Purpose: The poor retention and ambiguous differentiation of stem cells (SCs) within corpus cavernosum (CC) limit the cell application in erectile dysfunction (ED). Herein, the effects and mechanism of microRNA-145 (miR-145) gene modification on modulating the traits and fate of bone marrow-derived mesenchymal stem cells (BMSCs) were investigated. Materials and Methods: The effects of miR-145 on cell apoptosis, proliferation, migration, and differentiation were determined by flow cytometry, cell counting kit-8, transwell assays and myogenic induction. Then, the age-related ED rats were recruited to four groups including phosphate buffer saline, BMSC, vector-BMSC, overexpressed-miR-145-BMSC groups. After cell transplantation, the CC were harvested and prepared to demonstrate the retention and differentiation of BMSCs by immunofluorescent staining. Then, the target of miR-145 was verified by quantitative real-time polymerase chain reaction and immunohistochemical. After that, APTO-253, as an inducer of Krüppel-like factor 4 (KLF4), was introduced for rescue experiments in corpus cavernosum smooth muscle cells (CCSMCs) under the co-culture system. Results: In vitro, miR-145 inhibited the migration and apoptosis of BMSCs and promoted the differentiation of BMSCs into smooth muscle-like cells with stronger contractility. In vivo, the amount of 5-ethynyl-2′-deoxyuridine (EdU)+cells within CC was significantly enhanced and maintained in the miR-145 gene modified BMSC group. The EdU/CD31 co-staning was detected, however, no co-staining of EdU/α-actin was observed. Furthermore, miR-145, which secreted from the gene modified BMSCs, dampened the expression of KLF4. However, the effects of miR-145 on CCSMCs could be rescued by APTO-253. Conclusions Overall, miR-145 modification prolongs the retention of the transplanted BMSCs within the CC, and this effect might be attributed to the modulation of the miR-145/KLF4 axis. Consequently, our findings offer a promising and innovative strategy to enhance the local stem cell-based treatments.

Objective To evaluate the feasibility of the combination of amplification refractory mutation system (ARMS) and quantitative real-time polymerase chain reaction (PCR) method in fast detection of clarithromycin resistance of Helicobacter pylori (H.pylori) in gastric mucosa.Methods A total of 150 gastric mucosal specimens with positive H.pylori culture were collected from the H.pylori positive patients who failed in H.pylori eradication from January to August in 2013.The drug resistant gene mutation types of H.pylori in these samples were detected by quantitative real-time PCR based on ARMS.And the accuracy was confirmed by sequencing.The clarithromycin resistance of H.pylori was determined by E-assay.Chi-square test was used for statistical analysis.Results Among 149 gastric mucosal specimens (one specimens without wild type or mutation type had been eliminated),the results of quantitative real-time PCR based on ARMS of two samples were not consistent with the results of sequencing;the consistent rate was 98.7％ (147/149).Among 149 specimens with positive H.pylori culture,104 samples (69.8％) were clarithromycin resistance.In 101 samples the clarithromycin resistance was detected by quantitative real-time PCR based on ARMS;the consistent rate was 97.1％ (101/104).Both E-assay and clarithromycin resistant rate detected by E-assay or quantitative real-time PCR based on ARMS was 69.8％ (104/149) and 67.8％ (101/149),respectively,and the difference was not significant (x2 =0.141,P=0.932).Conclusion The combination of ARMS and quantitative real-time PCR method in fast detection of clarithromycin resistance of H.pylori in gastric mucosa is strongly feasible and highly consistent has high consistent rate with sequencing and E-assay.

ObjectiveTo check the isolated heart by coronary angiography to discover the location, na-ture and degree of the coronary artery lesions more accurately and increase the comprehensive evaluation ability of cardiovascular disease.MethodsTen fresh isolated hearts with different causes of death were extracted and injected with barium sulphate as contrast substance by ring injector, then developed under Xper FD20 angiography equipment. The obtained pictures and image data were handled by three-dimen-sional angiography images with the software attached to the angiography equipment. The coronary artery tissues were HE stained and observed by microscope. The HE staining results were compared with the angiographic results.ResultsThe imaging data obtained from the 10 cases for examination showed 8 cases without coronary artery stenosis and 2 cases with Ⅲ, Ⅳcoronary artery stenosis, which were consistent with HE staining results of coronary artery organization and the both results were confirmed. ConclusionIsolated coronary angiography has an unique advantage for accurate grading of classification of coronary artery stenosis, examination of vascular malformation and tiny lesions, which can provide reference for the localization of small lesions and basis during the autopsy for identification conclusion.

