OBJECTIVETo construct a multiple myeloma (MM)-specific APE1siRNA expression vector, and detect the specific knock-down effect of the siRNA on expression of APE1 protein.

METHODSAPE1siRNA cDNA sequence was designed, synthesized and inserted into pSilencer 2.0-U6 linear expression vector. pSilencer APE1siRNA was digested by enzyme EcoRI and BamHI, then linear vector and IgP fragments were conjugated by T4 DNA ligase. pSilencer IgP-APE1siRNA and pSilencer IE-IgP-APE1siRNA were digested by enzyme EcoRI or XhoI. Linear vector and IE or Kappa fragments were conjugated by T4 DNA ligase. Then a MM specific pSilencer K-IE-IgP-APE1siRNA was cloned. The recombinant products were identified by DNA sequencing and enzyme digestions at each step. pSilencer K-IE-IgP-APE1siRNA plasmid was transfected to KM3, HOS, MDA-231 cells by liposome. APE1 gene silence induced by RNAi was analysed by Western blot.

RESULTSAPE1 protein in KM3 cells could be knocked down effectively and specifically by pSilencer K-IE-IgP-APE1siRNA vector. After 2 days, the level of APE1 protein in KM3 cells transfected with siRNA was 0.118 +/- 0.047, while that transfected with plasmid only was 0.988 +/- 0.029. The efficiency of gene silence was 90%.

CONCLUSIONA MM specific APE1siRNA expression vector was successfully constructed.

Base Sequence ; Cell Line, Tumor ; Cloning, Molecular ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; genetics ; Genetic Vectors ; genetics ; Humans ; Molecular Sequence Data ; Multiple Myeloma ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

BACKGROUNDCurrently anti-inflammatory therapy with steroids for allergic rhinitis need long-term repeated administration, although it is effective. Gene therapy is being suggested to substitute it. The aim of this study was to investigate nonviral vector mediated exogenous gene expression in COS-7 cells in vitro and the effect of intranasal mouse interleukin (mIL)-12 transgene expression on allergen induced eosinophil infiltration of nasal mucosa in a murine model of allergic rhinitis.

METHODSIn vitro COS-7 cells were infected with Epstein-Barr virus (EBV)/lipoplex. The expression of IL-12 p70 in cell culture supernatant was examined by enzyme-linked immunosorbent assay (ELISA). In mice with ovalbumin (OVA) induced allergic rhinitis, EBV/lipoplex was administered by nasal drops before OVA challenge once a day from day 1 to day 10. The expression of IL-12 mRNA and protein, the change of eosinophil count in nasal mucosa and serum total IgE were measured 24 hours after the last challenge.

RESULTSEBV/lipoplex could effectively transfect COS-7 cells. The expression of IL-12 p70 in cell culture supernatant was significantly more than in blank control. IL-12 via EBV plasmid vector transduction could be overexpressed in vivo. In pGEG.mIL-12 treated models, the nasal mucosa revealed a high level of widespread mIL-12 transduction by immunohistochemistry and in situ hybridization. Histological evaluation revealed marked suppression of eosinophil infiltration in nasal mucosa. The eosinophil count in allergic rhinitis group [(26.5 +/- 9.8)/high-power field (HPF)] was significantly increased over control group [(0.40 +/- 0.52)/HPF] (F = 56.94, P < 0.01), while the count in IL-12 gene therapy group [(4.60 +/- 2.63)/HPF] was significantly less than that of allergic group (F = 56.9, P < 0.01). Serum total IgE between in gene therapy mice [(88.83 +/- 6.71) ng/ml] and allergic rhinitis mice [(103.1 +/- 5.7) ng/ml] showed a significant difference (F = 1216, P < 0.05).

CONCLUSIONSNonviral EBV plasmid vector, pGEG.mIL-12 was able to overexpress exogenous gene both in vitro and in murine nasal mucosa in vivo. IL-12 overexpression via EBV/lipoplex could stem allergen induced eosinophil infiltration in nasal mucosa in murine models of allergic rhinitis, which may suggest a new cytokine immunogenetic therapy for allergic rhinitis.

Administration, Intranasal ; Animals ; COS Cells ; Cercopithecus aethiops ; Eosinophilia ; therapy ; Genetic Therapy ; Herpesvirus 4, Human ; genetics ; Immunoglobulin E ; blood ; Interleukin-12 ; genetics ; Lipids ; administration & dosage ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Perennial ; therapy ; Rhinitis, Allergic, Seasonal ; therapy

OBJECTIVETo investigate the role of PGE(2) and cAMP in the postburn change in granulopoiesis in bone marrow in burned mice with endotoxemia.

