Objective To explore the effects of BCG and Poly I:C co-vaccination on the development of spleen T cell subsets of neonatal BALB/c mice. Methods Neonatal BALB/c mice were inoculated with BCG and/or Poly I:C intraperitoneally within 2-3 d after birth. Four weeks later, spleen cells of mice were isolated and the percentage of CD3+ CD8+ IFN-γ+,CD3+ CD8-IFN-γ+,CD3+ CD8+ IL-4+,CD3+ CD8- IL-4+,CD4+ Foxp3+ T cells,which represent Tc1,TH1,Tc2,TH2,Treg cells,respectively,were tested by flow cytometry at single cell level,and the ratios of TH 1/TH 2 and Tc1/Tc2 were calculated. Results The percentages of TH1 and Tc1 cells of BCG-vaccinated mice,Poly I:C-vaccinated mice and BCG plus Poly I:C-vaccinated mice were significantly higher than that of control mice(P＜0.05 or P＜0.01),and there was no difference among the three vaccinated group. The ratios of TH1/TH2 and total IFN-γ/IL-4 of the three vaccinated groups were higher than that of control group,but not the ratio of Tc1/Tc2. The TH1/TH2 ratio of BCG plus Poly I:C-vaccinated group was higher than that of BCG-vaccinated group(P＜0.05).The percentages of Trge cells showed no difference among the four groups(P＞0.05). Conclusion BCG and Poly I:C co-vaccination can significantly increase the number of Tc1 and TH 1 cells and TH 1/TH2 ratio in spleen cells. BCG and Poly I:C vaccination may have a synergistic effect on TH 1/TH2 ratio of spleen cells in neonataI mice. The percentage of CD4+ Foxp3+ T cells among four groups showed no significant difference.

Methods hMSCs were isolated from human bone marrow by density gradient fractionation and adherence to plastic flasks. Individual colonies were selected and cultured in tissue dishes. Packaging cells PT67 were transfected by PLEGFP-N1 retroviral vector , and hMSCs were transduced by viral supernatant infection. Meanwhile, hMSCs-EGFP were identified by immune phenotypes and whether it could differentiate into osteoblasts or adipocytes under conditioned media was investigated. Results The rate of stably transduced hMSCs-EGFP was up to 96% after being screened by G418. hMSCs-EGFP exhibited fibroblast-like morphological features. Flow cytometric analyses showed that hMSCs-EGFP were positive for CD73, CD105, CD166, CD90 and CD44, but negative for CD34 and CD45. In addition, it could functionally be induced into osteocytes or adipocytes under conditioned media. These biological features of hMSCs-EGFP were consistent with those of hMSCs.Conclusions hMSCs transduced by PLEGFP-N1 retroviral vector can be used in vivo securely because they can maintain their biological characteristics and differentiation. It is a simple and reliable way to trace the changes of hMSCs in vivo by EGFP during cell transplantation and gene therapy.

Bone Marrow Cells ; cytology ; Cell Cycle ; Cell Differentiation ; Cell Nucleus ; genetics ; Cells, Cultured ; Diploidy ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Immunophenotyping ; Mesenchymal Stromal Cells ; cytology ; metabolism ; ultrastructure ; Transfection

OBJECTIVETo investigate effects of the variation of immune function in high humidity environment in different time, and lay a foundation for further study of the related mechanism.

METHODThirty SD rats were divided into 3 groups (n = 10): 20 day group, 40 day group in 90% relative humidity chamber and control group in normal relative humidity. Peripheral blood and spleens were collected to detect the levels of T lymphocyte subsets by Flow Cytometery.

RESULTSIn peripheral blood of the 20 day group rats, the CD3+ %, CD4+ %, CD8+ % and CD4+/CD8+ were 52.91 +/- 6.27, 37.80 +/- 4.11, 14.85 +/- 3.73 and 2.72 +/- 0.82 separately. Expect CD3+ %, they all had significant differences (P < 0.05). In addition, the data of the 40 day group rats showed no diversity in statistics. In spleen, CD8+ % of the 20 day group rats was 6.23 +/- 2.87 with significant differences (P < 0.05) and IgG, IgA and IgM did not change a lot in blood serum of the high humidity groups except C3 of the 20 days group (P < 0.05).

CONCLUSIONIn high humidity environment, the immune function of the rats increased in the initial stage. As time went on, the immune function gradually went to normal level through the self adjustment.

