Objective To construct the recombined DNA pcDNA3.1-hBMP-2 and transfect into human marrow stromal stem cells (MSCs) in vitro, and to explore theeffects of transfection on cellular proliferation and expression of vascular endothelial growth factor (VEGF). Methods The expression of human bone morphogenetic protein 2(hBMP-2) in these cells after transfection was determined by in situ hybridization and immunohistochemical analysis and Western blot analysis. The changes of cell proliferation were observed by flow cytometry. The effects of BMP-2 gene transfection on expression of VEGF in the cells were analyzed by in situ hybridization of VEGF cDNA probe. Results Stable expressionof hBMP-2 in pcDNA3.1-hBMP-2 transfected MSCs was confirmed in the levels of mRNA and protein.Cellular proportion in S period increased, which indicated that the synthesis of cell DNA increased. The expression of VEGF in the cells increased obviously. Conclusion With the help of lipofectamine, the pcDNA3.1-hBMP-2 were transfected into human MSCs successfully. hBMP-2 plays an important role in promoting cellular proliferation and vascular generation during bone repair.
ObjectiveTo detect the expression of motilin in gastric cancer tissues and to explore the relationship between motilin protein expression and clinicopathologic characteristics of gastric cancer. MethodsThe immunohistochemical staining was used to detect the expression of motilin protein in gastric cancer, paracancerous tissues, and normal gastric mucosa tissues. The relationship between motilin protein expression and clinicopathologic characteristics of gastric cancer was analyzed. ResultsThe expression of motilin protein in gastric cancer tissues (1 206.43±631.67) was significantly higher than that in normal gastric mucosa tissues and paracancerous tissues, respectively (Plt;0.01). The difference of motilin protein expression between normal gastric mucosa tissues and paracancerous tissues was not significant (Pgt;0.05). The expression of motilin protein in gastric cancer was correlated with the site of tumor, differentiation degree, and lymph node metastasis (Plt;0.05). ConclusionMotilin may participate in the carcinogenesis of gastric cancer, and correlated with the invasion and metastasis of gastric cancer.
OBJECTIVE To determine the effect of basic fibroblast growth factor (bFGF) on the biological behaviour of ovarian epithelial neoplasm. METHODS Ten cases of normal ovarian tissues and eighty cases of ovarian epithelial tumor tissues were detected by immunohistochemical methods. Mias-2000 Picture Analysis System was used to study the relationship of bFGF expression intensity and microvessel count, FIGO stage, pathological grade and classification of ovarian epithelial neoplasm. RESULTS 1. Expression of bFGF was mainly in cytoplasm and nucleus in several cells of borderline and malignant tumor. 2. The expression intensity of bFGF was closely related to the malignant degree of ovarian epithelial neoplasm. The density of bFGF expression was (3.35 +/- 3.52)% in normal ovarian epithelium, (19.25 +/- 21.73)% in benign tumor, (33.78 +/- 10.86)% in borderline tumor and (48.18 +/- 12.93)% in malignant tumor. The results indicated that bFGF might play an important role in carcinogenesis of ovarian epithelial neoplasm. 3. The expression intensity of bFGF was increased with the FIGO stage of ovarian tumor. 4. The expression intensity of bFGF was increased accompanying with the decrease of differentiation degree in ovian neoplasm. 5. In borderline tumor, expression intensity of bFGF in serous cystadenoma was significantly higher than in mucinous cystadenoma, which indicated bFGF might be an important factor in canceration of ovarian epithelial neoplasm. CONCLUSION bFGF may play important roles in carcinogenesis, development, invasion and metastasis of ovarian epithelial neoplasm.
