Effect of Xintongtai Regulating p38 MAPK/AP-1 on Traditional Chinese Medicine Syndrome Score and Collagen Fibers in VSMCs of Rabbits with Atherosclerosis： An Exploration Based on Theory of Heart Receiving Qi From Spleen / 中国实验方剂学杂志

Objective:To explore the effects of Xintongtai （XTT） on traditional Chinese medicine （TCM） syndrome score and collagen fibers in vascular smooth muscle cells（VSMCs） of rabbits with atherosclerosis in the regulation of p38 mitogen-activated protein kinase （p38 MAPK）/activator protien-1 （AP-1）signaling pathway. Method:A total of 120 rabbits of SPF grade were randomly divided into the sham operation group， combined phlegm and blood stasis model group， rosuvastatin group， and low-， middle-， and high-dose XTT groups. The rabbit model of atherosclerosis due to combined phlegm and blood stasis was established by exposing them to high-fat diet and balloon injury. Following modeling， the corresponding drugs were administered by gavage for eight weeks （2.3， 4.6， 9.2 g·kg^-1 for low-， middle-， and high-dose XTT groups and 0.55 mg·kg^-1for rosuvastatin group）. At the end of medication， the abdominal aorta was isolated and stained with htoxylin-eosin （HE） for observing the vulnerable plaque. Matrix metalloproteinase-9 （MMP-9） and tissue inhibitor of metalloproteinase-1 （TIMP-1） were detected by immunohistochemistry （IHC）. The collagen fiber decomposition in VSMCs was observed after Masson staining. The protein expression levels of p38 MAPK and AP-1 in aorta was assayed by Western blotting. The combined phlegm and blood stasis syndrome was scored based on TCM syndrome scoring scale. Result:Compared with the model group， XTT at each dose and rosuvastatin significantly decreased MMP-9 content， increased TIMP-1， down-regulated p38 MAPK protein expression， and weakened the nuclear translocation of AP-1 （<italic>P</italic><0.01）. Compared with the low-dose XTT group， the middle- and high-dose XTT groups and rosuvastatin group exhibited obviously lowered MMP-9，elevated TIMP-1， down-regulated p38 MAPK protein expression， and diminished AP-1 nuclear translocation （<italic>P</italic><0.05，<italic>P</italic><0.01）. The TCM syndrome scores of the middle- and high-dose XTT groups and rosuvastatin group were significantly improved as compared with that in the model group （<italic>P</italic><0.05，<italic>P</italic><0.01）. The comparison with the low-dose XTT group revealed a remarkable improvement in TCM syndrome score of the middle- and high-dose XTT groups and rosuvastatin group （<italic>P</italic><0.01）. As demonstrated by Masson staining， the smooth muscle fibers in the model group were arranged in disorder， accompanied by enhanced collagen decomposition， thinned fibrous cap， and increased plaque vulnerability. Compared with the model group， the VSMCs in each XTT group and rosuvastatin group were orderly arranged， manifested as decreased collagen fiber decomposition and increased plaque stability. Conclusion:XTT down-regulates the expression of p38 MAPK and MMP-9， increases the level of TIMP-1， reduces the nuclear translocation of AP-1， diminishes the decomposition of collagen fibers in VSMCs， and improves the score of combined phlegm and blood stasis syndrome. XTT alleviates arteriosclerosis due to combined phlegm and blood stasis by regulating p38 MAPK/AP-1 signaling pathway and downstream cytokines and stabilizing vulnerable plaques.

