BACKGROUND:Articular cartilage regeneration can be regulatedbyautocrineorparacrinesecretionof various cytokines.
OBJECTIVE:To analyze biological properties of bone morphogenetic proteins and basic fibroblast growth factor in biological materials for repair of articular cartilage defect.
METHODS:Forty New Zealand white rabbits were used and equaly randomized intofourgroups: fibrin, basic fibroblast growth factor, bone morphogenetic protein, and combined treatment (basic fibroblast growth factor combined with bone morphogenetic protein) groups, respectively.Bioactivescaffolds with fibrin, basic fibroblast growth factor,bone morphogenetic protein, and basic fibroblast growth factor combined with bone morphogenetic protein were injected to repair the articular cartilage defect. Therapeutic effect andbiological properties of biological materials were compared.
RESULTS AND CONCLUSION:(1) Inthefibrin group,tworabbits appearedto havelimps. Inthebasic fibroblast growth factor group hand functionwaslimited inonerabbit. Inthebone morphogenetic protein group, one had a limpandonewasin a limitation of activity. Inthecombined treatment group,rabbitsrecovered wel andshowedno differencesintheknee joint before and aftersurgery (P< 0.05). (2) General observation: Inthe combined treatment group, soft solution cartilage defects disappeared,andangiogenesis and cartilage were similar with normal tissues. Inthebone morphogenetic protein group, fractured cartilage marginal existedand could not beclosely integrated with normal cartilage. The presence of chondrocytesin the periphery of the defectwas seen under light microscope. Inthefibrin group, defect site and surrounding tissues healed. Inthe basic fibroblast growth factor group, defect was repaired, but not smooth. (3) Results ofhematoxylin and eosin staining: Inthefibrin group, the bone defect was not repaired, obvious depression surface was seen. In basic fibroblast growth factor group, repair of cartilage defect was obvious. There were a lot ofchondrocytes. Inthe bone morphogenetic protein group, the bone defect was repaired; chondrocytes appeared, but irregular arrangement. Inthecombined treatment group, good bone defect repair and a large number of cartilage cels were seen. Taken together, biological materials with fibrin and basic fibroblast growth factor are ideal for repair of articular cartilage defect by promoting formation of cartilage by bone morphogenetic protein and enhancing chondrocyte proliferation by basic fibroblast growth factor.

OBJECTIVE:To investigate the effects of Ossotide tabletcombined with Xianling gubao capsule on the postoperative recovery effect oftrans-traumatic-verterbrae screw-setting combined with bone grafting in the treatment of osteoporotic thoracolum-bar vertebral fractures. METHODS:122 patients with osteoporotic thoracolumbar vertebral fractures who were treated with trans-traumatic-verterbrae screw-setting combined with bone grafting were randomly divided into control group and observation group.The two groups were treated by trans-traumatic-verterbrae screw-setting with bone grafting.Based on this,the control group was treated with 0.3-0.6 g Ossotide tablet,three times a day,taken orally after meals;observation group was additionally treated with 1.5 g Xianling gubao capsule,twice a day,taken orally after meals. Both of the two groups took medicine on the day of oper-ation. A course induded 4 weeks,and it lasted 4 courses. The indexes were observed after 3 and 6 months treatment. The changes of pain score,bone mineral density(BMD)levels,bone gla protein(BGP),bone alkali phsphatase(BALP),procollagen I ntermi-nal peptide (PINP) and other bone metabolism biochemical indexes in 2 groups before and after treatment were observed.Mean-while,the clinical efficacy,rates of thoracolumbar vertebral refractures and incidence of adverse reactions in the 2 groups were re-corded.RESULTS:After treatment,the total effective rate in observation group was significantly higher than control group,pain scores in 2 groups were significantly lower than before,and general decreased by time,observation group was lower than control group;BMD,BGP,BALP and PINP levels in 2 groups were significantly higher than before,and general increased by time,ob-servation group was higher than control group;after 6 months,BMD,BALP and PINP levels in control group were significantly higher than before,the differences were statistically significant(P<0.05);there were no significant differences in the incidence of adverse reactions,rates of thoracolumbar vertebral refractures between 2 groups(P>0.05). CONCLUSIONS:Ossotide tablet com-bined with Xianling gubao capsule can significantly improve the bone metabolism biochemical indexes of patients,promote bone mineral density and fracture healing,with good safety in short-term medication.

