68.1 years (51-83 years) and with mean follow-up period of 49.50 months (24-70 months) in the Control Group. All patients were active and lucid before they suffered fractures. Blood loss and operation duration in THR were compared. Hip function (Harris score) and health-related quality of life (HRQoL, KPS index score) were assessed during the follow-up after THR. Results Operative duration was (115.63 ±34.35) minutes in Study Group and (91.25 ±15.80) minutes in Control Group (P<0.05). Blood loss was (546.86 ±377.04) ml in Study Group and (320.00 ±155.94) ml in Control Group (P<0.05). At follow up, Harris score and KPS index score were (87. 25 ±7. 53) points and (95. 00 ±5. 16) points respectively in Study Group, and (90.20±5.46) points and (96.00 ±0.73) points respectively in Control Group (P>0.05). There were no infections or re-operations in two groups, but with one death in each group during the follow-up. Conclusions THR is the optimal treatment for displaced femoral neck fractures in the elderly patients.The secondary THR after failed internal fixation has higher risks in operation compared with the primary THR for a displaced femoral neck fracture in the elderly patient.
7.SIRT3:a potential target for CHF?
Zhong-Bao YUE ; Jia YOU ; Zhuo-Ming LI ; Shao-Rui CHEN ; Pei-Qing LIU
Chinese Journal of Pharmacology and Toxicology 2018;32(4):329-329
Pathological cardiac hypertrophy is a maladaptive response in a variety of organic heart disease(OHD),which is characterized by mitochondrial dysfunction that results from disturbed energy metabolism. SIRT3, a mitochondria-localized sirtuin, regulates global mitochondrial lysine acetylation and preserves mitochondrial function. However, the mechanisms by which SIRT3 regulates cardiac hypertrophy remains to be further elucidated. In this study, we firstly demonstrated that expression of SIRT3 was decreased in AngiotensionⅡ(AngⅡ)-treated cardiomyocytes and in hearts of AngⅡ-induced cardiac hypertrophic mice. In addition, SIRT3 overexpression protected myocytes from hypertrophy, whereas SIRT3 silencing exacerbated Ang II-induced cardiomyocyte hypertrophy.In particular,SIRT3-KO mice exhibited significant cardiac hypertrophy. Mechanistically, we identified NMNAT3 (nicotinamide mononucleotide adenylyltransferase 3), the rate-limiting enzyme for mitochondrial NAD biosynthesis, as a new target and binding partner of SIRT3.Specifically,SIRT3 physically interacts with and deacety-lates NMNAT3,thereby enhancing the enzyme activity of NMNAT3 and contributing to SIRT3-mediated anti-hypertrophic effects.Moreover,NMNAT3 regulates the activity of SIRT3 via synthesis of mitochon-dria NAD.Taken together,these findings provide mechanistic insights into the negative regulatory role of SIRT3 in cardiac hypertrophy.Sirtuin 3(SIRT3),a mitochondrial deacetylase that may play an impor-tant role in regulating cardiac function and a potential target for CHF
8.Efficacy of Chinese Herbs for Supplementing Qi and Activating Blood Circulation on Patients with Acute Coronary Syndrome and Type 2 Diabetes Mellitus after Percutaneous Coronary Intervention: a Clinical Observation.
Jian-peng DU ; Cheng-long WANG ; Pei-li WANG ; Shao-li WANG ; Zhu-ye GAO ; Da-wu ZHANG ; Hao XU ; Da-zhuo SHI
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(5):563-567
OBJECTIVETo observe the efficacy of Chinese herbs for supplementing qi and activating blood circulation (CHSQABC) on patients with acute coronary syndrome (ACS) and type 2 diabetes mellitus (DM) after successful percutaneous coronary intervention (PCI).
METHODSIn this ChiCTR-TRC-00000021, a total of 281 ACS patients complicated with type 2 DM after successful PCI were randomly assigned to the Western medicine treatment group (the control group, treated by routine Western medicine treatment) and the combined treatment group (the treatment group, treated by CHSQABC + routine Western medicine treatment). Patients in the combined treatment group took Xinyue Capsule (2 pills each time, 3 times per day) and Compound Chuanxiong Capsule (2 pills each time, 3 times per day for half a year and 1-year follow-ups). Primary endpoints covered incidence of death, nonfatal myocardial infarction (MI), ischemia-driven revascularization, and secondary endpoints included stroke, heart failure, and rehospitalization for ACS. At the same time scores for blood stasis syndrome (BSS) and the incidence of angina pectoris were evaluated before treatment, at month 1, 3, 6, 9, and 12 after treatment.
