AIM: To explore the effects of high D-glucose on the mRNA and protein expression of lectin-like oxidized LDL receptor-1 (LOX-1) in endothelial cells and to study the possible mechanism by which diabetes is easily accompanied by vascular diseases. METHODS: Cell morphology was observed with inverted phasecontrast microscope. LOX-1 mRNA was detected by quantitative reverse transcription polymerase chain reaction in vascular epithelial cells cultured with different concentration D-glucose (7 5, 15 or 30 mmol/L) or with the same concentration D-glucose (15 mmol/L) in different time (0, 12, 24, 48, 72 or 96 h), LOX-1 protein expression was also examined by immunohistochemisty. RESULTS: Cells, exposed to high concentration D-glucose, were crude with clear profiles and bright granules in the plasma. Different high concentration D-glucose could induce LOX-1 mRNA expression and the effect of 30 mmol/L glucose on LOX-1 mRNA was the most significant (P

Objective To assess the relationship between serum vitamin D and autoimmune thyroid disease.Methods Subjects included total 520 persons receiving regular health examination,and serum calcium,phosphorus,parathyroid hormone (PTH),thyroid peroxidase autoantibody (TPOAb) and 25-dihydroxy vitamin Ds was measured.The incidence of 25-dihydroxy vitamin D3 deficiency (≤30 μ g/L)was observed.The relationship between 25-dihydroxy vitamin D3 deficiency and autoimmune thyroid disease was analyzed.Results The serum 25-dihydroxy vitamin D3 of all the subjects was (24.47 ± 7.21) μ g/L,and the incidence of 25-dihydroxy vitamin D3 deficiency (≤30 μg/L) was 61.15％ (318/520),and the positive rate of TPOAb was 21.54％ (112/520).The proportion of TPOAb ＞ 50 kU/L or ＞ 100 kU/L in subjects with 25-dihydroxy vitamin D3≤30 μ g/L was higher than that in subjects with 25-dihydroxy vitamin D3 ＞ 30 μg/L [25.79％(82/318) vs.19.80％(40/202) and 9.43％(30/318) vs.4.46％(9/202)],and there was significant difference (P ＜ 0.05).The relationship between 25-dihydroxy vitamin D3 and TPOAb was assessed and showed significant inverse correlation (r =-0.13,P ＜0.05).Conclusions Vitamin D deficiency is very common in the population,and autoimmune thyroid disease is related with vitamin D deficiency,which may has impact on the body's immune regulation.Specific mechanism and whether vitamin D supplementation can intervene and treat autoimmune thyroid disease need further study.

Objective To study the effects of high volume hemofiltration (HVHF) according to pulse-indicated continuous cardiac output (PiCCO) on patients with acute respiratory distress syndrome (ARDS).Methods A prospective randomly controlled trial was conducted.163 patients with ARDS admitted to Taizhou People's Hospital,Medical College,Nantong University,between February 2011 and January 2014,were enrolled.The patients were randomly divided into conventional therapy group (n＝ 50),HVHF group (n =55),and PiCCO + HVHF group (n＝58) by random number table.The patients in conventional therapy group received routine treatment including mechanical ventilation and drug treatment according to ARDS treatment guideline.The patients in the HVHF group received HVHF treatment of 18 hours per day on 1,3,5,7 days on the basis of conventional therapy.Patients in the PiCCO + HVHF group received HVHF treatment according to PiCCO.The indexes of lung function and PiCCO monitoring were recorded at intensive care unit (ICU) admission (before) and 4 days and 7 days after treatment.The serum levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were determined by enzyme linked immunosorbent assay (ELISA),and the prognosis of patients was recorded.Results In three groups,oxygenation index (PaO2/FiO2),static lung compliance (Cs) were gradually increased,and respiratory rate (RR),lactic acid (Lac) were gradually decreased.The indicators in HVHF and PiCCO + HVHF groups were significantly improved compared with conventional therapy group.The indexes in PiCCO + HVHF group were significantly increased or decreased compared with those in HVHF group,and the statistical differences were found on the 7th day after treatment [PaO2/FiO2 (mmHg,1 mmHg=0.133 kPa):189.3 ± 36.8 vs.166.3 ± 36.1,Cs (mL/cmH2O):76.7 ± 18.9 vs.67.0 ± 18.2,RR (times/min):16.4 ±5.2 vs.19.2 ± 5.4,Lac (mmol/L):1.20 ±0.41 vs.1.41 ±0.43,all P＜0.01].In PiCCO +HVHF group,cardiac index (CI) was gradually increased,and extra vascular lung water index (EVLWI) and intra thoracic blood volume index (ITBVI) were gradually decreased.There were significant differences in the indexes 4 days and 7 days after treatment compared with those before treatment [CI (L·min-1·m-2):4.62 ± 1.13,4.83 ± 1.10 vs.4.01 ± 1.02,EVLWI (mL/kg):7.6 ± 2.7,6.5 ± 2.6 vs.12.4 ± 2.9,ITBVI (mL/m2):801.3 ± 120.9,785.4 ± 118.7 vs.980.1 ± 168.6,all P＜0.01].After treatment,the serum levels of TNF-α and IL-1β in three groups were gradually decreased.Compared with the conventional therapy group,the serum levels of TNF-α and IL-1β on 4 days and 7 days in the HVHF and PiCCO + HVHF groups were significantly decreased,and the statistical differences were found on 7 days [TNF-α (ng/L):68.35 ± 12.63,67.54 ± 12.90 vs.85.35 ± 13.70; IL-1β (ng/L):424.6 ± 142.9,412.2 ± 140.2 vs.895.2 ± 187.7,all P＜0.01].Compared with the HVHF group,the serum levels of TNF-α and IL-1β in the PiCCO + HVHF group were slightly decreased without statistical differences.Compared with the conventional therapy group,the number of organ failure,duration of mechanical ventilation,the length of stay in ICU and hospital mortality in HVHF group and PiCCO + HVHF group were lowered,and the statistical differences were found in PiCCO + HVHF group compared with HVHF group [number of organ failure:2.41 ± 0.79 vs.2.72 ± 0.80,duration of mechanical ventilation (days):4.8 ± 2.0 vs.5.7 ± 2.1,the length of stay in ICU (days):11.5 ± 3.4 vs.13.1 ± 3.6,hospital mortality:31.0％ (18/58) vs.41.8％ (23/55),all P＜0.05].Conclusions Levels of inflammatory factors in patients with ARDS could be reduced by HVHF.The oxygenation and compliance of lung can be improved,the number of organ failure can be lowered,the duration of mechanical ventilation and the length of stay in ICU can be shortened,and the hospital mortality could be declined by PiCCO guided HVHF.

