This article introduced the thoughts and practice of implementing microbiology course reform for stomatology majors in school of stomatology and department of microbiology in Tianjin medical university.To make the introductory courses more suitable and meet the needs of stomatology better,our microbiology classes included the basic knowledge of oral microoganism.

Objective To investigate the value of quantitative TB‐DNA test ,interferon gamma release test and the detection of tuberculosis antibodies for the diagnosis of active pulmonary tuberculosis .Methods 51 patients were diagnosed as tuberculosis from 2013 July to 2014 June in the hospital ,whose sputum smear microscopy for acid fast bacilli were positive .Then TB‐DNA quantitative test ,interferon gamma release test (T‐SPOT .TB)and tuberculosis antibody detection were performed for those pa‐tients .All the result were retrospectively analysed .Results The positive rate of T‐SPOT .TB was 90 .1% ,the positive rate of quan‐titative TB‐DNA test was 74 .5% and the positive rate of tuberculosis antibody detection was 37 .3% .Conclusion Because the re‐sult of T‐SPOT .TB is not affected by the process of specimen collection ,it is the most sensitive test of the three tests at present , and has higher value in the auxiliary diagnosis of active pulmonary tuberculosis than the other two .

Objective To investigate the expression and significance of Periostin,VEGF and MMP-9 in breast invasive ductal carcinoma.Methods Immunohistochemistry was employed to detect the expression of Periostin,VEGF and MMP-9 in breast invasive ductal carcinoma and normal breast tissue.Results In breast invasive ductal carcinoma and normal breast tissue,the positive rates of Periostin were 63.8 ％ (37/58) and 0 (x2 =24.272,P =0.000).The figures were 69 ％ (40/58) and 8 ％ (2/25) for the positive rates of VEGF (x2 =25.977,P =0.000),respectively,as well as 70.69 ％ (41/58) and 16.0 ％ (4/25) for the positive rates of MMP-9 (x2 =21.050,P =0.000),respectively.There were significant differences among the groups (P ＜ 0.05).In breast invasive ductal carcinoma,the expression of Periostin was correlated with clinical stage (x2 =4.835,P =0.028),whereas not correlated with age (x2 =1.155,P=0.282),histological grade (x2 =0.05,P =0.972),lymphatic metastasis (x2 =1.660,P =0.198).The expression of VEGF was correlated with clinical stage (x2 =4.230,P =0.040),lymphatic metastasis (x2 =9.667,P =0.002),whereas not correlated with age (x2 =0.506,P =0.477),histological grade (x2 =0.532,P =0.767).The expression of MMP-9 was correlated with clinical stage (x2 =8.456,P =0.004),lymphatic metastasis (x2 =5.494,P =0.019),whereas not correlated with age (x2 =0.153,P =0.695),histological grade (x2 =0.224,P =0.894).The expression of Periostin,VEGF and MMP-9 were positively correlated with each other in breast invasive ductal carcinoma (r =0.348,P =0.001; r =0.303,P =0.021; r =0.469,P =0.000).Conclusion Periostin,VEGF and MMP-9 are correlated closely with the occurrence and development of breast invasive ductal carcinoma,which might be valuable in evaluating the invasiveness,metastasis and prognosis.

Objective To study the gene expression profiles of ovarian cancer by cDNA microarray,and preliminarily analyze the function of part of differential expression genes.Methods BiostarH-40s gene microarray containing 4 097 genes was used to analyze gene expression patterns in tissue samples from 5 cases of human ovarian serous cystadenocarinoma(cy5-dUTP present ) and 5 cases of normal ovarian tissues(cy3-dUTP present).Results 163 genes of differential expression were found in more than 4 cases of ovarian cancer,the expression of 66 genes increased(up-regulated),the expression of 97 genes decreased(down-regulated). 37 differential genes with difinite gene function classification were found including three protocogene and tumor suppressor genes,one cyclin protein gene,three cytoskeletal and movement protein genes,one DNA binding,transcription and transcription factors gene,two cell receptor genes,five immune-related protein genes,seven metabolism genes,two protein translation and synthesis genes,three growth-related genes and seven other types of genes.Conclusion Gene expression profiles of ovarian cancer can be detected by cDNA microarray,and differential expression genes and their gene function classification of ovarian cancer are found.

