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.

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.

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

BACKGROUND:Ethyl linoleate has been proved to attenuate the inflammatory-cytokines release induced by lipopolysaccharide, but whether it can inhibit titanium-induced osteolysis and the underlying mechanism remain unclear. OBJECTIVE:To observe the effect of ethyl linoleate on the expression of inflammatory-related factors induced by titanium particles and explore its mechanism. METHODS:Forty-eight Kunming mice were randomly divided into blank control, titanium, dimethylsulfoxide (DMSO) and experimental groups. The back air pouch Inflammatory models were established in the mice of the titanium, DMSO and experimental groups, in which the 200 μL menstruum of DMSO (0.5%) and 200 μL ethyl linoleate (0.5%) were respectively administered into the pouch of the mice at 12 hours. Mice in the blank control group received no intervention. Fourteen days later, the inflammatory cel infiltration in the skin was examined through hematoxylin-eosin staining;the expression levels of inhibitorκB-α, nuclear factor-κB, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor-αand interleukin-6 as wel as ERK, p-ERK, JNK, p-JNK, p38 and p-p38 in MAPK signaling pathways were evaluated by western blot assay. RESULTS AND CONCLUSION:In the titanium group and DMSO group, there were numerous inflammatory cel s and vacuole-like necrotic tissues in the hair fol icle lacuna of dermis and loose connective tissues of hypodermis. The experiment group showed significant reduction in inflammatory cel infiltration and vacuole-like necrosis. Compared with the blank control group, the expression levels of inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-κB, tumor necrosis factor-α, interleukin-6, ERK, JNK and p38 in the DMSO and titanium groups were significantly increased, while inhibitorκB-αsignificantly decreased (P<0.05). Compared with the DMSO and titanium groups, there were significantly down-regulated levels of inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-κB, tumor necrosis factor-α, interleukin-6, ERK, JNK and p38, and up-regulated inhibitorκB-αlevel in the experimental group (P<0.05). In conclusion, ethyl linoleate can remarkably suppress the expressions of titanium-induced inflammatory factors associated with the inhibition of nuclear factor-κB and MAPK signaling pathway activation.

Objective The aim of the present study is to evaluate the reliability and validity of the Mandarin version of the PP (MPP) .Methods The first step in the establishment was to translate the original English version into mandarin version with the method of cross -culture translation .The reliability was performed with the internal consistency analysis and test -retest reliability .The validity was performed for the content validity and structure va-lidity .The samples were from 80 Chinese CI children ,and 43 parents answered this questionnaire again 1 month lat-er to evaluate the test -retest reliability .The average age at cochlear implantation were 26 ± 14 months ,ranging from 7 months to 68 months ,the average duration of CI use were 10 ± 7 months ,ranging from 0 month to 24 months .Results The reliability analysis indicates that the Cronbach'sαcoefficient was 0 .797 ,except for the well-being and happiness ,education ,whose coefficients are respectively 0 .303 ,and 0 .341 ,all of the other sundomainscoefficient were greater than 0 .5 ,indicating the internal consistency was good .Test -retest reliability of the scale Cronbach'sαwas satisfactory .All subdomains and total score of the scale coefficients were greater than 0 .70(P<0 .01) .The validity analysis indicated that the pearson correlation coefficients among the total scale and the 8 subdo-mains were 0 .395~0 .992 ,the correlation coefficients among each subdomains were 0 .09~0 .654 ,which confirmed with the psychological characteristics ,proving its good structure validity .Conclusion The Chinese version of the PP show s good reliability and validity and can be used to evaluation the quality of life in mandarin CI children.

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.