Background:Tissue microarray has been increasingly used in research of malignancies. It has been revealed that TGF-β signaling pathway contributes to the tumorigenesis and progress of malignancies. Aims:To determine the expressions of Runx3,Smad4,Cdk2 and p21,the key molecules in TGF-β signaling pathway by tissue microarray,and investigate their correlations with clinicopathological features and prognosis of gastric cancer. Methods:A total of 378 paraffin embedded tissue blocks,including 130 gastric cancer tissue and 248 para-cancer tissue from 130 patients undergoing radical resection of gastric cancer were obtained. Tissue microarray and immunohistochemistry were used to determine the expressions of Runx3,Smad4,Cdk2 and p21. Results:The aberrant expression rates of Runx3,Smad4, Cdk21 and p21 in gastric cancer tissue were significantly higher than those in para-cancer tissue(67. 7% ,35. 4% , 63. 8% and 70. 0% vs. 14. 1% ,12. 5% ,18. 1% and 37. 1% ,P < 0. 05,respectively). Aberrant expression of Runx3 was closely correlated with histological grade and lymph node metastasis of gastric cancer( P < 0. 05),while aberrant expressions of Smad4 and p21 were correlated with histological grade only(P < 0. 05);aberrant expression of Cdk2 was correlated with histological grade,lymph node metastasis and TNM staging(P < 0. 05). Pairwise correlations were seen among aberrant expressions of Runx3,Smad4 and p21 in gastric cancer,while Cdk2 was correlated with Runx3 only. Kaplan-Meier survival curve showed that 5-year survival rates in Runx3,Smad4 and p21 aberrant expression groups were significantly lower than those in normal expression groups(P <0. 05). Furthermore,Cox proportional hazard model indicated that Runx3 and Smad4 were independent prognostic factors for gastric cancer. Conclusions:Runx3,Smad4,Cdk2 and p21 might play pivotal roles in tumorigenesis and progression of gastric cancer. As interactions occurred among these four proteins,whether Runx3 and Smad4 could be used for predicting the prognosis of gastric cancer needs to be further studied.

Objective To study the expression of PIWIL1, PIWIL3 and PIWIL4 in human colon cancer and its clinical significance. Methods We collected cancerous tissues and its adjacent tissues of 106 patients with colon cancer, two tissue microarrays were constructed, with 62 and 150 points respectively. We studied the expression of PIWIL1, PIWIL3 and PIWIL4 through immunohistochemistry. Results The expression of PIWIL1, PIWIL3 and PIWIL4 were significantly higher in cancerous tissues than those in adjacent tissues (P＜0. 01). In cancerous tissues and its adjacent tissues, postive correlation were seen among PIWIL1, PIWIL3 and PIWIL4 expression (P＜0. 01). PIWIL1 expression was significant higher in low differentiation group than that in high differentiation group (t =- 2. 840, P＜0.01 ). PIWIL3 expression was higher in high clinical stage than that in low clinical stage (F= 3. 112, P＜0.05). The expression of PIWIL3 and PIWIL4 were significantly higher in patients with colon cancer with distant metastasis than those without distant metastasis (t= -3. 349, P＜0.01 ; t = - 2. 168, P＜0. 05). PIWIL3 and PIWIL4 expression were correlated with occurring of colon cancer (P＜0. 01). Conclusions The expressions of PIWIL1,PIWIL3 and PIWIL4 in colon cancer were correlated with the differentiation, clinical stage and distant metastasis of colon cancer. PIWIL3 and PIWIL4 expression were two independent related factors of occurring of colon cancer, which would be furtherly investigated to be served as novel markers for early diagnosis and promising molecular targets for colon cancer therapy.

Objective To investigate the microRNAs expression profile in human colorectal cancer with or without liver metastasis and try to screen miRNA associated with liver metastasis in colorectal cancer. Methods Twenty five surgical resected colorectal cancer specimens were collected and frozen in liquid nitrogen. Three without liver metastasis and three with liver metastasis were selected, from which total RNAs were isolated. The expressions of miRNAs in these two types of specimens were detected by illumine microRNA microarray, and the difference of miRNA expression was screened. The biochip results were verified with real-time RT-PCR in all colorectal caner specimens. Results The miRNA expression was significantly different in colorectal cancer with liver metastasis and without liver metastasis. Compared with colorectal cancer without liver metastasis, 28 miRNA expressions was different in colorectal cancer with liver metastasis, 4 up regulated and 24 down regulated. The quantity of miR-139-3p expression in colorectal cancer with liver metastasis was 1.75±0.40, up regulated compared with that incolorectal cancer without liver metastasis(0. 69 ±0.58,P＜0.05). The quantity of miR-19a expression in liver metastasis was 0. 39±0. 20, downregulated compare with no liver metastasis( 1.38 ± 0.98, P＜0. 05). The result of miRNA biochip was consistent with that of RT-PCR. Conclusion The difference of miRNA expression might relate to liver metastasis of colorectal cancer. The specific miRNAs expression profile might provide new target for diagnosis and treatment of colorectal cancer with liver metastasis.

