Objective To investigate the relationship between the expressions of TMPRSS4 and clinical pathologi-cal parameters in gastric carcinoma. To explore its significance in judging the prognosis of patients. To analyze the expression of TMPRSS4 in the epithelial-mesenchymal transition. Methods The expressions of TMPRSS4, E-cad-herin and Vimentin were detected by immunohistochemistry ( SABC) in 100 cases of gastric carcinoma specimens and corresponding adjacent normal tissue. Results In gastric carcinoma, TMPRSS4, E-cadherin and Vimentin positive rate was 47%,45%,37%. In adjacent tissues,their positive rate was 9%,30%,4%. The differences be-tween tumor tissues and adjacent tissues had statistical significance ( P<0.05 ) . High expression of TMPRSS4 cor-related significantly with lymph node metastases and TNM stage (P<0.05), but not with gender, age, size, tumor histological type,depth of invasion (P>0.05). The 5-year survival rate of patients with high TMPRSS4 expression was significantly lower than that in patients with low expression. The high TMPRSS4 expression was significantly correlated in gastric carcinoma accompanied by low expression of E-cadherin (rs = -0.207,P=0.038) and high Vimentin expression (rs=0.233,P=0.020). Conclusion The expression of TMPRSS4 is closely related to the biological characteristics in gastric carcinoma,detection the expression of TMPRSS4 is valuable in predicting tumor prognosis,invasion and metastasis. TMPRSS4 may promote invasion, metastasis of human gastric carcinoma through epithelial-mesenchymal transition by reduce the expession of E-cadherin.

Objective To acquire glucose transporter 4 (GLUT4) cDNA from tissues of human muscle by RT-PCR, and to clone and express GLUT4 cDNA in E.coli. Methods GLUT4 cDNA primers were designed first and ascertained by detecting through Genbank on NCBI and DNA Star, then by using specific primers, the subjective cDNA segment was acquired by RT-PCR from human tissue. After that, it was cloned into cloning vector of pGEM-3zf(-) and sequenced automatically. Finally the subjective cDNA was cloned into vector of pBV220 and expressed in E.coli. Results The desired DNA could be acquired from tissues of human muscle by RT-PCR using special primers, the acquired cDNA fragment could be cloned into vector of pGEM-3zf(-) and sequenced. The GLUT4 cDNA sequence was highly conserved. GLUT4 cDNA could be expressed in E.coli, too. Conclusion The entire GLUT4 cDNA can be acquired from human muscular tissue by RT-PCR, and it can be expressed in E.coli.

Objective To investigate the correlation between uric acid level and macrovascular disease in type 2 diabetic patients.Methods Sixty type 2 diabetic patients with lower limb atherosclerosis of carotid artery were randomly selected in study group who hospitalized in the First Hospital of Zibo from Mar.to Feb.2012.Sixty type 2 diabetes mellitus(T2DM) without carotid and lower limb athemsclerosis were served as control group.The blood pressure,blood lipid,blood glucose and other biochemical indexes,including blood uric acid,serum insulin (FNS),fasting blood glucose (FPG),apolipoprotein a (LP (a)),apolipoprotein A1,B (APO-A1,APO-B),glycosylated hemoglobin (HbA1c),high density lipoprotein cholesterol (HDL-C),low density lipopmtein cholesterol (LDL-C),triacylglycerol (TG) and total cholesterol (TC) were measured and determined.Results There was no significant difference in terms of blood pressure,blood lipid levels,APO-A1,APO-B,HbA1C,FNS and FPG in study group patiems (P ＞ 0.05).The level LP(a) in study group was (0.4 ± 0.2) g/L,significantly higher than that in control group ((0.2 ± 0.2) g/L; t =3.842,P ＜ 0.01).The blood uric acid level in study group was (362.3 ± 112.8)mmol/L,significantly higher than that of the control group((284.8 ±68.6)mmol/L;t =3.188,P＜0.01).Conclusion Uric acid and LP(a) are involved in the oocurrence and development of athemsclemsis,which is close related to the development of type 2 diabetic macmangiopathy.Therefore,in the process of preventing type 2 diabetes with macroangiopathy,we should pay attention to uric acid and LP (a) of the patient beside effective control of blood glucose,blood pressure,blood lipid level.