In order to obtain a virus-like particle vaccine both for porcine parvovirus (PPV) prevention and growth-promotion, VP2 gene of PPV NJ-a strain was amplified with PCR, and four copies of synthetic somatostatin gene were fused to the N-terminal of VP2 gene. The fused gene was cloned into pFast-HT A to construct the recombinant plasmid pFast-SS4-VP2, then the pFast-SS4-VP2 was transformed into DH10Bac competent cells and recombined with shuttle vector Bacmid, followed by identification with blue-white screening and PCR analysis for three cycles, and the positive recombinant was named as rBacmid-SS4-VP2. The positive Sf-9 cells were transfected with rBacmid-SS4-VP2 by Lipofectamine to produce recombinant baculovirus. When the cytopathic effect (CPE) was obvious, the transfected Sf-9 cell was harvested, and the positive recombinant virus was named as rBac-SS4-VP2. The insertion for the target gene into baculovirus genome was confirmed with PCR. SDS-PAGE and Western blotting revealed that the calculated protein of approximately 68 kDa was in the expressed in the insect cells. The Sf-9 cells infected with rBac-SS4-VP2 were stained positive against PPV antibody using the indirect immunofluorescence assay (IFA). Moreover, the virus particle self-assembly was observed under electron microscopy. 90 four-week-old mice were immunized by the recombinant protein coupled with different adjuvants alhydrogel, IMS and oil. VP2-specific ELISA antibodies, PPV-specific neutralizing antibody, somatostatin antibody and growth hormone levels were examined to evaluate the immunogenicity of this virus like particle. Results indicated that mice groups immunized rSS4-VP2 protein with alhydrogel and IMS developed similar humoral immune response comparing with inactived PPV vaccine. Mice group immunized with rSS4-VP2 generated higher level of SS antibody and growth hormone comparing with negative control, mice receiving rSS4-VP2 with alhydrogel developed the highest antibody titre than all other groups, while the oil group developed the lowest antibody level. This study provides not only a new rout for production of safe and effective virus like particle subunit vaccine, but also the foundations for peptide presentation and multivalent subunit vaccine design.
Animals ; Antigens, Viral ; biosynthesis ; genetics ; Artificial Gene Fusion ; Baculoviridae ; genetics ; Capsid Proteins ; biosynthesis ; genetics ; Mice ; Parvoviridae Infections ; prevention & control ; Parvovirus, Porcine ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Somatostatin ; genetics ; Swine ; Vaccines, Virus-Like Particle ; biosynthesis ; immunology ; Viral Vaccines ; biosynthesis ; immunology ; Virion ; genetics ; immunology

The aim of this study was to investigate the effects of miR-155 inhibitor transfection on the proliferation and apoptosis of THP-1 cells. The miR-155 inhibitor was transfected into THP-1 cells (THP-1I) by using X-treme GENE siRNA transfection reagent. Cells without transfection (THP-1C) and cells with negative transfection (THP-1IC) were used as controls. Quantitative real-time polymerase chain reaction (RT-PCR) was performed to detect the expression of miR-155 and relative expression of SHIP1 mRNA in the cells. Cell proliferation was assayed using CCK-8 method. Cell apoptosis were detected by flow cytometry. The expression of SHIP1, TAKT and pAKT in THP-1 cells were detected by Western blot. The results indicated that compared with THP-1C and THP-1IC, the expression of miR-155 in THP-1I cells was significantly reduced; miR-155 inhibition significantly increased apoptosis rate in THP-1 cells (P < 0.05) ; miR-155 inhibition in THP-1 cells caused no significant alteration in SHIP1 mRNA level but significantly increased its protein content, indicating some post-transcriptional modulations might exist underlying the modulation of miR-155 to SHIP1, the miR-155 caused significantly reduced protein level of pAKT (P < 0.05) without interfering TAKT protein content. It is concluded that the miR-155 inhibition may promote THP-1 cell apoptosis through increasing SHIP1 protein content and impairing its downstream PI3K/AKT signaling pathway. This study suggests that miR-155 inhibition may be a promising therapy strategy for treating acute myeloid leukemia (AML).
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Humans ; Leukemia ; genetics ; MicroRNAs ; genetics ; Phosphatidylinositol 3-Kinases ; RNA, Messenger ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; Transfection