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.

Objective To research on the applicational value of quantitative immunofluorescence assay of high-sensitivity C- reactive protein(HS-CRP) detection in surveillance of ankylosing spondylitis(AS) and Rheumatoid arthritis(RA).Methods Using quantitative immunofluorescence technique to detect the serum HS-CRP among 43 patients with AS and 62 patients with RA and 30 healthy controls.Results Within-run CV% of repeatability were 4.98% and3.68% for lower and higher concentration of HS- CRP,respectively;between-run CV% of the stability were 4.57% and 5.42% for lower and higher concentration of HS-CRP, respectively;There was a good correlation between results of quantitative immunofluorescence assay and BNP special protein analyzer method(r=0.997);Both the level and the positive rate of HS-CRP of patients with AS and patients with RA were higher than those of the controls in the active phase.The level and the positive rate of HS-CRP in patients with AS and patients with RA were obviously higher in the active phase than in the non-active phase(P0.05).Conclusion There were significant correlations between HS-CRP and the activity of AS and RA,So HS-CRP was useful tools for monitoring the changes of patient's condition.It doesn't have much significance in differential diagnosis of AS and RA.

Objective To detect the influence of fluoride on the expression TM9SF1 mRNA and Ras mRNA of osteoblasts. Methods The third generation of primary cultured osteoblasts were exposed to a series concentrations of 0,2.5,5.0, 10.0,20.0 mg/L fluoride for 10 days. The influence of different doses of fluorine on the expression of TM9SF1 mRNA and Ras mRNA of osteoblasts cultured in vitro was investigated by SYBR Green I methods. Results The osteoblasts of the control group and the 2.5 mg/L group were in the shape of long spindle, triangle or irregular polygon and had processes, and the cytoplasm was translucent, adjacent cells affixed to each other under light microscope. Those of the 20.0 mg/L group shaped as long spindle or irregular polygon, and some vacuolization and granular materials appeared in cytoplasm. The number of the cells decreased and the volume increased significantly. After exposed to fluoride for 10 days, osteoblasts of 2.5 mg/L group morphologically proliferated. There were statistical siguificances between each groups of TM9SF1 mRNA in human osteoblasts(F = 322.82, P < 0.01). The highest in the 2.5 mg/L group(9326.0 ± 115.97), the expression of TM9SF1 mRNA decreased along with the increasing dose of fluorine. There were statistical significances between 5.0, 10.0,20.0 mg/L groups(6495.0 ± 323.9, 4387.5 ± 545.2, 5962.5 ± 536.7) and control group(9221.0 ± 107.5, all P< 0.01). There was a statistical significance between each groups of Ras mRNA in human osteoblasts(F = 703.28, P < 0.01). The highest in the control group, the expression of Ras mRNA decreased along with the increasing of dose of fluorine. There were statistical significance between 2.5, 5.0, 10.0, 20.0 mg/L groups(6144.5 ± 270.82,5603.5 ± 88.39,3181.0 ± 159.81,4067.5 ± 37.4) and control group(6571.0 ± 196.58). Conclusion The influence on TM9SFI mRNA and Ras mRNA expression in osteoblasts correlates with the dose of fluorine.

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

Introduction:The 2010 targets of the China Hepatitis B Prevention Programme were a prevalence of hepatitis B surface antigen (HBsAg) less than 1.0% for children less than five years old and less than 6.0% for the total population. This survey assessed the prevalence of Hepatitis B infection in Lianyungang, Jiangsu province, China in 2009–2010.Methods:Multistage sampling was used with 2372 subjects among 17 selected villages. Blood specimen collection and testing by enzyme-linked immunosorbnet assay (ELISA) were completed using the following markers for hepatitis infection: HBsAg and antibody to HBsAg (anti-HBs); hepatitis B e antigen (HBeAg) and antibody to HBeAg (anti-HBe); and hepatitis B core antibody (total anti-HBc). The data were analysed with Epi Info, version 3.3.2.Results:The prevalence of HBsAg was 2.4% (95% Confidence Interval [CI]: 1.8–3.0; Adjusted Prevalence [AP] 2.9%); anti-HBs prevalence was 51.1% (95% CI: 49.1–53.1; AP 49.2%) and total anti-HBc prevalence was 41.7% (95% CI: 39.8–43.7; AP 45.5%). The prevalence of HBsAg and total anti-HBc positivity increased from young to older age groups, yet the prevalence of anti-HBs positivity decreased from young to older age groups (P< 0.001 for all). There was no difference in the prevalences of HBsAg and anti-HBs among females and males (P= 0.108 and 0.089), but females had a higher prevalence than males for total anti-HBc positivity (P< 0.001). Discussion: This survey showed that in 2010 the prevalence of HBsAg among children aged less than five years was lower than the national target of 1.0% and that the prevalence of HBsAg for the total population was lower than the national target of 6.0%.

