1.Study on LipofectinRM mediated murine SCF genetr ansduction into umbilical blood cells
Yubing CHEN ; Hongmei ZHANG ; Chunlei YU
Chinese Journal of Biochemical Pharmaceutics 2001;22(1):18-20
Purpose The aim is to study the transduction of m urine stem cell factor(SCF) into umbilical blood cells by LipofectinRMmediated and expression in them.Methods Murine SCF cDNA encoding extra cellu lar domain was isolated using PCR from plasmid pRC/CMV(containing SCF gene), and then recombined into the expression vector pcDNA3,and transferred into enrichme nt cultural umbilical blood cells. Semi-quantitative RT-PCR was used to examin e the expression on mRNA level.And the effects of supernatant of transfected umb ilical blood cells were investigated alone or in coordinate with GM-CSF by colo ny formation test of human bone marrow cells in semisolid culture.Results SCF mRNA was expressed in transfected umbilical blood cells.The su pernat ant of transfected umbilical blood cells could increase the number of CFU-GM, s ynergizing with GM-CSF.Conclusion The supernatant of umbil ical b lood cells transfected with vector containing mSCF gene can stimulate the colony formation of human bone marrow cells in combination with GM-CSF.
2.Application of arthroscope in diagnosis and treatment of knee synovitis
Yubing WANG ; Zixuan PANG ; Liang ZHANG ; Fajun KANG
Journal of Regional Anatomy and Operative Surgery 2014;(6):632-634
Objective To investigate the value of arthroscope in the diagnosis and treatment of knee synovitis. Methods 50 cases of knee synovitis diagnosed by arthroscopy and treated by endoscopic synovectomy were studied. There were 10 cases with rheumatoid arthritis, 11 cases with pigmented villonodular synovitis,5 cases with chronic infection of knee joint,12 cases with chronic non-specific synovitis,5 ca-ses with tuberculous synovitis of the knee,4 cases with meniscus injury,3 cases with unknown cause. The efficacy of the treatment was recor-ded. Results All these cases were clearly diagnosed by microscopic examination combined with synovial pathological examination,and 10 cases were corrected with clinical diagnosis post-operation. All cases received primary healing without serious complications. All cases were followed up,and 6 cases of pigmented villonodular synovitis,2 cases of rheumatoid arthritis,1 case of chronic non-specific synovitis and 1 case of tuberculous synovitis of the knee had recurred. The total effective rate was 80. 0%. Conclusion The application of arthroscopy and syno-vial biopsy was effective in diagnosis. Arthroscopic synovectomy had good effect on treatment with less trauma and complications.
3.The 483rd case: renal dysfunction, numbness of right hand
Yueyi ZHANG ; Gang CHEN ; Wei YE ; Yubing WEN ; Xuemei LI
Chinese Journal of Internal Medicine 2021;60(1):90-93
A 65-year-old woman presented with intermittent right hand numbness and elevated serum creatinine for more than 2 months. The histological examination of kidney biopsy showed renal arterioles occlusion and interstitial fibrosis. Pathological abnormality was originally considered as a part of systemic atherosclerosis. Thus, rosuvastatin 20 mg/d, fosinopril 10 mg/d, metoprolol 47.5 mg/d and aspirin 0.1g/d were administrated. No improvement of renal function was seen. Further Congo red staining was applied. Diffuse amorphous eosinophilic substance was deposited in interlobular artery and small arteriolar artery. Combined with the abnormal free light chain (FLC) level and ratio (serum κ 340 mg/L, κ/λ 10.932), the diagnosis of systematic light-chain amyloidosis was confirmed. The patient received 3 courses of chemotherapy regimen as BCD (bortezomib 2 mg d1, 8, 15, 22, cyclophosphamide 0.3 g d1, 8, 15, 22 and dexamethasone 40 mg d1, 8, 15, 22). A hematologic partial response was achieved and serum creatinine decreased to 180 μmol/L.
