Acute Disease ; Administration, Inhalation ; Animals ; Disease Models, Animal ; Hypertension, Pulmonary ; drug therapy ; Male ; Nitric Oxide ; administration & dosage ; therapeutic use ; Nitrites ; administration & dosage ; therapeutic use ; Rats ; Rats, Sprague-Dawley

The intracellular trafficking and subcellular distribution of exogenous gene is very important for gene delivery. A successful gene vehicle should overcome various barriers including endosomal membrane barriers to delivery gene to the target organelle. Traditional nonviral vehicle is unable to avoid endosomal pathway efficiently, so the efficiency of gene delivery is low and the application of gene drugs is limited. In order to achieve efficient nonviral gene delivery, a lot of researches based on endosomal escape have been carried out and some agents with the function of endsomal escape have been found. These agents facilitate the endsomal escape via various mechanisms, such as fusion into the lipid bilayer of endosomes, pore formation in the endosomal membrane, proton sponge effect and photochemical methods to rupture the endosomal membrane. In this review, various reported strategies for endsomal escape are described according to the escape mechanisms, and their applications in intracellular gene delivery are also discussed.
Cell Membrane ; metabolism ; Endosomes ; metabolism ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; Humans

To establish neonatal rat cardiac fibroblast inflammatory secretion model by using LPS 100 microg x L(-1) combined with ATP 5 mmol x L(-1), in order to study the inhibitory effect of quercetin on the secretion of inflammatory factors TNF-alpha, IL-1beta and IL-6 of cardiac fibroblasts, further investigate the effect of quercetin on the protein expression of p-NF-kappaB p65 (S276) and p-Akt (S473) by western blot, and discuss the inhibitory effect of quercetin on the inflammatory secretion of cardiac fibroblasts. According to the findings, quercetin with the concentrations between 51.74 micromol x L(-1) and 827.81 micromol x L(-1) had no significant effect on the activity of cardiac fibroblasts. Quercetin with the concentrations of 82.78, 41.39, 20.70 micromol x L(-1) could notably inhibit the increase of TNF-alpha and IL-1beta induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 36 h (P < 0.01 or P < 0.05). Quercetin with the concentrations of 82.78, 41.39 micromol x L(-1) could notably inhibit the increase of IL-6 induced LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 36 h (P < 0.05), without any notable effect of quercetin with the concentration of 20.70 micromol x L(-1). Quercetin with the concentrations of 82.78, 41.39, 20. 70 micromol x L(-1) could notably inhibit the NF-kappaB p65 (S276) activation induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 15 min, with the most significant effect in 20.70 micromol x L(-1). Quercetin with the concentrations of 82.78, 41.39, 20.70 micromol x L(-1) could notably inhibit the increase of p-Akt(473) expression induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 240 min (P < 0.05). Therefore, this study believes that quercetin could attenuate the secretion of inflammatory factors TNF-alpha, IL-1beta and IL-6 of cardiac fibroblasts by inhibiting the activation of NF-kappaB p65 (S276) and Akt (473).
Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; Endomyocardial Fibrosis ; drug therapy ; genetics ; immunology ; Female ; Fibroblasts ; drug effects ; immunology ; Heart ; drug effects ; Humans ; Interleukin-6 ; genetics ; immunology ; Male ; Proto-Oncogene Proteins c-akt ; genetics ; immunology ; Quercetin ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; genetics ; immunology

To identify acupuncture resources in six databases of Cochrane Library (CL) with computer retrieve. Seventy-two literatures were identified in Cochrane Database of Systematic Reviews (CDSR). Among them, 12 Cochrane systematic review (CSR) verified the effectiveness of acupuncture, 29 concerning the indeterminacy of the efficacy of acupuncture with 1 didn't support acupuncture for epilepsy and 31 remained as protocols; 121 literatures were found in Database of Abstracts of Reviews of Effects (DARE) with more types of diseases or symptoms and rich modality comparing to CSR; 4218 randomized controlled trials and clinical controlled trials were identified in Cochrane Central Register of Controlled Trials (CCRCT); 43 literatures in Cochrane Methodology Register Database (CMRD) which focused on blindness study, quality assessment of methodology of research and publication bias and so on; 25 literatures in Health Technology Assessment Database (HTAD) and 18 in NHS Economic Evaluation Database (NHS EED) which were centered on acupuncture analgesia. Consequently, acupuncture literatures in 6 databases of CL do provide good resources for acupuncture researchers due to its abundant content, concrete classification and high quality evidence.
Acupuncture Therapy ; Databases, Factual ; Health Resources ; Humans ; Libraries