OBJECTIVE: To determine the effect of human corticotrophin-releasing hormone (CRH) on the expression of connexin-43 phosphate (P-Cx43) in human myometrial smooth muscle cells (SMCs) and the function of cell gap junction intercellular communication in SMCs. METHODS: Human non-conceive myometial SMCs were cultured with different concentrations of CRH (0, 5.85, 58.5, 585 and 5850 pmol/L). Western blot was used to test P-Cx43 and Cx43 non-phosphate (NP-Cx43) of protein expression. Cell scratch was used to test cell gap junction intercellular communication opening status in human myometrial SMCs. RESULTS: Compared with the control group, the expression of P-Cx43 was higher in the CRH groups (P<0.01), and was concentration-dependent. There was no significant difference in NPCx43 between the control group and the CRH groups (P>0.05). The transmission of cell layers in the CRH groups was higher than that in the control group (P<0.01), and as the concentration of CRH increased, the time was concentration-dependent (P<0.01). CONCLUSION CRH can enhance the expression of P-Cx43 and the function of gap junction intercellular communication in the primary cultured myometrial SMCs.
Cell Communication ; drug effects ; Cells, Cultured ; Connexin 43 ; metabolism ; Corticotropin-Releasing Hormone ; pharmacology ; Female ; Gap Junctions ; drug effects ; Humans ; Myocytes, Smooth Muscle ; metabolism ; Myometrium ; cytology ; Phosphorylation ; drug effects

Objective To investigate the efficacy and clinical effectiveness of the steroid therapy with the all body and local application on hyperthyroid Graves ophthalmopathy.Methods A total of 57 patients (106 eyes)suffering from hyperthyroid Graves ophthalmopathy who hospitalized in the Department of Ophthalmology of the Second Affiliated Hospital of Harbin Medical University from Mar 2008 to Oct 2011 were enrolled in the study.Of the 57 patients,there were 21 males and 36 females,aged 18 to 57(average 37.28 ± 11.63).The clinical course was from 1 month to 5 years(average 29.63 ± 15.36 months).Based on the patient's age,sex and condition,the 57 patients were randomly divided into steroid pulse therapy with all body group(group SPT,29 patients,54 eyes)and peribulbar injection of long-term steraid group(group PI,28 patients,52 eyes).The patients in group SPT were treated with 0.5-l.0 g methylprednisolone by intravenous infusion daily for 3 days,and then stopped for 7 days,the treatment was repeated once.But the total application dose of methylprednisolone was not overruned 5.0-20.0 g.After that,the patients were treated with oral prednisone 40 mg/d.The dose of prednisone was reduced gradually to 5 mg/d.The total course of group SPT was 3 months.The patients in the group PI were treated with triamcinolone acetonide in periorbital injection of 20 mg once every two weeks for 3 months.After one month treated,all patients were visited.The patients' body weights of two groups were measured before the treatment and after one month treated.The ocular signs of the patients including visual acuity,diplopia,intraocular pressure,exophthalmos and horizontal visual palpebral fissure width were observed before and after the treatment.Results In the group SPT,the average body weight [(62.83 ± 7.23)kg]after the treatment was higher than that before the treatment[(57.03 ±7.13)kg],and also higher than that of the group PI[(59.89 ± 10.67)kg,t =25.25,2.08,all P＜ 0.05]after the treatment.In the group SPT,the average intraocular pressures before and after the treatment were (13.46 ± 1.78)and (13.69 ± 1.41)mm Hg(1 nm Hg =0.133 kPa),respectively.In the group PI,the average intraocular pressures before and after the treatment were (13.52 ± 2.19) and (13.77 ± 2.03)mm Hg,respectively.There was no significant differences before and after the treatment in both gr0ups(P＞ 0.05).In the group SPT and PI,the average exophthalmos after the treatment[(18.63 ± 2.22) and (14.85 ± 2.61)mm]were lower than those before the treatment[(20.78 ± 2.24) and (20.60 ± 2.81)mm,t =9.97,29.68,all P＜ 0.05],respectively.After the treatment,the average exophthalmos in the group PI was significantly lower than that of the group SPT(t =7.79,P＜ 0.05).In the groups SPT and PI,the average averagepalpebral fissure width after the treatment[(15.00 ± 1.80) and (11.65 ± 2.45)mm]were lower than those before the treatment[(17.22 ± 1.89) and (17.44 ± 2.37)mm,t =22.74,91.24,all P ＜ 0.05].After treatment of the group PI,the average exophthalmos was significantly lower than that of the group SPT(t =8.01,P ＜ 0.05).Conclusion Compared with the steroid pulse therapy with all body,the peribullbar application of long-term hormone for hyperthyroid Graves ophthalmopathy is more effective,with lcss side effects and worthy of promotion.