BACKGROUND:Restenosis after angioplasty severely limited the application and long-period therapeutic effects of percutaneous coronary intervention. Changes in smooth muscle cel phenotype and their proliferation are important mechanisms of restenosis after angioplasty.
OBJECTIVE:To use bal oon in vivo transduction of osteopontin short hairpin RNA (OPN-shRNA), to inhibit osteopontin expression at the injured blood vessels of a rabbit model of experimental atherosclerosis, and to prevent restenosis after angioplasty.
METHODS:A total of 20 rabbit models of atherosclerosis were established and randomly equal y assigned to empty plasmid group and OPN-shRNA plasmid group. The plasmid recombinant OPN-shRNA and empty plasmid were transferred to the ventral aorta by bal oon.
RESULTS AND CONCLUSION:After bal oon dilatation, specific green fluorescence was detected in the layer of vascular smooth muscle in the two groups. Moreover, with prolonged time of transfection, fluorescence intensity gradual y decreased. Compared with the empty plasmid group, the expanded artery lumen area obviously increased in the OPN-shRNA plasmid group, and plaque burden evidently reduced. Results indicated that bal oon catheter used in regional blood vessels in rabbit models of atherosclerosis could successful y transduce OPN-shRNA plasmid. The restenosis of the expanded blood vessels lessened, and thrombus burden relieved. It is of great importance to prevent the occurrence of restenosis after angioplasty in rabbit models.

OBJECTIVE: To explore the antitumor activities of kushen (Sophora flavescens) flavonoids (KS-Fs) in vivo and in vitro. METHODS: Cell proliferation was assayed by using methyl thiazolyl tetrazolium (MTT) method. H22 hepatocellular carcinoma and S180 sarcoma were induced in ICR mice. Lewis lung carcinoma was induced in C57BL/6 mice. H460 and Eca-109 tumor were induced in Balb/c nude mice by injecting 5x10(5) or 5x10(6) tumor cells in the right flank, respectively. RESULTS: KS-Fs could inhibit the growth of a variety of human tumor cell lines (A549, SPC-A-1, NCI-H460, etc.) in vitro. The antitumor efficacies were confirmed in the mice models of H22, S180 and Lewis lung tumors and the nude mice models of human H460 and Eca-109 xenografted tumors. The oral or intravenous maximum tolerated dose of KS-Fs was more than 2.8 g/kg or 750 mg/kg respectively, far more than the oral medial lethal dose of kushen alkaloids (< or = 1.18 g/kg). No adverse reactions were observed. CONCLUSION: These results suggest that KS-Fs or kurarinone may be developed as a novel antitumor agent.