Construction and expression of a recombinant protein with γ-aminobutyric acid type C receptor ρ2 subunits / 新乡医学院学报

Objective To construct prokaryotic expression vector of γ-aminobutyric acid type C receptor ρ2 (GABACR ρ2) gene,induce the expression of the recombination protein GABACR ρ2,and transfect the protein into SH-SY5Y cell line,achieve the transient expression of the recombination protein GABACR ρ2 in SH-SY5Y cell line.Methods The ρ2-Tat gene was inserted into plasmid pET30 to construct the recombinant plasmid pET-ρ2-GFP-Tat.The expression of GABACR ρ2 was detected by Western blot.The histidine-tagged recombination protein ρ2 was purified through immobilized Ni2+ absorption chromatographic column and the purity of GABACR ρ2 protein was detected by sodium dodecyl sulfate polyacrylamide gel elec trophoresis.The transduction and cellular localization of the GABAC R ρ2 was observed by fluorescence microscope.The SH-SY5Y cells were divided into normal group and hypoxia low glucose group.The cells in normal group were divided into normal control group which cultured in high glucose medium and without protein transfection and normal transfection group which cultured in high glucose medium and with protein transfection;the cells in hypoxia low glucose group were divided into treatment group which cultured under hypoxia low glucose condition and treatment transfection group which cultured under hypoxia low glucose and with protein transfection;the viability of SH-SY5Y cells in each group was evaluated by cell counting Kit-8.Results The recombinant plasmid pET30-ρ2-Tat was constructed successfully.The purified recombination protein ρ2-Tat successfully crossed the cytomembrane and transfected the SH-SY5Y cells.The viability of cells in normal control group,normal transfection group,treatment group and treatment transfection group was(100.0 ± 6.9)％,(89.3 ± 3.6)％,(51.4 ± 3.6)％and (66.1 ± 8.5) ％ respectively.There was no statistic difference in the cell viability between the normal control group and normal transfection group(P ＞0.05);the cell viability in treatment group was significantly lower than that in the normal control group (P ＜ 0.01);the cell viability in treatment transfection group was significantly higher than that in the treatment group (P ＜ 0.01).Conclusion The recombination protein ρ2 can through the membrane under the effect of Tat transduction peptide and can successfully establish a SH-SY5Y cell lines which transiently express recombinant protein ρ2,which provide a new research method for the study of ρ2 subunit function.

Correlation of interleukin-6 and C-reactive protien levels with left atrial thrombus and severe spontaneous echocontrast in atrial fibrillation patients / 天津医药

Objective To investigate the correlation of serum interleukin-6 (IL-6) and C-reactive protien (CRP) lev?els with left atrial thrombus and severe spontaneous echocontrast (SEC) in patients with atrial fibrillation (AF). Methods A case-control study of patients with atrial fibrillation (n=76) was carried out. All patients were divided into control group (n=45) and study group (n=31) according to their conventional echocardiography performance. Serum IL-6 and CRP were exam?ined in both groups. Results The levels of serum IL-6 in patients with thrombus and severe SEC were (324.13±42.86) ng/L and (332.29±53.17) ng/L, respectivly which is higher than that in patient without thrombus or without severe SEC (108.75± 25.43) ng/L and (93.59 ± 27.82) ng/L respectively. In parallel, CRP levels in patients of thrombus and severe SEC were (66.97 ± 17.65) mg/L and (71.81 ± 20.19) mg/L respectively which is higher than that in patients without thrombus or without severe SEC (17.28±6.52) mg/L and (16.76±8.73) mg/L respectively. All differences were of statistically significant(P<0.05). Conclusion Increase of serum IL-6 and CRP as well as high systemic inflammatory state correlate with left atrial thombus and severe SEC in patients with AF.

Evolution of Genetic Polymorphisms of Plasmodium falciparum Merozoite Surface Protein (PfMSP) in Thailand

Plasmodium falciparum malaria is a major public health problem in Thailand due to the emergence of multidrug resistance. The understanding of genetic diversity of malaria parasites is essential for developing effective drugs and vaccines. The genetic diversity of the merozoite surface protein-1 (PfMSP-1) and merozoite surface protein-2 (PfMSP-2) genes was investigated in a total of 145 P. falciparum isolates collected from Mae Sot District, Tak Province, Thailand during 3 different periods (1997-1999, 2005-2007, and 2009-2010). Analysis of genetic polymorphisms was performed to track the evolution of genetic change of P. falciparum using PCR. Both individual genes and their combination patterns showed marked genetic diversity during the 3 study periods. The results strongly support that P. falciparum isolates in Thailand are markedly diverse and patterns changed with time. These 2 polymorphic genes could be used as molecular markers to detect multiple clone infections and differentiate recrudescence from reinfection in P. falciparum isolates in Thailand.

