2.Expression and activities analysis of a fusion protein CREKA/tTF
Yi SU ; Jianghua YAN ; Shengyu WANG ; Jie HE ; Min YE
Chinese Journal of Biochemical Pharmaceutics 2010;31(2):94-97
Purpose To prepare a novel fusion protein of CREKA and tTF as a universal carrier targeting to cancer,and to analyze its activities.Methods CREKA and tTF gene were acquired by PCR,and inserted into plasmid pET22b(+)to construct recombinant plasmid CREKA/tTF/pET22b(+),and the fusion gene was expressed in E.coli BL21.The fusion protein Wag purified through Nickel-affinity chromatography column.After purifying,the fusion protein was refold by subsequent dialysis.The activities of the fusion proteins were measured by coagulation timing and quantitative fluorescence test in vitro.Results The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was obtained.The fusion protein was highly expressed in E.coli BL21.The coagulation of the fusion protein Was determined by the coagulation test.And the capability of the fusion protein effectively binding to clotted plasma proteins is identified in quantitative fluorescence test.Conclusion The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was built.The fusion protein CREKA/tTF with both TF and CREKA activity was successfully obtained.
3.Diallyl disulfide induces human leukemia HL-60 cells differentiation by up-regulating the expressions of p21,STAT1 and CAMTA1
Weiguo HUANG ; Hui TAN ; Lan YI ; Jie HE ; Qi SU
Chinese Pharmacological Bulletin 2010;26(4):513-516
Aim To investigate the molecular mechanisms of differentiation in human leukemia HL-60 cells induced by diallyl disulfide(DADS)using suppression subtractive hybridization(SSH).Methods In our privious study,the subtractive cDNA library was constructed successfully and efficiently. 30 clones were randomly analyed with restriction enzyme.The inserts of cDNAs were analyzed by restrictive enzyme EcoR I.Positive clones were sequenced and the homology of resulting cDNA sequences were analyzed through bioinformatics software Blastn.Results 18 clones contained 100~600 bp cDNA inserts.10 differantiation genes were obtained and involved in cell cycle,signal transduction,metabolism and RNA binding.And 3 of 10 genes,p21,STAT1 and CAMTA1 were up-regulated and detected by RT-PCR,the results matched with SSH.Conclusion sThere are tight correlation between the differentiation induced by DADS and three-upregulated gene:p21,STAT1 and CAMTA1.
4.Clinical analysis of 201 cases of childhood acute myelocytic leukemia.
Jun-jie FAN ; Yi-huan CHAI ; Hai-long HE
Chinese Journal of Pediatrics 2007;45(11):873-874
Adolescent
;
Child
;
Child, Preschool
;
Female
;
Humans
;
Infant
;
Leukemia, Myeloid
;
pathology
;
physiopathology
;
Male
5.Effects of Paired Box2,Proliferation Cell Nuclear Antigen and Cell Apoptosis in Nephrotic Syndrome with Steroid-Resistance
hui-qiong, ZHANG ; zhu-wen, YI ; xiao-jie, HE
Journal of Applied Clinical Pediatrics 2006;0(17):-
Objective To investigate the expression of paired box2(Pax2),proliferation cell nuclear antigen(PCNA) and cell apoptosis in steroid-sensitive and steroid-resistant groups with primary nephrotic syndrome(PNS) and to find out the action of Pax2 expression in PNS with steroid-resistance.Method The expressions of Pax2,PCNA were evaluated by immunohistochemistry and cell apoptosis by fluorescence micoscope.Results Pax2 expression in renal tubule had a positive correlation with PCNA expression in steroid-sensitive group.In steroid-resistant group,Pax2 expression had no correlation with PCNA.Pax2 had a negative correlation with cell apoptosis.Conclusions Pax2 proper expression facilitate PCNA expression and repair tubulointerstitial lesions in steroid-sensitive group.Renal tubular epithelial cell proliferation coordinated with cell apoptosis.Pax2 overexpression in steroid-resistant group lead to the decrease of cell proliferation and cell apoptosis and lead to the severe tubule lesions,which made to glucocorticoid resistance.
9.Characteristics of viral shedding in people infected with SARS-CoV-2 during difference stages
CHEN Xi ; ZHANG Yi-cheng ; ZHANG Jie ; ZHOU Min ; HE Qing ; LUO Jie ; XIAO Chong-kun ; ZHANG Zheng-dong
China Tropical Medicine 2023;23(3):310-
Abstract: Viral shedding of SARS-CoV-2 is a continuous dynamic process, which can be divided into latent stage, initial stage, peak stage and decreasing stage according to the characteristics of viral shedding. After being infected with SARS-CoV-2, the infected person generally stays in the latent period for 1-3 days, which is characterized by continuous negative nucleic acid test results and no infectiousness, and the risk of infection for close contacts is very low. At the initial stage of viral shedding is characterized by a rapid decline in the Ct value of nucleic acid tests in a short time, and clinical symptoms gradually appear. The infectiousness of the infected person gradually increases during this period, and the risk of infection for close contacts also gradually increases, but it is still in the early stage of infection, the possibility of viral shedding is low, and the risk of infection of secondary close contacts is low. The peak of viral shedding is characterized by low Ct value in nucleic acid test and obvious clinical symptoms; during this period, the infected person is the most infectious, and the risk of infection of the contact is the highest, so the scope of close contacts should be expanded appropriately. The decreasing period is characterized by the gradual increase of Ct value of nucleic acid test and the gradual disappearance of clinical symptoms; during this period, the infectiousness of the infected person gradually decreases to disappear. In an outbreak, an infected person in the decreasing phase is more likely to be an early infected person in the transmission chain. If infected individuals in the decreasing phase are found in an area without a SARS-CoV-2 epidemic, it suggests that the local outbreak epidemic has been spreading for some time and may be larger in scale. According to the characteristics of viral shedding, risk personnel can be determined more scientifically and accurately, so as to minimize the risk and reduce the waste of epidemic prevention resources.
10.Expression of proliferating cell nuclear antigen in renal tissues of children with primary nephrotic syndrome
zu-xiang, MA ; wei-ling, ZHAO ; xiao-jie, HE ; zhu-wen, YI
Journal of Applied Clinical Pediatrics 1992;0(05):-
Objective To investigate the expression of proliferating cell nuclear antigen(PCNA) in renal tissues of children with primary nephrotic syndrome(NS),and elucidate the relationship between PCNA expression and cell proliferation in renal tissues from the children with primary NS.Methods Paraffin-embedded renal biopsy tissue sections from 39 patients with primary NS were examined by immunohistochemical staining with anti-PCNA monoclonal antibody,normal renal tissue sections from 6 nephrectomized patients with nephroma were selected as control. Possible correlation between the percentage of PCNA positive cells and the pathologic type , histopathological score, clinical indices (serum albumin ,serum cholesterol ,serum creatinine and 24 hours urine protein ) before renal biopsy of NS were evaluated separately .Results The percentage of PCNA positive cells in glomeruli and tubulom terstitium of NS patients was significantly higher than that of the control (P