AIM: To study p42/p44 mitogen - activated protein kinases ( MAPK ) signal transduction pathway effect on vascular endothelial growth factor ( VEGF ) expression induced by elevated glucose concentration in cultured human retinal pigment epithelium ( hRPE) .
METHODS:hRPE cells were cultured and divided into four groups:normal glucose group (NG) (5. 6mmol/L), high glucose group ( HG1:15mmol/L D-glucose, HG2:20mmol/L D - glucose, HG3:30mmol/L D - glucose ), PD98059 group: hRPE cells were treated by an efficient and selective inhibitor PD98059 (20μmol/L) of p42/p44MAPK signal transduction pathway and solvent dimethyl sulfoxide group ( DMSO group) . The expression of VEGF and pigment epithelium derived factor ( PEDF ) mRNA was detected by RT-PCR. VEGF protein expression in cultured hRPE supernatants was detected by enzyme-linked immumosorbent assay ( ELISA) .
RUSULTS: VEGF mRNA and protein expression induced by elevated glucose concentration increased significantly. VEGF mRNA and protein expression were restrained in PD98059 group. Ratio of ( VEGF/β-actine)/( PEDF/β - actine ) in PD98059 group decreased significantly compare with that in high glucose group.
CONCLUSION: p42/p44MAPK signal transduction pathway might play a part in VEGF expression induced by elevated glucose concentration in cultured hRPE cells.

Objective To understand the relationship between clinical manifestations and Kashin-Beck disease(KBD) and their contribution to diagnosis of KBD and to construct the diagnosis model for KBD in adolescents.Methods A total of 2248 subjects under the age of 18 were collected from 6 KBD endemic and 1 non-KBD areas of the Shaanxi province in China.Analysis of 32 indicators,including gender,age,and KBD clinical indicators.Indicators of the distribution of measurement data between the two groups using t test and analysis of variance,x2 test with count data,multi-category ordered response variables Logistic regression analysis for model building.Results It showed the KBD prevalence rate in adolescent had an increasing tendency with age.Analysis of indicators between the two groups,in addition to the age factor(P ＜ 0.05),the difference of ankle pain,knee pain,wrist movement disorder and other 5 indicators(P ＜ 0.05) and the last bend,elbow movement disorder,syndactyly and other 9 indicators(P ＜ 0.01 ) were statistically significant.Sixteen clinical and radiographic features in the clinical manifestations were significantly related with the clinical severity grading with KBD(P ＜ 0.01 ).Four models on the diagnostic indictors were constructed by cumulative logit model for adolescent KBD (-21ogL,Score,Wald x2 test,P ＜ 0.01 ).Conclusions The establishment of the diagnostic model based on their contribution of the joint involvement in systemic performance-related indicators has an important role for clinical diagnosis of KBD.

OBJECTIVETo control the quality of the product, quantitative fingerprint was used to evaluate the composition of the amino acids in the Xingnao Tongluo injection.

METHODThe method of the quantitative fingerprint to the amino acids composition was established through AccQ Tag precolumn derivatization. The quality was evaluated by the quantitative test of the amino acids and the similarity in ten batches.

RESULTThe Xingnao Tongluo injection contained 12 amino acids and the contents of these amino acids were stable. All the ten batches of the samples had similarity of more than 0.90.

CONCLUSIONThe method was accurate, feasible and could be a simple and effective way to evaluate the quality of the traditional Chinese medicine.

Amino Acids ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control

The research aimed to investigate the therapeutic effects and mechanisms of Opuntia dillenii Haw polysaccharide (OPS) on atherosclerosis of rats. First atherosclerotic rat models were established by high-fat and high-calcium diet. Thirty days later, the rats were treated with low dosage of OPS (0.2 g x kg(-1) x d(-1)) or high dosage of OPS (0.4 g x kg(-1) x d(-1)) by intraperitoneal injection for 60 days continuously. At the end of treatment, thoracic aorta rings were prepared and vasorelaxation of rat thoracic aorta in different experiment groups were determined by using 620M multi wire myograph system in vitro. Blood and livers of rats were collected. Then plasma levels of total cholesterol (TC), triglyceride (TG) and low density lipoprotein (LDL) of rats were separately determined using whole automatic biochemical analyzer; protein level of hepatic apolipoprotein B (ApoB) and that of hepatic diglyceride acyltransferase (Dgat1) were measured by Western Blot technique. Results showed that the ability of rat thoracic aorta to relax decreased markedly in the model group compared with that in the normal group, and significant differences existed in vasorelaxation ratios induced by different concentrations of carbamylcholine chloride (Carb) between these two groups (P < 0.01). After OPS treatment, the ability of rat thoracic aorta to relax improved markedly, the vasorelaxation ratios induced by Carb at 5 and 10 μmol x L(-1) were respectively 0.34 ± 0.08 and 0.62 ± 0.15 in the group treated with low dosage of OPS, while the ratios induced by Carb at 1 and 5 μmol x L(-1) were respectively 0.54 ± 0.08 and 0.98 ± 0.02 in the group treated with high dosage of OPS, which were all significantly different with those in the model group (P < 0.01). Plasma contents of TC, TG and LDL reduced significantly by the treatments both with low and high dosages of OPS compared with those in the model group (P < 0.01). Protein level of hepatic ApoB and that of hepatic Dgat1 decreased significantly after the treatment with high dosage of OPS compared with those in the model group (P < 0.01). These results indicate that OPS can markedly improve the vasorelaxation of thoracic aorta of atherosclerotic rats and has significant anti-atherosclerotic effect; inhibiting the expression of ApoB and Dgat1 and thus decreasing the amounts of TC, LDL and TG serving as one of the molecular mechanisms of its antiatherosclerosis effect.
Animals ; Aorta, Thoracic ; drug effects ; Atherosclerosis ; drug therapy ; Cholesterol ; blood ; Lipoproteins, LDL ; blood ; Opuntia ; chemistry ; Phytotherapy ; Rats ; Triglycerides ; blood

