ABSTRACT
Objective To detect the expressions of Matrix Metalloprotein-11 ( MMP-11 ) and Cathepsin-D(Cath-D),and investigate their relationship and prognostic significance. Methods The study included 95 cases′ clinical date and postoperative specimens of gastric adenocarcinoma ( North China University of Science and Technology Affiliated Hospital,2010. 01-2013. 12) as observation group,70 cases of normal gastric tissue(from observation group) as control group. Expressions of MMP-11 and Cath-D were detected by IHC methods in two groups. Results The positive rate of MMP-11 was 51. 6%( 49/95) in observation group,5. 7%(4/70)in control group(χ2＝38. 884,P＜0. 05). The positive rate of Cath-D was 73. 7%(70/95) in observation group,28. 6%(20/70) in control group(χ2＝33. 082,P＜0. 05). The positive rate of MMP-11 was correlated with metastasis and vascular invasion(χ2＝7. 193、15. 566,P＜0. 05). The positive rate of Cath-D was correlated with maximal diameter,lymph node metastasis,vascular invasion and proliferation index(χ2＝7.431、5.654、6.569、6.801,P＜0.05).There was positive relationship between MMP-11 and Cath-D in observation group(r＝0. 46,P＜0. 05). The expressions of MMP-11 and Cath-D were correlated with prognosis in gastric adenocarcinoma ( P＜0. 05) . Conclusion The higher expressions and synergistic effect of MMP-11 and Cath-D may promote the occurrence and development in gastric adenocarcinoma. The joint detection of MMP-11 and Cath-D may be helpful to predict the prognosis of gastric adenocarcinoma.
ABSTRACT
Objective::To explore the effects of the biomimetic network membrane prepared by chitosan/gelatin/pectin on the proliferation and mineralization of mesenchymal stem cells (MSCs) , and to evaluate its feasibility of constructing tissue engineering bone.Methods:Chitosan, gelatin and pectin were made into a new biomimetic network membrane in a certain ratio by biomimetics.The experiment was divided into control group (MSCs+conventional medium) , material group (MSCs+network membrane+conventional medium) and material+OS group (MSCs+network membrane+OS medium) .The cell morphology was observed by inverted phase contrast microscope;the growth and secretion of extracellular matrix of the MSCs were observed under scanning electron microscope (SEM) .The proliferation of cells was determined by MTT assay (The MSCs were divided into negative control group and material group, and they were cultivated with blank medium and medium including materials) .The expression of calcium in MSCs was detected by Alizarin Red staining.Real-time polymerase chain reaction (RT-PCR) was used to determine the expression levels of osteocalcin (OC) mRNA and osteopontin (OPN) mRNA in the MSCs.Results:The network membrane was semitransparent thin film.The MSCs were short shuttle and clustered under inverted phase contrast microscope.After cultured for 7d, the MSCs were shuttle;after cultured for 14d, the number of MSCs was increased, with pseudo feet on the membrane;after cultured for21d, the MSCs clustered with a lot of neo-formed extracellular matrix.The MTT results showed that there was no significant difference in the proliferation level of MSCs between material group and negative control group (P＞0.05) .The Alizarin Red staining results showed that the MSCs in the network membrane were dyed orange red.The RT-PCR results showed that the expression levels of OC mRNA in the MSCs in material group and material+OS group were lower on the 7th and 14th days, but on the 21th day, the expression levels were significantly increased and reached the peak;the expression level of OC mRNA in the MSCs in material group was significantly increased on the 7th day, and the expression level reached the peak on the 14th day, then fell slightly on the 21th day;compared with control group, the expression levels of OC mRNA and OPN mRNA in the cells in material group and material+OS group at different time points were significantly increased (P＜0.01) , but there were no significant differences between material group and material+OS group (P＞0.05) .Conclusion:Chitosan/gelatin/pectin biomimetic network membrane has good biocompatibility, and MSCs can grow and proliferate well on the membrane.The membrane can induce the MSCs to express mineralization-related genes and proteins without inducers.
