BACKGROUND: Cytokine abnormality, nerve function abnormality and hormone levels may contribute to osteoporosis occurrence following spinal cord injury (SCI), many papers are about cytokine and hormone, but fewer is related to abnormal nerve function on bone accommodation.OBJECTIVE: To innovatively apply blood biochemistry and immunohistochemistry on the observation of change of calcitonin gene-related peptide distributing in trabecular bone of SCI rats, and to analyze its significance in the osteoporosis. DESIGN, TIME AND SETTING: Randomized control animal trials were performed from September 2008 to December 2008 at the laboratory of Orthopedic Institute in the Fourth Military Medical University of Chinese PLA,MATERIALS: Forty-eight Sprague Dawley rats aged 3 months, weighing (210:1:16) g, were divided into SCI group and control group equally. METHODS: Rats in the SCI group underwent spinal cord transection at the tenth thoracic vertebrae. Control rats underwent laminectomy without any spinal cord lesion. MAIN OUTCOME MEASURES: Each 8 rats were sacrificed at 1, 3, 6 weeks postoperatively. The serum concentrations of bone-specific alkaline phosphotase and serum cross-linked N-telopeptide of type Ⅰ collagen were determined. The stain intensity of calcitonin gene-related peptide in trabacular bone was determined with use of quantitative immunohistochemistry technique and computer image analysis system.RESULTS: The cross-linked N-telopeptida of type Ⅰ collagen significantly increased in SCI group at defferent interval compared with control group (P <0.05 or 0.01 ), serum concentrations of bone-specific alkaline phosphotase was lower than control group,without significant difference (P > 0.05). Immunoreactivity of calcitonin gene-related peptide in trabecular bone was strongly positive in control group, while weakened in SCI group (P < 0.05 or 0.01).CONCLUSION: Reduced calcitonin gene-related peptide in trabecular bone of SCI rats may be related to the occurrence of osteoporosis following SCI.

Objective To investigate the effects of substance P on anoxia/reoxygenation (A/R) injury to cardiomyocytes of neonatal rats.Methods Cardiomyocytes of neonatal rats were isolated from Sprague-Dawley rats,aged 1-3 days,and were cultured in 6-well plates for 72 h.The cardiomyocytes were then assigned into 4 groups (n =3 each) using a random number table:control group (group C),A/R group,substance P group (group SP) and substance P + D-SP group (group SP + D-SP).The cells were cultured routinely for 6 h in group C and the cells were exposed to 99.9 ％ N2 in an incubator at 37 ℃ for 3 h followed by reoxygenation for 2 h in the other groups.The cells were incubated with substance P with the final concentration of 10-7 mol/L for 1 h before anoxia in group SP.The cells were incubated for 1 h with substance P with the final concentration of 10-7 mol/L and D-SP (a specific antagonist of neurokinin-1 receptor) with the final concentration of 10-6 mol/L before anoxia in group SP + D-SP.The apoptosis rate and lactate dehydrogenase (LDH) activity were detected at the end of reoxygenation using TUNEL assay and LDH assay kit,respectively.Results Compared with C group,the apoptosis rate and LDH activity were significantly increased in A/R,SP and SP+ D-SP groups.Compared with A/R group,the apoptosis rate and LDH activity were significantly decreased in SP and SP + D-SP groups.Compared with SP group,the apoptosis rate and LDH activity were significantly increased in SP + D-SP group.Conclusion Substance P can attenuate A/R injury to cardiomyocytes of neonatal rats,and activation of neurokinin 1 receptors is involved in the mechanism.

Objective To evaluate the effect of calcitonin gene-related peptide (CGRP) on anoxiareoxygenation (A/R)-induced injury to neonatal rat cardiomyocytes incubated in high glucose medium.Methods Cardiomyocytes were obtained from 1-3-day old Sprague-Dawley rats and cultured in the culture medium containing 15％ bovine calf serum and then seeded onto 6-well plates at a density of 10 × 105/ml (3 ml/well).The cells were randomly divided into 5 groups (n =9 each):normal glucose medium control group (NG group),high glucose medium group (HG group),high glucose medium + A/R group (HG+ A/R group),high glucose medium + A/R + CGRP group (HG + A/R + CGRP group),and high glucose medium + A/R + CGRP+ CGRP8-37 group (HG + A/R + CGRP + CGRP8-37 group).The cells were incubated in normal glucose (5.5 mmol/L) medium for 72 h in NG group.In HG group,the cells were incubated in high glucose (25.0 mmol/L) medium for 72 h.In HG + A/ R group,the cells were incubated in high glucose medium for 72 h and then exposed to 3 h of anoxia followed by 2 h of reoxygenation.In group HG + A/R + CGRP,the cells were incubated in high glucose medium for 72 h,CGRP (final concentration 10-8 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.In HG + A/R + CGRP + CGRP8-37 group,the cells were incubated in high glucose medium for 72 h,CGRP (final concentration 10 8 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.In HG + A/R + CGRP + CGRP8-37 group,the cells were incubated in high glucose medium for 72 h,CGRP8-37 (final concentration 10-8 mol/L) and CGRP8-37 (CGRP receptor antagonist,final concentration 10-7 mol/L) was then added to the culture media and the cells were incubated for 1 h and then underwent A/R.Apoptosis in cardiomyocytes was detected using TUNEL and apoptosis index (AI) was calculated.The lactate dehydrogenase (LDH) activity in the culture medium was analyzed.Results AI and LDH activity were significantly higher in HG group than in NG group,and in HG + A/R group than in HG group.Compared with HG + A/R group,AI and LDH activity were significantly decreased in HG + A/R + CGRP group,while no significant changes were found in HG + A/R + CGRP + CGRP8-37 group.Compared with HG + A/R + CGRP group,AI and LDH activity were significantly increased in HG + A/R + CGRP + CGRP8-37 group.Conclusion CGRP attenuates A/R-induced injury to neonatal rat cardiomyocytes incubated in high glucose medium via combing with CGRP receptor.

