Objective To study the gene expression of Wnt signal transduction pathway in experimental liver fibrosis and to investigate its role in liver fibrosis. Methods Liver fibrosis model was induced with carbon tetrachloride in 8 SD rats. Another 8 healthy rats were served as control. The gene expression in liver tissues of models and controls were examined using real time PCR array. The differential gene expression was identified as either up- or down-regulated 2-fold. The expressions of smooth muscle actin (SMA), Wnt4, Frizzled2 and β-catenin in the tissues were examined by immunohistochemistry and Western blot. Results The examination confirmed that 36 genes were differentially expressed, including 25 genes up-regulated and 11 genes down-regulated. Compared with the controls, the expressions of Wnt4, Wnt5 a and W nt11 were up-regulated more than 13.9-, 16.5-and 2.17-fold respectively, while the expressions of Wntl and Wnt3 were down-regulated more than 2.32- and 2.15-fold respectively in fibrotic liver. Immunohistochemistry and Western blot showed that the expressions of SMA, Wnt4 and Frizzled2 in fibrotic liver were remarkably higher than those in normal controls. While the level of phosphorylated β-catenin was decreased. Conclusion Both canonical and noncanonical Wnt signal transduction pathway may involve in the mechanism of liver fibrogenesis.

Immuno-fluorescence technique can qualitatively determine certain nuclear translocation, of which NF-κB/ p65 implicates the activation of NF-κB signal pathways. Immuno-fluorescence analysis software with independent property rights is able to quantitatively analyze dynamic location of NF-κB/p65 by computing relative fluorescence units in nuclei and cytoplasm. We verified the quantitative analysis by Western Blot. When we applied the software to analysis of nuclear translocation in lipopolysaccharide (LPS) induced (0. 5 h, 1 h, 2 h, 4 h) primary human umbilical vein endothelial cells (HUVECs) , we found that nuclear translocation peak showed up at 2h as with calculated Western blot verification results, indicating that the inventive immuno-fluorescence analysis software can be applied to the quantitative analysis of immuno-fluorescence.
Active Transport, Cell Nucleus ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Fluorescent Antibody Technique ; Human Umbilical Vein Endothelial Cells ; Humans ; NF-kappa B p50 Subunit ; metabolism ; Software

Objective To investigate the inducing effects of selenium dioxide(SeO2 ) on the apoptosis in human cervical carcino-ma cell line Hela and its influence on the expression of apoptosis-related proteins caspase-3 and P53 .Methods Hela cells were trea-ted with different concentrations of SeO2 for 24 h in vitro ;the morphological changes of Hela cells were observed by the optical mi-croscope;the influence of SeO2 on the cell proliferation and vitality was examined by the MTT assay ;the flow cytometry was em-ployed to detect the cell apoptosis rate ;the expressions of caspase-3 and P53 proteins in Hela cells were determined by the Western blot analysis .Results Under the optical microscopy ,SeO2 generated the obvious influence on the cell growth morphology ,a large number of cells became rounded and shrunken ,and lost the normal form ,while the adherence cell number was evidently decreased and the proliferation was slowed down ;the MTT results showed that SeO2 markedly inhibited the cell proliferation and viability in a dose-dependent manner ,in which ,the cell apoptosis rates induced by the 0 ,1 .875 ,3 .750 ,7 .500 ,15 .000 and 30 .000 μmol/L con-centrations of SeO2 were 3 .12% ,30 .56% ,33 .42% ,37 .50% ,45 .43% and 69 .38% respectively ,which revealing the obviously in-creasing trend;the Western blot assay revealed that SeO2 could up-regulate the caspase-3 and P53 levels ,and reached the peak value at the concentration of 7 .500μmol/L .Conclusion SeO2 could induce the cervical cancer cell apoptosis possibly by up-regulating the expressions of caspase-3 and p53 in Hela cells .

