BackgroundThe favorable effect of bone marrow mesenchymal stem cells (BMSCs) on the reconstruction of injured corneas have been reported,but the mechanism remains unclear.ObjectiveThis study was to evaluate the anti-inflammatory and antiangiogenic effect after transplantation of BMSCs in chemically burned corneas.Methods BMSCs were isolated and extracted from the bone marrow.The cells were cultured and passaged and then were seeded on the amniotic membrane.Corneal alkali injury models were created in 18 clean SD rats by sticking the filter paper containing 1 mol/L NaOH at the central cornea for 40 seconds.The rats were then randomized into 3 groups.Amniotic membrane with BMSCs or amniotic membrane without BMSCs were transplanted in 1 week after the establishment of models,and the rats without transplantation were used as the control group.The severity of corneal lesion was graded,and angiogenesis area was measured 2 weeks after the transplantation.The expression of interleukin-2 ( IL-2 ),interferon-γ(IFN-γ),IL-10 and transforming growth factor-β ( TGF-β ) were examined by ELISA,and the mRNA of the matrix metalloproteinase-2 ( MMP-2 ),vascular endothelial growth factor (VEGF),epidermal growth factor(EGF) and basic fibroblast growth factor(bFGF) were analyzed by real-time PCR.ResultsThe positive rates of the cells were 99.78％ and 99.79％ for CD90 and CD29,7.90％,1.16％ and 1.28％ for CD34,CD45 and CDllba.The cells grew well on the amniotic membrane.The corneal inflammatory score and neovascularization area were similar among the three groups ( F =0.021,P-- 0.979 ; F =0.076,P =0.927 ).However,the corneal inflammatory score was significantly reduced and neovascularization area was significantly less in the amniotic membrane group compared with the BMSCs group and control group(P=0.011,0.001 ;P=0.005,0.000).The levels of IL-2 and IFN-γ secreted by Th1 cells were decreased (P =0.000,0.002;P =0.003,0.045 ) and the levels of IL-10 and TGF-β secreted by Th2 cells were increased in the BMSCs group compared with the amniotic membrane and control group ( P =0.000,0.000 ; P =0.000,0.021 ).No significant difference was found in VEGF expression among three groups( F=4.880,P =0.056).But the mRNA of the MMP-2 and bFGF were lower in the BMSCs group than the amniotic membrane group(P=0.009,0.003 ) and control group(P＜0.01 ).Conclusions BMSCs modulate the expression of inflammatory-related and angiogenic-related cytokines and therefore play the antiinflammatory and anti-angiogenic effects in the chemically burned cornea.

Adult ; Case-Control Studies ; Comet Assay ; DNA Mutational Analysis ; Female ; Humans ; Male ; Micronucleus Tests ; Middle Aged ; Occupational Exposure ; Vincristine

Objective To explore weaning dependent patients off mechanical ventilation through rehabilitative treatment. Methods Eighty-four patients dependent on mechanical ventilation were randomly divided into a rehabilitation group (n =44) and a control group (n =40). All were treated with routine medication, but in addition patients in the rehabilitation program followed a rehabilitation program. Three weeks later, the success rates and the time of weaning off mechanical ventilation were observed in the two groups. Results In the rehabilitation group,88.6% of the patients were successfully weaned off mechanical ventilation, versus 60% of the control group. The respective average weaning times were 6.8 ± 3.8 days and 14.3 ± 5.2 days. Both differences were statistically significant. Conclusions Rehabilitative treatment improves the weaning success rate and reduces the time needed for weaning mechanical ventilation-dependent patients.

