Objective To research the promoter methylation level of RAS association domain family 1A (RASSF1A) and RASSF1A gene mRNA expression level in cervical cancer tissue, and to analyze their relationships with clinicopathological parameters of cervical cancer and the clinical significance.Methods The RASSF1A gene promoter methylation and RASSF1A gene mRNA were detected respectively by methylation specific PCR and quantitative real-time PCR method in 40 cases of cervical cancer tissues and corresponding adjacent tissues.Results RASSF1A mRNA expression level in cervical cancer (0.26±0.05) was significantly lower than that in adjacent tissues (0.28±0.03), and the difference was statistically significant (t=2.27, P=0.026).The methylation rate of RASSF1A gene promoter region (0.71%±0.04%) was significantly higher than that in adjacent tissues (0.66%± 0.03%), and the difference was statistically significant (t=6.78, P=0.000).The expression of RASSF1A mRNA was significantly correlated with pathological differentiation (t=3.31, P=0.002), International Federation of Gynecology and Obstetrics (FIGO) stage (t=2.13, P=0.040), lymphatic metastasis (t=2.56, P=0.015).The promoter methylation level of RASSF1A gene was significantly correlated with pathological differentiation (t=2.08, P=0.045), FIGO stage (t=2.66, P=0.011), lymphatic metastasis (t=2.22, P=0.033), depth of invasion (t=2.12, P=0.041).Conclusion The RASSF1A gene promoter region methylation level and the RASSF1A gene mRNA expression level are associated with the malignant degree of cervical carcinoma.The RASSF1A gene promoter region methylation level may be used as a reference indicator for predicting the risk of metastasis of cervical cancer.

Test is an important way for getting feedback from students about teaching. The traditional examination is not suitable for the requirements of modernization teaching.Web-Based Course Examination of practice pathology simplifies the process of traditional examination greatly. It is one of the important means in modernization teaching.

Objective To investigate the micro-vessel density and vascular endothelial growth factor receptor 2 (VEGFR2) expression in mucosa and submucosa tissue of the patients with hemorrhoids. Methods Sixty hemorrhoids patients were randomly recruited and involved into experiment group and 20 cases of normal patients involved in control group. RASP, MASP, RSTV, RMTV, Expression of VEGFR2 and microvessel density in two groups were detected by immunohistochemistry. The expression abundance of VEGFR2 mRNA was detected by RT-PCR. Results Excessive angiogenesis was found in hemorrhoids tissue (5.59 ± 0.98) in experiment group as compared with that in control group (3.92 ± 0.81), the difference was statistically significant (P < 0.05). Staining score of hemorrhoid tissue and the control tissue were 5.95 ± 1.01, 2.32 ± 0.72, respectively (P < 0.05). Abundance of VEGFR2 mRNA expression in hemorrhoid tissue was significantly higher than that in control tissue (P < 0.05). Conclusion VEGFR2 plays an important role in angiogenesis during the development of hemorrhoids, which may be closely related to the occurrence of hemorrhoid disease and clinical symptoms.

Background and purpose:It has beenreported that miR-1284 is associated with gastric cancer lymph node metastasis in the research of microRNA microarray in human gastric cancer tissues. But the specific role of miR-1284 in gastric cancer has not been reported. The aim of this study was to investigate the effect of miR-1284 over-expression on the gene expression profiling and invasion/metastasis of human gastric cancer SGC-7901 cells. Methods:Gastric cancer SGC-7901 cells of LV-miR-1284 group were transfected with lentiviral vectors of miR-1284, cells of LV-NC-GFP group were transfected with lentiviral vectors without miR-1284, and cells of control group were not transfected with lentiviral vectors. The expression of miR-1284 was detected by the real-time fluorescent quantitative PCR. Differential expression genes were detected by the microRNA chip. Target genes of miR-1284 were predicted by the bioinformatics. Invasive ability was detected by the Transwell invasion assay. Metastasis ability was detected by subcutane-ously transplanted tumor model of nude mice.Results:Compared with LV-NC-GFP and control groups, the expressions of miR-1284 and 20 genes were up-regulated, and the expression of 17 genes was down-regulated in LV-miR-1284 group. One hundred and thirty-eight target genes of miR-1284 were predicted by the bioinformatics website. Compared with invasive cell number of LV-NC-GFP group (168.67±4.55) and control group (170.33±3.08), the ability of invasion ofcells was weakened in LV-miR-1284 group (70.00±2.37). Compared with the liver metastasis rate of LV-NC-GFP group (85.71%) and control group (85.71%), the ability of metastasis of cells was weakened in LV-miR-1284 group (14.29%). Conclusion:The ability of invasion and metastasis of SGC-7901 cells is suppressed by over-expression of miR-1284. The mechanism may be related to regulating the expression ofSUMO1 andJUNgenes.

