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.

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.

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.

A total of 209 type 2 diabetic patients were divided into three groups:without,with mild to moderate,or severe diabetic lower extremity arterial disease based on the ankle brachial index value.60 healthy subjects were used as a control group.Retinol binding protein4 (RBP4),cystatin C,and biochemical parameters were determined in all subjects.The results showed that RBP4 and cystatin C levels were progressively raised with the increasing severity of lower extremity arterial disease in various groups,being significantly higher in patients with type 2 diabetes mellitus with mild to moderate and severe arterial disease compared with control group (P＜0.05 or P＜0.01).RBP4 and cystatin C levels in groups with lower extremity arterial disease were significantly higher than those in the group without arterial disease (P＜0.01).Pearson correlation analysis showed that RBP4 level was positively correlated with total cholesterol,triglyceride,low density lipoprotein-cholesterol,fasting serum insulin,body mass index,cystatin C etc while negatively correlated with high density lipoprotein-cholesterol (P＜0.05 or P＜0.01).Logistic regression analysis showed that serum RBP4 and cystatin C were significantly associated with lower extremity arterial disease in type 2 diabetes mellitus.

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.

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.

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.

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.

BACKGROUND:The molecular mechanism of traumatic deep vein thrombosis is complex.Numerous studies focus on clinical observation and epidemiology,but its molecular mechanism has not been a new breakthrough.OBJECTIVE:By use of gene array technology,this study was aimed to study the expression changes of matrix metalloproteinases in rat models of traumatic deep vein thrombosis,and to explore the roles of matrix metalloproteinases in traumatic deep venous thrombosis.METHODS:A total of 150 SD rats,SPF grade,of 8-12 weeks old,body weight of 250-300 g,were divided at random into normal control group (n=10) and model group (n=140).Rat traumatic deep venous thrombosis models were set up by clamping the femoral vein and fixing the bilateral hind limbs,and the fixation of hip spica with plaster bandage was conducted in each group.Then rats were divided into 7 subgroups:post-traumatic 0.5 hours,post-traumatic 2.5 hours (initial period of thrombosis),post-traumatic 25 hours (thrombogenesis at thrombotic crest-time),post-traumatic 25 hours non-thrombogenesis at the thrombotic crest-time),post-traumatic 72 hours (thrombus resolution),post-traumatic 72 hours thrombus insolution) and post-traumatic 168 hours (nonthrombosis).At the corresponding phasess,the femoral vein tissues were incised,and total RNA of femoral vein was extracted using Trizol one-step method.Applying Genechip Rat Genome 430 2.0 genechips,the gene expressions in femoral vein were detected in different groups.The rate of traumatic deep venous thrombogenesis and non-thrombogenesis,the rate of thrombi solution and insolution were observed;the expressions of matrix metalloproteinases and tissue inhibitor of metalloproteinases at different time phases was detected by gene array data analysis.RESULTS AND CONCLUSION:Three model rats died and the remaining 147 rats were involved in the final analysis.At the post-traumatic 25 hours,the rate of thrombogenesis was 50.5% and nonthrombogenesis was 49.5%.To the post-traumatic 168 hours,the rate of thrombus solution was 56.7% and thrombus insolution was 43.3%.Both matrix metalloproteinases and tissue inhibitor of metalloproteinases exhibited differential expressions in the course of traumatic deep venous thrombosis.Under the thrombus insolution state,matrix metalloproteinases continued to show a high expression,tissue inhibitor of metalloproteinase expression was down-regulated in the thrombus formation,was significantly inhibited in the thrombus insoluUon process.In the process of traumatic deep vein thrombosis and insolution,matrix metalloproteinase was closely related to traumatic deep vein thrombosis,the matrix metalloproteinase/tissue inhibitor of metalloproteinases are likely to affect the biological state of thrombosis.

Objective To analyze the clinical information of a family with one patient who have systemic lupus erythematosus (SLE) and Fabry disease,as well as the enzymatic activity and gene mutation in these family members.Methods Clinical characteristics were collected from the proband and her family members.Peripheral blood samples from three members of this family were collected and the enzymatic activity was measured by fluorimetrie substrate assay.Genomic DNA was extracted from one male member with significantly decreased enzyme activity,the 7 exons and their flanking introns of GLA gene were amplified by PCR and directly sequenced.Results The enzyme activity of two family members was significantly decreased,the genetic analysis of the male member revealed a missense mutation in exon 2:c.334C＞T (CGC＞TGC)( p.R112C ).Family members except the proband had no definite evidence to support the presence of SLE.Conclusion The coexistence of SLE and Fabry disease is extremely rare.Immunological test,enzymatic activity and gene mutation analysis seem to be helpful for the differential diagnosis.