2.The effects of aerobic exercise on atherosclerotic plaques in apolipoprotein E gene deficient mice
Xu ZHANG ; Shixiu SHAO ; Tieshan LI ; Yong ZHANG ; Jianzhong BI
Chinese Journal of Physical Medicine and Rehabilitation 2011;33(8):561-565
Objective To investigate the influence of aerobic exercise on the vulnerability of atherosclerotic plaques and to observe the concentration of matrix metalloproteinase-9 ( MMP-9 ) and matrix metalloproteinase-14(MMP-14) in carotid atherosclerotic plaques in apolipoprotein E (APoE) gene deficient mice.Methods Eighty male, APoE gene deficient mice were divided into two equal groups: an aerobic exercise group and a limited action control group. Carotid atherosclerotic lesions were induced by perivascular constrictive collars placed on the right common carotid artery. Blood lipid levels in the exercise group were measured after 8 weeks of treadmill running and also in the control group. The morphological characteristics of carotid atherosclerotic plaques were observed in the two groups. Plaque area and fibrous cap thickness were measured. Lipid and collagen positive areas were quantified and the ratios correlated with the entimal areas were calculated. Plaque rupture rate and vulnerable index were calculated.Immunostaining was used to detect MMP-9 and MMP-14 expression in the atherosclerotic plaques. The mRNA levels of MMP-9 and MMP-14 mRNA in the fresh carotid plaques were quantified using a real-time reverse-transcriptase polymerase chain reaction (RT-PCR). Western blotting was performed for examining MMP-9 and MMP-14 protein expression in the fresh carotid plaques.Results No significant difference in serum lipid profiles or plaque area was found between the exercise and control groups. Compared with the control group, mean fibrous cap thickness, cap/core ratio and collagen content were all significantly higher in the exercise group, and lipid content was significantly lower. Plaque rupture rate and the vulnerable index were both significantly lower in the exercise group. Immunostaining showed that MMP-9 and MMP-14 expression were lower in the exercise group compared with the control group.The mRNA expression of MMP-9 and MMP-14 was also significantly lower.Conclusions Aerobic exercise can decrease the expression of MMP-9 and MMP-14 in carotid atherosclerotic plaques in ApoE gene deficient mice. Aerobic exercise may play a role in forestalling atherosclerosis by increasing the stability of plaque and decreasing plaque vulnerability.
3.Use of Green Fluorescent Protein to Study the Relationship between Legionella pneumophila and Its Protozoan Host
Shao-Song HUANG ; Run-Lin XU ; Yong-Jun LU ;
Microbiology 1992;0(02):-
Protozoans,the natural host of the facultative intracellular pathogen Legionella species,play an important role in survival,proliferation,virulence and stress resistance of Legionella species. By repeating transformation and selection,a spontaneous mutant of plasmid over expressing green fluorescent protein was obtained. This mutant replicates and is maintained stably in Legionella cells. The colonies of L. pneumophila harbouring the mutated plasmid were intense green in colour even under the daylight. After feeding BF1 strain of Tetrahymena thermophila with transformed L. pneumophila,the intracellular dynamic of changing of bacterial shape,bacterial proliferation and lysis of the host cell due to the bacterial proliferation were observed clearly under fluorescent microscopy. Thus,the present paper provides a simple and intuitionistic strategy for investigating the ecological and cellular relationship between L. pneumophila and its host.
4.Comparison of naveibine and cisplatin versus gemcitabine and cisplatin in refractory metastatic breast cancer
Zhong-Fu SHAO ; Gang XU ; Ming ZHOU ; Yong-Dong CHEN ;
Cancer Research and Clinic 1999;0(05):-
0.05).The degreeⅢ~Ⅳthrombocytopenia was more common in group A than in group B,but the degreeⅢ~Ⅳhypolekocytosis and phlebitis was more serious in group B.Conclusion NC and GC for treating refractory metastatic breast cancer have a high response rate and tolerable side effects.