Seasonal Changes of Protein in The Tissues of Male Catfish Mystus Cavasius (HAM) in Bhadra Reservoir, Karnataka, India.

Biochemical constituents of body of the freshwater fish Mystus cavasius were greatly influenced by the breeding activity. Protein was estimated from testes, liver and muscle at different breeding phases. There was an increase in protein levels of testes during maturation of testes was observed and it was related to simultaneous decrease in the liver and muscle. While in immature and preparatory stage, it was found to be the storage phase of protein in muscle and liver. Depletion of protein in muscle and liver and is transferred to testes during maturation has been found to be significant.

Survivin and livin in apoptosis of tumor cells / 国际肿瘤学杂志

The process of apoptosis runs through a variety of signal transduction, and is regulated by many apoptosis-related genes. Inhibitors of apoptosis protein (IAPs) are closely related to the infinite proliferation of tumor cells. IAPs including the recently discovered survivin, livin, are closely related to tumors. The anti-apoptotic mechanism is to be further studied.

Changes of expression of signaling pathway of NF-κB/IκB in rat lung tissue with diabetes / 中国综合临床

Objective To investigate the effect of activation of NF-κB signaling pathway on pathogenesis of lung in diabetes mellitus(DM) rat. Methods The experimental type 2 diabetic rats were built by injecting streptozotocin (STZ) and feeding with high fat and glucose food. At the 12nd and 24th week, we observed the alteration of morphology in the lung of rats in the control group(20 rats) ,the DM group(30 rats)using spectroscopic analysis. The collagen accumulation of lung was observed by masson trihrome staining, and alteration of NF-κB P65, IκBα, and PKC in lung was observed by immunohistochemistry. Results The tissue structure of lung in the DM rats distributed deranged in the light microscope, alveolar wall were thicken, extracellular matrixes increased and pulmonary fibrosis appeared. With the development of pathogenic condition, the expression increased obviously. The staining optical density value of NF-κB P65 in tissue of lung in the 12 w and 24 w DM group were 0. 20 ± 0. 01 and 0. 35 ± 0. 06 respectively, which was significantly higher than those of the control group at the corresponding time point ( 0. 12 ± 0. 02 and 0. 17 ± 0. 03, respectively, Ps ＜ 0. 0l ). The staining optical density value of IκB in tissue of lung in the 12 w and 24 w DM group were 0. 29 ±0. 02 and 0. 36 ± 0. 03, respectively, which were significantly higher than those of the control at the corresponding time point (0. 08 ± 0. 02 and 0. 22 ± 0. 08, respectively, Ps ＜ 0. 01 ). Conclusion The signaling pathway of NF-κB/IκB participate in the occurrence and development in the pathogenesis of lung in DM, and may be one of the mechanisms of lung injury.

Changes of the c-Jun N-terminal kinase in the brains of rats with chronicfluorosis / 中国地方病学杂志

