1.The conversion of human recombinant endothelial cell-derived IL-8 fusion proteins by thrombin
Bao-Hong ZHOU ; Da-Long MA ; ET AL ;
Chinese Journal of Immunology 1985;0(03):-
By using genetic engineering techniques,we expressed in E.coli fusion proteins which con-tained human endothelial cell-derived IL-8 (EDhIL-8)、MS2 protein and different length of ?- galactosidase segments,named MS2-hIL-8、lac-hIL-8, lac-T-hIL-8 respectively.The lac-T-hIL-8 has a synthesized thrombin recognition site.Because there is a natural thrombin recognition sitewithin the EDhIL-8,thrombin can hydrolyze lac-hIL-8 and MS2-hIL-8 into natural hIL-8 ex-hibited biological activity,but has no effect on lae-T-hIL-8 which contained two recognition sitesfor thrombin.These results here indicated that the recognition of thrombin dependents on notonly the amino acid sequences of the substrates,but also the conformation formed by these aminoacids.
2.Study on inhibition of lymphangiogenesis in gastric cancer by NM-3
Wei DA ; Jinshui ZHU ; Weixiong CHEN ; Long WANG ; Qun SUN
Chinese Journal of Digestion 2010;30(6):394-397
Objective To evaluate the inhibitory effect of 2-(8-hydroxy-6-methoxy-1-oxo-1-H-2-benzopyran-3-Y1) propionic acid (NM-3) on lymphangiogenesis in gastric cancer using orthotopic implantated tumor models of BALB/C nude mice. Methods A BALB/C nude mouse model of transplanted in situ human gastric cancer was established. Twenty-eight nude mice were divided into four groups with 7 each: control group, NM-3 treated group, carboplatin (10 mg/kg) treated group,and NM-3 combiantion group injected with normal saline, 5 mg/kg of NM-3, 10 mg/kg of carboplatin or 5 mg/kg of NM-3, + 10 mg/kg carboplatin, respectively, twice a week for 8 weeks. At the end of the 8th week, all mice were sacrificed for detection of lymphatic microvessel density (LMVD),lymphatic vessel endothelial hyaluranic acid receptor 1 (LYVE-1), podoplanin and Prox-1 byimmunohistochemistry with staining. Results In comparison with control group, the LYVE-1 level in other three groups was decreased with no significant difference (P> 0.05). The concentrations of podoplanin and Prox-1 in NM-3 group and combination group decreased significantly than those in control group and carboplatin group (P < 0.05). The number of LMVD in NM-3 group and combination group was 4.72±0.50 and 4.78± 0.38, respectively, which was significantly lower than that in control group (7.35±0.55)and carboplatin group (6.98i0.35, P<0.05). Conclusion The NM-3 can inhibit the growth of gastric cancer by interfering lymphangiogenesis of gastric cancer.
3.Effect of hydrogen on activation of caspase-3 in brain tissues during cerebral ischemia-reperfusion in rats
Feng JIN ; Long HE ; Yanqiu AI ; Da LI ; Wei ZHANG
Chinese Journal of Anesthesiology 2017;37(3):305-307
Objective To evaluate the effect of hydrogen on the activation of caspase-3 in brain tissues during cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-six healthy adult male Sprague-Dawley rats,weighing 220-250 g,were divided into 3 groups (n=12 each) using a random number table:sham operation (group S),I/R group and hydrogen group (group H).Cerebral ischemia was induced by occlusion of the middle cerebral artery followed by reperfusion in I/R and H groups.In group H,hydrogen-rich saline 5 ml/kg (0.6 mmol/L) was injected intraperitoneally at 3 days before establishment of the model and immediately after the onset of reperfusion.At 24 h of reperfusion,the rats were sacrificed,and hippocampal tissues were obtained for determination of neuroapoptosis (by TUNEL),apoptotic neuron count and expression of activated caspase-3 (by Western blot).The brain tissues in the ischemic area were obtained and stained with haematoxylin and eosin for examination of the pathological changes.Results Compared with group S,the expression of activated caspase-3 was significantly up-regulated,and the apoptotic neuron count was increased in I/R and H groups (P<0.05).Compared with group I/R,the expression of activated caspase-3 was significantly down-regulated,the apoptotic neuron count was decreased (P<0.05),and the pathological changes of brain tissues were significantly reduced in group H.Conclusion The mechanism by which hydrogen inhibits neuroapoptosis during cerebral I/R is probably related to inhibited activation of caspase-3 in brain tissues of rats.