Township and Village Health Services Integration Management (TVHSIM) is an essential form of China's two-tiered health service integration plan at the township and village level.Its main purpose,also one of the target goals in China's new healthcare reform,is to gradually integrate rural health services and appropriately allocate rural health resources.This study aims to assess the village doctors' satisfaction with the TVHSIM and provide scientific base to further improve TVHSIM.A cross-sectional study was carried out in which 162 village doctors from Qinghai,Inner Mongolia and Xinjiang in western China were interviewed.Descriptive analysis,independent t-test,one-way ANOVA,Spearman rank correlation and multiple linear regression were used to analyze the difference and relevance between village doctors' personal characteristics and their satisfaction with TVHSIM and six subscales.Village doctors with different years of practice,social insurance status and essential medical knowledge level showed statistically significant differences in their satisfaction levels (all P＜0.05).Age (P＜0.05) and years of practice (P＜0.01) were negatively correlated with Drug and Medical Device Management and Financing Management.Essential medical knowledge level (P＜0.05) was negatively correlated with Operations Management as well.However,social insurance status (P＜0.05) was positively correlated with Human Resources Management and Drug and Medical Device management.Gender,age and years of practice respectively had significant influence on village doctors' satisfaction with TVHSIM (P＜0.01).In conclusion,in order to further promote TVHSIM policy in rural China,a well-rounded social insurance model for village doctors is urgently needed.In addition,the development of TVHSIM is regionally imbalanced.Efficient and effective measures aiming at rationalizing gender and age structure and enhancing essential medical training should be carefully considered.

OBJECTIVETo clarify the gene type of Orientia tsutsugamushi (Ot) from Shandong province.

METHODSNested-polymerase chain reaction (nPCR) was used to identify the gene type of 23 isolated Ot strains, 2 pools of homogenized leptotrombidium (L.) scutellare, 10 blood specimens of scrub typhus patients, and at the same time to compare with the international reference strains Gilliam, Karp, Kato. Sequencing analysis of the Sta56 gene was also used to further identify the precise gene types.

RESULTSOf the 35 samples, 33 had the same products in the amplification of template Ot-DNA. They all belonged to Kawasaki strains endemic in Japan while 2 (FXS4 and LHGM2 strain) belonged to Karp strains. The Sta56 gene sequence homologies to Japan Kawasaki strain of the 2 representative strains (B-16 and FXS2 strain) of the 33 samples were 94.22%, 95.21% respectively, but they were less than 75.87% to other prototype strains; The homologies to Karp strain of FXS4 and LHGM2 strain were 83.03%, 96.45% respectively. B-16 and FXS2 strain were designated as of types strain Japan Kawasaki, FXS4 and LHGM2 as Karp strain.

CONCLUSIONThe results indicated that the dominant Ot strains in Shandong Province were similar to Kawasaki strains, but Karp strains also existed.

Animals ; DNA, Bacterial ; genetics ; Genotype ; Humans ; Mice ; Orientia tsutsugamushi ; genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Scrub Typhus ; epidemiology ; microbiology ; Sequence Homology ; Serotyping

OBJECTIVETo investigate the effect of inhaled nitric oxide (NO) on surfactant protein A (SP-A) and mannose binding ability (MBA) in neonatal rats with hyperoxia-induced lung injury.

METHODSSixty-four neonatal rats were randomly exposed to room air (Control group), >95% oxygen for 6 days (Hyperoxia group), 10 ppm NO for 24 hrs (NO group), and >95% oxygen for 6 days along with 10 ppm NO for 24 hrs (Hyperoxia + NO group). After 2 and 6 days of exposure, the lung pathologic changes, gene and protein expressions of SP-A and MBA were measured.

RESULTSThe rats from the Hyperoxia group presented with obvious lung injuries. The SP-A expressions of mRNA (0.81 +/- 0.04 vs 1.53 +/- 0.25) and protein (59.45 +/- 18.37 vs 89.77 +/- 16.41) in the Hyperoxia group decreased significantly 2 days after exposure but increased significantly 6 days after exposure (SP-A mRNA 0.81 +/- 0.02 vs 0.63 +/- 0.03; SP-A protein 93.57 +/- 13.71 vs 47.73 +/- 21.69) compared with those of the Control group (P < 0.05). NO treatment alleviated the hyperoxia-induced pathologic injuries 2 days after exposure. The SP-A mRNA expression (0.55 +/- 0.91) in the Hyperoxia + NO group was significantly reduced as compared to both the Control and Hyperoxia groups (P < 0.05), and the SP-A protein expression (55.12 +/- 17.53) in the Hyperoxia + NO group was noticeably lower than that of the Control group (P < 0.01) 2 days after exposure. The SP-A protein expression in the Hyperoxia + NO group (67.33 +/- 18.59) was significantly lower than that of the Hyperoxia group 6 days after exposure (P < 0.05). Two days after exposure, the NO group had significantly higher MBA than the Control group (0.821 +/- 0.133 vs 0.58 +/- 0.158); the Hyperoxia + NO group had significantly higher MBA than the Hyperoxia group (0.43 +/- 0.175 vs 0.738 +/- 0.141) (P < 0.05).

