3.Quantitative analysis of the patterns of heat shock protein 70 expression induced by Hantaan virus infection in vitro
Lu YU ; Heng MA ; Chun-Guang DUAN ;
Chinese Journal of Infectious Diseases 2001;0(06):-
Objective To explore the pattern and quantify the heat shock protein HSP)70 and HSP70 mRNA in Vero-E6 cells after infection with Hantann virus(HTNV).Methods The expres- sion of HSP70 and change of its mRNA level were detected by immunocytochemical staining,nucleic acid hybridization in situ and RT-PCR.Results In situ hybridization and RT-PCR were used to eval- uate the level of HSP70 mRNA during Hantaan 76-118 infection.HSP70 mRNA increased 0.5 h after infection,reached its peak by 12 h and gradually declined to steady state level by 72 h(vs.sham infec- ted group,P<0.05).The expression of HSP70 protein induced by Hantaan 76-118 infection was e- valuated by quantitative immunocytochemical staining.HSP70 increased 0.5 h after infection,reached its peak by 12 h and decreased at 72 h after infection(vs.sham infected group,P<0.05).Conclu- sions HSP70 can be induced directly by HTNV infection at both mRNA and protein levels,It pro- vides a basis for the further study of the pathogenesis,prevention and treatment of hemorrhagic fever with renal syndrome(HFRS).
4.The study of expression of matrix metaUoproteinases-2 in patients with colorectal cancer
Ya MA ; Guang-Ling CAI ; Guang-Yin YU ; Wei-Hua YIN ; Li CHEN ;
Chinese Journal of Primary Medicine and Pharmacy 2005;0(11):-
Objective Research the correlation between expression of MMP-2 in colorectal cancer and patho- logical factors,or biological behaviors in colorectal cancer.Methods The expression of MMP-2 in 68 cases with col- orectal cancer were detected by immunohistochemical staining.Results The expression of MMP-2 in colorectal can- cer was not correlated with tumor region and histologic type,but related with depth of tumor invasion and metasta- sis.The frequency of positive cases in patients with colorectal cancer with of lymph,node metastasis was significantly higher than that in patients without lymph node metastasis(P
5.Latent infection of human herpes virus in hematopoietic system.
Ke-Fu WU ; Xiao-Tong MA ; Guo-Guang ZHENG ; Yu-Hua SONG
Journal of Experimental Hematology 2008;16(6):1251-1256
Up to date, eight types of human herpes viruses have been identified, all of which are ubiquitous, and usually establish latent infection in the host after primary infection. Since most of the herpes viruses are maintained in an asymptomatic form, they are often neglected. However, under some circumstances, these herpes viruses can cause fatal or severe diseases. Furthermore, the association of herpes viruses with hematopoietic malignancies is attracting researchers' attention. With the extensive development of hematopoietic stem cell and organ transplantation, reports regarding transplantation failure and complication caused by infection of human herpes virus has been increasing. Cytokine storm was firstly suggested as the mechanism of graft-versus-host diseases. In recent years, which has also been applied in the pathogenesis research of inflammation, and is supposed to play an important role in severe virus infection. In this paper, through discussing the possible role of latent infection of human herpes virus in the failure or complication of bone marrow or hematopoietic stem cell transplantation, and in refractory leukemia, the function and significance of latent infection of human herpes virus and the cytokine storm it caused were investigated.