6.Significance of CCL20 expression in liver biopsies of chronic hepatitis B patients
Xianan SHAO ; Sidong XIONG ; Wei XU ; Fuhua YUAN ; Ruibin LI ; Yong WANG ; Zhihe CHEN ; Changjiang XU
Chinese Journal of Laboratory Medicine 2008;31(8):864-867
Objective To observe the correlation of histologicalactivity(HAI) of chronic hepatitis B (CHB) with CCL20 expression, and to investigate the impact of CCL20 expression in CHB infection. Methods On the basis of established competitive quantitative RT-PCR with an internal standard, the expression of the CCL20 in the hepatocytes in different infected patterns of HBV infected cells and liver biopsies were quantified and at the same time its correlation to HAI were explored. Results In the cell levels, the expression quantity of CCL20 in control cells (HepG2), persistent HBV infected hepatocytes( HepG2. 2. 15) are (2. 65 ± 0. 02) pg/106 cells, ( 1.22± 0. 04) pg/106 cells, respectively. There were significantly differences between them ( t = 39. 66, P < 0. 01 ). The expression of CCL20 was enhanced in hepatocytes stimulated by PMA but their expression pauern was not changed. Moreover, CCL20 expression in liver biopsies with CHB was (3.54 ± 0. 65 ) pg/20 mg and CCL20 expression in control groups was ( 8. 74±0. 56) pg/20 mg. The expression of CCL20 between two groups was different (t =30. 09,P <0. 01 ) and correlation lied in between HAI and CCL20 expression in liver biopsies of CHB patients ( r = 0. 675, P =0. 023 ). Conclusion CCL20 expression was down-regulated and it was correlated to HAI of liver biopsies in CHB patients.
7.Effect of Qiling Decoction combined HAART on expression levels of Treg cells and Th17 in HIV/AIDS patients.
Wen-Fang XU ; Yong WU ; Guo-Shao PAN ; Jian-Ping ZHONG ; Shao-Bo LAN ; Xue-Fang CHEN ; Qiu-Qiong LU
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(2):157-161
OBJECTIVETo explore the effect of Qiling Decoction (QD) combined highly active antiretroviral treatment (HAART) on expression levels of peripheral blood Th17 and Treg cells in HIV/AIDS patients.
METHODSTotally 55 HIV/AIDS patients were randomly assigned to the treatment group (28 cases) and the combination group (27 cases). Besides, 21 HIV negative patients were recruited as the healthy control group. Those in the treatment group received HARRT alone, while those in the combination group received HAART combined QD. The observation lasted for 24 weeks. Meanwhile, according to peripheral blood CD4+ T cell counts before treatment, HIV/AIDS patients were assigned to three subgroups. For patients in subgroup 1, 1 cells/microL < CD4+ T cell counts < or = 100 cells/microL; For patients in subgroup 2, 101 cells/microL < CD4+ T cell counts < or = 200 cells/lL; For patients in subgroup 3, 201 cells/microL < CD4+ T cell counts < or = 350 cells/microL. Expression of peripheral blood Th17 and Treg cells, and number of CD4+ T cell counts were detected using flow cytometry (FCM)in HIV/AIDS patients at the pre-treatment baseline, week 4, 12, and 24, as well as those in the healthy control group.
RESULTSCompared with the healthy control group, CD4+ T cell counts and the baseline expression level of Th17 cells in the peripheral blood of HIV/AIDS patients significantly decreased, the expression level of Treg cells significantly increased P < 0.01). Compared with before treatment in the same group, CD4+ T cell counts all increased at week 4, 12, and 24 in the two treatment groups, showing statistical difference (P < 0.05, P < 0.01). There was no statistical difference in the effective rate at various CD4+ T cell levels between the two groups (P > 0.05). There was no statistical difference in expression levels of Th17 and Treg cells between the combination group and the treatment group at any time point (all P >0.05). The Th17/Treg ration significantly increased in the combination group after 24 weeks of treatment, showing statistical difference when compared with the treatment group (U = 2.135, P = 0.038).
CONCLUSIONQD could improve the immune balance of Th17/Treg cells, which might be one of its mechanisms for improving HIV/AIDS patients' immunity.
Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adult ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; Case-Control Studies ; Drugs, Chinese Herbal ; therapeutic use ; Female ; HIV Infections ; drug therapy ; immunology ; Humans ; Male ; Middle Aged ; Phytotherapy ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology
8.Clinical research of percutaneous K-wires fixation after manipulative reduction combining with gypsum or splint fixation for treatment of the Barton fractures of aged people.
Jian-liang CHEN ; Long-jun ZHANG ; Yong XU ; Shao-bing ZHU ; Xiao-dong ZHENG
China Journal of Orthopaedics and Traumatology 2016;29(1):8-12
OBJECTIVETo investigate the feasibility of using percutaneous K-wires fixation after manipulative reduction combining with gypsum or splint fixation for treatment of the Barton fracture of aged people.
METHODSFrom June 2011 to May 2013, 91 patients with closed and fresh Barton fracture were analyzed retrospectively which information was complete. According to different methods these patients were divided into three groups: 33 patients in group A were treated by simplex fixation of gypsum or splint after manipulative reduction, including 20 males and 13 females with an average age of (63.27 ± 5.83) years old; 35 patients in group B were treated by percutaneous K-wires fixation after manipulative reduction combining with gypsum or splint fixation, including 23 males and 12 females with an average age of (64.25 ± 4.53) years old; 23 patients in group C were treated by open reduction and internal fixation including 15 males and 8 females with an average age of (64.04 ± 4.83) years old.