Objective To evaluate the impact of rosiglitazone (Ros) on liver expression of peroxisome proliferator-activated receptor γ (PPARγ),nuclear factor (NF-κB) and cyclooxygenase-2(COX-2) in treatment of nonalcoholic steatohepatitis (NASH) rats.Methods Thirty Sprague-Dawley rats were divided into normal group,model group and Ros treated group with 10 each.Except the normal group,the other two groups were given high fat diet for 12 weeks for NASH model.The rats in Ros treated group were gavaged 4 mg/kg of Ros daily at the 12th week for 8 weeks.All rats were sacrificed at the 20th week for blood sample and liver tissue.Biochemical parameters of liver function,lipid metabolism,glycometabolism and antioxidant enzyme activities were measured.The histological change of the liver were assessed with HE and Masson staining.The level of tumor necrosis factor (TNF)-α and prostaglandin E2 (PGE2) was measured using ELISA.The expression of PPARγ,NF-κB and COX-2 was detected with immunohistochemistry.The mRNA and protein expressions of COX-2 were tested by real-time PCR and Western blotting,respectively.Results In comparison with model group,Ros treated group showed significant improvement in hepatic steatosis,inflammation and fibrosis(all P value<0.05).In model group,the serum levels of fasting blood glucose,insulin and HOMA-insulin resistance index (HOMA-IRI),total cholesterol (TC),total triglyeride (TG),lowdensity lipoprotein cholesterol (LDL-C) and free fatty acids were increased,but HDL-C level was decreased.All above parameters markedly improved after Ros treatment.The levels of ALT and AST,total anti-oxidation competence,superoxide dismutase,catalase,glutathione peroxidase and malondialdehyde in Ros treated group were significantly ameliorated when compared with those in model group.Immunohistochemistry showed that the expression of NF-κB and COX-2 was significantly elevated,but PPARγ was decreased in model group.Real-time PCR and Western blot revealed that the mRNA and protein expressions of COX-2 were higher in the model group than those in normal group (0.57±0.08 vs 0.38±0.03;2.83±0.24 vs 1.00±0.03,P=0.000 and P=0.004,respectively),but significantly lower in Ros treated group (0.55±0.06 and 1.84±0.13,P<0.01).Conclusions Ros can reduce oxidative stress and insulin resistance in NASH rats by activing PPARγ expression and inhibiting expression of NF-κB and cyclooxygenases.

The main clinical symptoms of Parkinson's disease (PD) are resting tremor, muscle rigidity, bradykinesia, and so on. There is no effective treatment for PD yet, and dyskinesia symptoms affect the life qualities of PD patients. The therapy used for reinforcing kidney and activating blood circulation in treatment of PD can achieve good clinical effects.

Objective To investigate the whole genomic expression profiles of chronic atrophic gastritis with interleukin(IL)-1β-31CC/-511TT genotype as measured by oligonucleotide microarray technique.Methods Genomic RNA was extracted from gastric biopsies of 12 patients with chronic atrophic gastritis(6 with IL-1β-31CC/-511TT and 6 with IL-1β-31TT/-511CC).The genomic profiles of IL-1β gene polymorphisms 31CC/-511TT and 31TT/-511CC were compared and tested for differential expressed genes associated with 31CC/-511TT using Agilent human whole genomic oligonucleotide microarrays.The results were further analyzed in terms of gene ontology(GO).Results There were 200 differentially expressed genes associated with IL-1β-31CC/-511TT,159 of which were up-regulated and 41 were down-regulated.These genes mainly involved in macromolecule metabolic process,post-translational protein modification,ubiquitin cycle,and protein kinase cascade.Five genes had biological activities,one of which was down-regulated gene(PCSK5)and 4 were upregulated genes(PRKCA,NPLOC4,TRIB3 and MAPKAPK3).Conclusions The chronic atrophic gastritis with IL-1β-31CC/-511TT genotype has molecular phenotypes which is associated with malignance and inflammation.These individuals are needed more intensive preemptive treatment and dynamic surveillance.

Objective To prepare polyclonal antibodies against human PIWIL and to identify their property and tissue distribution of PIWIL.Methods PIWIL polypeptide was synthesized and conjugated to Keyhole limpet hemocyanin(KLH) as an immunogen.Then PIWIL-KLH conjugations were injected into rabbits subcutaneously to produce polyclonal antibodies.The specificity and sensitivity of antibodies were identified by ELISA and Western blot after purification by affinity chromatography.PIWIL were then stained on the tissue chip to study their distribution.Results Rabbit antibodies against PIWIL were prepared after injection of PIWIL-KLH conjugation.These antibodies specially recognized PIWIL peptides.Expression of PIWIL was found in the cytoplasm of epithelia cells of varied normal tissues and tumor tissues.Conclusion The successful preparation of the polyclonal antibody against PIWIL will provide an efficient reagent for further study of its role in the pathway of miRNA and RNA interference and in the pathogenesis of human disease.