During lymphocyte development antigen receptor genes undergo V(D)J recombination to obtain antigen binding specificity and diversity. This process is not only controlled by genetic factors, such as tissue- and stage- specificity of RAG-1/2 protein, germline transcriptional activity and ACEs, but also regulated at epigenetic level. The chromatin accessibility of recombinase is associated with the chromatin configuration around the targeted gene segments. Thus, activation of V(D)J recombination requires the recruitment of remodeling complexes for changing the accessibility in the localized chromatin. Moreover, docking of remodeling complexes, which serve for creating active chromatin environment, relies on certain patterns of chromatin modification. Some recent findings regarding epigenetic regulation mechanisms in V(D)J recombination, such as CpG methylation, histone modification, nucleosome remodeling and nuclear topology were reviewed.

To express human single chain antibody to collagenase IV as soluble proteins secreted by pichia pastoris, a recombinant vector scFv-pPIC9 was constructed, and then transformed into pichia pastoris GS115 by electuoporation, The transformants were selected by DNA hybridization and cultured in the media with methanlo. Soluble scFv were secreted into culturesupernatant and characterized by SDS-PAGE and Western Blot. Our results showed that the secreting of soluble scFv in pichia pastoris was as high as 20mg/L. The soluble scFv has similar immuno-activity to its counterpart produced in bacteria, and it is more easily and efficiently purified.

To express human single chain antibody to collagenase IV as soluble proteins secreted by pichia pastoris, a recombinant vector scFv pPIC9 was constructed, and then transformed into pichia pastoris GS115 by electuoporation, The transformants were selected by DNA hybridization and cultured in the media with methanlo. Soluble scFv were secreted into culturesupernatant and characterized by SDS PAGE and Western Blot. Our results showed that the secreting of soluble scFv in pichia pastoris was as high as 20mg/L. The soluble scFv has similar immuno activity to its counterpart produced in bacteria, and it is more easily and efficiently purified.

A mAb T2-2 was generated using hybridoma techniques, and its target was identified as a 46 ku-cytokeratin (CK), based on biochemical study and a completely overlapped binding pattern of mAb T2-2 with anti-pan-CKs antibodies. An epithelia-specificity of the mAb T2-2 was determined by screening 68 human normal and 65 tumor tissues using immunohistochemistry. Unlike most of anti-CKs antibodies, the mAb T2-2 recognized a mono-specific epitope only expressed on the 46 ku CK, suggesting that mAb T2-2 is superior to most anti-CKs antibodies that cross-reacted with many different kinds of CKs. In addition, it was found that the mAb T2-2 was multipurpose with a broad applicability to ELISA, immunohistochemistry, immunofluorescence, Western blotting, and was also compatible with various fixation reagents. These results strongly indicate that the mAb T2-2 has potential applications for studying CKs function and for diagnosis of tumor and other disorders.

Objective To investigate the characteristics of traditional Chinese medical syndrome types of fulminant and epidemic dengue fever patients admitted in Guangzhou and Xishuang banna in the year of 2013,and to ex plore the differences of etiology and pathogenesis, illness, and treatment for the patients in the two regions. Methods We collected the clinical data of 78 cases receiving integrative Chinese and western medicine from 255 patients admitted in Guangzhou Municipal Eighth People’s Hospital, and the clinical data of 39 cases receiving integrative Chinese and western medicine from 120 patients admitted in the People’s Hospital of Xishuangbanna Dai Nationality Autonomous Prefecture in the year of 2013. The traditional Chinese medical syndrome types and the syndrome scores of the total of 117 cases were investigated. The method of phenology was used for the analysis of epidemic time and epidemic region of dengue fever, and the theory of defense-qi-nutrient-blood syndrome differentiation of seasonal febrile diseases was used for the analysis of etiology and pathogenesis of dengue fever. Results ( 1) Dengue fever was epidemic in the first ten days of July and in the middle ten days of November of the year 2013 in Guangzhou region, and was epidemic in the middle ten days of August and the first ten days of October in Xishuangbanna region. The epidemicity of dengue fever in Guangzhou covered the end of summer and the whole autumn, and then disappeared before the coming of winter. In Xishuangbanna , the epidemicity of dengue fever was obvious in autumn, and disappeared in late autumn. ( 2) In the two hospitals, dengue fever patients were dominated by the syndromes of excessive heat in both Qifen and Xuefen, blood stasis blended with toxicity, excessive heat in Qifen, and lingering pathogens in order. (3) Before treatment, the scores of fever were higher in patients of Xishuangbanna hospital than those in patients of Guangzhou Eighth People’s Hospital ( P<0.01) . After treatment for 3 days, fever scores were improved in both hospitals (P<0.05 or P<0.01), but the differences between the two hospitals were insignificant (P>0.05) . After treatment for 6 days, fever disappeared in patients of both hospitals. (4) Before treatment, the scores of syndromes were higher in patients of Xishuangbanna hospital than those in patients of Guangzhou Eighth People’s Hospital ( P<0.05) . After treatment for 3 days, syndorme scores were improved in both hospitals ( P<0.01) , but the syndrome scores were higher in Xishuangbanna hospital than those in patients of Guangzhou Eighth People’s Hospital. After treatment for 6 days, syndrome scores were much improved in patients of both hospitals compared with those after treatment for 3 days (P<0.01) . Conclusion In dengue fever patients admitted in Guangzhou and Xishuangbanna region, the syndrome of excessive heat in both Qifen and Xuefen is the leading type, and then comes blood stasis blended with toxicity. The illness state of patients in Guangzhou region is milder than that of the patients in Xishuangbanna region, the time for symptom relief is about one week, and similar therapeutic effect can be achieved in the two regions .