ABSTRACT
Objective:To evaluate the anti-adhesion properties of xyloglucan(mXG)hydrogel in the L929 mouse fibroblasts,to establish the animal model of recurrent adhesion in the rats after adhesionlysis,and to investigate the prevention effect of mXG hydrogel in recurrent adhesion and its influence in the expressions of transforming growth factor-β1(TGF-β1)and connective tissue growth factor(CTGF).Methods:After seeding on the surface of mXG gel,the adhesion property of L929 mouse fibroblasts was detected with fluorescence staining.The rat models of recurrent adhesion were established after adhesionlysis.Fourty-eight SD rats were randomly divided into 3 groups respectively(n=16).In adhesion group,1 mL saline was injected into the abdominal wall and cecum of the rats;in commercial membrane group,the 2 cm×3 cm chitosan anti-adhesion membrane was used to cover the wound of abdominal wall of the rats;in mXG hydrogel group,1 mL 4% mXG hydrogel was injected into the abdominal wall and cecal wound of the rats,and abdominal surgery was ended after the complete formation of the hydrogel(3 min).Eight rats were killed in each group 7 and 14 d after the second operation,and the degrees of adhesion were evaluated and the histological changes were observed. Immunohistochemical staining was used to observe the expression levels of CTGF and TGF-β1.Results:A large number of L929 mouse fibroblasts proliferated well and exhibited a spherical morphology in control group;but in mXG hydrogel group,only very little L929 mouse fibroblasts were globular,showing the mXG hydrogel had good resistance to the adhesion of L929 mouse fibroblasts.Blunt separate adhesion surface formed in all of the rats after operation.7 d after the second operation,4-5 score adhesion with fibrous connective tissue hyperplasia formed in adhesion group;moderate adhesion formed in commercial membrane group with more connective tissue;most cecum and abdominal wall began to heal in mXG hydrogel group with less connective tissue.14 d after the second operation,more severe peritoneal adhesions presented in adhesion group with proliferated fiber connetive tissue and collegan;severe adhesions also presented in commercial membrane group;mild adhesion was found in two rats mXG group,the other rats had no adhesion,and the defects were almost completely recovered.On days 7 and 14 after the second operation,the mean adhesion score of the rats in mXG group was significantly lower than those in adhesion group and commercial membrane group(P<0.05).The expression levels of TGF-β1 and CTGF were increased with the increase of peritoneal adhesion scores and collagen deposition;the expression levels of TGF-β1 and CTGF were the highest in adgesion group and the lowest in mXG group.Conclusion:mXG hydrogels have good resistance to fibroblast adhesion,and mXG hydrogel is effective in reducing the formation of recurrent adhesion in the rats after adhesionlysis and can decrease the expression levels of adhesion-related factors TGF-β1 and CTGF.
ABSTRACT
Purpose To investigate the relation of EGFR and KRAS gene mutations with the pathological characteristics in non-small cell lung cancer (NSCLC).Methods The EGFR and KRAS gene mutations were detected and analyzed in 64 patients with NSCLC by capillary electrophoresis and fluorescent probe method.Results In 64 cases,the EGFR gene mutations were detected in 27 patients (42.2％);the KRAS gene mutations in 8 patients (12.5％).The EGFR and KRAS mutations synchronized in 4 patients (6.25％).The mutations rate of EGFR was related to gender,histology type and smoking condition (P ＜ 0.05).There was no association between mutation of EGFR gene with the age,differentiation,lymph node metastasis and TNM stages (P ＞ 0.05).The mutations rate of KRAS gene was higher in adenocarcinoma patients than that in squamous carcinoma (P ＜ 0.01).There was no relationship between mutation of KRAS gene with the gender,age,smoking condition,differentiation,lymph node metastasis and TNM stages (P ＞ 0.05).Conclusion In NSCLC,EGFR gene mutations rate is higher than KRAS gene mutation.The mutation rate of EGFR gene is higher in female,adenocarcinoma and never smokers;the mutations rate of KRAS mutations is higher in patients with adenocarcinoma.The mutations in EGFR and KRAS can exist at the same time.