METHODSOne hundred and seventy eight mice were randomly divided into burn with LPS administration, simple burn, simple LPS administration and control (injection of normal saline) groups. The COX-2 expression and the contents of PGE(2) and cAMP in myeloid cells in injured mice in all groups were determined by RIA (radioimmuno-assay) within 1 postburn week and immunohistochemistry methods. At the same time the change in granulopoiesis was dynamically observed.

RESULTSThe granulopoiesis was enhanced slightly at the early stage of burn and with endotoxin challenge, followed by suppression. The COX-2 expression in myeloid cells the contents of PGE(2) on supernatant of marrow cells and intracellular cAMP in the myeloid cells was increased at 12 postburn hour (PBH) up to 5 postburn day (PBD). Furthermore, the change in the cAMP was evidently and positively correlated with that of PGE(2) (r = 0.978, P < 0.01), but was negatively correlated with that of CFU-GM (r = -0.971, P < 0.01)

CONCLUSIONPGE(2) might play pivotal roles in the postburn granulopoiesis suppression in bone marrow during endotoxemia. This effect might be accomplished by its ligating to its special receptor and to activate adenylate cyclase so as to increase the intracellular content of cAMP in bone marrows.

Animals ; Bone Marrow Cells ; pathology ; Burns ; complications ; metabolism ; Cyclic AMP ; metabolism ; Cyclooxygenase 2 ; metabolism ; Dinoprostone ; metabolism ; Endotoxemia ; etiology ; metabolism ; Granulocytes ; pathology ; Male ; Mice ; Mice, Inbred Strains

Objective To investigate the effect of high power microwave(HPM) irradiation on neuropeptide Y (NPY) and neural nitric oxide synthase (nNOS) expression in the cerebral cortex and hippoeampus of Wistar rats. Methods A total of 110 Wistar rats were used for this study.Three groups of 30 Wistar rats were exposed to HPM irradiation at intensities of 3,10,30 and 100 mW/cm~2,respectively.Twenty rats served as controls and were ex- posed to sham HPM irradiation.At 6 h,and at 1,3,7,14 and 28 d after irradiation,five rats from each group were sacrificed,and their cerebral cortices and hippocampi were harvested.HE staining was used to highlight any change in the structure of the cerebral cortex or hippocampus.Immunohistochemistry techniques and image analysis were used to study the changes in NPY and nNOS expression.Results 10 to 100 mW/cm~2 HPM irradiation caused pyc- nosis and deep staining of some neurons in the cerebral cortex and hippocampus.The increase in nNOS expression and decrease in NPY expression observed were significant at 3 days after irradiation.Conclusion HPM irradiation can induce injury in neurons of the cerebral cortex and hippoeampus,and abnormal NPY and nNOS expression.

A new fungus, Ophiocordyceps alboperitheciata, parasitic on the larva of Noctuidae (Lepidoptera) was identified from a survey of entomopathogenic fungi in Kunming Wild Duck Forest Park, Yunnan Province, China. It can be primarily distinguished from relatives by its longer fertile parts, sterile tips, superficial perithecia, narrower asci, and smaller septa of ascospores. As revealed from phylogenetic analyses inferred from nrSSU, nrLSU, tef-1α, rpb1, and rpb2 sequence data, O. alboperitheciata belongs to the Hirsutella citriformis clade in the genus Ophiocordyceps of Ophiocordycipitaceae, and forms a separated clade from other related species. The uniqueness of the taxon is significantly evidenced by both molecular phylogeny and morphology. Furthermore, the interspecific relationships in the H. citriformis clade are discussed.

A new fungus, Ophiocordyceps alboperitheciata, parasitic on the larva of Noctuidae (Lepidoptera) was identified from a survey of entomopathogenic fungi in Kunming Wild Duck Forest Park, Yunnan Province, China. It can be primarily distinguished from relatives by its longer fertile parts, sterile tips, superficial perithecia, narrower asci, and smaller septa of ascospores. As revealed from phylogenetic analyses inferred from nrSSU, nrLSU, tef-1α, rpb1, and rpb2 sequence data, O. alboperitheciata belongs to the Hirsutella citriformis clade in the genus Ophiocordyceps of Ophiocordycipitaceae, and forms a separated clade from other related species. The uniqueness of the taxon is significantly evidenced by both molecular phylogeny and morphology. Furthermore, the interspecific relationships in the H. citriformis clade are discussed.

