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

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

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

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

Objective The immunotoxins generated by conjugating monoclonal antibody (mAb) and a certain toxin play an important and promising role in treating hematopoietic malignancies. However, most of the toxins used for the conjugation are toxic proteins, which are immunogenic in the patients. Norcantharidin (NCTD) is a small molecule toxin without immunogenicity, and thus has become a potential new drug for hematopoietic cancers. In this study, we prepared immunotoxin 2E8-NCTD by using the ZCH-4-2E8 cells produced in the laboratory of our hospital, and then detected its targeting effect against CD+19 lymphoid malignant Nalm-6 cells in vitro.Methods 2E8 mAb was obtained from mouse ascites and purified by gel chromatography. After its purity was checked by SDS-PAGE, immunotoxin 2E8-NCTD was generated by conjugating CD19 mAb with NCTD using activated ester method. The binding activity of the immunoconjugate to CD19 antigens on cell surface, and the expression levels of the CD19 antigens on Nalm-6 and K562 cells were determined by flow cytometry. The inhibitory effects of PBS, purified 2E8 mAb, NCTD, and immunotoxin 2E8-NCTD on the cell growth of either Nalm-6 or K562 cells were then compared.Results The purity of the 2E8 mAb was higher than 99% demonstrated by SDS-PAGE assay. 2E8 mAb was detected on the surface of 99.34% of the Nalm-6 cells, while on only 0.98% of the K562. The newly generated immunotoxin had a positive rate of 99.90% on the Nalm-6 with slightly reduced binding activity. Both 2E8-NCTD and NCTD significantly inhibited the growth of CD+19 Nalm-6 cells (P < 0. 001 ), while the purified 2E8 mAb did not show any significant influences on the growth of the same cell line ( P > 0.05 ). Meanwhile, no significant inhibitory effects on the CD-19 K562 cells were identified in the 2E8-NCTD, 2E8 mAb, or control groups, indicating a significant targeting effect of 2E8-NCTD against Nalm-6 cells.Conclusions The immunotoxin 2E8-NCTD can be synthesized by activated ester method. It has target killing effects on CD+19 Nalm-6 leukemia cells in vitro.

Aim: To study theapoptotic rate (AR) and the androgen and estrogen milieu in the proximal and distal ductal sys tems of prostate, in order to help exploring the effects of these factors on prostatic growth and the pathogenesis of be nign prostatic hypertrophy (BPH). Methods: The proximal and distal ends of the ductal system were incised from 20 normal prostate as well as the hypertrophic prostate tissue from 20 patients with BPH. The AR was determined by the DNA end-labeling method and dihydrotestosterone (DHT) and estrodiol (E2), by radioimmunoassay. Results:There was no significant difference in DHT and E2 density between the proximal and distal ends of the ductal systems in normal prostate. E2 appeared to be higher in BPH than in normal prostatic tissues, but the difference was statistically in significant. In normal prostatic tissue, the AR was significantly higher in the distal than in the proximal ends of the ductal system ( P ＜ 0.05), while the AR of the proximal ends was significantly higher ( P ＜ 0.01) than that in the BPH tissue. No significant correlation was noted between the DHT and E2 density and the AR both in the normal prostate and BPH tissues. Conclusion: The paper is the first time describing a difference in AR in different regions of the ductal system of normal prostate, while the hormonal milieu is similar, indicating a functional inhomogeneity of these regions. A low AR in the proximal duct, where BPH originates, and an even lower AR in the BPH tissue, sug gesting the participation of apoptosis in the BPH pathogenesis.

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 effects of wrist restrain gloves in elderly patients with restlessness on their unplanned extubation.Methods Totals of 80 cases of elderly patients with restlessness were divided into control group and observation group,40 cases in each group.And traditional wrist restrains were used to the patients in the control group,while the wrist restrain gloves (also know as wave plate) were applied in the observation group.The frequency of unplanned extubation and the effects of restrain were compared between two groups.Results It showed that the incidence of unplanned extubation in observation group was significantly lower than that in control group (5％ vs 45％),there was statistical difference between two groups (x2 =8.273,P ＜0.05).And patients' satisfaction of observation group was significantly higher than that of control group (100％ vs 30％),there was statistical difference between two groups (x2 =9.652,P ＜ 0.05).Conclusions Application of wrist restrain gloves could reduce the incidence of unplanned extubation in elderly patients with restlessness,and improve their satisfaction.