AIM:To study the effect and the molecular mechanism of CDX 2 over-expression on the prolifera-tion, growth and cell cycle of human gastric cancer cell line SGC-7901.METHODS:The SGC-7901 cells in LV-CDX2-GFP group were transfected with the recombinant lentivirus vector LV-CDX2-GFP, the cells in LV-GFP group were trans-fected with the negative control lentiviral vector for the negative control , and the cells in blank control group were without any treatment.The cell proliferation was detected by CCK-8 assay.The cell cycle distribution was analyzed by flow cytome-try.The expression of CDX2, Bax, Bcl-2, cyclin D1 and survivin was determined by semi-quantitative RT-PCR and Wes-tern blotting .RESULTS:Compared with LV-GFP group and blank control group , the proliferation activity of the SGC-7901 cells was significantly lower (P<0.05), the G0/G1 phase proportion increased (P<0.05), the mRNA and protein levels of Bcl-2, cyclin D1 and survivin were reduced (P<0.05), and the mRNA and protein levels of Bax were up-regula-ted (P<0.05) in LV-CDX2-GFP group.No statistically significant difference of the above indexes was observed (P>0.05) between LV-GFP group and blank control group .CONCLUSION:Over-expression of CDX2 mediated by lentivirus inhibits the proliferation and growth of human gastric cancer SGC-7901 cells and arrestes the cell cycle at G 0/G1 phase, which may be related to down-regulation of Bcl-2, cyclin D1 and survivin and up-regulation of Bax .

AIM:To evaluate the correlation between microRNA-1284 (miR-1284) and gastric cancer, and to investigate the underlying mechanism.METHODS: The expression of miR-1284 was examined by real-time PCR in 63 gastric cancer ( GC) tissue samples and 63 non-malignant adjacent tissue samples.The correlation between miR-1284 and the clinicopathological feature of GC was analyzed.Lentiviral vector containing miR-1284 was constructed and transfected into GC SGC-7901 cells.After transfection, the expression of miR-1284 was examined by real-time PCR.The cell activity was evaluated by CCK-8 assay.The cell cycle and apoptosis were determined by flow cytometry.The ability of cell migra-tion was measured by wound-healing assay.The potential target gene of miR-1284 was predicted by online bioinformatic softwares.The expression of JAG1 mRNA was examined by real-time PCR.The protein levels of JAG1, Notch1 and NF-κB were analyzed by Western blotting.RESULTS:Compared with non-malignant adjacent tissue samples, the results of real-time PCR showed significant downregulation of miR-1284 in 42 GC tissue samples ( P<0.05 ) .The expression level of miR-1284 was not significantly associated with age and gender of the patients, tumor size, TNM staging and lymph node metastases (P>0.05), but significantly associated with histologic grading (P<0.05).Compared with LV-NC-GFP group and control group, after transfection of miR-1284 in LV-miR-1284 group, the expression of miR-1284 was significantly in-creased (P<0.05), the percentages of apoptotic cells and the cells in G0/G1 phase were significantly increased (P<0.05), the cells activity and ability of migration were significantly decreased (P<0.05), and the expression of JAG1, Notch1 and NF-κB was significantly decreased (P<0.05).CONCLUSION:The inhibitory effect of miR-1284 on gastric cancer may be associated with the regulation of its targeting gene JAG1.

Objective To investigate the action of chondrogenesis differentiation of bone marrow stromal cells (BMSCs) transfected with adeno-hTGF-β1. Methods In the experiment group, replication-deficient a denoviruses carrying human hTGF-β1 complementary DNA (adeno-hTGF-β1 was constructed and applied to transfect to the first generation BMSCs. As a control, each BMSCs was transduced with 200 pfu of adeno-LacZ gene. One day after transfer, BMSCs were trypsinized, counted, and 5×105 cells aliuots were spun down at 500 rpm per minute in 15 ml polypropylene conical tubes and then cultured in a defined medium in an incubator at 37℃ for 21 days. The aggregates were harvested at time points to 21 days and assessed by gross observation, histological analyses and immunohistochemical localization of type Ⅱ collagen. Results When harvested at 21 days, each pellet shrinked to spheroid tissue with apearly opalescence in gross morphology and found to be relatively firm. H.E staining showed elongate dlining cells appeared as perichon drium-like cells at the surface. Some nests of cartilage were observed at the substrate of the tissue. Mature chon drocytes were embeded in the lacuna in the experiment group. In addition, Safranin'O staining confirmed the presence of sulfated proteoglycans in the ECM of chondrogenesis region. Immunohistochemical staining revealed the presence of type Ⅱ collagen in chondrogenesis region. By contrast, HE staining showed no evidence of cartilage formation in the control group. They were fibrous tissue with no architectural feature. Safranin'O staining and Immunohistochemical staining showed no evidence of sulfated proteoglycans or typeⅡ collagen expression. Conclusion BMSCs transfected with adeno-hTGF-β1 could induce its chondro-genesis when aggregate cultured in a defined medium in vitro, laying a foundation for the application of hTGFβ1 gene-transfected BMSCs in cartilage tissue engineering.