BACKGROUNDIntensive treatment such as autologous peripheral blood stem cell (PBSC) transplantation is an important therapeutic strategy in many hematologic malignancies. A number of factors have been reported to impact PBSC mobilization, but the predictive factors varied from one study to another. This retrospective study assessed our current mobilization and collection protocols, and explored the factors predictive of PBSC mobilization in patients with hematologic malignancies.

METHODSData of 64 consecutive patients with hematologic malignancies (multiple myeloma, n = 22; acute leukemia, n = 27; lymphoma, n = 15) who underwent PBSC mobilization for over 1 year were analyzed. Four patients with response to treatment of near complete remission or better were administered granulocyte colony-stimulating factor (G-CSF) to mobilize PBSCs. Sixty patients received G-CSF followed by chemotherapy mobilizing regimens. Poor mobilization (PM) was defined as when ≤ 2.0'10(6) CD34(+) cells/kg body weight were collected within three leukapheresis procedures.

RESULTSThe incidence of PM at the first mobilization attempt was 19% (12/64). The PM group was older than the non-PM group (median age, 51 vs. 40 years; P = 0.013). In univariate analysis, there were no significant differences in gender, diagnosis, and body weight between the PM and non-PM groups. A combination of chemotherapy and G-CSF was more effective than G-CSF alone as a mobilizing regimen (P = 0.019). Grade III or IV hematopoietic toxicity of chemotherapy had no significant effect on the mobilization efficacy. Supportive care and the incidence of febrile neutropenia were not significantly different between the two groups. In multivariate analysis, age (odds ratio (OR), 9.536; P = 0.002) and number of previous chemotherapy courses (OR 3.132; P = 0.024) were two independent negative predictive factors for CD34(+) cell yield. PM patients could be managed well by remobilization.

CONCLUSIONOlder age and a heavy load of previous chemotherapy are the negative risk factors for PBSC mobilization.

Adult ; Aged ; Female ; Granulocyte Colony-Stimulating Factor ; metabolism ; Hematologic Neoplasms ; metabolism ; pathology ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo research the optimal conditions for the callus induction of anther culture and the plant regeneration of Angelica dahurica var. formosana.

METHODCallus was induced from the anther of A. dahurica from Sichuan province on a MS medium. The effects of callus induction and plant regeneration of different pretreatment hours under low temperature (4 degrees C), different culturing conditions under darkness and illumination, and different culture with different hormone contents and ratios were studied.

RESULTThe results showed that A. dahurica anthers without low temperature pretreatment reached the highest induction rate then under the pretreatment under low temperature (4 degrees C) for two days. The optimal culturing condition was under the darkness. The culturing efficiency reached 38.89% on the medium of MS + 2.0 mg x L(-1) 2,4-D + 1.0 mg x L(-1) 6-BA. The optimum medium for differentiate anther callus was MS + 0.5 mg x L(-1) NAA + 1.5 mg x L(-1) KT + 10 mg x L(-1) AgNO3. 1/2MS medium supplemented with 0.5 mg x L(-1) IBA could well promote seedings to take roots.

CONCLUSIONAn efficient system for callus induction of anther culture and plant regeneration of A. dahurica was preliminarily established.

Angelica ; drug effects ; growth & development ; Culture Media ; chemistry ; pharmacology ; Flowers ; Tissue Culture Techniques ; methods

BACKGROUNDVanishing white matter disease (VWM), a human autosomal recessive inherited leukoencephalopathy, is due to mutations in eukaryotic initiation factor 2B (eIF2B). eIF2B is responsible for the initiation of protein synthesis by its guanine nucleotide exchange factor (GEF) activity. Mutations of eIF2B impair GEF activity at different degree. Previous studies implied improperly activated unfolded protein response (UPR) and endoplasmic reticulum stress (ERS) participated in the pathogenesis of VWM. Autophagy relieves endoplasmic reticulum load by eliminating the unfolded protein. It is still unknown the effects of genotypes on the pathogenesis. In this work, UPR and autophagy flux were analyzed with different mutational types.