Objective To investigate the expression of c-Jun-N-terminal kinase(JNK) in rat brains with chronic fluorosis and try to reveal the molecular mechanism for the neural impairment induced by the disease.Methods The rats were randomly divided into 3 groups, normal control group(drinking water containing less than 0.5 mg/L of sodium fluoride, NaF), lower fluoride exposed group(drinking water containing 5 mg/L NaF) and higher fluoride exposed group(drinking water containing 50 mg/L NaF), 24 in every group. The rats were examined at the sixth month after feeding. The concentration of fluorine in urine and blood was detected by F-ion selective electrode. The expression of JNK in brains was investigated by using Western blotting and immunohitochemistry staining, and analyze the correlation between activating of JNK and the concentration of fluorine in blood. Results The increased concentration of fluorine in urine(control: 0.92 ± 0.30, lower fluoride exposed group: 2.56 ± 0.91,higher fluoride exposed group: 5.73 ± 3.14, P ＜ 0.05) were observed when 6 months after the beginning of the experiment, and the amount of fluorine in blood was also higher in rats with fluorosis(control: 0.12 ± 0.07, lower fluoride exposed group: 0.36 ± 0.14, higher fluoride exposed group: 0.50 ± 0.18, P ＜ 0.05). The expression of phospho-JNK at protein levels were higher in the brains of rats with fluorosis than that of controls (control: 1.00 ± 0.37, lower fluoride exposed group: 1.20 ± 0.28, higher fluoride exposed group: 1.74 ± 0.69, P ＜ 0.05), whereas no change of total-JNK was found(F = 0.046, P ＞ 0.05). Furthermore, the expression of phospho-JNK in the parietal cortex(119.3 ± 14.1), occipital cortex(112.7 ± 5.4), hippocampus CA3(100.6 ± 8.9), dorsal thalamus (117.8 ± 10.4) and olivary nucleus( 112.6 ± 5.9) of rats in higher fluoride exposed group were higher than that in control( 104.1 ± 8.9,106.6 ± 9.6,106.6 ± 9.7,108.9 ± 6.4,100.3 ± 8.4, all P ＜ 0.05) and lower fluoride exposed group(96.7 ± 17.1,102.5 ± 8.3,106.4 ± 6.5,110.2 ± 9.3,102.4 ± 4.7,102.5 ± 9.8, all P＜ 0.05). The positive stained neurons of total-JNK also distributed in the same brain regions of rats, but no difference was detected between the rats with fluorosis and controls(all P ＞ 0.05). The increased level of phospho-JNK was positively correlated with the fluoride contents in blood of the rats with fluorosis (r = 0.677). Conclusions The expression of phospho-JNK in brains of rats with fluorosis was significantly increased with a correlation to fluoride content in blood, which might be connected to the mechanism of neural impairment induced by chronic fluorosis.

Influence of fracture fragment displacement distance on fracture healing / 中华创伤杂志

Objective To study the influence of the distance of displaced fragment on the union of diaphysis fracture. Methods A wedge-shaped bone fragment was taken from central radial of the right forelimb of 120 New Zealand white rabbits for estabhshment of experimental animal model. The bone fragment was fixed to the main bone with two Kirschner wires, with certain space between bone fragment and the main bone. Then, the rabbits were divided into five groups, ie, Group A (in situ fixation),Group B (the space was 1/5 diameter of the radial shaft), Group C (the space was 2/5 diameter of the radial shaft), Group D (the space was 3/5 diameter of the radial shaft), Group E (the space was 4/5 diameter of the radial shaft). The animals were killed at 2, 4, 6, 8 weeks after operation. X-ray photos were taken to observe the fracture healing and the improved Gary X-ray used for scoring. HE staining after tissue section was employed to observe the histomorphological changes of fracture healing. Immunohistochemical method was used to determine expression of BMP-2. Results X-ray findings showed insignificant statistical difference between Group A and Group B, delayed union in Groups C and D and nonunion of bone absorption in Group E. Morphological observation showed same change in fracture site in Groups A and B lout significant late in emergence, formation and remodeling of the callus in the other groups compared with Group A, mainly with delayed fracture union or nonunion. There was no statistical difference in expression of BMP-2 between Group B and Group A (P ＞ 0. 05), but there was statistical significance in Groups C, D and E compared with Group A at 2 weeks (P ＜0.01). There was statistical difference between Group E and Group A at 4 weeks (P ＜0. 01) but no statistical difference at 6 and 8 weeks between either two groups (P ＞ 0. 05). Conclusions The distance of displaced fragment will influence fracture healing. The larger distance of the displaced fragment will beget more obvious influence on fracture healing. When the distance is more than 2/5 diameter of the bone shaft, the fracture will present union disorder.