4.Effects of hydrogen-rich saline on expression of phosphor-p38MAPK during cerebral ischemia-reperfusion in rats
Da LI ; Yanqiu AI ; Feng JIN ; Long HE ; Wei ZHANG
Chinese Journal of Anesthesiology 2015;35(3):370-372
Objective To evaluate the effect of hydrogen-rich saline on the expression of phosphor-p38 mitogen-activated protein kinase (p-p38MAPK) during cerebral ischemia-reperfusion (I/R) in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 3 groups (n =20 each) using a random number table:sham operation group (group S),I/R group and hydrogen-rich saline group (group I/RH).Cerebral ischemia was induced in chloral hydrate-anesthetized rats by 2 h middle cerebral artery occlusion in I/R and I/RH groups.The artery was only exposed but not occluded in group S.At 3 days before operation and immediately after onset of reperfusion,hydrogen-rich saline (0.6 mmol/L) 10 ml/kg was intraperitoneally injected in group I/RH,while the equal volume of normal saline was given in S and I/R groups.Neurological deficits were blindly assessed and scored at the end of 24 h reperfusion.The animals were then sacrificed,and brains were removed for microscopic examination and for determination of the cerebral infarct size (by TTC),brain water content,cell apoptosis (by TUNEL),and expression of p38MAPk and phosphor-p38MAPK (p-p38MAPK) (by immunohistochemistry and Western blot).Apoptosis index was calculated.Results Compared with group S,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly increased,and the expression of p38MAPK and p-p38MAPK was up-regulated in I/R and I/RH groups.Compared with group I/R,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly decreased,and the expression of p38MAPK and p-p38MAPK was down-regulated in group I/RH.The pathological changes of cerebral tissues were significantly attenuated in group I/RH as compared with group I/R.Conclusion Hydrogen-rich saline can reduce cell apoptosis through inhibiting p-p38MAPK expression,thus attenuating cerebral I/R injury in rats.
5.Effects of simvastatin on expressions of uPA/PAI-1 in neutrophils in rats with septic shock
Xiaoling WU ; Li YU ; Ding LONG ; Xingwen DA ; Zhangyin MING
The Journal of Practical Medicine 2016;32(4):519-522
Objective To explore the effects of simvastatin on the protein expressions of urokinase-typeplasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1). Methods Male Sprague–Dawley rats were divided into saline group , LPS group and LPS plus simvastatin group , and were then pretreated with simvastatin (1 mg/kg) for 30 minutes before addition of LPS (8 mg/kg). Changes in left ventricular pressure were recorded. Ninety minutes after LPS injection, whole blood was collected from the inferior vena cava, and neutrophils were separated. The neutrophils were then lysed to detect levels of uPA and PAI-1. Results Left ventricular systolic pressure (LVSP: mmHg), maximal differential of left ventricular pressure (+dp/dtmax:mmHg/s), and heart rate (beats/min) were markedly decreased at different time points after administration of LPS, and maximal differential of left ventricular pressure increased in the rats receiving LPS as compared with those receiving saline, although the differences between the control and LPS groups were not statistically significant. LPS caused a great decline in uPA content and an elevation in PAI-1 content in neutrophils, but simvastatin diminished the impact of LPS on neutrophils. Conclusion Simvastatin plays a role in protection of cardiac function in rats with LPS-induced septic shock , and controls expressions of uPA and PAI-1 in neutrophils.
7.Quality classification standard of Scrophularia ningpoensis seedlings.