Objective: To study the compatibility and therapeutical basis of composite herbal medicines of Lingguishugan Decoction(LD)(Poria, Ramulus Cinnamomi, Rhizoma Atractylodis Macrocephalae, Radix Glycyrrhizae.)Methods: Ethanol extract test solutions of the different combinations were prepared according to the orthogonal layout L 16 (4 5). Pharmacologic experiments, such as the time of survival of mice in shortage of oxygen in normal pressure, the antagonistic effect on arrhythmia induced by chloroform and the diuretic effect were carried out with LD. Variance analysis, canonical correlation and stepwise regression analysis were applied to interelationship between the amount of each drug and the pharmacologic data. Then, the fingerprints of the solutions were tested by HPLC method and 50 chromatographic peaks were obtained. Correlation and regression analysis was applied to interelationship between the area of each peaks and the pharmacologic data.Results: The results confirmed the poria and cinnamon twig are the basis, while ovate atractylodes and licorice are the adjuvans. 17 peaks, which were established as the therapeutical basis of the decoction, were selected from all compounds, and cinnamic acid of cinnamomum cassia, glycyrrhizin of Glycyrrhiza uralensis and dehydrotumulosic acid of Poria cocos were selected as the markers to evaluate the quality of the decoction.Conclusion: This study provided a significant try of studying the compatibility of composite herbal medicines.

Objective To investigate the value of real-time three-dimensional echocardiography timing-excursion parametric index and 17 segment volume curves index in patients with restrictive cardiomyopathy.Methods Eight patients with restrictive cardiomyopathy (proven by biopsy),twenty eight subjects with normal left ventricular(LV) function were examined by Philips iE33 with X3-1 probe.Results Parameter index of the SD and maximum difference of Tmsv of 16 segments,12 segments,and 6 basal segments(Tmax 16SD,Tmsv-12SD Tmsv -6SD,Tmsv-16Dif,Tmsv-12 Dif,Tmsv-6Dif,Tmsv 16-SD%,Tmsv 12-SD%,Tmsv 6-SD%,Tmsv 16-Dif%,Tmsv 12-Dif%,Tmsv 6-Dif%) was significantly higher in patients with restrictive cardiomyopathy than that in subjects with normal LV function(all P＜0.001).Average and maximum value and minimum of excursion was significantly lower in patients with restrictive cardiomyopathy than that in subjects with normal LV function(P＜0.001,P＜0.005,P＜0.005).And parameter index of 17 segment volume curves index showed higher sensitivity,specificity,positive predictive value and negative predictive value for diagnose restrictive cardiomyopathy.Conclusions Real-time threedimensional echocardiography can evaluate and diagnose restrictive cardiomyopathy fastly.

Background Venous malformation damages the local tissue severely because of the progressive development and often presents with invasive biological behavior.Galectin-3 (Gal-3) is proved to be closely associated with local invasion of malignant tumor.Studying the role of Gal-3 on tissue invasion in venous malformation of ocular region is of important clinical significance.Objective This study was to explore the role of Gal-3 protein and mRNA expression in venous malformation of ocular region.Methods One hundred and eighteen pathological sections were collected from ocular venous malformation patients who received surgery in Department of Hemangioma Surgery,People's Hospital of Henan Province and Henan Eye Institute from June 2009 to June 2014.The specimens were further diagnosed by histopathological examination.Then the expressions of Gal-3 protein and mRNA in venous malformation of ocular region were detected by using immunohistochemistry and in situ hybridization and compared with 20 pieces of distal cutting edge specimens which were evidently normal.The associations of Gal-3 positive expressions with invasion and configuration of lesions were analyzed.Results Pathological examination showed that venous malformations tissues contain many big blood vessels lacuna, lined with fiat endothelial cells.Immunochemistry and in situ hybridization exhibited that Gal-3 protein and mRNA were expressed in the cytoplasm and nuclei.The positive expression rates of Gal-3 protein and mRNA in the venous malformation tissues were 55.93％ (66/118) and 59.32％ (70/118) , but those in the normal tissue were 15.00％ (3/20) and 20.00％ (4/20) ,showing significant differences between them (x2 =11.461, 10.633, both at P＜0.05).No significant differences were seen in the positive expression rates of Gal-3 protein and mRNA between the patients aged ≤ 12 years and ＞12 years or different genders (age: x2 =0.334,0.128;both at P＞0.05.gender:x2 =0.606,1.155;both at P ＞0.05).The incidence rate of invading ocular deep tissues was significantly higher in the Gal-3-positive groups than that in the Gal-3-negative groups of protein and mRNA (protein :x2 =32.688, P＜0.05;mRNA : x2 =23.695, P＜0.05).In the Gal-3-negative groups,96.15％ (Gal-3 protein negative group) and 97.92％ (Gal-3 mRNA negative group) lesions showed the spherical shape with clear boundaries.The lesions texture with the fuzzy boundaries and the incidences of vague structure in lesions were significantly higher in the Gal-3-positive groups than that in the Gal-3-negative groups of protein and mRNA (protein :x2 =28.255, P＜0.05;mRNA : 28.186, P＜0.05).Conclusions Gal-3 expression rate is raised in the deep tissue-invaded and texture disorder ocular venous malformation.These results suggest that invasion and damage of ocular venous malformation are associated with the up-regulation of Gal-3.