Cytokines
;
immunology
;
Hematopoietic System
;
immunology
;
virology
;
Herpesviridae Infections
;
Humans
;
Virus Latency
6.Expression, purification and detection of activities of AprA and its inhibitor AprI from Pseudomonas aeruginosa
Run CAO ; Yaping GAO ; Yu LIU ; Chenghua LIU ; Peng JIN ; Yuanfang MA ; Guang YANG
Military Medical Sciences 2016;40(12):974-977,1012
Objective To clone and express the alkaline protease AprA , one important virulence factor secreted by Pseudomonas aeruginosa(PAE)in Escherichia coli, to clone and express the inhibitor of AprA (AprI) and its substrate flagellin , and to detect the function of AprA and the inhibitory function of AprI .Methods The genes encoding AprA ,AprI and flagellin gene were amplified respectively by PCR using PAE PAO 1 genome DNA as the template .The expression vec-tors (pET-28a-AprA, pET-28a-AprI and pET-28a-Flagellin) were constructed and transformed into E.coli BL21(DE3) respectively.The recombinant AprA protein was expressed by IPTG induction and purified via denaturing and renaturation. The recombinant AprI and flagellin were expressed and purified by Ni 2+affinity chromatography .The cleavage activities of AprA on flagellin were detected by SDS-PAGE.Results Recombinant AprA , AprI and flagellin protein were expressed and purified .It was demonstrated that AprA cleaved flagellin , which was blocked by AprI .Conclusion Recombinant AprA could cleave its substrates as an alkaline protease , and its inhibitor AprI inhibits the activities of AprA .This study will contribute to further investigations on the role of AprA in the pathogenesis of PAE .
7.Recent advance in predicting oral bioavailability.
Acta Pharmaceutica Sinica 2006;41(10):917-920
9.Internal fixation combined with bone grafting through the injured pedicle for the treatment of thoracolumbar burst fractures via a paraspinal intermuscular approach under MAST Quadrant retractor.
Sheng-hua HE ; Du-jun MA ; Zhi-tao SUN ; Wei-ji YU ; Ye-guang WANG
China Journal of Orthopaedics and Traumatology 2015;28(1):17-20
OBJECTIVETo study clinical effects of short-segment fixation and injured vertebra bone grafting through injured pedicle for the treatment of thoracolumbar burst fractures under MAST Quadrant retractor via a paraspinal muscle approach.
METHODSThe data of 42 patients with thoracolumbar burst fractures treated from June 2009 to September 2012 were reviewed. There were 19 males and 23 females, with an average age of (55.2±11.9) years old. The mean injury time was (5.8±4.3) days. Fracture segments included T10 in 3 cases, T11 in 6 cases, T12 in 13 cases, L1 in 9 cases, L2 in 7 cases, and L3 in 4 cases. According to Denis classification, 9 patients were type A, 21 patients were type B, 5 patiens were type C, 5 patients were type D, and 2 patients were type E. All the patients were treated with short-segment pedicle screw-rod system fixation under MAST Quadrant via the paraspinal muscle approach. The operative time, blood loss, complications and the height of vertebra, kyphosis Cobb angle, VAS scores, JOA scores were measured before and after treatment.
RESULTSAfter treatment, the vertebral height and kyphosis Cobb angle were restored. Compared with preoperative results, postoperative vertebral height and kyphosis Cobb angle, VAS scores and JOA scores were all improved. But there was no statistically significance in vertebral height, kyphosis Cobb angle between postoperative at 1 week and 1 year.
CONCLUSIONInternal fixation combined with injured vertebra bone grafting through the injured pedicle for the treatment of thoracolumbar burst fractures via the paraspinal intermuscular approach under MAST Quadrant is a safe, minimally invasive, effective and satisfactory method.
Adult ; Aged ; Aged, 80 and over ; Bone Transplantation ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; surgery
10.Two-step Tandem Chromatography Purification of Anti-human CD80 Monoclonal Antibody 4E5 from Mouse Ascites
Hong-Bing MA ; Yu-Hua QIU ; Ran TAO ; Wen-Xiang LI ; Ying XU ; Xue-Guang ZHANG ;
China Biotechnology 2006;0(08):-
A two-step method for the purification of blocking-type anti-human CD80 monoclonal antibody 4E5 from mouse ascites was developed using anion exchange and gel filtration in combination. The ascites was first purified by anion exchange after centrifugation and filtration. The experimental parameters of sample loading and elution were optimized. The optimized loading condition was pH 8.0,50 mmol/L Tris-HCl and satisfactory results were obtained using a 0~0.5mol/L NaCl step elution. The fraction containing the protein of interest was directly loaded on gel filtration column and eluted using a 20 mmol/L phosphate buffer at pH 7.2. The purity of the obtained monoclonal antibody was up to 95% with a recovery of 61%. The purity of mAb could efficiently inhibit the growth of Daudi cells. The amplification of the method was also studied using a Bio-Scale Q5 column and the result was satisfied.