In order to study the bladder carcinoma cell growth suppression by introduction of foreign retinoblastoma (Rb) gene and explore a gene therapy approach for bladder cancer, a replication-deficient adenovirus vector encoding a wild-type Rb, AdCMVRb, was constructed and transfected into the cultured human bladder carcinoma cell line EJ. The efficiency of gene transfection and expression was detected by immunochemical staining, Western blotting and polymerase chain reaction. The role of Rb in suppressing EJ growth was observed by cell-counting, [3H]thymidine incorporation and flow cytometry. The results showed that wild-type Rb gene could be transfected effectively into cultured EJ with Ad-CMVRb and could arrest the cells at GO/Gl phases of the cell cycle, leading to inhibition of DNA synthesis. The results demonstrated the potential of adenovirus-mediated Rb gene therapy for bladder cancer.

Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; Societies, Medical ; United States

Objective To construct the pGEX-4T-2-CblN/Grb2, express and purify the corresponding chimeric protein; To investigate whether the chimeric CblN/Grb2 possesses ubiquitin ligase activity. Methods Total RNA of SK-BR-3 cells were isolated and reversely transcribed into cDNA, which were used as templates to amplify Grb2 SH2 by PCR. The gene fragment encoding the N-terminal part of Cbl protein (named as CblN) was amplified by PCR using pEFHACbl plasmid encoding human Cbl as templates. BamH I and EcoR V restriction enzyme digestion sites were introduced into both flanks of SH2 by overlapping extension PCR and the modified gene were cloned into pcDNA3.1(+). The pcDNA3.1(+)-CblN/Grb2 was obtained by replacing SH2 of CblN with Grb2SH2 and then used as templates to amplify CblN/Grb2 by PCR. The pGEX-4T-2-CblN/Grb2 was constructed by subcloning CblN/Grb2 into the prokaryotic expressing vector pGEX-4T-2. The GST-CblN/Grb2 fusion protein was expressed in E. coli of DH5? under IPTG induction and further purified with Glutathione Sepharose 4B. In vitro ubiquitination assay was performed to investigate whether the GST-CblN/Grb2 fusion protein is able to mediate auto-ubiquitinating reaction, namely whether it possesses ubiquitin ligase activity. Results The fusion expressing vector of pGEX-4T-2-CblN/Grb2 was successfully constructed; The GST-CblN/Grb2 fusion protein was correctly expressed and purified; In vitro ubiquitination assay indicated that GST-CblN/Grb2 fusion protein is able to mediate auto-ubiquitinating reaction and therefore possesses ubiquitin ligase activity. Conclusion Expression, purification of GST-CblN/Grb2 and identification of its′ activity have laid the foundation for further study of chimeric ubiquitin ligase′ effects on the growth of HER2 positive tumor cells.

The association of polymorphisms of calcium channel?1 subunit ( Cav1.1 ) gene ( - 1551T/C at exon 11 ) with thyrotoxic periodic paralysis (TPP) was investigated by PCR-RFLP.The distributions and frequencies of - 1551TC + CC genotype and C allele in TPP group were significantly higher than those in hyperthyroidism (HT) and normal control (CON) groups.There was no statistic difference between HT and CON groups.Cavl.1 gene - 1551TC + CC genotype and allele may contribute to the development of TPP in male HAN population from North China.

Zoonoses are a class of infectious diseases that are transmitted from animals to humans. More than 200 known types of zoonoses have been reported across the world until now. Among 1 400 pathogens of human infectious diseases, approximately 61% are zoonotic origin, and 75% human emerging infectious diseases are zoonoses. These zoonoses pose a great threat to human and animal health and decrease livestock production. To effectively tackle the persistent challenges resulting from zoonoses, WHO collaborates with member governments, academia, non-governmental and charitable organizations, and regional and international partners to prevent and manage zoonotic threats and their public health, social and economic impacts. Although great success has been achieved in the management of zoonoses, there are still multiple challenges for zoonoses control in China due to environmental, climate, socioeconomic factors and antimicrobial resistance. Based on the One Health concept, the integration of modern biological, information, artificial intelligent and big data tools through multidisciplinary and multi-sectorial collaborations may facilitate the containment and elimination of zoonoses.

OBJECTIVETo study the effect of ethyl acetate fraction of Blumea balsamifera (BBE) residue on treating rats of adjuvant arthritis (AA) and its mechanism.