An efficient modified route based on the targeting mechanism of antibacterial fluoroquinolones for the shift from the antibacterial activity to the antitumor one was further developed. Using a fused heterocyclic ring, s-triazolothiadiazine as a carboxyl bioisostere of ciprofloxacin, the title compounds, 1-cyclopropyl-6-fluoro-7-piperazin-1-yl-3-(6-substituted-phenyl-7H-[1, 2, 4]triazolo[3, 4-b][1, 3, 4]thiadiazin-3-yl)-quinolin-4(1H)-ones (5a-5e) and their corresponding N-acetyl products (6a-6e), were designed and synthesized, separately. Meaningfully, a ring-contraction of fused six-membered thiadiazine occurred by a sulfur extrusion reaction gave new tri-acetylated fused heterocycles related to pyrazolo[5, 1-c][1, 2, 4] triazoles (7a-7e). The in vitro antitumor activity against L1210, CHO and HL60 cell lines was also evaluated for the synthesized fifteen heterocycles compared to parent ciprofloxacin by methylthiazole trazolium (MTT) assay. Interestingly, the results displayed that fifteen fused heterocyclic compounds showed more significant growth inhibitory activity (IC50 < 25.0 micromo x L(-1)) than that of parent ciprofloxacin (IC50 > 150.0 micromol x L(-1)), and the active order decreased from 7a-7e to 5a-5e to 6a-6e, respective.

Bones are stained into red color with feeding madder, but we do not know whether the fed madder can change the bone biomechanical properties and bone mineral contents in animals. In this research, we established a rat model with feeding madder. The bone biomechanical properties were detected by universal material mechanics, bone mineral contents were detected by inductively coupled plasma mass spectrometry and spectrometer, and red color material in bone was analyzed by high performance liquid chromatography. The results showed that bone biomechanical parameters in femur diaphysis in the 10% and 15% group rats were significantly higher than those in the control group after feeding madder for 6 months. The level of calcium, magnesium and zinc in femur diaphysis in 10% and 15% group rats were higher than those in the control group after feeding madder for 6 months. However, it was shown that the kidney congestion and hyperemia and the level of blood urea nitrogen and creatinine in the 15% group rats were significantly different compared to those in the control group rats after feeding madder for 6 months. The red colored material in bone is related to alizarin analyzed with high-performance liquid chromatography. The conclusion could be drawn that feeding 10% madder in diet was not toxic to the rats fed for 6 months, and it could improve bone biomechanical properties and increase bone mineral elements.
Animals ; Anthraquinones ; toxicity ; Biomechanical Phenomena ; Bone Density ; Bone and Bones ; drug effects ; physiology ; Calcium ; Femur ; Magnesium ; Rats ; Zinc

Objective:To explore the relationship between proactive personality, resilience and achievement motivation of military cadets, and to explore the mediating role of resilience between proactive personality and achievement motivation, as well as the moderating effect of cadets’leading ability on this mediating role.Methods:In this study, 109 military cadets were measured with proactive personality scale, Connor-Davidson resilience scale (CD-RISC) and achievement motivation scale. SPSS 26.0 was used for descriptive statistical analysis and correlation analysis.Results:①There was a significant correlation among the total scores of proactive personality, resilience and achievement motivation ( Ps<0.01). ②Proactive personality could significantly predict the level of achievement motivation ( a=0.454, P<0.001), and resilience could significantly predict the level of achievement motivation ( b=0.231, P=0.019). ③Resilience significantly mediated the relationship between proactive personality and achievement motivation ( c′=0.3, P=0.003). ④Cadets' leading ability moderated the effect of resilience on achievement motivation (index=0.338, 95% CI: 0.057 to 0.881). The mediating effect of resilience between proactive personality and achievement motivation was significant for cadets' leaders (Effect=0.381, 95% CI: 0.085 to 1.005), but not significant for other students (Effect=0.043, 95% CI: -0.069 to 0.252). Conclusion:The proactive personality of military cadets can affect the level of achievement motivation through psychological resilience, which is also moderated by cadre identity, suggesting that military education should improve the level of achievement motivation in many ways.