2.Dendritic cells modified with interleukin 23 acquire antigen from apoptotic pancreatic carcinoma cell and induce CTLs immune response
Guang TAN ; Zhongyu WANG ; Shuo YIN
Chinese Journal of Immunology 2001;0(10):-
Objective:To study the immune response to murine pancreatic carcinoma by bone marrow-derived dendritic cells(BM-DCs) modified with interleukin-23 after acquiring antigen from apoptotic pancreatic carcinoma cell.Methods:The murine IL-23 cDNA was sub-cloned into dual-expression vector.DCs were pulsed with apoptotic tumor cell antigen after transduced with interleukin-23 gene.The immune preventative and immunotherapeutic effects of DC vaccines on mice with pancreatic cancer were assessed.Results:IL-23 protein could apparently increase the antigen presenting ability of DC.After the vaccination of DC vaccines.IFN-? production in treatment group were significantly more than that of control group(P
3.Preparation of Bone Marrow Dendritic Cells with TNF-? and The Immune Response Against Malignant Pancreatic Cell by Dendritic Cell Vaccine
Guang TAN ; Jinshuai REN ; Zhongyu WANG ; Shuo YIN
Chinese Journal of Bases and Clinics in General Surgery 2003;0(06):-
Objective To study the method of obtaining a large number of dendritic cells (DC). To study the specific cytotoxicity T lymphocyte (CTL) effect against tumor cells initiated by DC pulsed with peptide of cancer cell. Methods Development of cells with cytologic features of DC in bone marrow cultures supplemented with granulocyte macrophage-colony stimulus factor (GM-CSF) and IL-4. Determining the DC phenotype and the specific structure by electronic microscopy. The CTL effect against pancreatic carcinoma leading by the DC pulsed with tumor cells lysate in vitro was observed. Results A large number of typical DC was proliferated by supplementing with GM-CSF and IL-4 cytokines. DC had specific cell appearance and structure, and highly expressed various cell surface molecules. TNF-? had the ability of stimulating DC mature, the mature DC had the enhancing abilities of antigen presenting and IL-12 self-secreting, as well as, expressed higher levels of CD54, MHC-Ⅱ and CD86 molecules than control group (P
4.Role of Spry1 in osteogenic differentiation of human bone marrow mesenchymal stem cells under miR-21 regulation
Nan YANG ; Wei ZHOU ; Guang WANG ; Yin DING ; Yan JIN
Chinese Journal of Tissue Engineering Research 2014;(32):5085-5090
BACKGROUND:Previous studies have found that miR-21 expression is increased during osteogenic differentiation of bone marrow mesenchymal stem cells, but the action and molecular mechanism of miR-21 are stil unclear. OBJECTIVE:To verify the target gene of miR-21, Spry1, and to explore the role of Spry1 in osteogenic differentiation of human bone marrow mesenchymal stem cells. METHODS:Luciferase report was used to verify Spry1 gene targeted by miR-21, and western blot assay was used to detect the expression of Spry1 in the osteogenesis of human bone marrow mesenchymal stem cells. Spry1 expression vector was established and transfected into human bone marrow mesenchymal stem cells. Osteogenesis ability of human bone marrow mesenchymal stem cells was analyzed after Spry1 high expression by alkaline phosphatase, alizarin red staining, RT-PCR and western blot. RESULTS AND CONCLUSION:Luciferase report suggested that Spry1 was a target gene of miR-21. The expression level of Spry1 was decreased in the osteogenesis of human bone marrow mesenchymal stem cells. Increasing expression of Spry1 could inhibit osteogenic differentiation of human bone marrow mesenchymal stem cells. These results indicate that Spry1 as a target gene of miR-21 negatively regulates osteogenic differentiation of human bone marrow mesenchymal stem cells, and plays an important role in bone formation process.
