Objective To explore the methods of extraction, isolation, purification, and biological activities of ray cartilage glycosaminoglycans (RCG). Methods RCG was purified by guanidine hydrochlorid extraction, acetone fractional precipitation, ultrafiltration, and Sephadex column chromatography. The purity and molecular mass of RCG were measured by means of HPLC. The model of mouse with Lewis lung carcinoma was made, the experimental mice were randomly divided into normal saline group, RCG (500, 250, and 125 mg/kg) groups, and CTX (60 mg/kg) group. Tumor growth states of mice were observed, tumor growth curve was described, inhibitory rates of primary tumor and number of lung metastasis focus were measured; microvessel density (MVD) was quantitated by immunohistochemistry using monoclonal antibodies of CD31; the expression of vascular endothelial growth factor (VEGF) mRNA was determined with RT-PCR. Results Using HPLC, a single glycosaminoglycans with molecular mass 9.7?104 was collected and its purity exceeded ninty-nine percent. Tumor growth curves in RCG groups were smooth compared with saline group. There were significant differences of inhibitory rates of primary tumor, number of lung metastasis focus and MVD between RCG groups and saline group. VEGF mRNA expression levels in RCG groups were reduced significantly compared with saline group. Conclusion RCG could effectively inhibit the growth and metastasis of primary Lewis lung carcinoma in C57BL/6 mouse and angiogenesis.

Adult ; Aged ; Anesthesia ; methods ; Bronchoalveolar Lavage ; methods ; Female ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; surgery ; Retrospective Studies

Adolescent ; Adult ; Child ; Female ; Fistula ; surgery ; Humans ; Male ; Recurrence ; Reoperation ; Retrospective Studies ; Thyroglossal Cyst ; surgery ; Young Adult

Objective To investigate the clinical efficacy of fluoroscopy-guided intestinal adhesion lysis, as a new non-surgical method, in treating incomplete adhesive small intestinal obstruction in order to improve the therapeutic results of adhesive intestinal obstruction. Methods A total of 93 patients with incomplete adhesive small intestinal obstruction were enrolled in this study. The patients were divided into study group (n=49) and control group (n=44). Fluoroscopy-guided intestinal adhesion lysis together with restoration of inter-intestinal loop enterocele was carried out for the patients of the study group , while traditional conservative surgical therapy was employed for the patients of the control group. The study group was comparable with the control group in patients’ age, gender, medical history, disease course, X-ray findings, etc. Results Of the 49 cases in the study group, complete cure was obtained in 40 with a cure rate of 81.6%. The mean hospitalization day was 0.3 day, and the average operation time was 3.25 hours. Among the 44 patients in the control group, complete cure was obtained in 37 with a cure rate of 84.1%. The mean hospitalization day was 7.6 days, and the average therapeutic time was 183.26 hours. Conclusion For the treatment of incomplete adhesive small intestinal obstruction , the therapeutic efficacy of fluoroscopy-guided intestinal adhesion lysis together with restoration of inter-intestinal loop enterocele is better than that of traditional conservative surgical therapy.

Aim The m_5AChR-G_ 11? fusion protein was expressed by baculovirus-Sf9 cells system, then using it to identify the specific agonists and antagonists for m_5AChR via detecting the affinity of GDP and m_5AChR-G_ 11?. Methods The m_5AChR-G_ 11? fused cDNAs were generated via a two-step PCR protocol and inserted into pBacPAK9 virus vector. We expressed m_5AChR-G_ 11? fusion protein and m_5AChR protein using baculovirus-Sf9 cell system. [ 3H]QNB and [ 35S]GTP?S binding tests were performed to detect the expressional level of receptor proteins and determine the affinity of GDP and m_5AChR-G_ 11? fusion protein. Results The expression level of m_5AChR-G_ 11? was (47.6?3.2) nmol?g -1 protein. The affinity of GDP to G_ 11? partner changed in the presence of different muscarinic ligands. IC_ 50 values of GDP in the presence of ACh, YM796, Oxotremorine, Methixene, Dextimide and atropine were 128.0, 72.1, 68.5, 16.2, 14.9 and 9.7 ?mol?L -1 respectively, and that in the absence of muscarinic ligand was 20.8 ?mol?L -1. Conclusion The m_5AChR-G_ 11? fusion protein has the pharmacological specificity of M_5 receptor and the efficient coupling interaction of the two partner. Affinity of GDP to ligand bound m_5AChR-G_ 11? fusion protein represents the species of muscarinic ligands. ACh is a full agonist for m_5AChR-G_ 11? fusion protein, YM796 and oxotremorine are partial agonists, while methixene, dextimide and atropine are antagonists.

Adamantinoma ; diagnostic imaging ; metabolism ; pathology ; surgery ; Adult ; Diagnosis, Differential ; Female ; Femur ; Follow-Up Studies ; Humans ; Humerus ; Ilium ; Keratins ; metabolism ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Retrospective Studies ; Sarcoma, Ewing ; pathology ; Sarcoma, Synovial ; pathology ; Tibia ; Tomography, X-Ray Computed ; Young Adult

Objective To observe the effects of chronic arsenic exposure on estrogen receptor-binding fragment-associated gene 9 (Ebag9) and estrogen-responsive finger protein (efp) mRNA expression in female rat' s myocardium.Methods Fifty female Wistar rats were randomly divided into five groups according to arsenic (As2O3) concentrations in drinking-water:0.00(control),0.05,0.10,0.20,0.40 mg/L groups and RT-PCR was used to detect Ebag9 and efp mRNA expression of myocardium at the 32 weeks of experiment.Results Ebag9 and efp mRNA expression levels in 0.00,0.05,0.10,0.20,0.40 mg/L groups were respectively as follows:0.54 ±0.14,0.52 ± 0.10,0.48 ± 0.24,0.58 ± 0.13,0.45 ± 0.19 and 0.85 ± 0.14,0.86 ± 0.12,0.87 ± 0.09,0.99 ±0.10,0.86 ± 0.19.Compared to the control group,Ebag9 mRNA level of the 0.20 mg/L group was increased,and decreased in other groups,but the difference between two groups was not significant(all P ＞ 0.05).Compared to control group,the efp mRNA level of 0.20 mg/L group increased significantly(P ＜ 0.05),and showed increased tendency in other arsenic groups,but the difference between two groups was not significant (all P ＞ 0.05).Conclusions Ebag9 and efp mRNA expression have changed in myocardium of rats exposed to chronic arsenic.Arsenic may has endocrine disruptor effect to female rat's myocardium.

Objective To evaluate the effect of multiplex polymerase chain reaction(Multiple-PCR) in identification of Brucella strains.Methods Six standard Brucella strains (Brucella abortus,Brucella melitensis,Brucella suis,Brucella canis,Brucella ovis,Brucella neotomae) were used as positive controls and Escherichia coli O∶157 and Yersinia enterocolitica O∶9 were used as negative controls.A total of 29 Brucella strains were tested.Brucella strains were amplified by BCSP31-PCR and the species of Brucella with positive results were identified with Multiple-PCR method.Results The results of all 29 amplified Brucella strains were positive with BCSP31-PCR.The identified results of all Brucella strains were positive with Multiple-PCR,including 20 strains of Brucella melitensis,5 isolates of Brucella suis,3 strains of Brucella abortus and 1 strain of Brucella canis.Conclusion Multiple-PCR method is a rapid,specific,simple and low risk method for identification of Brucella species.

OBJECTIVETo explore the effect of Ray cartilage glycosaminoglycans (RCG) on the expression of MMP-9 in Lewis lung carcinoma of mice.

METHODThe model of mice with Lewis lung carcinoma was induced. The experimental mice were randomly divided into normal saline group, RCG groups at varied concentrations and CTX group. Tumor growth state was observed, and tumor inhibitory rate of primary tumor and number of lung metastasis focus were measured. The expression of MMP-9 mRNA and protein in Lewis lung carcinoma was determined with RT-PCR and Western blot.

RESULTAs compared with normal saline group, tumor growth curves in RCG groups were smooth, there were significant differences of inhibitory rates of primary tumor and number of lung metastasis focus between RCG groups and normal saline group, and MMP-9 mRNA and protein expression levels in RCG groups were reduced significantly.

CONCLUSIONRCG can inhibit effectively the growth and metastasis of implanted Lewis lung carcinoma in C57BL/6 mice, which is probably attributed to reducing the expression of MMP-9 mRNA and protein.

Animals ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Carcinoma, Lewis Lung ; enzymology ; pathology ; Cartilage ; chemistry ; Cell Line, Tumor ; Glycosaminoglycans ; isolation & purification ; pharmacology ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Skates (Fish)

10.3969/j.issn.2095-4344.2013.25.014