Objective To construct the RNA interference(RNAi)recombinant adenovirus vector targeting at human hypoxia-inducible transcription factor 1?(HIF-1?)and to evaluate its effect on human lung adenocarcinoma cell line SPCA-1.Methods The recombinant adenovirus Ad was constructed.HIF-1? inserted with HIF-1? RNAi fragment via AdEasy system.The virus was purifed by CsCl gradient centrifuge.The functional titer of recombinant adenovirus was measured by transfection test in HEK 293 cells.SPCA-1 cells were transducted with 2 multiplicity of infection(MOI)Ad.HIF1? in vitro,the expression rate of green fluorescence protein(GFP)was recorded by flow cytometry,HIF-1? mRNA and protein level was measured by Real-Time RT-PCR and flow cytometry.ResultsThe recombinant shuttle plasmid PAdTrack.HIF-1? and adenovirus plasmid Ad.HIF-1? were all correct shown by enzyme digestion confirmation.The plasmid pAd.HIF-1? was transducted into HEK293 cells,15%GFP expressionwere seen after 3 days.The final titers of recombinant adenovirus were 5.0?1010 TU/mL.SPCA-1 cells was transducted by Ad.HIF-1? in vitro for 48 h,GFP expression rate was 92%,HIF-1? mRNA and protein level decreased 89% and 87%,respectively.Conclusion RNAi adenovirus vector of human HIF-1? gene has been successfully constructed,which could facilitate the research onHIF-1? gene related gene therapy for lung cancer.

Objective To study the changes of serum endotheline(ET),D-dimer and fibrinogen(Fbg) in infants with pneumonia complicated with heart failure(HF) and explore the changes of blood coagulation,fibrinolysis,and endothelial cell function.Methods Eighty patients with infant pneumonia complicated with HF and 20 controls were studied.Serum ET,D-dimer,Fbg,activated partial thromboplastin time(APTT),prothrombin time(PT) and thrombin time(TT) were measured.Results ET,D-dimer,Fbg,APTT in every group had differences.With the disease deteriorating,the values of ET,D-dimer,Fbg increased,but the values of APTT decresed gradually.There were positive relations between ET and D-dimer(r=0.42 P

Anti-Arrhythmia Agents ; therapeutic use ; Cardiomyopathies ; drug therapy ; etiology ; Child, Preschool ; Female ; Humans ; Moricizine ; therapeutic use ; Tachycardia ; complications ; drug therapy ; Treatment Outcome

Air ; analysis ; Chromatography, Gas ; methods ; Ethylene Oxide ; analysis ; Furans ; analysis ; Methylene Chloride ; analysis ; Workplace

AlM:To evaluate the efficacy of combined pranoprofen eye drops and artificial tears on the treatment of mild to moderate dry eye syndrome after trabbeculectomy.METHODS:This prospective case control study included 63 cases (63 eyes) of patients with mild to moderate dry eye syndrome after trabbeculectomy in our hospital from November 2013 to June 2013. All subjects were randomly divided into two groups. Observation group was treated with combined pranoprofen eye drops and artificial tears and control group received simple artificial tears marking the eyes at 1, 2, 4wk. The patient's symptoms, signs, BUT, S▏t, and FL were observed before treatment and 1, 2, 4wk after treatment.RESULTS: After 2wk, the symptoms of observation group were improved, there was statistically significant difference (P<0. 05). FL difference of each group was statistically significant ( P<0. 05 ); After 4wk, symptoms and signs were improved. There was statistically significant difference ( P < 0. 05 ). The BUT of the observation group and corneal FL scores of two groups showed significant differences (P<0. 05). CONCLUSlON: Artificial tears joint pranoprofen eye drops has good curative effect in the treatment of mild to moderate dry eye syndrome after trabbeculectomy.

Migrations of orthopedic wires to cardiovascular system are uncommon and rarely reported. We report a case of right ventricle embolization with the Kirschner wire that was used for right 2nd rib osteosynthesis 2 years and 8 months previously in a 50-year-old man. The patient was asymptomatic and migration of the Kirschner wire was discovered by routine chest X-ray. An 8 cm-long Kirschner wire was successfully retrieved from the right ventricle. The treatment strategy for Kirschner wire removal from right ventricle is discussed.
Bone Wires ; adverse effects ; Embolism ; etiology ; Foreign-Body Migration ; Heart Ventricles ; Humans ; Male ; Middle Aged ; Rib Fractures ; surgery ; Ribs

OBJECTIVETo analyze and predict the structure and function of mosquito densovirus (MDV) nostructual protein1 (NS1).

METHODSUsing different bioinformatics software, the EXPASY pmtparam tool, ClustalX1.83, Bioedit, MEGA3.1, ScanProsite, and Motifscan, respectively to comparatively analyze and predict the physic-chemical parameters, homology, evolutionary relation, secondary structure and main functional motifs of NS1.

RESULTSMDV NS1 protein was a unstable hydrophilic protein and the amino acid sequence was highly conserved which had a relatively closer evolutionary distance with infectious hypodermal and hematopoietic necrosis virus (IHHNV). MDV NS1 has a specific domain of superfamily 3 helicase of small DNA viruses. This domain contains the NTP-binding region with a metal ion-dependent ATPase activity. A virus replication roller rolling-circle replication(RCR) initiation domain was found near the N terminal of this protein. This protien has the biological function of single stranded incision enzyme.

CONCLUSIONThe bioinformatics prediction results suggest that MDV NS1 protein plays a key role in viral replication, packaging, and the other stages of viral life.