METHODSERS tolerance, reflected by apoptosis and cell viability, was detected in human oligodendrocyte cell line transfected with the wild type, or different mutations of p. Arg113His, p. Arg269FNx01 or p. Ser610-Asp613del in eIF2Bε. A representative UPR-PERK component of activating transcription factor 4 (ATF4) was measured under the basal condition and ERS induction. Autophagy was analyzed the flux in the presence of lysosomal inhibitors.

RESULTSThe degree of ERS tolerance varied in different genotypes. The truncated or deletion mutant showed prominent apoptosis cell viability declination after ERS induction. The most seriously damaged GEF activity of p. Arg269FNx01 group underwent spontaneous apoptosis. The truncated or deletion mutant showed elevated ATF4 under basal as well as ERS condition. Decreased expression of LC3-I and LC3-II in the mutants reflected an impaired autophagy flux, which was more obvious in the truncated or deletion mutants after ERS induction.

CONCLUSIONSGEF activities in different genotypes could influence the cell ERS tolerance as well as compensatory pathways of UPR and autophagy. Oligodendrocytes with truncated or deletion mutants showed less tolerable to ERS.

Cell Line ; Endoplasmic Reticulum Stress ; genetics ; physiology ; Eukaryotic Initiation Factor-2B ; genetics ; Humans ; Mutation ; genetics ; Oligodendroglia ; metabolism ; Unfolded Protein Response ; genetics ; physiology

Background: Feline mammary carcinoma is the third most common cancer that affects female cats. Objectives: The purpose of this study was to screen differential serum proteins in feline and clarify the relationship between them and the occurrence of feline mammary carcinoma. Methods: Chinese pastoral cats were used as experimental animals. Six serum samples from cats with mammary carcinoma (group T) and six serum samples from healthy cats (group C) were selected. Differential protein analysis was performed using a Label-free technique, while parallel reaction monitoring (PRM) was performed to verify the screened differential proteins. Results: A total of 82 differential proteins were detected between group T and group C, of which 55 proteins were down regulated and 27 proteins were up regulated. Apolipoprotein A-I, Apolipoprotein A-II (ApoA-II), Apolipoprotein B (ApoB), Apolipoprotein C-III (ApoC-III), coagulation factor V, coagulation factor X, C1q, albumen (ALB) were all associated with the occurrence of feline mammary carcinoma. Differential proteins were involved in a total of 40 signaling pathways, among which the metabolic pathways associated with feline mammary carcinoma were the complement and coagulation cascade and cholesterol metabolism. According to the Label-free results, ApoB, ApoC-III, ApoA-II, FN1, an uncharacterized protein, and ALB were selected for PRM target verification. The results were consistent with the trend of the label-free. Conclusions This experimen is the first to confirm ApoA-II and ApoB maybe new feline mammary carcinoma biomarkers and to analyze their mechanisms in the development of such carcinoma in feline.

Objective To investigate the relationship between white blood cell count,neutrophils ratio and erythrocyte sedimentation rate and short outcomes among patients with acute ischemic stroke at admission to the hospital.Methods A total of 2675 acute ischemic stroke patients were included in this study.Data on demographic characteristics,life style,history of disease,white blood cell count( WBC),neutrophils ratio(NEUR),erythrocyte sedimentation rate(ESR) and clinical outcomes were collected for all the participants.Poor clinical outcome was defined as neurologic deficiency (NIHSS≥5) at discharge or death during hospitalization.Results White blood cell count,neutrophils ratio and erythrocyte sedimentation rate were higher in patients with poor outcome than m those without clinical outcome.According to the quartile range,WBC,NEUR and ESR were divided into four levefs at admission.After adjustment for multivariate,compared with WBC≤5.6 × 109/L,the odds ratio (95％ confidence intervals) of poor outcome with ≥8.7 × 109/L was 1.883 (1.306-2.716).When compared with NEUR≤0.56,the odds ratio (95％ confidence intervals) of poor outcome with 0.57-0.64 and with ≥0.74 were 1.572 (1.002-2.466) and 2.577 ( 1.698-3.910),respectively.When compared with ESR≤4 mm/h,the odds ratio (95％ confidence intervals) of poor outcome with ≥17 mm/h was 2.426 (1.233-4776).Elevated WBC count and NEUR at admission were significantly and positively associated with poor clinical outcomes among patients with acute ischernic stroke (trend test P＜0.05).Elevated ESR was not significantly or positively associated with poor clinical outcomes among patients with acute ischemic stroke (trend test P＞0.05).Conclusion There appeared associations between WBC,NEUR,ESR and poor outcome among patients with acute ischemic stroke at admission to the hospital.Both elevated WBC count and NEUR showed significantly positive association with poor clinical outcomes among patients with acute ischemic stroke at admission.