Aim To investigate the effect of ribonucleic acidⅡon apoptosis in human leukemia cell lines K562 and KG1 a.Methods Cell counting kit-8(CCK-8)as-say was performed to detect proliferation activity of K562 and KG1 a cells treated with ribonucleic acidⅡ. Apoptosis index was assessed by flow cytometry(FCM) and fluorescent Hoechst 33258 staining was used for observing morphologic changes of apoptosis.Expres-sion levels of p53,Bax,Bcl-2 and cleaved caspase-3 were analyzed by Western blot.Results The prolifera-tion of K562 and KG1 a cells was significantly inhibited by ribonucleic acid Ⅱ treatment for 12 h,24 h,48 h at concentrations of 100~300 mg·L-1 ,which indica-ted the inhibitory effect of ribonucleic acid Ⅱ was in dose-dependent and time-dependent manners.FCM re-sults displayed a dose-dependent increase in cell apop-totic rate.Hoechst 33258 staining showed the typical apoptotic morphology in some leukemic cells treated with ribonucleic acid Ⅱ,including increased nuclear chromatin concentration and edge accumulation.West-ern blot analysis showed the increased expression of p53,Bax,cleaved caspase-3 and decreased expression of Bcl-2 in K562 and KG1 a cells treated with ribonu-cleic acid Ⅱ.Conclusions Ribonucleic acid Ⅱ can induce apoptosis of leukemia K562 and KG1 a cells by up-regulating p53,which mediates Bcl-2/Bax balance and activates caspase-3 .

According to the current situations and development of (TCMIs), the author of the article reveals the scientific connotation of TCMIs in theory, preparations and clinic application, and points out that TCMIs are an innovative and breakthrough of conventional dosage forms of traditional Chinese medicines, the combination of traditional theory and modern technology as well as a type of modern dosage form with the characteristics of traditional Chinese medicines, which conforms to the principle of including the essence and excluding the wastes for traditional Chinese medicine preparations, meets the demands for quick-acting of traditional Chinese medicines and guides one of the development orientation of traditional Chinese medicines. In the meantime, an analysis was also made on key issues, such as adverse reactions of TCMIs, modern clinical application, special drug delivery route and diversity of components and ingredients.
Drug Delivery Systems ; methods ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Exanthema ; chemically induced ; Humans ; Injections ; adverse effects ; Medicine, Chinese Traditional ; adverse effects ; methods ; trends ; Nausea ; chemically induced ; Product Surveillance, Postmarketing ; methods ; statistics & numerical data ; Vomiting ; chemically induced

With improved overall survival of cervical cancer patients, the importance of the quality of life (QOL) is increasingly recognized. This study was conducted to compare the QOL of women with different stage cervical cancer before and after treatment to facilitate improved cervical cancer prevention and treatment. We used the generic Medical Outcomes Study Short Form-36 (MOS SF-36) to collect QOL information. Based on SF-36, we interviewed cervical cancer patients at West China Second Affiliated Hospital and Sichuan Cancer Hospital between May 2010 and January 2011. A total of 92 patients with precancerous lesions, 93 with early cancer, and 35 with advanced cancer responded to our survey. Average physical component summary (PCS) scores were significantly different between the three groups at every time point (P < 0.05). Average mental component summary (MCS) scores were significantly different between the three groups after treatment (P < 0.05). Average PCS and MCS scores increased gradually from the pretreatment to posttreatment period for patients with precancerous lesions. However, they reached the lowest at 1 month after treatment for patients with early and advanced cancers and rebounded between 1 and 6 months after treatment. Our results indicate that patients with precancerous lesions and early cervical cancer show better overall QOL than do those with advanced cervical cancer. Additionally, patients with early cancer recover more quickly than do those with advanced cancer in terms of both physical and mental functions. Thus, early detection and treatment initiatives may improve the QOL for patients with precancerous lesions and cervical cancer.
Adult ; Aged ; Carcinoma in Situ ; pathology ; therapy ; Cervical Intraepithelial Neoplasia ; pathology ; therapy ; Chemoradiotherapy ; China ; Female ; Follow-Up Studies ; Humans ; Hysterectomy ; methods ; Lymph Node Excision ; Middle Aged ; Neoplasm Staging ; Precancerous Conditions ; pathology ; therapy ; Quality of Life ; Surveys and Questionnaires ; Uterine Cervical Neoplasms ; pathology ; therapy ; Young Adult

OBJECTIVETo investigate the interaction on smoking and the lung cancer related genes miR-196a2 rs11614913, miR-146a rs2910164, miR-300 rs12894467, miR-26a-1 rs7372209, miR-27a rs895819 in Fujian Han population.