CONCLUSIONSInhaled low dose NO may decrease SP-A protein expression and increase MBA of the lung tissue. This lessens the pathologic lung injury in neonatal rats with hyperoxia.

Administration, Inhalation ; Animals ; Animals, Newborn ; Hyperoxia ; pathology ; Lung ; drug effects ; metabolism ; pathology ; Mannose ; metabolism ; Nitric Oxide ; administration & dosage ; Pulmonary Surfactant-Associated Protein A ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the development of the sphincter muscle complex(SMC)and defecation function in pediatric patients with congenital anorectal malformations(ARM).Methods A total of 64 children underwent MRI,among whom 39 were patients with ARM,and the others were patients without ARM undergoing MRI because of other dieases.The dimensions of the SMC in different planes were evaluated with different sequences and coils.The relationship between the SMC development and the defecation function was investigated.Results In control group,the absolute value of SMC width was (3.63?0.22)mm,which had a high correlation with age(r=0.998,P0.05).The SMCs in intermediate ARM patients[muscle index(MI)=0.47?0.05]and low ARM patients(MI=0.49? 0.05)were well developed.The SMCs in a portion of patients with high ARM(MI=0.28?0.06)were poorly developed,when MI≤0.18,anorectal contraction pressurewas significantly lower(t=3.55, P0.18[(0.85?0.20)vs(2.24?1.02)kPa].The length of anal canal with high-pressure[(10.88?3.64)vs(20.26?4.34)mm]was shorter(t=5.18,P0.18,the anorectal angle was less than 90 degrees,and normal continent function was found in 21 of 23 cases(91%).Conclusion MRI can be employed to evaluate the development of SMC in patients with ARM,MI was an objective criteria to evaluate the development of SMC.When MI≤0.18, maldevelopment of SMC will be highly suspected.

OBJECTIVETo assess the value of fluorescent in situ hybridization (FISH) for detecting common chromosome aneuploidies in interphase nuclei of amniotic fluid cells.

METHODSEighty two uncultured amniotic fluid samples and supernatants from 2 successfully and 5 unsuccessfully cultured amniotic fluid samples were analyzed with FISH. Results from standard cytogenetic analysis of 79 uncultured amniotic fluid samples and 2 successfully cultured amniotic fluid samples were compared with FISH results.

RESULTSAll of the 89 samples were succeeded analyzed with FISH. Positive findings included 3 cases with trisomy 21, 1 case with 47, XYY and 1 case with 69, XXX, which were consistent with results of karyotype analysis.

CONCLUSIONFISH is a rapid and accurate method for prenatal diagnosis, and can also provide a remedy to failed amniotic fluid cells culture.

Adult ; Amniotic Fluid ; cytology ; Cell Culture Techniques ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Karyotyping ; Pregnancy

OBJECTIVETo investigate the effect of cytochrome P450 isozymes on aristolochic acid induced cytotoxicity on renal proximal tubular epithelial cell (cell line HK-2).

METHODHuman renal tubular cells (cell line HK-2), were treated with aristolochic acid (AA) alone or in combination with cytochrome P450 isozymes inhibitors, including alpha-naphthoflavone (CYP450 1A1 and 1 A2 inhibitors), ketoconazole (CYP450 3A4 inhibitor), sodium diethyldithiocarbamate (CYP450 2A6 and 2E1 inhibitors), quinidine (CYP450 2D inhibitor), alpha-lipoic acid (NADPH: P450 reductase inhibitor), sulfaphenazole (CYP450 2C inhibitor) in the presence or absence of liver microsome(S9). The inhibition of cell proliferation rate was studied by MTT assay and the lactate dehydrogenase release rate was determined with continuous monitoring method.

RESULTAA inhibits cell proliferation and promotes the release of LDH over the range of 12.5-100 mg x L(-1), in a dose-dependent manner. Addition of S9 into the culture system reduced AA cytotoxicity, with the cell proliferation inhibition reducing and the release of LDH decreasing (AA + S9 group vs the same concentration of AA alone group, P < 0.05). In the absence of S9, ketoconazole or alpha-naphthoflavone has no obvious effect on AA cytotoxicity, however,under the conditions of adding S9, ketoconazole or alpha-naphthoflavone enhances AA cytotoxicity. Other inhibitors of CYP450 isozymes has no distinct effect on AA cytotoxicity.

CONCLUSIONThe microsomal enzyme of Liver can reduce the AA cytotoxicity, and CYP450 3A, CYP450 1A may be the major cytochrome P450 isozymes which impact AA cytotoxicity.

Animals ; Aristolochic Acids ; toxicity ; Cell Line ; Cell Proliferation ; drug effects ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Cytotoxicity, Immunologic ; drug effects ; Enzyme Inhibitors ; pharmacology ; Humans ; Kidney Tubules ; drug effects ; enzymology ; immunology ; Male ; Rats ; Rats, Wistar