METHODThe rats were immunized with the Freund's complete adjuvant (FCA). After modeling, 28 days' treatment with BBE was performed. During the experimental process, rat mass, toe girth, arthritic index (AI), proliferation of immune organs and pathological section were measured. After treatment, blood samples were collected through fossa orbitalis vein for detection of serum SOD, MDA, GSH, NO, OH*, ALP, AST, ALT, NAG and SA content using colorimetric method and IL-1, IL-6, TNF-α content using ELISA method.

RESULTAdministration with BBE (high dose) could significantly ameliorate joint swelling and arthritis index, effectively inhibit synovial hyperplasia, down-regulate the levels of MDA, NO, OH*, ALP, AST, ALT, NAG, SA, IL-1, IL-6, TNF-α and up-regulate the SOD and GSH levels in serum.

CONCLUSIONThe results suggested BBE possesses substantial anti-arthritis and antioxidant activities.

Acetates ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; isolation & purification ; Arthritis, Experimental ; blood ; drug therapy ; metabolism ; Asteraceae ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; Humans ; Interleukin-1 ; blood ; Interleukin-6 ; blood ; Male ; Malondialdehyde ; blood ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; blood

Antineoplastic Agents ; administration & dosage ; therapeutic use ; Cladribine ; administration & dosage ; therapeutic use ; Humans ; Leukemia, Hairy Cell ; drug therapy ; Male ; Middle Aged

Objective To explore the effect of fibroblast growth factor receptors 1-dominant negative strategy (FGFR1-DN) on alkaline phosphatase (ALP) activity of bone marrow stromal stem cells (BMSCs) after osteogenic induction.Methods BMSCs were transfected with eukaryotic expression plasmid pcDNA 3.1 (+)-DN FGFR1 and pcDNA3.1 (+)-FGFR1.The experiment was conducted in 4 groups:FGFR1-DN transfection group,FGFR1 transfection group,pcDNA3.1(+) empty vector transfection group and non-transfection group.The ALP activity of BMSCs was detected in logarithmic growth phase after osteogenic culture.The qualitative detection of ALP activity was carried out immunohistochemically while the quantitative detection by cALP kit.The ALP activity was compared between the 4 groups at 7 and 14 days after osteogenic induction.Results Compared with 7 days,the ALP activity at 14 days was significantly increased in the 4 groups,and the increase in FGFR1-DN transfection group was significantly higher than in the other 3 groups (P ＜ 0.05).At both 7 and 14 days,the ALP activity in FGFR1-DN transfection group was the highest while that in FGFR1 transfection group was the lowest (P ＜ 0.05).Conclusions FGFR1-DN can promote the ALP activity of BMSCs during osteogenesis.This may provide an experimental basis for the joint application of local gene therapy and tissue engineering and for construction of tissue engineered bone with better biocompatibility.

BACKGROUND: Full-thickness skin graft was a base for bum and plastic surgery, while uniform pressure and regional brake were key factors to ensure skin graft survival and avoid from necrosis. Traditionally, package and pressurized fixation were performed after skin transplantation; however, it induced residual dead space and unclear skin graft fixation, as well as suturing scar.OBJECTIVE: To study the effect of vacuum sealing drainage applied to full-thickness skin graft.METHODS: A total of 8 New Zealand rabbits were used to establish full-thickness skin graft models in three regions of bilateral spine. Vacuum sealing drainage, traditional pressurized suture and common wrapping were performed in the three regions, respectively. The skin graft survival was observed, and survival rate was calculated at 14 and 21 days. On the 3rd, 7th, and 14th days, samples were selected from skin graft and stained. The morphology was observed under light microscope and transmission electron microscope. The time to remove the drain vessel was that when the fluid was not increased or the fluid was clear.RESULTS AND CONCLUSION: The survival rate of vacuum sealing drainage group was significantly higher than that in other two group (P < 0.05), while the survival rate of traditional pressurized suture group was significantly higher than that in the common wrapping group (P< 0.05). Morphology examination demonstrated that regional cuticular layer was necrotic in the vacuum sealing drainage group after early skin transplantation, while inflammatory cell infiltration, fibroblast degeneration, and mitochondrial swelling were also observed. At later skin transplantation, fibroblast and basal cell were proliferated, and function of mitochondria and plasmid was active. This suggested that vacuum sealing drainage promoted survival rate of skin graft.

Endoscopy ; Female ; Humans ; Mesenchymoma ; surgery ; Nose Neoplasms ; surgery ; Young Adult