Conventional medical experiments can hardly simulate cardiac excitation propagation and observe the evolvement of cardiac electrical activities firsthand as is possible with computer simulation. Based on the anatomic structure of the heart, simulation of cardiac electrical activity mainly consists of the emulation of the excitation process among the cardiac cells and calculation of the electrical activities of individual cardiac cells. In this study we establish a geometric ventricular structure model demonstrating the direction of the cardiac muscle fibers and the layers of the ventricular cells, and endow different action potential models to the ventricular cells of different layers, and observe the activation process of the ventricular parts in view of the three-dimensional anatomy. This method gives attention to both enough calculation amounts and efficiency, which achieves satisfactory simulation results of ventricular electrical activity based on the anatomic structure and cell electrophysiology through an improved algorithm on personal computer.
Algorithms ; Computer Simulation ; Electrophysiology ; Heart ; anatomy & histology ; physiology ; Humans ; Models, Anatomic ; Models, Cardiovascular

Objective To evaluate the correlation between lesions of dorsal artery of foot and type 2 diabetes on CDFI. Methods Dorsal artery of foot was examinated in 97 cases with type 2 diabetes and 46 cases without diabetes mellitus. Results There were variable changes in intima-media of dorsal artery of foot in type 2 diabetes patients.And the patients with hyperlipidemia and hypertension showed serious lesions in the dorsal artery of foot and bifurcation of the blood vessel .Simple arteriosclerosis showed not only lesions in intima-media of dorsal artery of foot but also relative mild lesions in bifurcation of the blood vessel. Blood vessel lesion incidence in type 2 diabetes was significantly higher than that of the control group (P<0.01). Conclusions There was a significant correlation between lesions of dorsal artery of foot and type 2 diabetes patient's condition. CDFI is an effective method in evaluating patient's condition, degree and prognosis of type 2 diabetes, and has an important clinical value in early diagnosis, prognosis and treatment.Simple arteriosclerosis showed focal lesions in bifurcation of the blood vessel while dorsal artery of foot showed relatively mild lesions .

OBJECTIVETo evaluate the bowel control of the anus-preserving operation for elderly patients over 75 years with low rectal cancer.

METHODSThirty-nine elderly patients over 75 years with low rectal carcinoma (4-7 cm from anal verge) were treated during the study period. The patients were divided into different groups according to the surgical procedures and anastomotic locations. The bowel control and patients satisfaction were compared.

RESULTSThe time of recovering normal defecation frequency was (9.8+/- 2.9) months. There were no differences in bowel control and anorectal manometric findings between the lower anastomosis group and super-lower anastomosis group, the lower anastomosis group and anorectal anastomosis group. The patients in anorectal anastomosis group displayed significantly better bowel control and anorectal manometric findings than those in the super-lower anastomosis group (P< 0.05). The time of recovering normal defecation frequency in colonic J-pouch-anal anastomosis group was (7.7+/- 1.7) months, shorter than (10.6+/- 2.8) months in direct anastomosis group (P< 0.01). The complication rate of I degree incontinence was 36.1%, but there was no difference between the two groups. The anorectal manometric findings were better in J-pouch-anal anastomosis group than those in direct anastomosis group (P< 0.05).

CONCLUSIONColonic J-pouch-anal anastomosis for lower rectal carcinoma can significantly improve the bowel control in a short term without increasing the complication rate.

Aged ; Aged, 80 and over ; Anal Canal ; surgery ; Anastomosis, Surgical ; Defecation ; Fecal Incontinence ; etiology ; Female ; Humans ; Male ; Postoperative Period ; Rectal Neoplasms ; physiopathology ; surgery

OBJECTIVETo study the association of ORMDL3 single nucleotide polymorphisms (SNP) with lysophosphatidylcholine (LysoPC) and apolipoprotein B (apoB) levels.

METHODSA total of 300 children diagnosed with bronchial asthma between January 2010 and December 2012 were selected for the asthma group, and 298 children diagnosed with upper respiratory tract infection in the same period were selected for the control group. Serum LysoPC and apoB levels were measured using enzyme-linked immunosorbent assay. Genotype analysis was performed using the TaqMan probe.

RESULTSLysoPC and apoB levels were significantly higher in the asthma group than in the control group (P<0.01). Among children with various genotypes of ORMDL3 gene at locus rs12603332, the asthma group had significantly higher LysoPC and apoB levels than the control group (P<0.01). Among the children with asthma, those with CC genotype had significantly higher LysoPC and apoB levels than those with CT and TT genotypes (P<0.01).

CONCLUSIONSLysoPC and apoB may intervene in the pathological process of asthma. Pro-inflammatory gene ORMDL3 SNP rs12603332 may be associated with high LysoPC and apoB levels, which leads to the occurrence of childhood asthma.

Apolipoproteins B ; blood ; Asthma ; blood ; genetics ; Child ; Child, Preschool ; Female ; Humans ; Lysophosphatidylcholines ; blood ; Male ; Membrane Proteins ; genetics ; Polymorphism, Single Nucleotide