Objective To explore the expression of the vascular cell adhesion molecule 1 (VCAM1) in the acellular dermal matrix grafting in pigs. Methods Experimental models were established with 15 Inbred Strain mini pigs, 6 full-thichness skin defect wounds, 6 cm × 6 cm in size, were produced on both-side backs of the each pig, and then the pigs were randomly divided into 3 groups. In Group A (n=5, control) the thin auto-skintransplantation alone was made; in Group B (n=5), the grafting was performed in the acellular allo-dermal matrix combined with the thin auto-skins; in Group C (n=5), the grafting was performed in the acellular xeno-dermal matrixcombined with the thin auto-skins. The areas of the wounds were measured and the survival condition of the grafted skins was observed at 3, 9, 21 and 30 days after the grafting. The histological samples were harvested from the grafting area at 3, 6, 9, 12, 21 and 30 days after the procedure. The flow cytometry was employed to analyze the changes in the VCAM1 level in the sample at different time points after the grafting. Results In the 3 groups, the transplanted skin base was easily separated at 3 days after transplantation; the areas of the wound healing accounted for 94%±12%，92%±9%, and 91%±11%, respectively, at 21 days; good wound healing was achieved at 30 days. At 9 and 12 days after transplantation, there was an evidentlyincreased level of the VCAM-1 expression in the tissue samples in the composite skin grafting groups. Compared with the control group, the difference was significant (Plt;0.05); however, the VCAM-1 expression at 3 days was not statistically different between the composite skin grafting groups and the control group after transplantation. In contrast, the level of the VCAM-1 expression was significantly higher at 6 days in the control group than in the composite skin grafting groups (Plt;0.05). The levels of the VCAM-1 expression were significantly lower at 30 days than at 3 days after transplantation in all the 3 groups (plt;0.01). Conclusion The highest level of the VCAM-1 expression can be delayed in the composite skin grafting when compared with that in the thin auto-skins alone, which implies that the VCAM-1 expression may be correlated with angiogenesis and composite skin survival. The VCAM-1 expression is not different between the acellular allo-dermal matrix composite skin grafting groups and the acellular xeno-dermal matrix group.
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.
【摘要】 目的 观察不同种培养基中重组人色素上皮衍生因子(rPEDF)融合蛋白的表达。 方法 将前期研究已构建的pET28aPEDF原核表达重组体转化E.coli BL21大肠杆菌表达宿主菌,酶切鉴定阳性菌落后,分别在M9和LB培养基中用异丙基βD硫代半乳糖(IPTG,IsopropylbetaDthiogalactoside)诱导表达,SDSPAGE电泳检测表达的PEDF蛋白, 美国ImagePro Plus 分析系统进行蛋白定量分析。结果 LB和M9培养基中均获得相对分子质量约54×103的rPEDF融合蛋白。但LB培养基获得的是rPEDF融合蛋白的包涵体,目的蛋白占总蛋白含量为21046%,M9培养基获得的是可溶性的rPEDF的融合蛋白,目的蛋白占总蛋白含量的1231%。结论 不同种培养基中均有rPEDF 融合蛋白的表达。【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.
Objective To observe the expression of heat shock protein 70 (HSP70) in human liver after hepatic transplantation, and to study its correlation with the occurrence and progression of acute allograft rejection.Methods Fifteen biopsy specimen of allograft liver after transplantation were collected and divided into three groups according to their pathological changes: control group (no rejection), mild acute rejection group, and moderate/serious acute rejection group. The expressions of HSP70 in grafts were detected by using immunohistochemical method and imaging analysis. Results HSP70 was expressed in all 3 groups, and appeared mainly in hepatocellular cytoplasm. The immunohistochemical imaging analysis of HSP70 showed: integral optical density (IOD) which was 30.99±11.14 in the control group was lower than that in the mild acute rejection group (68.84±21.37) and that in the moderate/serious acute rejection group (71.82±19.99), P＜0.01； and the IOD in the moderate/serious acute rejection group was higher than that in the mild acute rejection group (P＜0.05). Conclusion HSP70 plays a role in cellular protection for allograft liver, and the continuously increasing expression of HSP70 in graft maybe closely relates to the occurrence and progression of acute allograft rejection.
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.
OBJECTIVE: To observe the protein expression of phosphorylated form of P38 mitogen-activated protein kinase(P38MAPK) and c-Jun in hypertrophic scar skin and to explore their influences on the formation and maturation of hypertrophic scar. METHODS: The expression intensity and distribution of phosphorylated form of P38MAPK and c-Jun were examined with immunohistochemistry and pathological methods in 16 cases of hypertrophic scar skin and 8 cases of normal skin. RESULTS: In normal skin, the positive signals of phosphorylated form of P38MAPK mostly distributed in basal lamina cells of epidermis, while c-Jun was mainly located in epidermal cells and endothelial cells. The positive cellular rates of two proteins were 21.3% +/- 3.6% and 33.4% +/- 3.5% respectively. In proliferative hypertrophic scar skin, the particles of phosphorylated P38MAPK and c-Jun were mainly located in epidermal cells and some fibroblasts. The positive cellular rates of two proteins were significantly elevated to 69.5% +/- 3.3% and 59.6% +/- 4.3% respectively (P lt; 0.01). In mature hypertrophic scar, the expression of these proteins decreased but was still higher than that of normal skin. CONCLUSION: The formation and maturation of hypertrophic scar might be associated with the alteration of phosphorylated P38MAPK and c-Jun protein expression in hypertrophic scar.
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.
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 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.
Objective To investigate the expression levels of osteoprotegerin (OPG) and receptor activator of NF-κB l igand (RANKL) mRNAs in bone tissues of the femoral head of the patients suffering glucocorticoid-induced osteonecrosisof the femoral head (ONFH), and to discuss the relationship between OPG/RANKL and ONFH. Methods Between March2007 and March 2008, bone tissues of the femoral head were collected as the experimental material from 35 patients suffering ONFH (experimental group) and from 21 patients suffering fracture of femoral neck (control group). The ratio of men to women in both groups was 4 ∶ 3, whose age was 41-70 years old (55.34 on average in the experimental group and 55.33 on average in the control group). The experimental group received over 3 weeks’ glucocorticoid treatment or more than 1 week’ s high-dose glucocorticoid treatment in recent 2 years, while the control group never received more than 1 week’s hormone treatment. In the two groups, the microstructure of bone tissues of the femoral head was detected by HE staining and the bone tissue total RNA was extracted, and then the expression levels of OPG mRNA and RANKL mRNA were examined by realtime quantitative PCR (RTQ-PCR) for each sample. Results HE staining: bone trabeculae and bone units were replaced by interrupted bone fragments, which were surrounded by many inflammatory granulation tissues and few osteocytes were seen in bone lacunae in the experimental group. In the control group, bone trabeculae and bone units were made by complete lamellar bones which surrounded blood vessels and osteocytes were seen in lacunae. RTQ-PCR testing: in the experimental group, OPG mRNA and RANKL mRNA were 1.35 ± 0.42 and 4.36 ± 1.35, respectively, while in the control group they were 1.78 ± 0.63 and 3.49 ± 1.02, respectively. The expression level of OPG mRNA in the experimental group was significantly lower than that in the control group, and the expression level of RANKL mRNA of the former was significantly higher than the latter. The OPG mRNA/ RANKL mRNA ratio in the xperiment group (0.34 ± 0.16) was significantly lower than that in the control group (0.54 ± 0.20), and there was significant difference (P lt; 0.05). Conclusion The glucocorticoid-induced ONFH may be related to the expression levels of OPG mRNA/RANKL mRNA in bone tissues.