Objective To explore the methylation status of Rap1 GTPase activating protein (Rap1GAP) promoter in colon cancer, and to provide the oretical basis and research direction for the early diagnosis, targeted therapy, anti-multidrug resistance of colon cancer and so on. Methods The paraffin embedded specimens of 33 patients with colonic adenocarcinoma diagnosed by pathology were analyzed from Department of Pathology of Xinzhou City People′s Hospital from January 2010 to September 2014, including 19 males and 14 females, and aged 41-72 years old. The paraffin embedded specimens of 16 patients with colonic adenoma were enrolled, including 9 males and 7 females, and aged 34-58 years old. 13 normal tissues from the tumor distal margin (from the tumor > 15 cm) were selected. Quantitative methylation specific PCR (q-MSP) was applied to detect methylation level of Rap1GAP gene promoter. The methylation level differences of Rap1GAP gene promoter region among 3 groups or between different clinicopathologic factor subgroups were compared. Results The methylation rates [median (interquartile range)] of Rap1GAP promoter were 65.43 % (50.35 %), 21.37 % (8.39 %) and 17.43 % (15.71 %) in colonic adenocarcinoma group, colonic adenoma group and adjacent normal tissue group, respectively. The methylation rate of colonic adenocarcinoma group was significantly higher than that of colon adenoma group or that of adjacent normal tissue group (P< 0.05). The methylation rates of Rap1GAP promoter in colonic adenocarcinoma were not correlation with age, sex, differentiation and the stage of TNM [ male vs. female: 42.74 % (70.44 %) vs. 21.98%(80.00%);≤60yearsoldvs.>60yearsold:36.26%(62.62%)and26.23%(76.42 %);well-differentiated vs. moderately/poorly-differentiated: 21.98 % (40.32 %) vs. 42.74 % (74.20 %); TNM Ⅰ-Ⅱ vsⅢ-Ⅳ: 25.31 % (48.27 %) vs. 36.26 % (75.55 %); all P> 0.05]. Conclusion The methylation status of RAP1GAP promoter maybe associate with genesis and development of colon cancer, which might be used as a target for early diagnose of colon cancer.

Objective To establish a suitable extraction and purification process line for industrial production of liquiritin. Methods With the extraction rate of liquiritin as index, orthogonal test was used to determine the optimum conditions; with the retention rate of liquiritin and impurity removal rate as the indexes, orthogonal test was used to optimize the best ultrafiltration process parameters. Results The optimum extraction conditions were: 24 times 0.75%ammonia water, extracted three times, each time under 60 min. The liquiritin average extraction rate was 72.3%. The best ultrafiltration process parameters were: 10 nm inorganic ceramic membrane, pressure of 0.12 MPa, temperature of 25 ℃. The liquiritin average retention rate was 98.9%, and the average removal rate of impurity was 23.3%. Conclusion This process has low production cost and good safety, and is suitable for industrial application.

Objective To investigate the expression of signal-induced proliferation-associated gene 1 (SIPA1), Ezrin and E-cadherin (E-cad), and their relationship with clinical patterns in epithelial ovarian carcinoma.Methods Immunohistochemistry was used to detect the expression of SIPA1, Ezrin and E-cad in normal ovarian tissue, benign epithelial ovarian tumor, borderline epithelial ovarian tumor and epithelial ovarian carcinoma,respectively. Results The positive rate of SIPA1 expression was 44.2 % (23/52), 64.5 %(20/31), 93.3 % (28/30) and 100.0 % (15/15) in epithelial ovarian carcinoma, borderline epithelial ovarian tumor, benign epithelial ovarian tumor, and normal ovarian tissue, respectively, and there was a statistical difference (χ2 = 29.159, P= 0.000). The corresponding rates were 57.7 % (30/52), 61.3 % (19/31), 90.0 %(27/30) and 93.3 % (14/15) for the positive rate of Ezrin expression (χ2= 14.555, P= 0.002), as well as for 23.1 % (12/52), 58.1 % (18/31), 86.7 % (26/30) and 0 (0/15) for the positive rate of E-cad expression, respectively (χ2= 45.731, P= 0.000). In patients with epithelial ovarian carcinoma, the expression of SIPA1 was correlated with tumor differentiation (χ2=3.895, P=0.048), but not with histological type and clinical stage (all P>0.05). The expression of Ezrin was not correlated with histological type, tumor differentiation and clinical stage (all P>0.05). There was a positive correlation between expression of E-cad and SIPA1, Ezrin in epithelial ovarian carcinoma, respectively (r= 0.339, P= 0.014; r= 0.284, P= 0.041), but no correlation between the expression of SIPA1 and Ezrin (r= 0.214, P= 0.128). Conclusions SIPA1, Ezrin and E-cad play important roles in the occurrence and development of epithelial ovarian carcinoma. They cooperate in the progression and their combined detection can better evaluate the prognosis of epithelial ovarian carcinoma.