Xue ZHANG ; Da-xia CHEN ; Jun TAN ; Long-yun LI
China Journal of Chinese Materia Medica 2015;40(6):1079-1085
The morphological indexes of the Scuophularia ningpoensis seedlings including the longth, diameter and weight were measured, clustering analysis was used to set up the standard quality grading of seedlings of S. Ningpoensis by SPSS. Field experiment was carried out to measure the indicators of plants growth and development, the yield and the quality. The results showed that the growth and yield of class I seedlings were better than those of class II and III. The content of main active ingredients was affected barely by seedlings classification. To ensure the quality, class II seedlings or above should be used for plantation. The established quality classification standard of S. ningpoensis seedlings was scientific and feasible, and provides the basis for the standardized cultivation of S. ningpoensis.
Drugs, Chinese Herbal
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analysis
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standards
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Quality Control
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Scrophularia
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chemistry
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classification
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growth & development
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Seedlings
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chemistry
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growth & development
8.The Establishment of an Anti-Trypanosoma Drug Screening System with Leucyl-tRNA Synthetase as an Inhibition Target
Guang-Wei GAO ; Ying YAO ; Da-Zhong DING ; Long YE ; Hu-Chen ZHOU ; Da-Wei LI ;
China Biotechnology 2006;0(12):-
Trypanosoma is a human parasite severely affecting poor tropical areas.However,current frontline drugs for Trypanosoma treatment have severe side-effects with decreased effectiveness.Based on the fact that aminoacyl-tRNA synthetase is a bonafide drug target for several microorganisms,including bacteria and fungi,it is plausible that it may also be effective target of Trypanosoma.The Trypanosoma brucei leucyl-tRNA synthetase(tbLeuRS)was cloned,expressed and purified to develop an in vitro enzymatic assay system.The assay conditions were further optimized for the effective screening of tbLeuRS inhibitors thus establishing an anti-Trypanosoma drug screening system targeting tbLeuRS.The results indicated that this system can be employed for the effective screening of anti-Trypanosoma drugs with satisfactory specificity.In addition,this system can also be used for compound optimization,as well as IC50 testing.Using this system a series of compounds are identified that are effective Trypanosoma inhibitors without toxicity to human cells.Therefore,targeting tbLeuRS may represent a new venue for the development of anti-Trypanosoma drugs.
9.Relationship between Cytochrome P450 1A2 Gene Polymorphism and Susceptibility to Chronic Obstructive Pulmonary Disease
Long ZHAO ; Ying LI ; Da CHEN ; Tao MA ; Linhui XU ; Shuyue XIA
Chinese Journal of Rehabilitation Theory and Practice 2014;(9):873-876
Objective To investigate the relationship between cytochrome P450 1A2 (CYP1A2) gene polymorphism and susceptibility to chronic obstructive pulmonary disease (COPD). Methods CYP1A2 gene polymorphisms in 100 COPD cases and 100 healthy controls were tested with polymerase chain reaction- restriction fragment length polymorphism (PCR- RELP). Results Genotype frequencies of 4 SNPs in both the COPD and control groups were in accordance with Hardy-Weinberg equilibrium (P>0.05). There was significant difference between the COPD and control groups in genotype and allele frequencies of 1D and 1F (P<0.05), but not of 1C and 1E (P>0.05). Conclusion CYP1A2*1D and CYP1A2*1F polymorphisms may play an important role in the development of COPD.
10.Clinical Research of Blood CsA C2 Level Monitoring in Renal Allograft Recipients
da, XU ; xiang-hui, WANG ; pei-jun, ZHOU ; chen-long, CHU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(06):-
Objective To decrease the incidence of acute rejection in renal allograft recipients by monitoring of cyclosporine A (CsA) concentration at 2-hour after dosing(C2). Methods The CsA C2 and CsA trough concentration(C0) were assayed in renal allograft recipients.All patients were followed up for at least 1 year.The correlation of C0 and C2 monitoring with clinical outcomes was analyzed. Results At 1 week and 1 month post-transplantation,the incidence of acute rejection in patients with C2 in target level was 4.41% and 10.29%, respectively,but the incidence of acute rejection in patients with C2 in lower level was 42.37% and 36.20%,respectively. ConclusionBy reflecting the drug exposure of CsA more accurately,C2 monitoring is beneficial for decreasing the incidence of acute rejection after renal allograft transplantation.