OBJECTIVETo observe the influence of Pilose antler polypeptide on the glycosaminoglycan and type II collagen in the articular cartilage in experimental knee osteoarthritis.

METHODSTotally 64 New Zealand white rabbits of 6 months old were randomly divided into 2 groups:normal group (n = 8) and model group (n = 56). Model group was surgically induced into osteoarthritis model by method of Hulth. After successful modeling, the rabbits of model group were further divided into 2 groups: Pilose antler polypeptide-treatment group and control group, 24 rabbits in each group. Pilose antler polypeptide-treatment group received 0.5 ml intra-articular injection of Pilose antler polypeptide dilution liquid once in per 2 days for 30 days, while control group received 0.5 ml intra-articular injection of physiological saline. On days 7, 15 and 30 after intervention, articular cartilage samples were collected respectively. The content of glycosaminoglycan in articular cartilage was observed by toluidine blue staining and the expression of type II collagen in cartilage matrix was detected by immunohistochemical staining.

RESULTSAlong with the prolonging of time, the content of glycosaminoglycan and type II collagen in cartilage matrix of the Pilose antler polypeptide-treatment group and control group decreased gradually. On days 7, 15 and 30 after intervention, integrated optical density of the type II collagen positive area in cartilage matrix of the Pilose antler polypeptide-treatment group were (312.06 +/- 14.12), (273.31 +/- 12.42) and (248.34 +/- 10.41), which had statistically significant differences. Integrated optical density of the type II collagen positive area in cartilage matrix of the control group were (253.47 +/- 15.53), (215.67 +/- 9.72) and (160.01 +/- 13.23), which had statistically significant differences. At the same period, integrated optical density of the type II collagen positive area in cartilage matrix of the Pilose antler polypeptide-treatment group was higher than that of control group, which had statistically significant difference.

CONCLUSIONPilose antler polypeptide can inhibit reduction of the glycosaminoglycan and type II collagen in cartilage matrix and delay the degeneration of articular cartilage.

Animals ; Antlers ; chemistry ; metabolism ; Collagen Type II ; metabolism ; Disease Models, Animal ; Female ; Glycosaminoglycans ; metabolism ; Humans ; Male ; Osteoarthritis, Knee ; drug therapy ; metabolism ; Peptides ; metabolism ; pharmacology ; Rabbits

Autopsy ; Female ; Humans ; Reye Syndrome ; pathology ; physiopathology ; Young Adult

Objective To explore the influence of vascular endothelial growth factor(VEGF) gene-transfected bone marrow mesenchymal stem cells (MSCs) on pulmonary alveolar structure and microvascular in rats with bronchopulmonary dysplasia.Methods Bone marrow MSCs were isolated from SD rats by way of density gradient centrifugation,purified,and transfected with pcDNA3.1 (-)/VEGF164 and blank plasmid pcDNA3.1 respectively using the liposome mediated method.After placing newborn SD rats in the oxygen box of 950 mL/L for 14 days,they were randomly divided into the transfected group(MSCs/VEGF group),the control group(MSCs group),and the blank group(serumfree medium group),with 10 rats in each group respectively,and they were injected respectively with 1 × 105 MSCs transfected by VEGF,MSCs and the same amount of simple serum-free medium by airway.After transplantation for 1 week and 4 weeks,lung tissue was observed by means of hematoxylin eosin staining to study lung structure and radial alveolar counts(RAC),and VEGF protein expression and angiogenesis densities were evaluated by immunohistochemistry,and finally VEGF164 protein was detected using Western blot.Results After transplantation for 1 week,4 weeks,the RAC,VEGF expression,vascular density by immunohistochemistry in transfected group were significantly more than those in the control group and the blank group(all P ＜0.05).After transplantation for 1 week,4 weeks,the VEGF164 protein level in transfected group was significantly more than that in the control group and the blank group (all P ＜0.05).Conclusions Eukaryotic expression vector pcDNA3.1 (-)/VEGF164 can effectively be expressed in MSCs.Transplantation of vascular endothelial growth factor gene by means of transfected MSCs brings better improvement in pulmonary alveolar structure and microvascular regeneration.VEGF is closely related to lung development in newborn rats.