Effect of AngiotensinⅡ on Myocardial Fibroblasts Proliferation and Their Signal Transduction Mechanism / 实用儿科临床杂志

Objective To study the effect of angiotensin Ⅱ(AngⅡ) on myocardial fibroblasts(MFs) proliferation,the expression and transposition of protein kinase C epsilon(PKC?) and alpha(PKC?),and to find out the mechanism of AngⅡpromoting proliferation and signal trarsduction.Methods The primary culture neonate rat's MFs was used depending on the different time of cell adherence,by the method of immunohistochemical method identifying MFs,2-4 generations MFs were divided into experimental group and control group,experimental group was added with AngⅡ 10-6 mol/L,and nothing was added to control group.Colorimetric method of metrazolium salt(MTT) was used to detect the MFs proliferation; indirect immunofluorescence was used to detect the distribution and location of PKC? and PKC?,then Image-Pro-Plus 4.0 was used to add up fluorescence intensity.Results 1.The number of MFs in experimental group increased much more than that in control group and there was obviously statistical significance(P

The expression and significance of the multidrug resistance-related proteins P-gp, MRP, LRP and GST-? in lung cancer using tissue microarray / 中国癌症杂志

Background and purpose:Resistance to anticarcinogen is one of the key factors that affect the treatment efficiency in lung cancer. The purpose of this study was to investigate the clinical significance of the multidurg resistance-related proteins P-gp, multidrug resistance-related proteins(MRP),lung resistance associated protien(LRP) and GST-?by detecting their expression in lung cancer and to investigate the mechanism of resistance to anticarcinogen. Methods:S-P immunohistochemistry was used to examine the expression level of proteins P-gp, MRP, LRP and GST- ?in 226 samples of lung cancer and 23 samples of normal lung tissues. Results:The positive rates of P-gp, MRP, LRP and GST-? in lung cancers were 46.0%, 42.0%, 54.4%, 62.4% respectively. Significant difference existed between tumorous tissue and normal lung tissue (17.4%, 13.0%, 17.4%, 21.7%). The positive rates of P-gp, MRP, LRP and GST-? in poorly differentiated-type of NSCLC were 33.3%, 22.8%, 33.3%, 47.4%, compared with differentiated-type of NSCLC (59.7%, 58.1%, 73.6%, 79.1%) (P

Expression and significance of p53 and p21~(WAF1)proteins in hepatocellular carcinoma / 肿瘤研究与临床

Objective To explore the expression and significance of p53 and p21_(WAF1)proteins in hep- atocellular carcinoma(HCC).Methods Immunohistochemical(S-P)method was used to detect the expression of p53 and p21~(WAF1)proteins in the 41 patients with HCC and 30 cases of paracancerous tissues.Results The positive rates of p53 and p21~(WAF1)proteins were 43.9 % and 75.6 % respectively.The expression of the proteins was significantly higher in tumor than that in corresponding paracancerous tissue(P0.05). p53 ex- pression showed significant difference in different pathologic grades and cases with or without intrahepatic metastasis and thrombus in the portal veins(P

A comparision between ?-AP and INS in the patient with Alzheimer disease by different treatment methods / 中国基层医药

Objective To check the change of content of serum beta-apoliprotien(?-AP) and insulin(INS) in the patient with Alzheimer disease, and to reflect the effect of using different drug. Methods 60 cases of this disease and 30 cases of the control group were been detected by radioimmunoassay(RIA). Results The content of ?-AP in Alzheimer disease(AD) was obviously higher than that in control groups(P

The Immunohistochemical Study of C-kit Gene on Apoptosis in Spermatogenesis / 대한비뇨기과학회지

