OBJECTIVE: To study the effect of simvastatin on the expression of bone morphogenetic protein-2 (BMP-2) and alkaline phosphates (ALP) activity in the primary cultured bone marrow stromal cells, and to elucidate the mechanism of the anabolic osteogenetic effect of simvastatin. METHODS: Bone marrow stromal cells in femur and tibia of adult mouse were cultured in vitro. after treated with different concentrations of simvastatin (0, 0.1, 0.2, 0.5 and 1.0 mumol/L) or recombinant human BMP-2 for 72 hours, ALP activity of bone marrow stromal cells was determined. BMP-2 expression of bone marrow stromal cells was analyzed by using immunocytochemistry and Western blotting. RESULTS: After treated with simvastatin for 72 hours, BMP-2 expression increased, while little BMP-2 expression could be observed in the control group. ALP activity also increased in a dose-dependent manner; t-test showed that ALP activity in the group which concentrations of simvastatin were 0.5 mumol/L (t = 2.35, P = 0.041), 1.0 mumol/L (t = 2.348, P = 0.041) had significant difference when compared with control group. CONCLUSION: Simvastatin lead to high expression of BMP-2 in bone marrow stromal cells, via the increased auto- or para-crine of BMP-2, and ALP activity increased. These may be parts of the mechanism on the anabolic osteogenetic effect of simvastatin.
Objective To evaluate the effect of composite (bFGF/PDPB) of basic fibroblast growth factor（bFGF） and partially deproteinized bone (PDPB) on the repair of femoral head defect. Methods Forty-eight femoral heads with defect derived from 24 New Zealand rabbits were divided into 3 groups at random, which were implanted with bFGF/PDPB(group A), PDPB(group B) and nothing(group C) respectively.The rabbits were sacrificed at 2,4,and8 weeks after operation, and then the femoral heads were obtained. The specimens injected with Chinese ink were created. Then X-ray examination, histopathological and morphological examination of blood vessel, and image analysis were made. Results The bone defects healed completely 8 weeks after operation in group A. The implants in the repaired tissue were not substituted completely in group B. The bone defects did not heal completely in group C. Two weeks after operation, affluent newly formed vessels were seen in repaired areas in groupA. No significant difference between group A and group B was observed 8 weeks after operation. In group C, newly formed vessels were scarce 2, 4, and 8 weeks after operation. There were 3 sides rated excellent, 2 good and 1 fair in group A; 1 excellent, 2 good, 2 fair and 1 poor in group B; and 1 fair and 5 poor in group C according to the X-ray evaluation 8 weeks after operation. Eight weeks after operation, the volume fraction of bone trabecula in repaired tissue was higher in group A than that in group B (Plt;0.05), and the fraction in group C was thelowest among the 3 groups (Plt;0.05). Conclusion The composite ofbFGF and PDPB can effectively promote the repair of femoral head defect of rabbit.
Objective To study the relationship between metalloproteinases (MMPs) and breast cancer. Methods The literature in recent years on the relationship between the expression of MMPs and breast cancer was reviewed. Results The balance between MMPs and tissue inhibitors of metalloproteinases (TIMPs) is keeping normally kept in human body. Many of the studies showed that the expression of MMPs is increased in breast cancer. Conclusion The growth, invasion and metastasis of breast cancer is closely related with the increased expression of MMPs. This suggests that MMPs is a valuable prognostic marker and TIMPs would be a novel drug against cancer.
【Abstract】ObjectiveTo explore the effects of RECK gene on the biological behaviors of hepatocellular (HepG2). MethodsThe RECK cDNA was transfected to HepG2 with lipofectamine 2000. Detected its protein expressions with Western blot before and after transfection, analyzed the effects of RECK on MMP-9 activity using gelatin zymography, observed the effects on proliferation ability by MTT assay and plate clone formation assay, compare the changes of invasion ability by cell adhesion assay and in vitro invasion experiment. Results RECK protein was expressed steadily in transfected HepG2 cells and the amount of activated MMP-9 were decreased significantly. Their proliferation abilities weren’t different before and after transfection but their invasion abilities decreased sharply. ConclusionRECK gene can transfect HepG2 cells by liposome efficiently. It can inhibit the activity of MMP-9 and the invasion ability of HepG2.