Objective To investigate the clinical outcomes of patients undergoing microsurgery for cerebellar medulloblastoma. Methods This retrospective analysis of the clinical and follow-up data involves 57 patients who received microsurgery for pathologically confirmed cerebellar medulloblastorna, and the microsurgical techniques for medulloblastoma were discussed. Results Among the 57 patients, 42 had total tumor resection, 13 had subtotal and 2 had partial resection of the tumors. Patency of the midbrain aqueduct was achieved in all the cases after the surgery. Hydrocephalus was found in 43 patients before the operation and only in 16 patients after the operation. Tumor relapse occurred in 19 patients 2 years after the operation, including 8 with implantation metastasis compromising the central nervous system and 1 patient with frontal lobe metastasis who received reoperations. The earliest tumor relapse occurred 20 days after the surgery. The 2- and 5-year postoperative survival rates in these patients were 68.4% and 49.1%, respectively. Conclusion Good therapeutic effects can be achieved with total resection of the tumors and postoperative whole brain and spinal cord radiotherapy in patients with medulloblastomas.

OBJECTIVETo report a novel deafness-causing mutation c.465T>A, p.Y155X in connexin 26 (CX26) (also called gap junction protein beta-2, GJB2), and perform functional analysis of the mutated protein p.Y155X in Hela cells to explore the underlying mechanism on deafness.

METHODSMutations in CX26 gene of the proband in an autosomal recessive inherited deafness family were tested by direct DNA sequencing method. Mutant p.Y155X, which was found in the deafness family, and wild type CX26 (wtCX26), were directionally subcloned into the pEGFP-N1 plasmid to construct the recombinant fusion protein expression vector of CX26 p.Y155X-EGFP and wtCX26-EGFP, followed by transfecting into HeLa cells. The expression of the mutated and wild type proteins was analyzed using Western blot analysis. The intracellular localization of proteins and the formation of gap junction-like plaques at plasma membrane were observed under confocal microscope. Gap junction coupling was tested by calcein-AM dye transfer experiment.

RESULTSA novel nonsense mutation c.465T>A, p.Y155X in the CX26 gene was found in the autosomal recessive deafness family. The molecular weight of protein p.Y155X was smaller than that of wtCX26 in transiently expressed HeLa cells. The mutated protein failed to reach the cell surface to form gap junction plaques, and displayed cytoplasmic accumulation. Also, no calcein-AM dye was transferred from the donor cells to the recipient cells when both were transfected with CX26 p.Y155X. The wtCX26 protein localized at the cell membrane to form gap junction plaques with permeability to fluorescent dye calcein AM.

CONCLUSIONCX26 p.Y155X could not be targeted to the plasma membrane and there was no formation of gap junction channels between the adjacent cells. The mutation c.465T>A, p.Y155X in CX26 gene was responsible for the autosomal recessive hearing impairment in this family.

Amino Acid Sequence ; Child ; Codon, Nonsense ; genetics ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; Deafness ; genetics ; Female ; HeLa Cells ; Humans ; Male ; Molecular Sequence Data ; Pedigree ; Sequence Homology, Amino Acid

AIMTo investigate whether mitochondrial calcium uniporter participates in the cardioprotection of tumor necrosis factor alpha (TNFalpha) pretreatment in isolated rat hearts subjected to ischemia/reperfusion.

METHODSIsolated perfused rat hearts were subjected to 30 min regional ischemia (occlusion of left anterior descending artery) and 120 min reperfusion. The infarct size, coronary flow (CF) and lactate dehydrogenase (LDH) release during reperfusion were measured. The mitochondria of the heart were isolated and suspended in the swelling buffer for measurement of absorbance at 520 nm.

RESULTSPretreatment with TNFa at 10 U/ml for 7 min followed by 10 min washout reduced the infarct size and LDH release, and improved the recovery of CF during reperfusion. Administration of spermine (20 micromol/L), an opener of mitochondrial calcium uniporter, for 10 min during early reperfusion attenuated the reduction of infarct size and LDH release, and improvement of CF induced by TNFalpha. In isolated mitochondria of the heart pretreated with TNFalpha, the absorbance at 520 nm decreased less than that of mitochondria without TNFalpha pretreatment. Administration of spermine (50 micromol/L) attenuated the change of the absorbance induced by TNFalpha.

CONCLUSIONThe findings indicate that TNFalpha protects myocardium against ischemia/reperfusion injury via inhibiting mitochondrial calcium uniporter opening as well as mitochondrial permeability transition pore opening.

Animals ; Calcium Channels ; drug effects ; metabolism ; Cardiotonic Agents ; pharmacology ; In Vitro Techniques ; Ischemic Preconditioning, Myocardial ; methods ; Male ; Mitochondrial Membrane Transport Proteins ; drug effects ; Myocardial Reperfusion Injury ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Spermine ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology