Aged ; Cardiovascular Diseases ; diagnosis ; etiology ; Cerebrovascular Disorders ; diagnosis ; etiology ; Diagnostic Errors ; Female ; Humans ; Laryngitis ; complications ; diagnosis ; Middle Aged

Alprostadil ; administration & dosage ; adverse effects ; therapeutic use ; Hearing Loss, Sudden ; drug therapy ; Humans ; Injections, Intravenous ; Lower Extremity ; Male ; Middle Aged ; Pain ; etiology

Objective To construct mouse granulocyte-macrophage colony stimulating factor (mGM-CSF) gene eukaryotic expressing plasmid pcDNA3-GM-CSF, to transfect the recombinant into erythroleukemia cell line FBL-3, and identify their biological activity.Methods GM-CSF gene eukaryotic expressing plasmid was constructed by subclone and recombinant was transfected into FBL-3 cells by electroporation. After screening by G418 and cloning by limiting dilution,we obtained positive cell clones(FBL-3-GM-CSF). PCR and RT-PCR were used to identify the integration and stable expression of GM-SF gene in FBL-3-GM-CSF cells. The biological activity was confirmed by the hematopoietic progenitor cell proliferative assay and hematopoietic progenitor cell colony formation assay. Results Mouse GM-CSF cDNA was amplified from the prokaryotic expressing plasmid PET-30a(+)-GM-CSF by PCR firstly and BamH Ⅰ and EcoRⅠrestriction sites were introduced. The inserted fragment was cut by BamH Ⅰ and EcoR Ⅰ digestion and ligated into pcDNA3 vector. The pcDNA3-GM-CSF eukaryotic expressing plasmid was constructed. The recombinant was cleared with appropriate endoneucleases and sequenced. The findings showed that the orientation of the insert was correct, while no rearrangement or mutation was found. PCR and RT-PCR assay showed that GM-CSF gene had integrated into FBL-3-GM-CSF cells and stably expressed. The hematopoietic progenitor cell proliferative assay and hematopoietic progenitor cell colony formation assay demonstrated that the cultured supernatant of FBL-3-GM-CSF cells of expressing GM-CSF should obviously stimulate proliferation of murine marrow mononuclear cells, and could stimulate hematopoietic progenitor cell colony formation. The number of colony formation was 54.67?4.83. The rate of colony formation was 0.547 %.Conclusions GM-CSF gene eukaryotic expressing plasmid is constructed successfully. A cell clone, which can express stably GM-CSF gene and possess biological activity,is obtained. Our studies have founded the base for the preparation of GM-CSF gene-modified vaccine of tumor cell and the study of feasibility of immune therapy of leukemia.

Amnion ; surgery ; transplantation ; Animals ; Humans ; Neurons ; transplantation ; Spinal Cord ; transplantation ; Spinal Cord Injuries ; surgery ; Tissue Transplantation ; methods

Anemia, Refractory ; Humans ; Male ; Middle Aged ; Thrombocytosis

As the major commercial source of natural rubber,Hevea brasiliensis attracts much attention.However,the heterozygous nature,long breeding cycle are strong limitations for conventional breeding.While genetic engineering,which can be used to widen the germplasm base and produce desirable agronomic traits quickly and efficiently,offers a viable alternative approach to complement traditional breeding.Comprehensive analysis indicates that in the past two decades,with calli derived from immature anther or integumental tissues of immature fruit as receptors,both biolistic and Agrobacterium-mediated transformation methods were employed for developping rubber genotypes with improved latex yield,tolerance to tapping panel dryness syndrome,producing high-value recombinant proteins,etc.Being recalcitrant to tissue culture,the transformation efficiency of Hevea is comparatively low,and the procedures are still needed to optimize.Finally,breeding objectives and strategies to improve transformation efficiency were also proposed in the review.

To study the role of anti- C. albicans IgY and serum in protection of C. albicans infection of several animal models. To develop three animal models of C. albicans infection: a burned rat model of C. albicans infection, a mouse model of vaginal candiasis and a immunosuppression mouse model of C. albicans infection. And we compared the contribution of anti- C. albicans IgY and serum to the clearance of the C. albicans in three animal models of C. albicans infection. Anti- C. albicans IgY can protect against C. albicans infection in a burned rat model of C. albicans infection and a mouse model of vaginal candidiasis. The serum can effectively protect the mice from disseminated candidiasis in a immunosuppression mouse model. Humoral immunity component involving anti- C. albicans IgY and serum protect against C. albicans infection in a burned rat model of C. albicans infection ,a mouse model of vaginal candidiasis and a immunosuppression mouse model of C. albicans infection.

Objective To investigate the influence of citalopram on the fast response action potential,slow response action potential,in vitro electrocardiogram(ECG) and in vivo ECG of cardiac myocytes,and explore its mechanism of adverse cardiac effects. Methods Conventional microelectrode technique was employed to record the fast and slow response action potentials of the isolated papillary muscles of guinea pigs.In vivo and in vitro ECG were recorded from anesthetized animals and Langendorff-perfused hearts,respectively. Results Citalopram could prolong the RR interval and QRS duration of in vivo ECG.The premature ventricular contraction and atrial ventricular block were induced by 12.5?10-6 mol/L citalopram.The maximum ascending velocity of 0 phase(Vmax),action potential amplitude(APA) and action potential duration(APD50 and APD90) were dose-dependently decreased by citalopram in the fast and slow response action potentials of guinea pigs,respectively. Conclusion Citalopram can inhibit sodium and calcium channels effectively,which may be the ionic mechanism that citalopram induces arrhythmia in the clinical practice.

Objective To analyze plague epidemic tendency in the Three-River Region of Qinghai.Methods Using retrospective study,the Three-River Region during 1954-2006 year pestis epidemic focus were investigated and analyzed.Result Pestis prevailed mainly in Yushu,Chindu,Qumalai,Nangqian,Zhiduo and the Geermu.Tanghla Township.It was first found that the nature plague focus of miefitus existed in Chengduo County.There are 1 5 kinds of 12 branches in 8 trees infected plague animals were founded,336 Yersinia pestis were separated from the driven objects.Among them there were 291 Himalayas marmot body,account for 86.60%of the total,13 of Tibet sheep,accounts for 3.87%.10 of Qinghai field-mouse,accounts for 2.98%,Also there were 114 Yersinia pestis which were separated from each kind of vector insect in vivo.And,46 pestis strains came from the axe shape of flea in vivo account for 40.35%(46/114),38 pestis strains separated from Xie mountain flea,account for 33.33% (38/114).During 1960-2006 years there were 85 human plague cases were founded,238 occurred,134 died,the case fatality rate wero 56.30%(134/238),the popular seasons were started from May to November,the peak season happened in Aug and Sep.After Oct mainly due to Tibet sheep pestis which will cause as the origin of infection.The majority of sickness was pulmonary plague,account for 49.58%(117/238),whereas the first round case caused by the gland bubonic plague,account for 77.12%(91/118).Conclusions There are two pestis strains natural epidemic focus places in Three-River Source Region of Qinghai including the Himalayas marmot pestis strain and the Qinghai field-mouse pestis strain.The case of human pestis strain causes by the marmot strain,the fiehl-mouse mold mushroom spawn causes human pestis strain has not yet discovered,Three-River Source Region of Qinghai is a pestis strain key popular area in Qinghai Province.

Adult ; Humans ; Lipomatosis, Multiple Symmetrical ; Male ; Middle Aged