In order to study the expression change of insulin-like growth factor-1 (IGF-1) and its receptor genes in different generations of tendon cell in culture, Dig-labeled synthesized oligonucleotide probes were used to detect the mRNA expression in primary, 6th and 13th generation of tendon cell. The results showed that IGF-1 receptor mRNA was expressed in all of the 3 above generation tendon cells. IGF-1 mRNA was expressed only in primary and 6th generation cells. Tendon cell of 13th generation did not express IGF-1 mRNA. It might suggest that the absence of IGF-1 mRNA expression be one of the causes which led to the decrease of reproductive ability of 13th generation tendon cell.
OBJECTIVE: To explore the expressive characteristics of epidermal growth factor (EGF) and its receptor (EGFR) in tissues of fetal and adult intestines. METHODS: The expression intensity and distribution of EGF and EGFR were detected with pathological and immunohistochemical methods in 6 specimens of adult (16-54 years) intestines and 18 specimens of fetal intestines with different gestational ages (13-31 weeks). RESULTS: Positive protein particles of EGF and EGFR could be detected in tissues of fetal and adult intestines. The protein expressions of EGF and EGFR were elevated progressively with the gestational age. EGF was mainly located in the cytoplasm and extracellular matrix of intestinal villus cells, endothelial cells and tunica serosa epithelial cells, while EGFR chiefly distributed in the cellular membrane of these cells. CONCLUSION: The endogenous EGF and EGFR might be involved in the intestinal development at embryonic stage, in the structural and functional maintenance at adult stage, and in the wound healing after injury.
The expression and rearrangement of bcl-2 gene in 64 cases with colorectal carcinoma were studied by immunohistochemical technique and semi-nest PCR respectively. The results showed the abnormal changes of the expression and rearrangement of bcl-2 gene had emerged in the early stage of colorectal carcinoma. The tumors with the expression of bcl-2 were associated with a higher incidence of metastasis to lymphatic node. The rearragement of bcl-2 was significantly higher in late-stage than that in early-stage. These suggest that bcl-2 gene involves in the regulation of the development of colorectal carcinoma. The state of the changes of bcl-2 gene in colorectal carcinoma may predict the therapeutic effect and prognosis of colorectal carcinoma.
ObjectiveTo systematically review the association between expression of osteopontin (OPN) and Chinese population with hepatocellular carcinoma (HCC) and its clinical pathological characteristics. MethodsSuch databases including CBM, CNKI, VIP and WanFang Data were searched from inception to July 2014, for studies about the association between expression of OPN and Chinese population with HCC and its clinical pathological characteristics. Two reviewers independently screened literature according to the exclusion and inclusion criteria, extracted data, and assessed methodological quality of included studies. Then, meta-analysis was performed using RevMan 5.2 software. ResultsA total of 10 case-control studies (involving 723 HCC cases and 102 controls) were included. The results of meta-analysis showed that:OPN expression was higher in HCC group than normal control group (OR=10.25, 95%CI 6.13 to17.14); and higher in imperfect capsular infiltration group than perfect capsular infiltration group (OR=2.71, 95%CI 1.58 to 4.64). However, no significant difference was found in OPN expression between isolated tumour group and multiple tumours group (OR=0.95, 95%CI 0.56 to 1.62); between high differentiation group and low differentiation group (OR=0.60, 95%CI 0.36 to 1.01); and between clinical stages I-Ⅱ group and clinical stages Ⅲ-IV group (OR=0.93, 95%CI 0.53 to 1.63). ConclusionCurrent evidence shows that OPN may take part in the whole course (occurrence and advance) of HCC in Chinese population, but the problem whether it can be used as a factor to evaluate prognosis needs to be further studied.
