1.Effects of substance P on anoxia/reoxygenation injury to cardiomyocytes of neonatal rats
Xin ZHAO ; Lu CHEN ; Zheng GUO
Chinese Journal of Anesthesiology 2014;34(7):873-875
Objective To investigate the effects of substance P on anoxia/reoxygenation (A/R) injury to cardiomyocytes of neonatal rats.Methods Cardiomyocytes of neonatal rats were isolated from Sprague-Dawley rats,aged 1-3 days,and were cultured in 6-well plates for 72 h.The cardiomyocytes were then assigned into 4 groups (n =3 each) using a random number table:control group (group C),A/R group,substance P group (group SP) and substance P + D-SP group (group SP + D-SP).The cells were cultured routinely for 6 h in group C and the cells were exposed to 99.9 % N2 in an incubator at 37 ℃ for 3 h followed by reoxygenation for 2 h in the other groups.The cells were incubated with substance P with the final concentration of 10-7 mol/L for 1 h before anoxia in group SP.The cells were incubated for 1 h with substance P with the final concentration of 10-7 mol/L and D-SP (a specific antagonist of neurokinin-1 receptor) with the final concentration of 10-6 mol/L before anoxia in group SP + D-SP.The apoptosis rate and lactate dehydrogenase (LDH) activity were detected at the end of reoxygenation using TUNEL assay and LDH assay kit,respectively.Results Compared with C group,the apoptosis rate and LDH activity were significantly increased in A/R,SP and SP+ D-SP groups.Compared with A/R group,the apoptosis rate and LDH activity were significantly decreased in SP and SP + D-SP groups.Compared with SP group,the apoptosis rate and LDH activity were significantly increased in SP + D-SP group.Conclusion Substance P can attenuate A/R injury to cardiomyocytes of neonatal rats,and activation of neurokinin 1 receptors is involved in the mechanism.
2.Effect of calcitonin gene-related peptide on anoxia-reoxygenation induced injury to neonatal rat cardiomyocytes incubated in high glucose medium
Lu CHEN ; Xin ZHAO ; Zheng GUO
Chinese Journal of Anesthesiology 2014;34(10):1185-1188
Objective To evaluate the effect of calcitonin gene-related peptide (CGRP) on anoxiareoxygenation (A/R)-induced injury to neonatal rat cardiomyocytes incubated in high glucose medium.Methods Cardiomyocytes were obtained from 1-3-day old Sprague-Dawley rats and cultured in the culture medium containing 15% bovine calf serum and then seeded onto 6-well plates at a density of 10 × 105/ml (3 ml/well).The cells were randomly divided into 5 groups (n =9 each):normal glucose medium control group (NG group),high glucose medium group (HG group),high glucose medium + A/R group (HG+ A/R group),high glucose medium + A/R + CGRP group (HG + A/R + CGRP group),and high glucose medium + A/R + CGRP+ CGRP8-37 group (HG + A/R + CGRP + CGRP8-37 group).The cells were incubated in normal glucose (5.5 mmol/L) medium for 72 h in NG group.In HG group,the cells were incubated in high glucose (25.0 mmol/L) medium for 72 h.In HG + A/ R group,the cells were incubated in high glucose medium for 72 h and then exposed to 3 h of anoxia followed by 2 h of reoxygenation.In group HG + A/R + CGRP,the cells were incubated in high glucose medium for 72 h,CGRP (final concentration 10-8 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.In HG + A/R + CGRP + CGRP8-37 group,the cells were incubated in high glucose medium for 72 h,CGRP (final concentration 10 8 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.In HG + A/R + CGRP + CGRP8-37 group,the cells were incubated in high glucose medium for 72 h,CGRP8-37 (final concentration 10-8 mol/L) and CGRP8-37 (CGRP receptor antagonist,final concentration 10-7 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.Apoptosis in cardiomyocytes was detected using TUNEL and apoptosis index (AI) was calculated.The lactate dehydrogenase (LDH) activity in the culture medium was analyzed.Results AI and LDH activity were significantly higher in HG group than in NG group,and in HG + A/R group than in HG group.Compared with HG + A/R group,AI and LDH activity were significantly decreased in HG + A/R + CGRP group,while no significant changes were found in HG + A/R + CGRP + CGRP8-37 group.Compared with HG + A/R + CGRP group,AI and LDH activity were significantly increased in HG + A/R + CGRP + CGRP8-37 group.Conclusion CGRP attenuates A/R-induced injury to neonatal rat cardiomyocytes incubated in high glucose medium via combing with CGRP receptor.
3.Characteristics of human bocavirus infection in children in Chongqing area.
Yuan DING ; Xiao-dong ZHAO ; Xin CHEN
Chinese Journal of Pediatrics 2009;47(6):466-467
Child
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China
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epidemiology
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Female
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Human bocavirus
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Humans
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Infant
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Infant, Newborn
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Male
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Parvoviridae Infections
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epidemiology
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virology
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Respiratory Tract Infections
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epidemiology
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virology
4.Electrochemical Analysis of Azodicarbonamide in Hot Solution by Using Nation Film Electrode
Changzhi ZHAO ; Li ZHANG ; Chen XIN
Chinese Journal of Analytical Chemistry 2017;45(2):169-174
Based on the high solubility of azodicarbonamide (ADC) in hot solution and its electrostatic interaction with Nafion film,a new electroanalytical method was developed for the determination of ADC by using Nafion film electrode.The effect of temperature on the solubility of ADC and the mechanism of the reduction reaction of ADC on Nafion film electrode were investigated.Under the experimental conditions such as water bath at a constant temperature of 80 ℃,pH 6.0 and optimal test parameters,the differential pulse voltammetric response was proportional to the concentration of ADC in the range of 0.93-10.5 μg/L,and the detection limit was estimated to 0.58 μg/L.The relative standard deviation was less than 5.86 % and the recovery was 95.8%-104.0% for the determination of the ADC in flour samples.The semicarbazide and nitrofurazone did not interfere with the determination of ADC.