Objective To evaluate the effects of fluticasone propionate (FP)on Foxp3 expression in CD4+T cells, to explore the possible mechanisms of childhood asthma. Methods Thirty asthmatic children, 15 with inhaled FP and 15 without inhaled FP, and 16 healthy children were recruited. Peripheral blood mononuclear cells (PBMC)labeled for CD4 and intracellular Foxp3 were analyzed using flow eytometry. The levels of IL-2 and IL-6 in serum and supernatant before and after stimulation by Phytohemagglutination (PHA)were measured by ELISA. The expression of phosphorylated signal transducer and activator of transcription 5 (STAT5)in PBMC was detected by Western-blot.Results Compared with healthy control, the percentage of CD4+ Foxp3+ cells in PBMC in asthmatic children without inhaled FP was significantly decreased. After inhaled FP and in remission stage, the percentage of CD4+ Foxp3+ cells in asthmatic children was up regulated with a decreased serum IL-6 level and an increased phosphorylated STATS expression. Conclusions Decreased Foxp3 protein expression in peripheral CD4+ T regulatory cells (Treg)is characterized in childhood asthma. Inhaled glucoeorticoid therapy of childhood asthma might be attributed to its ability of increasing Foxp3 expression by upregulation of phosphorylated STAT5 to balance the T cell response.

Benzoates ; pharmacology ; Cells, Cultured ; Cellular Senescence ; drug effects ; radiation effects ; Gene Expression Regulation, Enzymologic ; drug effects ; radiation effects ; Humans ; Infant, Newborn ; Male ; Matrix Metalloproteinase 1 ; genetics ; Matrix Metalloproteinase 3 ; genetics ; Methoxsalen ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Retinoids ; pharmacology ; Skin ; cytology ; metabolism ; Time Factors ; Tissue Inhibitor of Metalloproteinases ; metabolism ; Ultraviolet Rays

Objective To develop an interlocking-style vascular shunt device for the treatment of distal limb ischemia resulting from vascular disconnection and defect.Methods A one-way interlocking buckle was designed with the space between the clamping teeth being 0.5 mm,which prevented the device from moving backwards and fixed the vessel and shunt tube conveniently.The interlocking buckle combined with silicone tube was used to connect the two ends of the defected vessel,which was compared with conventional method by suture ligation and silicone tube by the tests on vessel bursting pressure and tensile biomechanics.Results The vessel repaired with the developed device behaved better than that by the conventional method in the tests on vessel bursting pressure and tensile biomechanics (P＜0.05).Conclusion The vascular shunt device can be used for the treatment of distal limb ischemia resulting from vascular disconnection and defect,and thus facilitates the vascular graft in rear hospital after evacuation.

This study aimed to analyze the etiology of the encephalitis outbreak in Longyan, Fujian Province, China in 2010, in order to provide valuable information for this prevention and control of this disease. Pathogens were confirmed from cerebrospinal fluid samples with fluorescent RT-PCR, virus isolation (RD cells), and neutralization tests. Then, the VP1 fragments or whole genome nucleotide sequences were determined for four virus strains using PCR. Homology was assessed using the MegAlign software, and a phylogenetic evolutionary tree was drawn using Mega 4.0 software. The results confirmed that the etiology of the outbreak was the ECHO6 intestinal virus, and the nucleotide sequence of the VP1 segment indicated that the C2 subtype was responsible. The genome sequence consisted of 7407 nucleotides, and resembled the genome of other ECHO and CoxB viruses with homology levels of 78.5%-87.3%. The encephalitis outbreak in Longyan in 2010 was caused by the ECHO6 C2 subtype intestinal virus, and its complete genome sequence length is similar to the standard strain (U16283) with a sequence homology of 80.4%.
Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Echovirus 6, Human ; classification ; genetics ; isolation & purification ; Echovirus Infections ; epidemiology ; virology ; Encephalitis ; epidemiology ; virology ; Female ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny