The B Phycoerythrin(B PE) could both be separated after crude phycobiliproteins from Porphyridium cruentum were precipitated in ammonium sulphate and dialyzed,then chromatographed on a hydroxylapatite (HA) Column and a Sephadex G 100 column respectively.The purity of B PE separated by two methods was 4.92 and 3.78 (A 545 nm/A 280 nm) respectively.When the purified B PE was electrophoresed under polyacryiamide gradient gel electrophoresis condition,only one band was observed.The B PE possessed an absorption maximum at 545nm and a room fluoresence emission at 574.5nm.

Objective: To study the effect of methylene blue on the metabolism of reactive oxygen and acid production of Streptococcus mutans . Methods: Streptococcus of cricetusa, rattusb, mutansc, sobrinsd and mutans e,f and g were exposed to methylene blue at 2.5?10 -4 g/L for 1 h. The metabolism of reactive oxygen and acid production were detected by electron spin trapping method and gas chromatography respectively. Results: Electron spin resonance (ESR) spectrums of DMPO-O - ? 2 and DMPO-OH ? were observed in Streptococcus mutans cultured in glucose fluid, but were not found in those exposed to methylene. The total of acid production by Streptococcus mutans incubated with glucose + methylene blue were significantly lower than that with single glucose. Conclusion: Methylene blue may inhibit oxygen species metabolism and acid production of Streptococcus mutans , it may be used as a kind of dental caries prevention agent.

Objective To explore the proliferation-promoting effect of bovine mammary gland epithelial cells (BMECs) co-cultured with umbilical cord mesenchymal stem cells (UC-MSCs) in serum-free culture mediuum.Methods Bovine UC-MSCs and BMECs were selected for co-culturing in direct or indirect contact.In the direct contact culture groups, UC-MSCs and BMECs were co-cultured at concentration ratios of 2∶1, 1∶1, 1∶2, 1∶3, 1∶4, 1∶5, and 1:10, respectively.In the indirect contact culture group, the supernatant of UC-MSCs was used as the conditioned medium to re-suspend BMECs.In the control groups, UC-MSCs and BMECs were cultured alone.The cell growth status in each group was observed at 0, 4, 8, 12, 24, 36, 48, 60, 72 h after culture, and cell proliferation was detected by cell counting kit-8 (CCK-8) assay.Results At 48 h, the optical density of the conditioned medium-BMECs group was significantly higher compared with the control groups (P<0.05).Meanwhile, the optical density in the direct contact group at a concentration ratio of 1∶2 reached the peak, which was extremely significantly higher compared with the control groups (P<0.01) and significantly higher compared with the other direct contact culture groups and the conditioned medium-BMECs group (P<0.05).Conclusions Co-culture of UC-MSCs and BMECs in serum-free culture medium is capable to promote the proliferation of BMECs, and the co-culture by cell-to-cell contact has a better effect.The optimal concentration ratio of UC-MSCs to BMECs is 1∶2, and the optimal culture time is 48 h.

Objective To evaluate ascending venography in the diagnosis of iliac vein compression syndrome.Methods From April 2011 to April 2013,we have had 556 patients with varicose veins suspected of Cockett syndrome.The degree of varicose veins by the International Union of Venous Clinical Classification (CEAP classification) was as following[1]:shallow varicose veins of lower limb (C2) in 190;varicose veins with limb swelling (C3) in 149 cases ; with body skin changes,such as pigmentation,eczema or lipid hard skin disease (C4) in 130; with healed ulcers (C5) in 17; with active ulcer(C6) in 70.Deep vein anterograde contrast and femoral venous cannula angiography were performed on 760 times.Results Iliac vein compression syndrome (Cockett) was detected by ascending venography in 154 patients,the diagnosis was established by following femoral venous cannula angiography.In the other 48 patients in whom Cockett syndrome was suspected by ascending venography,final diagnosis was reached by femoral venous cannula angiography.Altogether there were 202 iliac vein compression syndrome cases,with a positive rate 38.19％ (202/529).The narrowness was larger than 50％ in 173 cases.In 145 cases there were visible collateral vessels.Conclusions Deep vein ascending angiography is a useful screening method in the diagnosis and treatment of Cockett syndrome.

Objective: To observe the expression of long non-coding RNA myocardial infarction associated transcript (LncRNA-MIAT) in tumor necrosis factor-α (TNF-α) induced endothelial cells (ECs) inflammation in vitroand to study the impact of LncRNA-MIAT on inflammatory regulation. Methods: LncRNA-MIAT expression in ECs was induced by TNF-α at different time and concentration. Expressions of intercellular adhesion molecule-1 (ICAM-1) and LncRNA-MIAT in inflammatory ECs were examined by quantitative real time polymerase chain reaction (qRT-PCR) and Western blot analysis. Moreover, ECs was transfected by siRNAMIAT to observe the effect of LncRNA-MIAT knock-down on ICAM-1 expression. Results: LncRNA-MIAT expression showed the increasing trend by elevated time and concentration of TNF-stimulation. Compared with TNF-α stimulation at 0h, 6h and 12h, LncRNA-MIAT expressions were increased at 24h and 48h of TNF-αstimulation respectively, allP<0.05; compared with TNF-α concentration at 0ng/ml and 0.125ng/ml, LncRNA-MIAT expressions were elevated by TNF-α stimulation at 1.000ng/ml and 10.000ng/ml respectively, allP<0.05. With siRNAMIAT knock-down, TNF-α induced ICAM-1 protein expression was significantly reduced in ECs,P<0.05. Conclusion: LncRNA-MIAT might be involved in ECs inflammatory response and it may play a role to promote inflammation.

Objective To analyse the efficacy of microvascular decompression for hemifacial spasm (HFS) caused by vertebral basilar artery compression. Methods A total of 141 patients with HFS treated by microvascular decompression in our hospital were collected in this study. The improvement of the symptoms after operation was compared between patients with HFS caused by vertebral basilar artery compression (28 cases) and patients with HFS caused by non-vertebral basilar artery compression (113 cases). Results There was no significant difference in the effective rate between the two groups of HFS (96.43%vs. 98.23%,P=0.49) with mean following-up 13.81 ± 1.57 months. And there was no significant difference in the delayed cure rate after surgery between two groups (37.04%vs. 20.72%,χ2=1.38, P>0.05). Conclusion Microvascular decompression is a safe and effective method for the treatment of HFS caused by compressed vertebral basilar artery.

BACKGROUND:Osteoporosis caused by chemotherapy has become one of the serious side effects that impact the skeletal system. Icarin shows a strong anti-osteoporosis activity, which can have protective effect on osteoporosis induced by chemotherapy. OBJECTIVE: To study the protective effect and mechanism of icarin against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cels differentiating into osteoblasts. METHODS:MTT assay and alkaline phosphatase (ALP) staining were used to determine the optimal protective concentration of icarin against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cels differentiating into osteoblasts. mRNA expressions of osteoblast-specific transcription factors, OC, ALP, Runx2, and Wnt/β-catenin signaling pathway target genes, β-catenin, C-Myc, cyclin D1, were determined using RT-PCR method at different time after intervention with the optimal concentration of icarin. Expressions of Runx2, β-catenin, c-Myc, cyclin D1 regulated by the optimal concentration of icarin were detected using western blot assay at the protein level. RESULTS AND CONCLUSION:Cel viability and ALP activity decreased significantly in the cyclophosphamide group compared with the control group, but there was no significant difference in cel viability between icarin group and cyclophosphamide group. Icarin at 100 μmol/L showed the best protective effect against cyclophosphamide-induced obstacle of osteogenic differentiation of bone marrow mesenhymal stem cels. Compared with the control group, cyclophosphamide chemotherapy reduced the expressions of ALP, OC, Runx2 at mRNA level and Runx2 at protein level, weakened the expressions ofβ-catenin, cyclin D1 at mRNA level and active β-catenin, Cyclin D1, c-myc at protein level, and increased the expression of DKK1. Compared with the cyclophosphamide group, 100 μmol/L icarin increased the expression of osteoblast-specific transcription factors and Wnt/β-catenin signaling pathway genes at mRNA and protein levels, and reduced the expression of DKK1 protein. These results show that cyclophosphamide can lead to osteogenic differentiation disorder of mouse bone marrow mesenchymal stem cels, and in contrast, icarin shows a protective effect and its optimal intervention concentration is 100 μmol/L. Additionaly, the protective roleof icarin is probably related to activation of Wnt/β-catenin signal pathway.