Objective To investigate the effect of hepatitis C virus (HCV) F protein on proliferation and collagen expression of hepatic stellate cells. Methods After pcDNA3.1-f plasmid containing HCV f gene or empty pcDNA3.1 plasmid was transfected hepatic stellate cells LX2 by liposome, LX-f or LX-p cells were obtained by G418 screening. The proliferation of LX-f or LX-p cells was analyzed by MTT, and the contents of collagen type Ⅰand Ⅲ secreted by LX-f or LX-p cells were detected by ELISA. Results After 24 h cultivation, the proliferation rate of LX-f cells was higher than that of LX-p cells at each time point （Plt;0.01）. After 48 h cultivation, the contents of collagen typeⅠand Ⅲ secreted by LX-f were (25.89±0.42) ng/ml and (18.21±0.49) ng/ml, which was significantly higher than those of LX-p cells 〔(22.65±0.49) ng/ml and (15.29±0.62) ng/ml〕, Plt;0.01. Conclusion HCV F protein is able to promote proliferation of hepatic stellate cells, and up-regulate the excretion of collagen type Ⅰand Ⅲ in those cells, which induces hepatic fibrosis.
Objective To investigate the expressions of monocyte chemoattractant protein-1 (MCP-1) and vascular endothelial growth factor (VEGF) in gastric cancer tissues and normal gastirc mucosa tissues and the situation of helicobacter pylori (HP) infection, and detect their relationships and clinicopathologic significances. Methods Expressions of MCP-1 and VEGF were detected by immunohistochemistry in gastric cancer tissues and normal gastric mucosa tissues (5-10 cm from the mass), and HP was detected in specimen from gastric antrum by Giemsa dyeing method. Results MCP-1 and VEGF expressions in gastric cancer tissues were significantly higher than those in normal gastric mucosa tissues (P＜0.05), but there was no difference in HP positive and negative tissues included the cancer and the normal tissues （P＞0.05）. The expressions of MCP-1 and VEGF in carcinoma with tumordiameter ＞5 cm, poorly differentiated， lymph node metastasis， distant metastasis and Ⅲ+Ⅳ stage of TNM were significantly higher than those with tumor diameter ≤5 cm, well and moderately differentiated， non-lymph node metastasis， non-distant metastasis and Ⅰ+Ⅱ stage of TNM (P＜0.05). Conclusion The high expressions of MCP-1 and VEGF in gastric cancer may relate to tumor angiogenesis and metastasis, but HP infection may be irrelevant.
ObjectiveTo investigate the relationship between lipoprotein-associated phospholipase A2 (Lp-PLA2) level and in-hospital prognosis in patients with acute type A aortic dissection within 24 hours of admission.MethodsFortysix patients diagnosed with type A aortic dissection were included in our hospital and their Lp-PLA2 levels within 24 hours of admission were measured between January 2017 and June 2019. According to their Lp-PLA2 levels within 24 hours of admission, 23 patients were classified into a high Lp-PLA2 group (Lp-PLA2 > 200 μg/L, 16 males and 7 females at age of 52.0±14.0 years) and 23 patients were into a low Lp-PLA2 group (Lp-PLA2 ≤200 μg/L, 15 males and 8 females at age of 53.0±11.0 years). The relationship between Lp-PLA2 level and clinical outcome was analyzed.ResultsThe incidences of bleeding, hospital infection, multiple organ dysfunction and mortality in the high Lp-PLA2 group were higher than those in the low Lp-PLA2 group (P<0.05). Seven (15.2%) patients died during 3 months of follow-up. The 3-month survival rate of patients with an increase of Lp-PLA2 was significantly lower than that of the patients with normal Lp-PLA2 (P<0.01), which was an independent predictor of adverse outcomes at 3 months of onset (P<0.01).ConclusionLp-PLA2 may be a predictor of disease progression in the patients with acute type A aortic dissection, and the patients with significantly elevated Lp-PLA2 have a higher 3-month mortality than the patients with normal Lp-PLA2.
ObjectiveTo investigate the expressions of type Ⅰ and type Ⅲ collagen protein in hepatic alveolar echinococcosis tissues, and to explore its relationship with the biological behavior in progress of hepatic alveolar echinococcosis (HAE). MethodsTwenty samples of normal liver tissues and liver tissues at the edge of the lesion with HAE in our hospital from Jan. 2012 to Dec. 2014 were collected, and HE and Masson staining were performed. The pathological changes and the degree of fibrosis of liver tissues around HAE lesion were observed under light microscope. The expressions of type Ⅰ and type Ⅲ collagen protein in liver tissues were detected by immunohistochemical staining. ResultsThe degree of liver fibrosis of liver tissues at the edge of the lesion with HAE was grade Ⅱ, and the degree of fibrosis of normal liver tissues was grade 0, the difference between the two was statistically significant (P < 0.05). The color index of type Ⅰand type Ⅲ collagen protein in the liver tissues at the edge of the lesion with HAE was 7.45±1.85 and 8.00±1.62, respectively, which were higher than those of normal liver tissues (3.10±1.02 and 3.50±0.89), the difference were statistically significant (t=-9.21, P=0.001;t=-10.88, P=0.001). ConclusionsThere is liver fibrosis around the lesion in the patients with HAE. HAE may promote the expressions of type Ⅰ and type Ⅲ collagen and then induce the occurrence of liver fibrosis.