Animals ; Computational Biology ; Culicidae ; virology ; Densovirus ; chemistry ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Protein Structure, Secondary ; Protein Structure, Tertiary ; Viral Nonstructural Proteins ; chemistry ; genetics

Objective To study interleukin-23 (IL-23) levels in serum and dendritic cells of acute-on-chronic liver failure (ACLF) patients with chronic hepatitis B (CHB) and to explore its relationship with the prognosis.Methods Peripheral blood mononuclear cells and serum were collected from 40 ACLF patients with CHB (including survival group 27 cases and non-survival group 13 cases) and 26 healthy controls.Monocytes were induced to immature dendritic cell in vitro and TNF-α was added to induce dendritic cell maturation.IL-23 mRNA of dendritic cells was detected by real time polymerase chain reaction (PCR), and serum IL-23 level was measured by enzyme-linked immuno sorbent assay (ELISA).Differences among the parameters with normal distribution were compared using t test, those with non-normal distribution were compared using non-parametric Mann-Whitney U-test, and the relationship between two variables was assessed by Spearman′s rank correlation.Results International normalized rate (INR) and model for end-stage liver disense (MELD) scores in non-survival group of ACLF were higher than those in survival group (INR: 2.32 vs 1.64, U=69.00, P=0.002 2;MELD:36 vs 30, U=64.50, P=0.001 4).However, there were no significant differences between two groups at gender, age, alanin aminotransferase (ALT), aspartate aminotrans ferase (AST), bilirubin, creatinine, hepatitis B virus (HBV) DNA, hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and serum IL-23.IL-23 mRNA level in dendritic cells at baseline in non-survival group of ACLF was significantly higher than that in survival group (76 vs 43, U=71.50, P=0.002 8).After treatment, serum IL-23 was significantly declined in survival group ([160±75] ng/L vs [91±49] ng/L, t=4.012, P=0.000 2), but not in non-survival group.Significant positive correlation was observed between IL-23 mRNA level in dendritic cells and MELD score at baseline (r=0.7198,P<0.01).Conclusions Persistent high serum IL-23 level suggests poor prognosis in ACLF patients with CHB.IL-23 mRNA expression in dendritic cells has good consistency with MELD score and the patients with high IL-23 mRNA expression has poor outcome.

Objective: To observe the clinical efficacy of acupuncture plus MOTOmed intelligent motor training in treating children with spastic cerebral palsy, and analyze the effects on lower limb motor function, intelligence development level, immune function and cerebral hemodynamics.Methods: A total of 42 children with spastic cerebral palsy were selected as the observation objects, and enrolled into the observation group. Another 42 cases treated in the same period were selected as the control group. Both groups received MOTOmed intelligent motor training, and the observation group was given additional acupuncture therapy, and the control group was given additional conventional rehabilitation treatment. After 2 consecutive treatment courses, the psychomotor development index (PDI) and mental development index (MDI) of Children's Developmental Center of China (CDCC) scale, the scores of gross motor function measure (GMFM) scale and modified Ashworth scale (MAS), and the changes in CD3+, CD4+, CD8+ and CD4+/CD8+ were observed. The peak systolic velocity (PSV) and mean flow velocity (MFV) of the anterior cerebral artery (ACA), middle cerebral artery (MCA) and posterior cerebral artery (PCA) were observed and measured. The clinical efficacy was evaluated. Results: Compared with the same group before treatment, the scores of GMFM, PDI and MDI, levels of CD3+, CD4+ and CD4+/CD8+, PSV and MFV levels of ACA, MCA and PCA in both groups were significantly increased after treatment (all P<0.05), while the CD8+ level had no significant change (both P>0.05). After treatment, the total effective rate of lower limb spasm in the observation group was 90.5％, significantly higher than 71.4％ in the control group (P<0.05). The scores of GMFM, PDI and MDI, the levels of CD3+ and CD4+, PSV and MFV, and the levels of ACA, MCA and PCA in the observation group were all significantly higher than those in the control group (all P<0.05). There were no significant differences in the CD8+ level and CD4+/CD8+ between the groups (all P>0.05). Conclusion: Acupuncture plus MOTOmed intelligent motor training has a better clinical efficacy than conventional rehabilitation plus MOTOmed intelligent motor training in treating children with spastic cerebral palsy, and is also superior in improving lower limb motor function and the level of intellectual development. And the mechanism may be related to the improvement of cerebral hemodynamics.

OBJECTIVETo construct an eukaryotic expression plasmid containing gp120 gene of HIV-1 subtype B and obtain gp120 gene expression in HepG2 cells.

METHODSAccording to the published gp120 gene sequence in Genbank, a pair of primers was designed and synthesized. The PCR amplification product of gp120 gene was cloned into pMD-18T vector using TA cloning followed by BamHI and XhoI digestion and sequence analysis. The target gene was then subcloned into a highly efficient eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid was sequenced and identified by restrictive endonuclease digestion, and transfected into HepG2 cells via liposome. The expression of gp120 gene was analyzed by RT-PCR and Western blotting, respectively.

RESULTSRestriction endonuclease digestion and sequence analysis verified successful construction of the recombinant vector pcDNA3.1(+)/gp120. The target fragment gp120 was identical with U26942 in Genbank, and the expression of gp120 gene was detected in the lysate of the transfected HepG2 cells by RT-PCR and Western blotting.

CONCLUSIONThe eukaryotic expression plasmid for gp120 has been constructed successfully, which is capable of stable expression in HepG2 cells.

AIDS Vaccines ; biosynthesis ; genetics ; Base Sequence ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cloning, Molecular ; Eukaryotic Cells ; metabolism ; Gene Expression ; HIV Envelope Protein gp120 ; biosynthesis ; genetics ; HIV-1 ; genetics ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Molecular Sequence Data ; Plasmids ; genetics ; Transfection ; Vaccines, DNA ; biosynthesis ; genetics