ABSTRACT
Objective: To investigate the effect of cartilage tissue engineering scaffold PVA/ι-CA on the biological behavior of the ATDC-5 cells,and to evaluate its feasibility on constructing tissue engineering cartilage. Methods:The polyvinyl alcohol (PVA)and carrageenan were used to make the composite scaffold material PVA/ι-CA according to a certain proportion by physical blending technology and repeated freezing thawing method,and the porosity and pore size of PVA/ι-CA were detected.The ATDC-5 cells were seeded into the composite scaffold and its growth was observed; the expressions of collagen type Ⅱ in the ATDC-5 cells were tested by immunohistochemical staining and immunofluorescence staining; the morphology of the ATDC-5 cells was confirmed by Toluidine blue staining.The growth and secretion of extracellular matrix of the ATDC-5 cells were observed under scanning electron microscope (SEM);the proliferative rates of ATDC-5 cells in composite scaffold materials in negative control group (added with DMEM culture media)and experimental group (added with DMEM contain scaffold)were determined by MTT assay.The composite scaffolds were implanted subcutaneously in the SD rats.The histocompatibility and vascularization in vivo of the composite scaffolds were evaluated.Results:The average porosity of cartilage tissue engineering scaffold PVA/ι-CA was (86.88±3.88)%,and the average pore size was 20-40 μm.The HE staining results showed that the ATDC-5 cells grew well with the polygon and plumpness morphology. All the samples were stained positive for collagen type Ⅱ by immunohistochemistry and immunofluorescence staining,which verified the normal phenotype of chondrocytes on the scaffolds. All the sample were stained positive for toluidine blue staining,which verified ECM deposition of the ATDC-5 cells on the scaffolds.The number of the positive cells was significantly increased with the prolongation of time.After cultured for 7 d,few of the ATDC-5 cells presented polygonal;after cultured for 14 d,the ATDC-5 cells distributed more densely,and contacted with each other on the scaffold;after cultured for 21 - 28 d,the ATDC-5 cells filled the interconnected pores of the scaffolds,synthesizing a significant amount of neo-formed ECM.The proliferation of ATDC-5 cells in PVA/ι-CA grew fast during 7-14 d,and it became slow during 21-28 d;the difference was not statistically significant compared with control group (P >0.05).The subcutaneous implantation results showed the inflammatory reactions were slight at the early stage and eviated gradually,there was an increasing angiogenesis at the late stage,and the degradation and absorption of the meterial were slight.Conclusion:PVA/ι-CA composite material will be an ideal material for the cartilage tissue engineering.
ABSTRACT
BACKGROUND:Polyvinyl alcohol (PVA) hydrogel with similar porous structure and mechanical properties to the natural cartilage is very suitable for the repair of articular cartilage. However, the pure PVA hydrogel after lyophilization wil be accompanied by the shrinkage of the polymer network and the col apse of the pores, leading to the inhomogeneous performance of the material even in the state of re-swel ing. Addition of the active polymer wil increase the cel adhesion ability of PVA hydrogel. OBJECTIVE:To construct PVA/lota-carrageenan (l-CA) composite materials with different mass fractions of l-CA and evaluate the biocompatibility with vascular endothelial cel s. METHODS:PVA/l-CA composite films with different contents of l-CA were fabricated and then co-cultured with vascular endothelial cel s. Attachment, proliferation and morphological changes of vascular endothelial cel s on the composite were observed by scanning electron microscope and MTT assay to evaluate its biocompatibility. PVA/l-CA three-dimensional scaffold with different contents of l-CA were constructed, and hemolysis experiment was conducted according to the biological evaluation standards of medical devices, and the porosity and pore size were observed using scanning electron microscope. RESULTS AND CONCLUSION:In vitro experimental results showed that the addition of l-CA could significantly increase the biological activity of PVA hydrogel, and promote the cel attachment and proliferation on the scaffold. The hemolysis rate of each experimental group was less than 5%(the accepted safety standard), suggesting that the composite materials were in accordance with the standard of medical devices for hemolysis experiment. These findings indicate that the composite scaffolds with 20%-30%l-CA possess the pore size suitable for cel growth and proliferation and the porosity beneficial for transportation of nutrients and metabolites, which can serve as an excel ent scaffold for tissue engineering.
ABSTRACT
Objective To research the expression of mitofusin-2 (mfn2) and tumor metastasis suppressor genes (nm23)in bladder cancer cells and its correlation with clinical pathological feature. Methods Immunohistochemistry was used to measure the expression of mfn2 and nm23.Sixty-five cases of bladder cancer were sampled,which include fif-ty cases of male and fifteen cases of female. TNM stage:Forty-seven cases were in stage I;Ten cases were in stage II; Five cases were in stageⅢ; Three cases were in stageⅣ. Other fifteen cases were sampled from normal bladder or benign tumor of bladder as control . All cases were collected from department of pathology,affiliated Hospital of hebei union university. Results The positive expression rate of mfn2 in bladder cancer tissues was significantly higher than those in normal blad-der tissues and benign tumor of bladder(χ2=32.528,P<0.05);The positive expression rate of nm23 in bladder cancer tis-sues was significantly lower than those in normal bladder tissues and benign tumor of bladder (χ2=19.719,P<0.05);the high expression level of mfn2 in bladder cancer was associated with tumor differentiation and TNM stage(P<0.05),but not corre-lated with age,sex,lymph node metastasis and clinical grade. The low level of nm23 was associated with TNM stage,clinical grade and LN metastasis. Conclusion The positive expression rate of mfn2 was increased in bladder cancer. It indicated that there was a close relationship between mfn2 and the occurrence and development of bladder cancer;The expression rate of nm23 was decreased in bladder cancer,it may be a predictor for metastasis and prognosis of bladder cancer.