Background::Psoriasis is a chronic systemic inflammatory disease, and hyperuricemia is a common comorbidity in patients with psoriasis. However, there are limited reports on the relationship between serum uric acid levels and biological treatment efficacy. The purposes of this study were to compare the differences in serum uric acid levels between patients with psoriasis and healthy controls and analyze the risk of hyperuricemia.Methods::A total of 196 patients with psoriasis and 191 age- and sex-matched healthy controls were enrolled in this retrospective cohort study. One hundred and twenty-seven patients with severe psoriasis were treated with biologics. Sixty-eight patients received adalimumab, and 59 patients received secukinumab. Serum uric acid levels were measured at baseline, week 24, and week 48 of treatment.Results::Patients with psoriasis had higher serum uric acid levels than healthy controls (6.4 ± 1.7 mg/dL vs. 5.7 ± 1.5 mg/dL, P < 0.001). Hyperuricemia was found in 33.7% (66/196) of patients with psoriasis, which was significantly higher than that in healthy controls (13.1% [25/191], P < 0.001). Serum uric acid levels and hyperuricemia were not related to the severity of psoriasis ( P > 0.05). No significant changes in serum uric acid levels and hyperuricemia were observed following adalimumab treatment ( P > 0.05). The serum uric acid level in patients treated with secukinumab was 6.7 ± 1.6 mg/dL at week 24, which was not statistically different from that at baseline (6.6 ± 1.4 mg/dL, P = 0.885). Serum uric acid levels were significantly decreased at week 48 (6.3 ± 1.5 mg/dL vs. 6.6 ± 1.4 mg/dL, P = 0.007) in patients treated with secukinumab. Secukinumab had no significant effect on hyperuricemia either ( P > 0.05). Conclusions::The serum uric acid levels and prevalence of hyperuricemia in patients with psoriasis were significantly higher than those in healthy controls. Secukinumab treatment for 48 weeks successfully decreased serum uric acid levels in patients with psoriasis, whereas adalimumab had no significant effect on serum uric acid levels.

OBJECTIVETo identify the type of CTGAATCA from -nt.199 to -nt.192 of the cytokeratin 13(CK13) gene 5' flanking region and determine its transcriptional effect on CK13 gene expression.

METHODSThe CAT systems were used to assess the effects of different motifs of CK13 gene 5' flanking region on transcription. The clones of pCAT-enhancer with the total length, -nt.207 to +nt.63 and the same length of -nt.207 to +nt.63, but the T, G of -nt.198, -nt.197 being changed to A, T of the CK13 gene 5' flanking region, were constructed and transferred to HeLa cells with the help of lipofectin. Then work was done to detect the instant CAT expression of different clones and evaluate the effects of CTGAATCA of the 5' flanking region on CK13 gene expression. The type of the cis-element of CTGAATCA was identified with electrophoretic mobility shift assay (EMSA) and competition-EMSA.

RESULTSCTGAATCA in the CK13 gene 5' flanking region is an AP1 cis-element by EMSA and competition-EMSA, it promotes CK13 gene expression.

CONCLUSIONCTGAATCA from -nt.199 to nt.192 of the CK13 gene 5' flanking region is an AP1 reaction element, not a cAMP reaction element. It promotes transcriptional activity of CK13 gene 5' flanking region.

5' Flanking Region ; genetics ; Base Sequence ; Binding Sites ; genetics ; Binding, Competitive ; Chloramphenicol O-Acetyltransferase ; genetics ; metabolism ; DNA ; genetics ; metabolism ; Electrophoretic Mobility Shift Assay ; Gene Expression Regulation ; HeLa Cells ; Humans ; Keratins ; genetics ; Mutation ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transcription Factor AP-1 ; metabolism ; Transcription, Genetic ; genetics ; Transfection

OBJECTIVE: To explore the regulative effect of expression of VEGF gene in nasopharyngeal carcinoma, and to discuss the future application of microRNA in the gene therapy for nasopharyngeal carcinoma. METHOD: We constructed the recombination miRNA plasmid vectors which target VEGF gene and plasmids were transfected into CNE-2 cells by using Lipofectamine 2000 Reagent. The VEGF mRNA and VEGF protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting respectively. WST-8 assay was used to determine the inhibitory effect of microRNA on cell growth. Stable cell lines and wild type CNE-2 cell line were inoculated to subcutis of nude mice to establish animal models. The tumor growth and volume were observed. RESULT: After the transfection of CNE-2 cells , the expressions of VEGF mRNA and VEGF protein were down-regulated at different degree. Whereas, CNE-2 cell growth showed no change by observation of fluorescence microscopy, and cell proliferation was not inhibited in WST-8 assay. However, in vivo, growth of xenograft was inhibited in preliminary experiments of nude mice. CONCLUSION By miRNA plasmid constructed artificially, miRNA can effectively interfere nasopharyngeal carcinoma cells by down-regulating the expressions of VEGF gene, therefore can inhibit the growth of tumor xenografted in vivo. Future application of microRNA in the gene therapy of nasopharyngeal carcinoma might be expected.
Animals ; Cell Line, Tumor ; Female ; Genetic Therapy ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; MicroRNAs ; genetics ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; Plasmids ; Vascular Endothelial Growth Factors ; genetics ; metabolism