Objective To obtain recombinant human stanniocalcin 1 ( STC1 ) with biological activity in Escheri.coli cells expression.Methods The gene was cloned into pET32b( +) vector by fused with thioredoxin and His tag .E.coli BL21(DE3) competent cells were transfomed by the recombinant vector .After renaturation, the fusion protein was digested with thrombin and intact STC1 protein was purified from the digested protein using Ni ion affinity chromatography .Recombi-nant humanSTC1 protein was confirmed by Western blot analysis using goat anti-STC1 antibody.The biological activity of STC1 in rat was assayed using standard method for assessment of renal function .Results The recombinant human STC 1 fu-sion protein is successfully expressed in Escherichia coli, the fusion protein was purified by affinity chromatography from the inclusion body and renaturated .Intact hSTC1 protein was released by thrombin digestion and purified by Ni ion affinity col-umn.The intact STC1 proteins was confirmed by Western blot analysis .Rat bioassay revealed that STC1 boosted phosphate reabsorption.Conclusion Recombinant STC1 protein was successfully expressed and has native biological activities .This protein could be used as an antigen for the preparation of monoclonal antibody against humanSTC 1.

Objective To investigate value of real-time three-dimensional echocardiography timingexcursion parametric index and 17 segment volume curves index in patients with restrictive cardiomyopathy and constrictive pericardits.MethodsSeventeen patients with restrictive cardiomyopathy (proven by biopsy) ,six patients with constrictive pericardits (proven by CT or surgical),twenty subjects with normal left ventricular(LV) function were examined by Philips iE33 with X3-1 probe.Results Parameter index of Tmsv 16-SD,Tmsv 12-SD Tmsv 6-SD,Tmsv 16-Dif,Tmsv 12-Dif,Tmsv 6-Dif,Tmsv 16-SD(%) ,Tmsv 12-SD(%),Tmsv 6-SD(%),Tmsv 16-Dif(%),Tmsv 12-Dif(%),Tmsv 6-Dif(%) was significantly higher in patients with restrictive cardiomyopathy than that in subjects with normal LV function(all P ＜0.05).Average and maximum value and minimum of excursion was significantly lower in patients with restrictive cardiomyopathy than that in subjects with normal LV function (all P ＜0.005).Whereas,compare with subjects with normal LV function,the parametric indexes of timing-excursion and 17 segment volume curves were not significantly difference in patients with constrictive pericardits(all P ＞0.05).Conclusions Realtime three-dimensional echocardiography can evaluate and diagnose fastly restrictive cardiomyopathy and constrictive pericardits.