PURPOSE: Information has been limited on the genetic control of germ cell apoptosis and its role in the pathogenesis of male infertility. The object of this study was to investigate the effects of c-kit gene expression and apoptosis on human spermatogenesis. MATERIALS AND METHODS: Testicular specimens were obtained from 90 infertile males with nonobstructive azoospermia(NOA) due to primary testicular failure and from 9 healthy volunteers. The specimens of infertile men were divided into 4 groups according to hitopathologic findings: Sertoli cell only(SCO) syndrome(A), maturation arrest(B), hypospermatogenesis(C) and disorganization with sloughing(D). C-kit gene expression and apoptosis were detected with immunohistochemical stain. RESULTS: The frequency of c-kit expression was lower(p<0.05) and that of apoptosis was higher(p<0.01) in infertility groups B, C, and D compared to that of the control group. A significant inverse correlation was observed in the c-kit gene expression and apoptosis in groups B, C and D(p<0.05). A similar relationship was also observed in the Sertoli cells in group D and the control group. CONCLUSIONS: With the result, we suggest that variation in c-kit gene expression and apoptosis are associated with abnormal spermatogenesis and play a role in the pathophysiology of male infertility.

Relationship between pluse pressure and serum levels of C-protein in elderly patients with HBP / 中国医师杂志

Objective To investigate the relationship between pluse pressure(PP) and serum levels of C-reactive protien(Hs-CRP) in elderly patients with HBP.Methods To measure serum levels of Hs-CRP,total cholesterol(TC),triglyceride(TG),high density lipoprotein-cholesterol( HDL-C),low density lipoprotein-cholesterol(LDL-C) and FBG in 160 patients. Patients were divided into four groups according to the levels of their PP,100mmHg whose serum levels of Hs-CRP were compared.Results Hs-CRP rose with the levels of PP,and were significantly higher in four groups(P

Study on the Correlation among the p53 Protein Expression, Tumor Biological Behavior, p53 Gene Mutation and EBV Infection in NPC / 中国医师杂志

Objective To investigate p53 protein expression in nasopharyngeal carcinoma (NPC) and its correlation with biological behavior, p53 gene mutation and EBV infection in 43 cases of NPC. Methods Immunohistochemical staining was applied to detect p53 protein,PCR-SSCP was used to detect p53 gene mutation, and EBV infection was determined with PCR. Results The positive rate of p53 protein expression was 76 7%(33/43)in the NPC. The positive rate of p53 protein expression in stages Ⅲ and Ⅳ was significantly higher (80 6%) than that in stages Ⅰ and Ⅱ(66 6%) (P

Liver injury of acute pancreatitis:analysis of 214 cases / 中国实用内科杂志

Objective To study clinical characteristics and prognosis of patients with liver injury of acute pancreatitis.Methods 290 cases of acute pancreatitis admitted between January 2001 to October 2003 were reviewed and analyzed retrospectively.Results Through comparing different types of AP with liver injury and C-reactive protein (CRP) changes,it was found that the more severe AP was,the more significant liver injury was;and liver injury of severe AP had some connection with CRP(.P.

Inhibitory effect of P195 and its antibodies on the invasion of merozoite of Plasmodium falciparum into human erythrocyte / 中华传染病杂志

Objective To map out the binding site of P195,which is the major protein on the surface of P.falciparum merozoites,to human erythrocytes,and offer a basis for designing malaria vaccine to blockade invasion of merozoites into human erythrocytes.Methods Eight proteins derived from P195 were expressed in E.coli,and purified by Ni-chelare affinity chromatography.There after,the eight fragments and rabbit serums immunized by which were added into culture medium of P.fatciparum in vitro respectively.Twenty-four hours later,the invasion of merozoite to erythrocyte was observed.Results The antibodies which were induced by three fragments of P195,M6(Amino Acid,AA384～595),M7(AA 595～897)and M11(AA 1397～1563)could inhibit the invasion of P.falciparum merozoite into human erythrocytes.Especially,one fragment of P195,M6,had the ability to inhibit the invasion of P.falciparum merozoite into human erythrocytes.Conclusion M6,a fragment of P195 on the merozoite of P.falciparum may contain a domain thought to be involved in the recognition of human erythrocyte.The domain can be used as a candidate antigen for a malaria vaccine.