Objective To observe the expressions and distribution of transient receptor potential cation channel 6 (TRPC6) and integrin-linked kinase (ILK) in the glomeruli of renal biopsytissue of patients with proteinuric kidney diseases,and to investigate the effect of TRPC6 over-expression on ILK in vitro.Methods The archival histological specimens of patients admitted to Tangdu hospital from 2012 to 2013,with 24-hour urinary protein over 1 g,were collected.The expressions and distribution of TRPC6 and ILK in the glomeruli of renal biopsy tissue were observed by immunohistochemistry.MPC5 podocytes were cultured in vitro and they were stimulated with 10-7 mol/L ADR for 12,24 and 36 h.The pcDNA3.1(+)-TRPC6 plasmid and pcDNA3.1(+) were transfected into MPC5 podocytes by liposome 2000 reagent to establish the TRPC6 overexpression group and the negative control group respectively.Western blotting was used to detect the expressions of TRPC6 and ILK protein.Results There were 14 cases of membranous nephropathy,13 cases of focal segmental glomerulosclerosis (FSGS),15 cases of membranoproliferative glomerulonephritis,12 cases of mesangial proliferative glomerulonephritis,10 cases of hyperplastic sclerosis nephritis,15 cases of IgA nephropathy,13 cases of purpura nephritis,15 cases of lupus nephritis,13 cases of hypertensive renal injury,14 cases of diabetic nephropathy and 9 cases of normal renal tissue included.In glomerulus,TRPC6 was expressed mainly in podocytes,and the expressions of TRPC6 in these renal tissues were higher than that in normal renal tissues (all P ＜ 0.05),except for hypertensive nephropathy.ILK was expressed in podocytes and the mesangial areas.The expressions of ILK in FSGS,lupus nephritis and diabetic nephropathy were higher than that in normal kidney tissue (all P ＜ 0.05),while the other renal tissues was high but showed no statistical difference with normal kidney tissue (all P ＞ 0.05).The expressions of TRPC6 and ILK were positively correlated in renal tissues of FSGS and diabetic nephropathy (r=0.906,P ＜ 0.001;r=0.783,P=0.001 respectively).The expressions of TRPC6 and ILK protein in 24 and 36 h stimulating with ADR were significantly higher than that in the control group (all P ＜ 0.05).The expression of ILK in the TRPC6 overexpression group was significantly higher than that in the normal control group (P ＜ 0.05).Conclusions The expressions of TRPC6 and ILK increase in the glomeruli of patients with kidney diseases with proteinuria being the main manifestation,especially in FSGS and diabetic nephropathy.The up-regulation of TRPC6 can increase the expression of ILK protein,which may be involved in podocyte injury.

The use of rnicroarrays of oligonucleotides or cDNA is considered to be a promising approach for DNA and RNA sequence analysis, diagnostics of genetic diseases, gene polymorphism studies and analysis of gene expression. To manufacture cDNA microarrays the samples were printed onto glass microscope slides treated with poly-L-lysine, and then the slides were processed by heat and UV light treatment to attach the cDNA sequence to the glass surface. But the immobilization efficiency of cDNA on the glass surface was low. A simple procedure for manufacture cDNA microarrays on a slide treated with 3-aminopropyltrimethoxysilane is described. The efficiency for attaching cDNA to the amino-modified slides is greater than that to the slides treated with poly-L-lysine. The cDNA microarray made by the amino-modified slides is stable for use in 80℃, 75 % humidity, 3 600Lx light, exposure in air, respectively.

Objective To observe the preventive effects of puerarinon development of deep vein thrombosis (DVT) in elderly women with osteoporosis after hip replacement. Methods 100 elderly women with osteoporosis who scheduled for femoral head arthrolastybetween January 2012 and January 2014 in Cangzhou Central Hospital were selected and then randomly divided into a study group (n=50) and a control group (n=50). The control group received routine anti?inflammatory and anticoagulant therapy ,while the study group received intravenous drip of puerarinof 400 mg mixed with 5%glucose liquid 500 ml daily for 15 days as a treatment course. After the treatment, DVT incidence rate ,change of early diagnostic indexes and hemorheology were compared with their baselines. Results DVT incidence rate was significantly lowerin the study group than in the control group (8.0%vs. 24.0%, P<0.05). Ascompare with the control group,thromboxane A2/prostacyclin,plasma homocysteine and hemorheology?were significantly reduced (P < 0.05). Conclusions Puerarin has a preventive effect for deep vein thrombosis in elderly women with osteoporosis after hip replacement.