RESULTSAll the patients were followed up for 10 to 16 months with an average of 12 months by the method of recheckging in outpatient clinic. According Dienst scoring system: in group A, the result was excellent in 15 cases, good in 7 cases, fair in 11 cases; in group B, excellent in 25 cases, good in 6 cases, fair in 4 cases; in group C, excellent in 16 cases, good in 5 cases, fair in 2 cases. The excellent and good rate of group C was better than that of group B and A, further more group B is better than group A. According to X- ray standard, in group A, 26 patients were acceptable, 7 patients were unacceptable; in group B,28 patients were acceptable, 7 patients were uacceptable; in group C, 19 patients were acceptable,4 patients were unacceptable; there were no statistical significant difference (P > 0.05). There were 18 patients with emerging recrispation in group A, 10 patients in group B, 6 patients in group C, the result of group B and C was better than that of group A, and there was no statistically significant between group B and C (P > 0.05).
CONCLUSIONUsing percutaneous K-wires fixation after manipulative reduction combining with gypsum or splint fixation for treatment of the Barton fracture of aged people is an effective method, it has advantages of micro-invasived, reliable fixation, less complication, not need to fix wrist joint and early functional exercise.
Aged ; Bone Wires ; Female ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Radius Fractures ; therapy ; Retrospective Studies ; Splints
9.Construction and prokaryotic expression of His-tagged expression vector of human IP-10 and identification of its activity
Ziyun SHAO ; Zhifeng LIU ; Yi PENG ; Jia XU ; Qinghe QIN ; Peng DENG ; Yong JIANG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To construct prokaryotic expression vector of His-tagged human IP-10 for further study of its biological function in the inflammatory response. METHODS: The coding sequence of IP-10 lacking signal peptide was amplified from human lung cDNA library by polymerase chain reaction (PCR) and the fragment was cloned into pET-14b plasmid for the construction of His-tagged fusion protein expressing vector, pET-14b/IP-10. After being identified by enzyme digestion and sequencing, the recombinant vector was transformed into a strain of E. coli, BL21 (DE_3). The expression of His-tagged fusion protein was induced with IPTG and purified with Ni+-NTA affinity chromatography. Then the chemotactic activity of IP-10 was determined by transwell migration assay on THP-1 cells. RESULTS: The construction of pET-14b/IP-10 recombinant vector was proved by enzyme digestion and sequencing. The fusion protein IP-10, which was purified by a routine Ni+ affinity method, had an activity on the induction of cell migration of THP-1. CONCLUSION: We successfully construct IP-10 fusion protein expressing vector and get the fusion protein with high bioactivity, which provides essential materials for the future studies on IP-10.
10.Application of in vivo electroporation technology in plasmid mediated gene transfer and DNA vaccination
Zhou ZHANG ; Zhiyong XU ; Pei CHEN ; Fang ZHOU ; Xiansheng ZHANG ; Yong LIU ; Yiming SHAO
Chinese Journal of Microbiology and Immunology 2010;30(6):551-554
Objective To investigate the effects of in vivo electroporation on plasmid mediated reporter gene expression and immunogenicity of DNA vaccine. Methods Luciferase expression plasmid was administered intramuscularly to BALB/c mice at 8μg and 40μg dosage level through injection with or without eletroporation Luciferase expression level in murine muscle was detected by IVIS imaging system 24 h after injection. DNA vaccine plasmid p1.0-gp1455m carrying codon-optimized env gene of CN54 strain ( HIV-1 CRF07_BC) was administered to mice at dosages of 8μg and 40μg through the two approaches mentioned above. Mice were immunized at week 0,2 and 4. Env-specific immune responses were detected at two weeks post the second and the third vaccinations. Env-specific antibody immune responses were determined by ELISA. Euv-specific cellular immune responses were determined by IFN-γ ELISPOT. Results Luciferase expression level in murine muscle was significantly increased as much as 35 folds through in vivo eletroporation. Results of ELISA and ELISPOT revealed that in vivo eletroporation could significantly enhance both the humoral and cellular immune responses induced by DNA vaccination. The responses induced by electrodelivered p1.0-gp1455m at 8 μg dosage were better than those induced by simple intramuscular injection with 40 μg of plasmid DNA. On the other hand, 2 injections followed by electroporation elicited comparable level of humoral and cellular immune responses with those induced by 3 injections without electroporation. Conclusion In vivo electroporation was capable of enhancing both the plasmid-mediated gene expression and immunogenicity of DNA vaccine.