Age of Onset ; Cystadenocarcinoma, Serous ; genetics ; metabolism ; pathology ; Female ; Genes, BRCA1 ; Genes, BRCA2 ; Humans ; Immunohistochemistry ; Neoplasm Staging ; Ovarian Neoplasms ; genetics ; metabolism ; pathology ; Receptors, Progesterone ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Adenocarcinoma, Clear Cell ; metabolism ; pathology ; Adenocarcinoma, Mucinous ; metabolism ; pathology ; Carcinoma, Endometrioid ; metabolism ; pathology ; Carcinoma, Transitional Cell ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Humans ; Neoplasms, Glandular and Epithelial ; classification ; metabolism ; pathology ; Ovarian Neoplasms ; classification ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; metabolism ; WT1 Proteins ; metabolism

Objectives: To investigate the effects of adhesion and proliferation of bone mesenchymal stem cells (BMSCs) in the surface of lactic acid/glycolic acid/asparagic acid-co-polyethylene glycol PLGA-[ASP-PEG] tri-block polymer scaffolds, try to find a new biomaterial to induce seed cells in vitro for bone tissue engineering. Methods: Modified PLGA with polyethylene glycol (PEG) and asparagic acid (ASP) that has many ligands, and synthesis PLGA-[ASP-PEG] polymer material. BMSCs were cultured in PLGA-[ASP-PEG] polymer material and PLGA used as control group. Through precipitation method, MTT assay and total cellular protein detection to test the adhersion and proliferation of BMSCs. Scanning electron microscope is used to observe cells appearance. Results: BMSCs on the surface of PLGA-[ASP-PEG] polymer scaffolds are adherention to the culture flask, the number of cells is much higher than PLGA’s. The precipitation method suggest that adhesion and proliferation of BMSCs on the surface of PLGA-[ASP-PEG] is much higher than the control group(P

Carcinoma in Situ ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Fallopian Tube Neoplasms ; metabolism ; pathology ; Fallopian Tubes ; metabolism ; pathology ; Female ; Humans ; Neoplasm Invasiveness ; Ovary ; pathology ; Tumor Suppressor Protein p53 ; metabolism

Objective To construct wtp53/junB fusion gene and its eukaryotic expression vector in order to provide the basis for further application of polygene union therapy in hepatocellular carcinoma. Methods Polymerase chain reaction (PCR), reverse transcription-PCR (RT-PCR) and gene recombination techniques were used to construct the eukaryotic vector of pEGFP-C1-wtp53/junB fusion gene, which carries the enhanced green fluorescent protein (EGFP). The transfection of pEGFP-C1-wtp53/junB in hepatoma HepG2 cells was detected by the location of green fluorescence. Results The DNA sequence of wtp53/junB fusion gene was successfully cloned into the pEGFP-C1 plasmid and the sequence was the same as what we expected. Green fluorescence located on cell nucleus proved that pEGFP-C1-wtp53/junB was transfected into HepG2 cell line successfully. Conclusion We successfully constructed the eukaryotic vector of pEGFP-C1-wtp53/junB fusion gene, which carries the EGFP, and transfects it into human hepatoma cell nucleus. It may lay the basis for studying the synergetic effect of wtp53 and junB in hepatocellular carcinoma.

Objective To explore the effects of surface modification of PLGA-[ASP-PEG] scaffold with typeⅠcollagen on the adhesion,proliferation and osteogenic differentiation of rabbit bone marrow stromal cells (BMSCs).Methods After PLGA-[ASP-PEG] materials were modified with typeⅠcollagen chemically,the collagen was coated onto the materials physically.The BMSCs obtained from rabbits were cultured on the modified PLGA-[ ASP-PEG] and on the unmodified PLGA-[ ASP-PEG] as control.The adhesion and proliferation behavior of the cells was analyzed and the expressions of osteogenie marker alkaline phosphatase,osteocalcin,osteopontin,typeⅠcollagen and core binding factor al were also detected.Results X-ray photoelectron spectrometry(XPS) confirmed that TypeⅠcollagen was grafted onto the surface of PLGA-[ASP-PEG] successfully and the collagen content on the materials modified chemically and physically was significantly increased.The abilities of adhesion and proliferation and the expressions of osteogenie makers of the BMSCs were significantly greater than those in the control group(P＜0.05).Conclusion Since Type collagen I can improve the biocompatibility of PLGA- [ASP-PEG] scaffold materials,it can be used as a new way to optimize scaffolds in tissue engineering.