4.A comparative study on preoperative ICGR15 dye excretion detection and liver function indexes in patients with primary liver cancer
Yubing XU ; Meng FANG ; Jie ZHANG ; Jiabin SHEN ; Chunmei RAO ; Chunfang GAO
Chinese Journal of Laboratory Medicine 2015;38(11):733-736
Objective To study the correlation between preoperative dye exclusion test and liver function in patients with primary hepatic carcinoma.Methods This was a cross sectional survey.A total of 192 cases of primary liver cancer patients were recruited from May 2014 to March 2015 at the Second Military Medical University Affiliated Eastern Hepatobiliary Surgery Hospital.Hereinto, 160 cases were male and 32 females, the male to female ratio was 5: 1.The age of the patients ranged from 26 to 72 years old, and the average age was 50.5 years old.ICG 15 minutes retention rate of ICG clearance test was determined by PDD method in 192 cases of primary liver cancer patients.ICGR15 value was stratified into three stages: ICGR15 < 10% , ICGR1510%-20% , and ICGR15 > 20%.The ICGR15 stage of patients with different ChildPugh grades was analyzed.The biological liver function indexes of patients were simultaneous detected including TBIL, TBA, TP, ALB, PA, ALT, AST, PT-INR, HA, LN, Ⅲ, Ⅳ, APRI, PLT etc.The correlations of ICGR15 and biological indexes of liver function were analyzed using Spearman nonparametric correlation analysis.Results (1) ICGR15 was positively correlated with Child-Pugh grade (r =0.477, P < 0.01) in the 192 cases of HCC.The hierarchical analysis showed that there were significant differences between ICGR15 and different Child-Pugh grades (P < O.05).(2) Child-Pugh classification and ICGR15 comparison further showed that, ICGR15 increased with Child-Pugh grade.While ICG plasma clearance rate (ICGK) and effective hepatic blood flow (EHBF) reduced (P < 0.05).(3) The correlation analysis between ICGR15 and biological indexes of liver function showed that: ICGR15 was positively correlated with TBIL,TBA, ALT, AST, AFU, GGT, PT-INR, HA, LN, Ⅲ, Ⅳ and APRI index [(AST/ULN) × 100/ PLT (× 109/L)] (r =0.422, 0.389, 0.219, 0.301, 0.219, 0.244, 0.325, 0.652,0.403, 0.523, 0.519, 0.434, P < 0.05);and was negatively correlated with TP, ALB, PA, SOD, WBC, PLT (r =-0.290,-0.532, 0.546, 0.531, 0.256, 0.327, P< 0.05).Conclusions ICGR15 as a indicator for liver reserved and dynamic function can comprehensively reflect the liver reserve function is associated with the existing Child-Pugh grades and liver function biochemical indexes.Therefore, ICGR15 could be served as a sensitive index reflecting the preoperative liver reserve function.
5.Enhanced uptake of human mesangial cell line to oxidized low density lipoprotein stimulated by interleukin-1β partly through the lectin-like oxidized low-density lipoprotein receptor pathway
Hua LIU ; Hang LI ; Jianling TAO ; Yubing WEN ; Guojuan ZHANG ; Xuewang LI
Chinese Journal of Nephrology 2010;26(7):520-524
Objective To analysis if intedeukin-1β (IL-1β) can regulate human mesangial cells (HMC) to uptake oxidized low density lipoprotein (Ox-LDL) and if the effect of IL-lβ be changed through the lectin-like oxidized low-density lipoprotein receptor 1 (LOX-l)pathway. Methods The uptake of HMC to Ox-LDL stimulated by IL-1β was observed using Oil Red "O" and flow cytometry. The level of LOX-1 in HMC induced by IL-1β and Ox-LDL was examined using real-time PCR and Western blotting. Results Uptake of Ox-LDL and Dil-Ox-LDL by HMC was up-regulated upon stimulation with IL-1β in a dose- and time-dependent manner. Intracellular mean fluorescence density of Dil-Ox-LDL with LOX-1 blocker in IL-1β stimulation group was decreased compared to that without blocker. The peak level of LOX-1 mRNA reached after 6 h of stimulation and was as high as 6.87-fold of control. IL-1β could induce LOX-1 mRNA expression in a dose-dependent manner. Treated with 10 μg/L IL-1β for 12 h, the upregulation effect on LOX-1 mRNA was as high as 6.57-fold of control. IL-1β could induce LOX-1 protein expression in a time- and dose-dependent manner. The peak level of LOX-1 protein reached after 24 h of stimulation of 5 μg/L IL-1β and was as high as 1.88-fold of control. Treated with 10 μg/L IL-1β for 24 h, the up-regulation effect on LOX-1 protein reached peak and was as high as 2.57-fold of control. IL-1β could induce LOX-1 mRNA and protein expression in a dosedependent manner. Conclusion The expression of LOX-1 can be up-regulated by IL-1β in a dose-dependent manner and the enhanced uptake of HMC to Ox-LDL stimulated by IL-1β partly through the LOX-1 pathway, which means the dyslipidemia of HMC can be enhanced by inflammatory cytokines.