Objective To explore the enhancement effects and mechanisms of IL-32β on human cervical carcinoma cells C33A to hypoxic/hypoglycemic condition.Methods After cultured in hypoxia/hypoglycemic circumstance and normal circumstance for 20 hours respectively, the mRNA and protein expression of IL-32β in C33A cells were detected by real time-polymerase chain reaction (RT-PCR) and Western blotting respectively.Trypan blue stain was used to detect C33A cells viability in hypoxia/hypoglycemic circumstance and adding 10, 100,500 ng/ml IL-32β circumstance.The xenografted tumor of nude mice was established by intraperitoneal injection, and their volumes were tested for a given time after injecting 0, 1.0 mg/kg IL-32β.siRNA was used to construct IL-32β knockdown cells and detect the expression of VEGF.Results Under the hypoxia/hypoglycemic circumstance, the expressions of IL-32β mRNA were (6.12 ± 0.03) times of the normal circumstance (F =43.16, P ＜ 0.05), the expressions of IL-32β protein were (2.23 ± 0.04) times of the normal circumstance (F =22.32, P ＜ 0.05).The C33A cells viability in hypoxia/hypoglycemic circumstance was (51.92 ± 3.41) ％, whereas, viability in 10 ng/ml IL-32β group was (55.23 ± 3.92) ％ (F =14.25, P ＜ 0.05), viability in 100 ng/ml IL-32β group was (62.52 ± 4.14) ％ (F =35.53, P ＜ 0.01), viability in 500 ng/ml IL-32β group was (69.14 ± 2.45) ％ (F =56.28, P ＜ 0.01).After 28 days, the volume of xenografted tumor of 0 mg/kg IL-32β group was (578 ± 64)mm3, and 1.0 mg/kg IL-32β group up to (1 402 ± 142) mm3 (F =27.84, P ＜ 0.01).In addition, compared with control group, the expression of VEGF in IL-32β knockdown C33A cells was significantly decreased (F =36.85, P ＜ 0.05).Conclusion IL-32β can enhance the resistance to hypoxic/hypoglycemic condition of C33A cells, which is associated with the increase of VEGF.

Objective To investigate the expression of Rap1 GTPase-activating protein 1 (Rap1GAP),matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9),and their relation with clinical patterns in colorectal carcinoma.Methods Immunohistochemistry was used to detect the expression of Rap1GAP,MMP-2 and MMP-9 in colorectal carcinoma,villous adenoma,tubular adenoma and normal colorectal tissue,and their relationship with clinicopathological parameters was analyzed.Results The positive rate of Rap1GAP expression was 30.4 ％ (14/46),77.8 ％ (14/18),69.6 ％ (16/23) and 95.2 ％ (20/21) in colorectal carcinoma,villous adenoma,tubular adenoma,and normal colorectal tissue,respectively (x2 =30.659,P=0.000).The figures were 71.7 ％ (33/46),55.6 ％ (10/18),52.2 ％ (12/16) and 9.5 ％ (2/21) for the positive rate of MMP-2 expression (x2 =22.459,P =0.000),as well as for 76.1％ (35/46),61.1％ (11/18),56.5 ％ (13/23) and 14.3 ％ (3/21) for the positive rate of MMP-9 expression,respectively (x2 =22.643,P =0.000).In patients with colorectal carcinoma,the expression of Rap1GAP was correlated with tumor differentiation (x2 =5.275,P =0.022),but not sex,age,or lymphatic metastasis (all P ＞ 0.05).The expression of MMP-2 and MMP-9 were correlated with lymphatic metastasis (x2 =6.661,P =0.010;x2 =8.475,P =0.040),but not sex,age or tumor differentiation(all P ＞ 0.05).There was a negative correlation between expression of Rap1GAP and MMP-2,MMP-9 in colorectal carcinoma,respectively (r =-0.424,P =0.003;r =-0.294,P =0.048),but no correlation between the expression of MMP-2 and MMP-9 (r =0.101,P =0.505).Conclusions Rap1GAP,MMP-2 and MMP-9 play important roles in the malignant biological behavior of colorectal carcinoma,and the expression of Rap1GAP is negatively correlated with MMP-2 and MMP-9.The interactions among the three affect the occurrence and development of colorectal carcinoma.

Objective To study the effects of ultrafiltration process on activating blood and removing blood stasis efficacy ofShentong Zhuyu Decoction; To investigate the feasibility of applying ultrafiltration technology in purifying Shentong Zhuyu Decoction.Methods The mice micro artery and vein diameter, clotting time and opening capillary of auricle microcirculation of mice were used as indexes to observe the effects of different ultrafiltration process on activating blood and removing blood stasis efficacy ofShentong Zhuyu Decoction.ResultsShentong Zhuyu Decoction showed satisfying efficacy of activating blood and removing blood stasis. There was no significant difference between the non-ultrafiltration process and ultrafiltration processed by 20 and 50 nm ultrafiltration membranes.Conclusion Ultrafiltration technology can be applied to purifying Shentong Zhuyu Decoction, and the membrane pore size must be more than 20 nm.