RESULTSThe incidence of ischemia-driven revascularization was obviously less in the treatment group than in the control group (P < 0.05). No patient had nonfatal MI in the treatment group, while 5 patients in the control group had it. The incidence of non-fatal MI showed an obvious lowering tendency in the treatment group, but with no statistical difference when compared with that in the control group (P > 0.05). Four patients readmitted to hospital in the treatment group, while 12 patients readmitted. There existed obvious tendency in the treatment group, but with no statistical difference when compared with that in the control group (P > 0.05). The incidence of angina was significantly lower in the treatment group at month 6, 9, and 12 than that at month 1 , but it was lower in the control group at 9 months (P < 0.05). The incidence of angina was 15. 4% in the treatment group, obviously lower than that in the control group (26.2%, P < 0.05). Compared with before treatment, scores for BSS were obviously lowered in the treatment group at 1, 3, 6, 9, and 12 months of treatment and in the control group at 3, 6, 9, and 12 months of treatment (P < 0.05). It was obviously lower in the treatment group than in the control group at 3, 6, 9, and 12 months of treatment (P < 0.01).
CONCLUSIONAdministration of CHSQABC combined routine Western medicine treatment could reduce the event of revascularization and post-PCI recurrent angina, and improve scores for BSS of ACS patients complicated with DM after PCI.
Acute Coronary Syndrome ; complications ; surgery ; therapy ; Angina Pectoris ; Combined Modality Therapy ; Diabetes Mellitus, Type 2 ; complications ; therapy ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Incidence ; Medicine, Chinese Traditional ; Myocardial Infarction ; Percutaneous Coronary Intervention ; Qi
9.Expression of apoptosis inducing factor and apoptosis of nervous cells in infectious brain injury rat models induced by pneumolysin and their significance
NG-Yue WA ; Huai-Li WANG ; Zhi-Hong ZHUO ; Guo-Dong LI ; Pei-chao N TIA ; LUO-Qiang
Chinese Journal of Neuromedicine 2011;10(10):1009-1013
Objective To explore the expression ofapoptosis inducing factor (AIF) and the apoptosis ofnervous cells in infectious brain injury rat models induced by pneumolysin (PLY) and their significance.Methods A total of 80 infant SD rats were randomly divided into normal saline treatment group (NS group,n=40) and PLY treatment group (PLY group,n=40).Each group was divided into 4 subgroups:NS observation subgroups at the 6,12,24 and 48 h of treatment and PLY observation subgroups at the 6,12,24 and 48 h of treatment (n=10).Five rats in these subgroups were injected with Evan' s blue (EB) to determine the damage of BBB by measuring the content of EB in the brain tissues;the other 5 rats in these subgroups were not given EB,and their brain tissues were obtained to detect the protein expressions of NSE,GFAP and AIF by immunohistochemistry,and their apoptosis was examined by TUNEL staining.Results As compared with those in the NS treatment subgroups,the brain EB content and the expression levels of NSE,GFAP and AIF in the PLY treatment subgroups were significantly higher,and the number of apoptotic cells was significantly increased (P<0.05).The number of apoptotic cells was positively correlated to the protein expression level of AIF in the PLY treatment group (r=0.959,P=0.000).Conclusion Apoptosis plays a role in the formation and development of infectious brain injury of rats and AIF might involve in the apoptosis ofneurocyte.
10.Effects of estrogen and tibolone on bladder histology and estrogen receptors in rats.
Xin YANG ; Ya-zhen LI ; Zhuo MAO ; Pei GU ; Ming SHANG
Chinese Medical Journal 2009;122(4):381-385
BACKGROUNDEstrogen deficiency causes atrophic changes within the urogenital tract, and is associated with urinary symptoms. The purpose of this study was to investigate the effects of estrogen and tibolone on bladder histology, and the changes of estrogen receptor alpha and beta (ERalpha and beta) protein expression in the detrusor muscle.
METHODSForty female rats were separated into four groups of ten each. They received a sham operation (Sham), ovariectomy (Ovx), ovariectomy plus estrogen replacement (Ovx + E), or ovariectomy plus tibolone treatment (Ovx + T). After 12 weeks each rat was anesthetized and the bladders were removed. The bladders' ultra structure, collagen fiber (CF) to smooth muscle (SM) ratio and ER subtypes were studied. Statistical analyses were performed using the one-way analysis of variance test.
RESULTSOvx resulted in significant degeneration in bladder ultra structure; however, estrogen and tibolone reversed those changes. Ovx increased the CF/SM ratio, estrogen and tibolone resulted in an increase. Two estrogen receptors (ERs) were expressed in the bladder detrusor, with ERbeta the main subtype. Ovx resulted in up-regulation of ERalpha and down-regulation of ERbeta. With estrogen and tibolone treatment, ERbeta showed a significant increase but ERalpha showed no significant difference compared with Ovx.
CONCLUSIONSEstrogen deficiency deteriorates bladder ultra structure and histology. Supplementary estrogen can improve bladder function which may be due to inhibition of collagen hyperplasia and increased SM density. ERbeta has an important role in mediating estrogen function in the bladder. Tibolone has a mild estrogenic action and has an effect on bladder function and structure to some degree.
Animals ; Estrogen Receptor Modulators ; pharmacology ; Estrogens ; pharmacology ; Female ; Immunohistochemistry ; Microscopy, Electron, Transmission ; Norpregnenes ; pharmacology ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; metabolism ; Urinary Bladder ; drug effects ; metabolism ; ultrastructure