6.Quality Study on Cinnabar-analysis of Dyeing Status for Cinnabar
Xiao XING ; Zhimin LIU ; Tongtong XU ; Da GU ; Yubing LIANG ; Xin ZHANG ; Jianlong YANG
China Pharmacy 2016;27(27):3864-3866
OBJECTIVE:To analyze the dyeing status of cinnabar and its pieces,and provide reference for its quality clinical safetey appicaton. METHODS:TLC was used for the qualitative identification of amaranth,carmine,erythrosine,acid red 73, 808 udan and indirubin. HPLC-MS was used to detect the 808 udan :HPLC conditions were as follows,column was Acquity UPLC BEH C18 with mobile phase of cetonitrile-0.1% formic acid(70∶30,V/V)at a flow rate of 0.3 ml/min,the detection wave-length was 520 nm;MS conditions were as follows,ion source was electrospray ionization source,scanning mode was positive ion scanning with full scanning tandem mass spectrometry,nebulizer pressure was 30 psi,drying gas was nitrogen,ion spray voltage was 4 000 V,collision energy was 30 V,and the injection volume was 5 μl. The volumetric method was used for content determi-nation of HgS. RESULTS:TLC spots of amaranth,carmine,erythrosine,acid red 73,808 udan and indirubin were clear and well-separated. 4 batches of 808 udan dyeing were included in the 18 batches of samples,6 batches had non-compliance contents (including 3 batches of 808 udan dyeing). CONCLUSIONS:Dyeing-doped and other quality problems exist in the cinnabaris in markets,which should be noticed.
7.Practical skills promotion of students by competition of acupuncture and moxibustion
Yan ZHONG ; Qianyun YANG ; Jing SHEN ; Liang PENG ; Xiangming ZHANG ; Yubing LI ; Hao WANG
International Journal of Traditional Chinese Medicine 2010;32(4):381-
Objective This article introduced the concrete methods and benefits of promoting and holding competition of acupuncture and moxibustion in TCM universities or colleges. A regular skill competition will not only stimulate students' passion for TCM study and expand teaching methods, but also serve as a effective way to enhance practical skills of students.
8.Expressions of anticoagulant genes KLF2 and KLF4 in blood for prediagnosing deep venous thrombosis in rats*☆
Liqing YAO ; Yaojun DAI ; Xueling ZHAO ; Yubing ZHANG ; Ya NING ; Hongkun LI ; Wen LI
Chinese Journal of Tissue Engineering Research 2011;15(7):1327-1330
BACKGROUND:There is lack of an effective measuring means to diagnose deep venous thrombosis (DVT) in clinic.KLF2 and KLF4 are down-expressed at prethrombotic state,which may be served as predictive molecular markers to diagnose DVT.OBJECTIVE:To explore the feasibility of KLF2 and KLF4 as molecular markers to prediagnose DVT in rats.METHODS:Totally 90 rats were obtained from 100 rats to establish traumatic DVT models and divided into the prethrombotic,thrombosis crest-time and non-thrombosis groups.The remained 10 rats served as control group.Rat blood was collected at each time point,and the expressions of KLF2 and KLF4 were detected by real-time PCR.RESULTS AND CONCLUSION:The KLF2 and KLF4 mRNA expressions in the prethrombotic group and thrombosis crest-time group were lower than that of the control group.However,the KLF2 and KLF4 mRNA expressions in the non-thrombosis group was higher than that of the control group.Therefore,KLF2 and KLF4 may be candidate molecular markers for prediagnosis of DVT in rats.