5.A study of urinary tryptase expression in patients with diabetic nephropathy and its pathological significance
Jingmin ZHENG ; Guang YIN ; Shifeng YUN ; Jianping WANG ; Zhihong LIU
Journal of Medical Postgraduates 2014;(11):1176-1179
Objective Increased renal mast cells have been found in significant correlation with clinicopathological features of diabetic nephropathy ( DN) .However, the exact pathological significance of mast cells still remained to be elucidated.As one of the most abundant serine proteases, tryptase is specifically expressed by mast cells.The study was to observe the expression of tryptase in the urine of patients with diabetic nephropathy and realize the pathological significance of urinary tryptase and renal mast cells in the development of diabetic nephropathy. Methods Urine samples from 103 patients with diabetic nephropathy who were hospitalized at the clinical unit of the nephrology centre of Jingling Hospital from January 2012 to June 2013 were collected.According to concentra-tion of urinary albumin excretion rate, the patients were conducted to early-stage DN group (microalbuminuria,n=10), middle-stage DN group (proteinuria stage, n=31) and end-stage DN group (renal insufficiency stage,n=62).Meanwhile, urine samples from 20 normal persons were taken as the normal control group.Tryptase level in each urine sample was measured by using an ELISA kit.The average tryptase levels were compared among different groups and the correlation between urinary tryptase level and clinical indices was analyzed. Results ①In most cases, tryptase was not detected in the urine samples from normal persons(0.7 ±1.4 ng/mg creati-nine).However, the urinary trypatse level increased significantly with the development of diabetic nephropathy (11.6 ±10.5 ng/mg creatinine for early stage, 29.7 ±19.2 ng/mg creatinine for middle stage, and 44.6 ±43.4 ng/mg creatinine for late stage group).②The urinary tryptase level was found in significant correlation with ser-um creatinine (r=0.325, P<0.01), serum cystatin C (r=0.391, P<0.01), serum urea nitrogen (r=0.27, P<0.01), 24 hour uri-nary protein (r=0.389, P<0.01), urine C3 (r=0.276, P<0.05), urine retinol-binding protein (r=0.365, P<0.01), urine lysozyme (r=0.461, P<0.01), urine N-acetyl-β-glucosaminidase level (r=0.568, P<0.01), urinary kidney injury molecule-1 (r=0.434, P<0.05) and interleukin-18 level (r=0.375, P<0.05).No significant correlation was found between urinary tryptase level and age, gender, fasting blood sugar, postprandial blood sugar, HbA1c, triglyceride, high density lipoprotein or low density lipo-protein. Conclusion Mast cells play an important role in the development of diabetic nephropathy and might be a novel and effective target for the treatment of DN.
7.Clinical application of fat granule auto-graft in facial soft tissue depression
Lianbo ZHANG ; Bin WANG ; Qingguo GAO ; Guang ZHANG ; Weitian YIN
Chinese Journal of Medical Aesthetics and Cosmetology 2008;14(2):91-93
Objective To evaluate the clinical application of fat granule auto-graft in facial soft tissue depression reconstruction.Methods Autologous subcutaneous fat granules were obtained by syringe aspiration from donating site.then washed with normal saline.Small amounts of fat granules were injected into the facial sites with soft tissue depression by means of multiple passes immediately.Results We performed such fat iniection in a total of 18 cases,all of the procedures were safe and successful.In most cases,single injection were enough,only one underwent two sessions of fat iniection.All members were followed-up for 1.5 months to 24months,the average were 14 months.All facial tissue depression were reconstructed for difierent degrees.The rate of fullness and symmetry.fullness and pretty symmetry and fullness with little asymmetry were 77.8%,16.7%and 5.5%.respectively.No infection,fat necrosis or liquefaction occured.Conclusion Being satisfled in correction of deformity of facial depression.the implantation of autologous fat globules iS safe and effective with less side-effects.
9.High-level Secretion Expression of Human ScFv Against Botulinum Neurotoxin A in Pichia pastoris*
Hui WANG ; Jun YIN ; Xiao-Jun HOU ; Hong-Guang XING ;
Microbiology 1992;0(02):-
The specific ScFv gene against botulinum neurotoxin A (BoNTa)was cloned into pPIC9k. Positive integrators were screened by increasing the dose of G418 in culture and expressed in Pichia pastoris GS115. As a result, engineered recombinant clone were obtained. 26 kD product of interest was seen easily in SDS-PAGE. Expression of human ScFv got the highest level 15% of total secreted proteins during 72~84 h after 1% methanol inducing. Purification of ScFv was finished by two steps: gel filter and ion exchange. Competing ELISA showed that recombinant ScFv could compete with antiserum to specific bind BoNTa.
10.Reaction Time in Patients with Depression
Yin-hua WANG ; Ning LI ; Xu-guang GAO
Chinese Journal of Rehabilitation Theory and Practice 2006;12(11):983-986
ObjectiveTo investigate whether subjects with major depression display attention deficits.Methods35 antidepressant-free,non-elderly patients with unipolar depression diagnosis according to ICD-10 and CCMD-Ⅲ as well as 35 health control subjects matched for education,age and sex were administered three tests for attention(sustained attention,Selective attention and Attention Blink).ResultsOn the Continuous Performance Test(CPT),depressed participants needed more reaction time and made more mistake than nondepressed participants.There were no differences between two block in reaction time in the CPT of patients and control subjects.The performance on the Stroop Color-Word Test was impaired in depression.Compared with normal subjects,depressed participants presented increased choice reaction times(CRT).Stroop interference was higher in depressed patients than in controls.On Attention Blink(AB) paradigm,both groups performed equally well at reporting the first target appearing in a rapidly presented stream of stimuli.All groups showed an AB,and AB for depressive participants were larger and longer.Correlations were observed between the HAMD score and the reaction time in CPT.ConclusionThe depressive participants showed a decrement tendency in vigilance,a higher Stroop interference and a larger and longer AB.