Objective To investigate the expression of COX-2 in human cervical cancer and explore their relationship between the COX-2 expression and the clinicopathologic characteristic of cervical cancer. Methods The published studies were searched in the CBMdisc (1979 to 2009), CNKI (1979 to 2009), VIP (1989 to 2009) and WANFANG Database (1982 to 2009), and other relevant journals were also hand searched, to identify all the relevant case-control trials. The quality of the included studies was assessed. The Cochrane Collaboration’s software RevMan 4.2.10 was used to test the heterogeneity, overall effect and publication bias of the combined studies. Results A total of 9 studies were recruited. As for the positive rate of COX-2 expression, significant differences was tested between cervical cancer vs. normal cervical tissues, lymph node metastasi vs. non-lymph node metastasi, clinical stages I-II vs. clinical stages III-IV, cell differentiation G1 vs. cell differentiation G2-G3 and cervical squamous cell carcinoma vs. adenocarcinoma with OR (95%CI) at 28.03 (9.53 to 82.50), 5.16 (3.36 to 7.93), 0.53 (0.33 to 0.84), 3.11 (1.86 to 5.22) and 5.00 (2.68 to 9.35) respectively. Conclusions According to the domestic evidence, higher COX-2 expression might be associated with cervical cancer. However, more high quality case-control studies are expected for further study.
Objective To investigate the expression of enhancer of zeste homolog 2 （EZH2） mRNA in colorectal cancer tissues， and to explore the relationship with clinicopathological features of it. Methods PCR was used to examine the expression of EZH2 mRNA in colorectal cancer tissues， paracancerous tissues， and normal epithelium tissues of 27 patients with colorectal cancer. Meanwhile， the relationship between the expression of EZH2 mRNA and clinicopathological features of colorectal cancer were analyzed. Results Expression level of EZH2 mRNA in colorectal cancer tissues， paracancerous tissues， and normal epithelium tissues were 3.01±1.29， 1.20±0.69， and 0.87±0.37 respectively， and the expression value of colorectal cancer tissues at the top （P＜0.01）， but there were no significant difference between the paracancerous tissues and normal epithelium tissues （P=0.403）. Expression of EZH2 mRNA were related to TNM stages （P=0.002） and degree of differentiation （P=0.005）， but were not related to age （P=0.388）， gender （P=0.963）， diameter of tumor （P=0.579）， histological type （P=0.945）， location of tumor （P=0.611）， and lymph node metastasis （P=0.449）. Conclusions EZH2 mRNA expresses highly in colorectal cancer tissues， so it may play an essential role in the emergence and development of colorectal cancer. EZH2 mRNA can be used as one of the referenced indexes to determine the malignant degree and process of colorectal cancer.
【摘要】 目的 觀察不同種培養基中重組人色素上皮衍生因子(rPEDF)融合蛋白的表達。 方法 將前期研究已構建的pET28aPEDF原核表達重組體轉化E.coli BL21大腸桿菌表達宿主菌,酶切鑒定陽性菌落后,分別在M9和LB培養基中用異丙基βD硫代半乳糖(IPTG,IsopropylbetaDthiogalactoside)誘導表達,SDSPAGE電泳檢測表達的PEDF蛋白, 美國ImagePro Plus 分析系統進行蛋白定量分析。結果 LB和M9培養基中均獲得相對分子質量約54×103的rPEDF融合蛋白。但LB培養基獲得的是rPEDF融合蛋白的包涵體,目的蛋白占總蛋白含量為21046%,M9培養基獲得的是可溶性的rPEDF的融合蛋白,目的蛋白占總蛋白含量的1231%。結論 不同種培養基中均有rPEDF 融合蛋白的表達。【Abstract】 Objective To observe the express of recombinant pigment epithelial derivative facto (rPEDF) in the different medium. Methods The pET28aPEDF was transformed into E.coli BL21. After the colonies were positive identification which were induced by IsopropylbetaDthiogalactoside in medium M9 and LB. The PEDF protein were detected by SDSPAGE and analyzed by American ImagePro Plus system. Results LB and M9 medium obtained the relative molecular mass about 54×103 rPEDF fusion protein. But LB medium obtained the inclusion bodys of rPEDF fusion protein，the purpose protein account for 21.046%；LB medium obtained the soluble rPEDF fusion protein，the purpose protein account for 12.31%. Conclusion The rPEDF protein was expressed in the different medium.