Objective To analyze 5 patients suptered heart failure and ventricular tachycardia((VT).)Methods The Implantable Biventricular pacing cardioverter defibrillator was used to analyzed 5 patients(4 with primary dilated cardiomyopathy and 1 with coronary heart disease) suffered from heart failure and ventricular tachycardia(VT).Results 2 cases had history of syncope,4 patients were implanted INSYNC 7272 implantable cardioverter defibrillator(ICD) and 1 patient was implanted V-350 ICD.During the 1~14 month follow-up period,all the cardio functions were improved without VT or syncope.1 patient with VT and VF was detected and terminated by ICD.The patient was saved.Conclusions Implantable biventricular pacing cardioverter defibrillator is an effective approach to treat sudden cardiac death,mortal ventricular arrhythmia and heart failure.

AIM: To investigate the mechanism of the AP-1 signal transduction pathway inhibited by JIP in nasopharyngeal carcinoma cells. METHODS: AP-1 activity was triggered by Dox-induced LMP1 expression in Tet-on-LMP1-HNE 2 cells (L7). The retention of phospho-JNK in the cytoplasm caused by JIP was examined with immunofluroscence assay. RESULTS: 24 h after transfection of L7 cells with the JIP expression plasmid, the translocation of activated JNK was inhibited, which resulted in the retention of phospho-JNK in the cytoplasm and down-regulation of the AP-1 activity. CONCLUSION: JIP down-regulates the activity of AP-1 through the inhibition of the translocation of JNK.

FXYD6, FXYD domain containing ion transport regulator 6, has been reported to affect the activity of Na(+)/K(+)-ATPase and be associated with mental diseases. Here, we demonstrate that FXYD6 is up-regulated in hepatocellular carcinoma (HCC) and enhances the migration and proliferation of HCC cells. Up-regulation of FXYD6 not only positively correlates with the increase of Na(+)/K(+)-ATPase but also coordinates with the activation of its downstream Src-ERK signaling pathway. More importantly, blocking FXYD6 by its functional antibody significantly inhibits the growth potential of the xenografted HCC tumors in mice, indicating that FXYD6 represents a potential therapeutic target toward HCC. Altogether, our results establish a critical role of FXYD6 in HCC progression and suggest that the therapy targeting FXYD6 can benefit the clinical treatment toward HCC patients.
Animals ; Antibodies, Monoclonal ; pharmacology ; Antineoplastic Agents ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Female ; HEK293 Cells ; Humans ; Ion Channels ; antagonists & inhibitors ; metabolism ; Liver Neoplasms ; drug therapy ; metabolism ; Mice, Inbred BALB C ; Mice, Nude ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays

CD146 is a newly identified endothelial biomarker that has been implicated in angiogenesis. Though in vitro angiogenic function of CD146 has been extensively reported, in vivo evidence is still lacking. To address this issue, we generated endothelial-specific CD146 knockout (CD146(EC-KO)) mice using the Tg(Tek-cre) system. Surprisingly, these mice did not exhibit any apparent morphological defects in the development of normal retinal vasculature. To evaluate the role of CD146 in pathological angiogenesis, a xenograft tumor model was used. We found that both tumor volume and vascular density were significantly lower in CD146(EC-KO) mice when compared to WT littermates. Additionally, the ability for sprouting, migration and tube formation in response to VEGF treatment was impaired in endothelial cells (ECs) of CD146(EC-KO) mice. Mechanistic studies further confirmed that VEGF-induced VEGFR-2 phosphorylation and AKT/p38 MAPKs/NF-κB activation were inhibited in these CD146-null ECs, which might present the underlying cause for the observed inhibition of tumor angiogenesis in CD146(EC-KO) mice. These results suggest that CD146 plays a redundant role in physiological angiogenic processes, but becomes essential during pathological angiogenesis as observed in tumorigenesis.
Animals ; CD146 Antigen ; genetics ; metabolism ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Female ; Fibrosarcoma ; metabolism ; pathology ; Male ; Melanoma, Experimental ; metabolism ; pathology ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; NF-kappa B ; metabolism ; Neovascularization, Physiologic ; drug effects ; Phosphorylation ; drug effects ; Proto-Oncogene Proteins c-akt ; metabolism ; Retinal Vein ; growth & development ; pathology ; Signal Transduction ; drug effects ; Transplantation, Homologous ; Vascular Endothelial Growth Factor A ; pharmacology ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

Animals ; Blood-Brain Barrier ; metabolism ; Ferritins ; metabolism ; Horses ; Mice ; Receptors, Transferrin ; metabolism ; Spleen ; chemistry