5.The Effect of Laboratory Mental Stress on Serum Levels of Glucose and Insulin
Xin CHEN ; Ying ZHAO ; Baotang YANG
Chinese Mental Health Journal 1991;0(05):-
Objective: To study the effect of laboratory menta l stress on serum levels of glucose and insulin Methods: 48 heal thy male college students were recruited Fasting serum glucose test and insuli n measurement were done before and after mental arithmetic test Result s: Glucose level increased after arithmetic test (434?079/477?0 70, t=381, p
6.The Effect of Laboratory Mental Stress on Plasma Levels of Angiotensin II,Aldosterone and Atrial Natriuretic Peptide in Different Salt-Sensitivity Individuals
Xin CHEN ; Aitao HE ; Ying ZHAO
Chinese Mental Health Journal 1988;0(06):-
Objective:To study the influence of laboratory mental stress on plasma levels of angiotensin II (Ang II), aldosterone(ALD)and atrial natriuretic peptide (ANP) in individuals with different salt sensitivity Method:48 healthy male undergraduates were divided into salt sensitive group (SS group, n=14) and non-salt-sensitive group (NSS group, n=34) Their blood pressure, plasma levels of Ang II, ALD and ANP were measured before and after mental arithemtic test Result:Blood pressure, plasma levels of Ang II and ALD increased significantly in both groups, with SS group having greater increase Plasma level of ANP had no change after the test Conclusion:Laboratory mental stress increases the plasma levels of Ang II and ALD in male students Salt-sensitivity can magnify this effect
7.Mutation analysis of atlastin gene in patients with hereditary spastic paraplegia
Xin CHEN ; Beisha TANG ; Guohua ZHAO
Journal of Clinical Neurology 2001;0(05):-
Objective To investigate the mutation characteristics of atlastin gene in Chinese patients with hereditary spastic paraplegia(HSP) and establish the base of gene diagnosis of HSP.Methods Mutation analysis of atlastin gene was made by use of polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP) combined with DNA direct sequencing in 30 unrelated affected HSP individuals in China in which 20 cases were from autosomal dominant families and ten cases were sporadic HSP patients.Results No abnormal SSCP bands were found in the 30 individuals and the results of DNA direct sequencing were also normal.Conclusion Mutation of atlastin gene may be rare in Chinese HSP patients.
8.Mutation analysis of paraplegin gene in Chinese patients with hereditary spastic paraplegia
Xin CHEN ; Beisha TANG ; Guohua ZHAO
Journal of Clinical Neurology 1995;0(04):-
Objective To investigate the mutation characteristics of paraplegin gene in Chinese patients with hereditary spastic paraplegia (HSP) and establish the base of the gene diagnosis of HSP.Methods Mutation analysis of paraplegin gene was carried out by use of polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) combined with DNA direct sequencing in 22 unrelated affected HSP individuals in China, in which 8 probands were from autosomal recessive families and 14 cases were sporadic.Results All of the exons could be detected by PCR. 2 probands were found to have abnormal SSCP bands in exon 15 and 2 substitutes (G2063A, G2066A in exon 15) were found by DNA direct sequencing. But there were no changes in other patients of families.!The same abnormal SSCP bands and G→A substitutes were revealed in control individuals. So these changes were two polymorphisms, in which G2066A was not reported previously.Conclusion Mutations of paraplegin gene may be rare in Chinese patients with HSP. G2063A and G2066A are two polymorphisms, in which G2066A has not been reported previously.
9.Factors regulating the adipo-osteogenic differentiation of bone marrow mesenchymal stem cells
Xin ZHOU ; Jiajia ZHAO ; Lili CHEN
Chinese Journal of Tissue Engineering Research 2017;21(17):2759-2765
BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) with multiple differentiation potential can be induced into osteogenic or adipogenic differentiation under certain conditions. OBJECTIVE: To review the related factors regulating the adipo-osteogenic differentiation of BMSCs. METHODS: A computer-based search of CNKI and PubMed databases was performed for literature concerning the related factors regulating the adipo-osteogenic differentiation of BMSCs published from January 2006 to August 2016. The search terms were bone marrow mesenchymal stem cells, osteogenic differentiation, adipocyte differentiation in Chinese and English, respectively. RESULTS AND CONCLUSION: Signaling pathways, transcription factors, and smal molecule compounds that are interacted are key factors in the regulation of BMSCs differentiation, so the techniques to intervene BMSCs differentiation based on these key molecules may correct bone or fat abnormality and can be applied to tissue engineering and regenerative medicine in the future. Additionally, the biological clock is also one of the most important factors for adipo-osteogenic differentiation of BMSCs by regulating signaling pathways or transcription factors.
10.Over-expression and Purification of Formate Dehydrogenase form Pichia pastoris in Escherichia coli
Shao-Xin CHEN ; Bing-Zhao SHI ;
Microbiology 1992;0(01):-
Formate dehydrogenase(FDH)coding gene was amplified from genomic DNA of Pichia pastoris by polymerase chain reaction, and the codon TAG(bases 649-651)was mutated to GAG using site-directed mutagenesis.The recombinant plasmid pET-FDH was con- structed by inserting the mutated DNA fragment into expression vector pET-22b(+),and transformed into E.coli BL21(DE3).FDH was expressed as a form of soluble prutein fused with 6?His tag at high level through IPTG induction.The amount of FDH was up to about 30% of the total cell protein.The cells-free crude extract was purified by one affinity chromatographic step,and resulting enzyme preparation revealed a specific activity of 6.45 U/mg.