9.Effects of cathepsin B and cathepsin C gene on vascular endothelial cells in a rat model of traumatic deep venous thrombosis
Liqing YAO ; Ya NING ; Xueling ZHAO ; Yubing ZHANG ; Hongkun LI ; Wen LI
Chinese Journal of Tissue Engineering Research 2011;15(11):2074-2078
BACKGROUND: Deep venous thrombosis (DVT) always occurs after orthopedic surgery. At present, clinical diagnosis of DVT has been lack of an effective measuring means for a long time. Cathepsin may be an effective biological marker of DVT. OBJECTIVE: To study the expression change of cathepsin B and cathepsin C in the rat blood cells before and after DVT and to investigate the feasibility of cathepsin B and cathepsin C as candidate molecular markers for early diagnosis of DVT. METHODS: Totally 100 Sprague Dawley rats were randomly divided into normal control group (n=10) and model group (n=90). Rat traumatic deep vein thrombosis models were established by clamping the femoral vein and fixing the bilateral hind limbs. According to observation time points and the different situations of thrombosis, rat models were assigned to three subgroups: pre-thrombosis, intra-thrombosis, and non-thrombosis. Blood RNA of each group was extracted and reverse transcribed into cDNA. The expression of cathepsin B and cathepsin C in blood cells was detected using real-time fluorescence quantitative PCR. RESULTS AND CONCLUSUON: Expression of cathepsin B and cathepsin C in the blood cells was obviously expressed in the intra-thrombosis subgroup. There was no significant difference in cathepsin B and cathepsin C expression between pre-thrombosis, non-thrombosis groups and normal control group. These findings suggest that cathepsin B and cathepsin C are closely related to DVP and they can be used as the candidate molecular markers for early diagnosis of DVT.
10.Effect of cathepsin L/G on venous vascular wall in traumatic deep vein thrombosis rat models
Wen LI ; Jihong HU ; Xingguo LI ; Hongkun LI ; Yubing ZHANG ; Xueling ZHAO ; Bing WANG
Chinese Journal of Tissue Engineering Research 2011;15(24):4525-4529
BACKGROUND: At present, the basic molecular etiological mechanism and core regulatory network of deep vein thrombosis (DVT) remains uncertain, and there is not an ideal measure for early diagnosis of DVT. OBJECTIVE: To study the underlying impact of cathepsin L/G in DVT rat model. METHODS: DVT rat models (n = 50) were established by clamping both femoral vein in three different positions within 3 seconds with mosquito forceps and fixing with cast. According to different observation phases and biological situations of the femoral vein thrombosis, model rats were divided into thrombogenesis group, pre-thrombogenesis group and non-thrombogenesis group. An additional 10 normal rats served as control group. Femoral vein was obtained at corresponding time points to exact total RNA. After a gene chip-based screening, the data of gene expression were further dissected by real-time PCR. RESULTS AND CONCLUSUON: Gene chip hybridization analysis results demonstrated that differential expression of cathepsin L/G gene was significant among groups, and the expression was greatest in the thrombogenesis group, followed by pre-thrombogenesis and non-thrombogenesis groups, which was significantly greater than the control group (P < 0.05). Real-time PCR analysis results were consistent with gene chip hybridization analysis results. These indicate that DVT is associated with an increase in expression of cathepsin L/G in local venous vascular wall, and they may be candidate molecular markers for early diagnosis of deep vein thrombosis.