AIM:To investigate the effects of angiotensin II ( Ang II) on the immune maturation and the oxi-dized low-density lipoprotein (Ox-LDL)-uptaking capacity of human monocyte-derived dendritic cells (DCs).METH-ODS:Human peripheral blood mononuclear cells were isolated by density gradient centrifugation , and the monocytes were purified by positive selection with anti-CD14 magnetic beads.After cultured with rhGM-CSF (100 μg/L) and rhIL-4 (50μg/L) for 5 d, the monocytes differentiated into immature DCs .On the 6th day of the culture, the cells were treated with various concentration levels of Ang II or pretreated with losartan .The immunophenotypic expression of HLA-DR and CD83 was analyzed by flow cytometry .The secretion levels of IL-12 and IFN-γin the culture supernatants were measured by ELISA.Furthermore, DCs were incubated with DiI-labelled Ox-LDL.The DiI-Ox-LDL-incorporated fraction was investiga-ted by flow cytometry .The mRNA expression of 3 scavenger receptors , scavenger receptor A ( SR-A) , CD36 and lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1), was examined by real-time PCR.RESULTS: Ang II induced the maturation of human monocyte-derived DCs, stimulated the expression of CD83 and HLA-DR, and promoted the secre-tion of IL-12 and IFN-γ, which were suppressed by losartan .Furthermore, Ang II increased the Ox-LDL-uptaking capacity of DCs, which was partially reduced by losartan .The incubation of DCs with Ang II enhanced the mRNA expression of LOX-1 in a dose-dependent manner , which was reduced by losartan .However, the expression of SR-A and CD36 was not changed .CONCLUSION:Ang II promotes the immune maturation of human monocyte-derived DCs and increases the up-take of Ox-LDL probably through the up-regulation of LOX-1 expression.

Objective To investigate the combined inhibition effect and the potential mechanism of rapamycin (mammalian target of rapamycin inhibitor) and PD98059 (mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) inhibitor) on mouse colorectal cancer (CRC). Methods S-ICR mice were subcutaneously injected with 20 mg/kg of 1,2-dimethylhydrazine dihydrochloride in the nape for 20 weeks to induce CRC. From the 16th week, the mice were treated with alone or combined injection with 0.25 mg/kg rapamycin and 2.5 mg/kg PD98059. The drugs were administered for 8 weeks. Subsequently, the animals were sacrificed and dissected, the tumor sizes were measured, and the tumors were harvested for pathological assay. Furthermore, the phosphorylation of mTOR, p70S6K, and 4E-BPl proteins was detected by using immunohistoehemistry. Results The mice treated with rapamycin (44. 44 %) or PD 98059 (either alone (38.89%) or combination treatment (6.67%) were significantly less likely to develop cancer compared with mice receiving none of them (77.78%, P<0. 05). The average size of tumors was (6.15±2. 192), (8.85±3. 983), (2.917±0. 191), (16.36±6.855) mm3 respectively (P<0.05).The anti-cancer effect of the combination treatment was substantially significant. The proteins of phospho-mTOR, phospho-p70s6K and phospho-4E-BPl were significantly down-regulated after treatments (all P values < ,0.05). Conclusions Combined treatment was more effective than single-drug treatments of rapamycin or PD98059 alone for the prevention and treatment of mouse CRC. The mTOR signal pathway might be involved in the inhibitory mechanism.

Objective To evaluate regional blood flow occlusion (RBFO) in hepatectomy for liver neoplasms. Methods In this study, hepatic tumors were resected under RBFO in 28 cases (RBFO group), and under Springle's technique (control group) in 24 cases. The Child-Pugh classifications of liver function were grade A in all patients. The ligature ribbon was put in liver parenchyma around tumor to block the blood supply before resecting the tumor under guiding of B sounography in RBFO group. Anesthesia time, blood loss and transfusion, hospitalization, change of liver function and complications were compared between the two groups. Results Blood loss, anesthesia time and postoperative hospital stay were (340±92) ml, (98.4±25.0) min, ( 10.2±2.3 ) d in RBFO group and (620±124) ml, ( 135.8±47.5 ) min, (16.5±5.1 ) d, respectively, in control group, differences were all significant between the two groups (P <0.01, t = 9.222,9.328 and 5.875, respectively). On post-op day 2, ALT (U/L) was (378.4±35.2) vs. (539.2±115.4) (t=7.012, P<0.01), TBIL (37.5±11.2) vs. (51.8±29) mmol/L(t=8.818, P<0.01),PT (17.4±2.4) vs. (20.4±2.8) see(t =4.16, P<0.01) in RBFO group and control group, respectively. ALT was (57.1±15.5) vs. (98.1±21.2) U/L(t =8.039),TBIL (25.4±4) vs. (46.3±13) mmol/L(t=8.085),PT (13.2±4.2) vs. (15.7±2.2) see (t=2.621)on post-op day 7 respectively, again the differences were all significant between the two groups (all P<0.01). Conclusion Regional blood flow occlusion is an effective technique to control blood loss during hepatectomy for liver neoplasms.