METHODSFrom January 2006 to January 2012, by using a hospital-based case-control study, 1 053 cases were pathologically diagnosed as primary lung cancer from the Department of Thoracic Surgery and 1 058 controls were randomly selected from the visiting relatives of patients and visiting people of Cangxia community health service of Fuzhou city according to match with age and genders. They were recruited for questionnaires survey and genotyping detection. Research objects of genders, height, weight, cultural degree, marital status, family history of cancer, lung disease history, smoking, drinking tea, drinking, and so on. After informed consent, we collected 5 ml fasting venous blood from every object, used MALDI-TOF-MS to analysis genotyping of polymorphic loci. Logistic regression model was constructed by using SNP as independent variable, and the multiple factors were constructed with different loci. The possible association between SNP and cigarette smoking was analyzed by using the crossover analysis. The relative excess risk of interaction (RERI) were used to analyze on smoking and SNP loci additive interaction of dominant and recessive genetic models.

RESULTSSmokers in case group who smoked P50(P25-P75)30.00 (0.00-56.00) packages in a year were higher than control group (0.00(0.00 - 20.48) pack years) (Z=14.57,P<0.001). Passive smoking index for non-smokers was 11.40(0.00-25.00), higher than the controls (0.00(0.00-13.11)) (Z=10.71,P<0.001). Site detection rate of rs11614913, rs2910164, rs12894467, rs7372209 and rs895819 in cases was 95.82%(1 009/1 053), 97.72%(1 029/1 053), 97.82% (1 030/1 053), 97.15% (1 023/1 053) and 96.01% (1 011/1 053) respectively. The controls were 98.11% (1 038/1 058), 98.96% (1 047/1 058), 98.30% (1 040/1 058), 98.68% (1 044/1 058) and 98.02% (1 037/1 058) respectively. rs11614913 dominant genetic model, TT genotype and smoking could increase the risk of primary lung cancer (OR=4.04, 95%CI: 2.67 -6.12). Recessive genetic model, CC genotype and smoking increased the incidence of primary lung cancer risk (OR=4.76, 95%CI: 3.16 -7.17). rs12894467 dominant genetic model, TT genotype and smoking could increase the risk (OR=2.98, 95%CI: 2.28 -3.90) in primary lung cancer. In recessive genetic model, CC genotype and smoking increased the incidence of primary lung cancer risk (OR=1.94, 95% CI: 1.10-3.43). Dominant genetic model of rs2910164, CC genotype and smoking could increase the risk (OR=2.18, 95% CI: 1.60 -2.98) in primary lung cancer. Recessive genetic model, GG genotype and smoking increased the incidence of primary lung cancer risk (OR=3.29, 95% CI: 2.16 -5.03). Especially rs12894467 dominant and recessive gene model and genders, smoking and there were combined effects(χ(2)=8.58, P=0.003; χ(2)=4.76, P=0.040).

CONCLUSIONRs11614913, rs12894467 and rs2910164 polymorphism were potentially associated with primary lung cancer in Fujian Han population.

Case-Control Studies ; China ; Genetic Predisposition to Disease ; Genotype ; Humans ; Lung Neoplasms ; genetics ; MicroRNAs ; genetics ; Polymorphism, Single Nucleotide ; Risk Factors ; Smoking ; adverse effects

OBJECTIVETo explore the feasibility of IGF2 imprinting system in target gene therapy for tumors.

METHODSThe mouse H19 enhancer, DMD and promoter H19 were amplified by PCR from mouse genomic DNA and then cloned into the plasmid pDC312. The EGFP and DT-A fragments were amplified by PCR and cloned into the recombinant plasmid, and then the shuttle plasmid were transfected into HEK293 cells together with the adenoviral vector Ad5, namely, Ad-EGFP and Ad-DT-A. Adenovirus hexon gene expression was applied to confirm the presence of adenovirus infections. The effect of the IGF2 imprinting system was tested by fluorescence microscopy. RT-PCR and Western blotting after transfection of the recombinant adenoviral vectors into cancer cells were used to show loss of IGF2 imprinting (LOI) and maintenance of IGF2 imprinting (MOI), respectively. The anti-tumor effect was assessed by MTT and flow cytometry after the HCT-8 (LOI). Human breast cancer cell line MCF-7 (MOI) and human normal gastric epithelial GES-1 (MOI) cell line were transfected with Ad-DT-A in vitro. The anti-tumor effect was detected by injecting the Ad-DT-A in nude mice carrying HCT-8 tumors.