Relative deficiency in production of glycoprotein hormone erythropoietin (Epo) is a major cause of renal anemia. This study planned to investigate whether the hypoxia-regulated system of Epo expression, constructed by fusing Epo gene to the chimeric phosphoglycerate kinase (PGK) hypoxia response elements (HRE) in combination with cytomegalovirus immediate-early (CMV IE) basal gene promoter and delivered by plasmid intramuscular injection, might provide a long-term physiologically regulated Epo secretion expression to correct the anemia in adenine-induced uremic rats. Plasmid vectors (pHRE-Epo) were synthesized by fusing human Epo cDNA to the HRE/CMV promoter. Hypoxia-inducible activity of this promoter was evaluated first in vitro and then in vivo in healthy and uremic rats (n = 30 per group). The vectors (pCMV-Epo) in which Epo expression was directed by a constitutive CMV gene promoter served as control. ANOVA and Student's t-test were used to analyze between-group differences. A high-level expression of Epo was induced by hypoxia in vitro and in vivo. Though both pHRE-Epo and pCMV-Epo corrected anemia, the hematocrit of the pCMV-Epo-treated rats exceeded the normal (P < 0.05), but that of the pHRE-Epo-treated rats didn't. Hypoxia-regulated system of Epo gene expression constructed by fusing Epo to the HRE/CMV promoter and delivered by plasmid intramuscular injection may provide a long-term and stable Epo expression and secretion in vivo to correct the anemia in adenine-induced uremic rats.
Anemia/blood/*therapy ; Animals ; Base Sequence ; Blood Urea Nitrogen ; Cell Hypoxia ; Creatinine/blood ; Erythropoietin/biosynthesis/*genetics/secretion ; Gene Expression Regulation ; Genes, Reporter ; Genetic Therapy ; HeLa Cells ; Humans ; Injections, Intramuscular ; Kidney/pathology ; Luciferases, Firefly/biosynthesis/genetics ; Molecular Sequence Data ; Plasmids/*genetics ; Promoter Regions, Genetic ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins/biosynthesis/genetics/secretion ; Response Elements ; Transcriptional Activation ; Uremia/blood/*therapy

Objective: Non-small-cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR)-activating mutations have higher response rate and more prolonged survival following treatment with single-agent EGFR tyrosine kinase inhibitor (EGFR-TKI) compared with patients with wild-type EGFR. However, all patients treated with reversible inhibitors develop acquired re-sistance over time. The mechanisms of resistance are complicated. The lack of established therapeutic options for patients after a failed EGFR-TKI treatment poses a great challenge to physicians in managing this group of lung cancer patients. This study evaluates the in-fluence of EGFR-TKI retreatment following chemotherapy after failure of initial EGFR-TKI within at least six months on NSCLC pa-tients. Methods:The data of 27 patients who experienced treatment failure from their initial use of EGFR-TKI within at least 6 months were analyzed. After chemotherapy, the patients were retreated with EGFR-TKI (gefitinib 250 mg qd or erlotinib 150 mg qd), and the tumor progression was observed. The patients were assessed for adverse events and response to therapy. Targeted tumor lesions were as-sessed with CT scan. Results:Of the 27 patients who received EGFR–TKI retreatment, 1 (3.7%) patient was observed in complete re-sponse (CR), 8 (29.6%) patients in partial response (PR), 14 (51.9%) patients in stable disease (SD), and 4 (14.8%) patients in progres-sive disease (PD). The disease control rate (DCR) was 85.2%(95%CI=62%-94%). The median progression-free survival (mPFS) was 6 months (95%CI=1-29). Of the 13 patients who received the same EGFR-TKI, 1 patient in CR, 3 patients in PR, 8 patients in SD, and 2 patients in PD were observed. The DCR was 84.6%, and the mPFS was 5 months. Of the 14 patients who received another EG-FR-TKI, 0 patient in CR, 6 patients in PR, 6 patients in SD, and 2 patients in PD were observed. The DCR was 85.7%, and the mPFS was 9.5 months. Significant difference was found between the 2 groups in progression-free survival but not in response rate or disease control rate. Conclusion:Retreatment of EGFR-TKIs can be considered an option after failure of chemotherapy for patients who were
previously controlled by EGFR-TKI treatment.