Objective To construct a replicationdefective recombinant adinovirus including the target gene human bone morphogenetic protein 4(fragment hBMP-4). Methods The hBMP-4 gene fragment was cut down from pCS2(+)/hBMP-4, cloned into the eukaryotic expressive vector pcDNA 3.1(+), then subcloned into pShuttle-CMV and transformed into the competent E. coli BJ5183/p by electroporation. The resulting recombinant plasmid pAdE/hBMP-4 was transformed into the packaging of thecell lines HEK293 to produce the replication-defective recombinant adenovirusescontaining the hBMP-4 gene. These replication-defective recombinant adinoviruses were transfected into HEK293 and HeLa cells. Then, total RNA and total protein were detected by RT-PCR and the Western-blot assay. Results The pAdE/hBMP-4 was confirmed by the restrictional endonuclease digestion. In HEK293 and HeLa cells, the specific transcription of the hBMP-4 gene was confirmed by RT-PCR, and the expression of the hBMP-4 protein was confirmed by theWestern-blot assay. Conclusion The replication-defective recombinant adinovirus expression vector containing the hBMP-4 gene can be constructed and expressed successfully, which has laid a foundation for the further research on the genetherapy of hBMP-4.
Objective To observe the expression and relationship of high-mobility group A(HMGA)1, HMGA2, MIB-1 labeling index (LI) and let-7 in retinoblastoma (RB). Methods Forty-four RB samples were studied, including 11 poorly-differentiated samples, 33 well-differentiated samples; eight invasive and 36 non-invasive samples. The expression of HMGA1, HMGA2 and MIB-1 LI in RB were analyzed by immunohistochemitry. The HMGA1, HMGA2 were scored on a scale of 0 to high expression. 0: no expression; low: 1%-10%; medium: 11%-50%; high: ＞50%. The MIB LI were scored on a scale of 0 to high expression. 0: no expression; low: 1%-40%; high: ＞40%. Semiquantitative reverse transcription-polymerase chain reaction was used to assay the let-7 expression level: ge;80% showed no significantly decreased expression; 60%-79% showed medium decrease in expression; ＜60% highly decreased in expression. ResultsIn 44 RB samples, there were 14 cases with no HMGA1 expression (32%), 11 cases with low expression (25%), 10 cases with medium expression (23%), and nine cases with high expression (20%). Expression level of HMGA1 was significantly higher in poorly differentiated RB than in well-differentiated RB (chi;2=11.3,P＜0.01); however, no statistically significant difference was found between invasive tumors and noninvasive tumors (chi;2=5.9,P＞0.05). There were 11 cases with no HMGA2 expression (25%), 11 cases with low expression (25%), nine cases with medium expression (20%), and 13 cases with high expression (30%). Expression level of HMGA2 was significantly higher in poorly differentiated and invasive RB than in well-differentiated and noninvasive RB respectively (chi;2=20.9, 8.7;P＜0.05). There were 4 cases with no MIB-1 LI expression (9%), 18 cases with low expression (41%), and 22 cases with high expression (50%). Expression level of MIB-1 LI was significantly higher in poorly differentiated RB than in well-differentiated RB (t=5.2,P＜0.05). Higher expression of MIB-1 LI was found in invasive tumors than in noninvasive tumors, with no significant difference (t=-1.1,P＞0.05). Twenty-seven cases had no significantly decreased expression of let-7 (61%). There were eight cases with medium decreased expression (18%) and nine cases with highly decreased expression (21%). Correlation analyses revealed that MIB-1 LI expression significantly correlated with HMGA1and HMGA2 proteins (r=0.327, 0.602;P＜0.05). A significantly inverse correlation existed between let-7 expression and HMGA1, HMGA2 proteins and MIB-1 LI respectively (r=-0.247,-0.310,-0.392;P＜0.05). Conclusions Overexpression of HMGA1, HMGA2 and MIB-1 LI and down regulation of let-7 were demonstrated in RB. Supplying let-7 to RB cells can possibly inhibit HMGA1 and HMGA2 expression.