RESULTSThe expression of EGFP protein, DT-A mRNA and DT-A protein were seen to be positive only in the HCT-8 tumor cell line. Infection with Ad-DT-A resulted in obviously growth inhibition in HCT-8 cells (75.4 ± 6.4)% compared with that in the control group, and increased the percentage of apoptosis in the HCT-8 cells (20.8 ± 5.9)%. The anti-tumor effect was further confirmed by injecting the recombinant adenoviruses in HCT-8 tumor-bearing nude mice, and the results showed that the Ad-DT-A inhibited the tumor growth, with on inhibition rate of 36.4%.

CONCLUSIONSThe recombinant adenoviruses carrying IGF2 imprinting system and DT-A gene have been successfully constructed, while Ad-DT-A can effectively kill the tumor cells showing loss of IGF2 imprinting. It might play an important role in future target gene therapy against malignant tumors based on loss of IGF2 imprinting.

Adenoviridae ; genetics ; Animals ; Apoptosis ; Breast Neoplasms ; genetics ; pathology ; Colonic Neoplasms ; genetics ; pathology ; therapy ; Diphtheria Toxin ; biosynthesis ; genetics ; Female ; Genetic Therapy ; methods ; Genetic Vectors ; Genomic Imprinting ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Insulin-Like Growth Factor II ; genetics ; metabolism ; MCF-7 Cells ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Peptide Fragments ; biosynthesis ; genetics ; Plasmids ; RNA, Messenger ; metabolism ; Random Allocation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

OBJECTIVETo explore the operative method and its clinical effects of pronation and supination external rotation trimalleolar fractures.

METHODSFrom March 2000 to July 2006,42 patients of the pronation and supination external rotation trimalleolar fractures treated with open reduction and internal fixation. Thirty-one were males and 11 were females,with an average age of 40.5 years (from 19 to 76 years). Four cases were open fractures and 38 cases close fractures. The fractures were classified as pronation-external rotation (grade IV) injury in 18 cases and supination-external rotation (grade IV)in 24 cases according to the system of Lauge-Hansen. The time of injury to operation was 2 hours to 27 days. The medial, lateral and posterior malleolus were exposed by standard anteromedial and Gatellier-Chastang approaches. The reduction and internal fixation started with the posterior,then the medial and the lateral malleolus and distal tibiofibular syndesmosis in sequence. The anteroposterior, lateral and mostise X-ray films were taken after operation.

RESULTSAll the patients were followed up for an average time of 13.5 months(from 6 to 24 months). The time of union was from 12 to 16 weeks. The results were excellent in 20,good in 16, fair in 4 and poor in 2 cases according to Baird-Jackson ankle scoring system based on pain, stability, walking ability,range of motion and radiological manifestations. The excellent and good rate was 85.7%. There were no infection,malunion and nonunion of the fractures except that the inserted screw to distal tibiofibular syndesmosis was broken in 1 case.

CONCLUSIONThe key of operative treatment is to restore the anatomy of ankle and to regain the ankle function maximally.

Adult ; Aged ; Ankle Injuries ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Pronation ; Supination

Objective To explore the effects of potassium iodate (KIO3) on the proliferation, cell cycle, and the mRNA/protein levels of cyclin D 1 and Ki67 of SW579 cells, a human thyroid squamous carcinoma cell line . Methods The effects of different doses of KIO3(0,10-6,10-5,10-4,10-3,10-2,10-1 mol/L)on SW579 cell prolifer-ation were assessed by CCK8 method.SW579 cells were then treated with 0 (control), 10-6 or 10-2 mol/L KIO3 for 48 h.The cell cycle was measured by flow cytometry .The mRNA and protein levels of cyclinD 1 and Ki67 were re-spectivelyanalyzedbyreal-timePCRandWesternblot.Results 10-6and10-2mol/LofKIO3respectivelyexhibi-ted the most promoting and suppressive effect on SW579 cell proliferation.G1 phase of cells in 10-6 group short-ened significantly( P<0.05) , and S phase prolonged significantly( P<0.05); while each phase of cells in 10-2 mol/L group changed in the opposite way( P<0.05) .KIO3 at the dose of 10-6 mol/L significantly up-regulated the mRNA levels of cyclin D1 and ki67 in cells( P<0.05) ;whereas, the mRNA and protein levels of cyclin D1 and Ki67 were significantly down-regulated in cells treated with 10-2 mol/L KIO3( P<0.05) .Conclusions Different doses of KIO3 affect the proliferation and cell cycle of SW579 cells probably by regulating the levels of cyclin D1and Ki67 .