Objective To compare the applicability of different occupational health risk assessment methods in small furniture manufacturing industry. Methods American EPA inhalation risk model, Singapore semi-quantitative risk assessment model of occupational exposure to chemical substances and Australian Occupational Health and Safety Risk Assessment model, were used to assess the occupational health risk in a small furniture manufacturing industry from a city of Zhejiang Province. Results The results of American EPA model showed that the workers who exposed to benzene and formaldehyde had low risk of carcinogens, and who exposed to benzene and xylene had very high risk of non-carcinogens. According to Singapore semi-quantitative risk assessment model, there were low and medium health risk caused by toluene and xylene, and high risk caused by wood dust in preparation and polishing jobs. Similar to the results of other models, Australia qualitative risk assessment model showed that there were medium health risk caused by toluene and xylene, and high risk caused by benzene, wood dust and noise. All of the three methods could found the key risk control point in furniture manufacturing industry. The risk ratios of the three methods were higher than the toxic work classification ratio (P<0.01), and the risk ratio of EPA model were higher than the results of Singapore model and Australia model (P<0.05) . Conclusion All of the three methods can be applied to assess the occupational health risk in furniture manufacturing industry, and the combined application of multiple risk assessment methods can be used as one of the risk assessment strategies.

BACKGROUND:The immunomodulatory ability of bone marrow mesenchymal stem cells(BMSCs)gives it a promising future in treating graft-versus-host disease(GVHD),especially with previous success in treating patients with acute GVHD.However,there are fewer reports concerning BMSCs in treating chronic GVHD,particularly for sclerodermatous chronic graft-versus-host disease(ScGVHD).OBJECTIVE:To evaluate the efficacy and safety of treatment of BMSCs for ScGVHD,and to primarily explore the immunological mechanism of clinical efficacy.METHODS:Four ScGVHD patients at the Affiliated Hospital of Academy of Military Medicine Science,between September 2006 and August 2008,were enrolled for this trial.The median patient age was 41 years,1 female and 3 male.The patients received BMSCs infusion at a dose of(1.0～2.0)×10~7 cells every time by intrabone marrow injection from the anterosuperior iliac spine and BMSCs from the same donor for the same patient were infused more than once.Concomitant medications for ScGVHD were individualized for each patient,but all were current standard medicines and the doses were significantly tapered.RESULTS AND CONCLUTION:After BMSCs infusion,the ratio of Th1 to Th2 was dramatically overturned,with an increase of Th1 and a decrease of Th2 reaching at a new balance.Correspondingly,symptoms of all the four patients gradually improved.During the course of BMSCs treatment,the life signs and laboratory results from the recipients remained normal.By the time of this report,there has been no recurrence of leukemia in the four patients.Although this study alone cannot guarantee the application of BMSCs in ScGVHD,the results are strongly in favor of the idea that the BMSCs treatment for ScGVHD patients is therapeutically practical without any detectable side effects,which may provide a new insight into the matter of treating ScGVHD clinically,thus will greatly increase the survival rate of leukemia after allogeneic bone marrow transplantation.

Objective To study the distribution of human papillomavirus type 16 (HPV16) variants and their clinical significance in Han women with Cervical Intraepithelial Neoplasia (CIN). Methods Randomly making a collection of DNA samples of cervical cells from 77 Han out-patients infected with HPV16, PCR amplification of HPV16 DNA fragments containing E6 and E7 genes and sequenced. To study the HPV16 variants types in these out-patients and explore the relationship between the HPV16 variants and CIN by comparing the E6 genes sequenced with the reference strains downloaded from the GenBank. Results Among 77 patients, the minimum age is 21 years old, the maximum age is 56 years old, and the average age is 36.39±6.86 years old. 61 patients (accounting for 79.2% ) were diagnosed as CIN Ⅱ and higher grade lesions while 16 patients (accounting for 20.8% ) as CIN Ⅰ. In this research, only European variant and Asian variant were found by Parsimony analyses of the sequences. There are 38 Asian variants and 39 European variants. With χ~2 test, χ~2=0.0034, P=0.9535 > 0.05, it suggested that there was no enough evidence to support Asian- and European- variants had the different risk in the cause of cervical intraepithelial neoplasia and cervical cancer. Conclusion It was not found Asian- and European- variants of HPV16 had different effect on the cervical cancer, but found only two major variants-Asian- and European-variants in Han people in this research. So we have reason to speculate that there are two major HPV16 variants(Asian- and European- variants) in China's Han women, while other variants, especially high cancer-causing Asian/American variant are not common.

Objective To set up a ultraviolet-visible spectrophotometry for quantitative determination of the total saponins from Flos Hostae Plantagineae. Methods The ultraviolet-visible spectrophotometry was used to detect the absorbance with the wavelength of 450 nm taking vanillin-glacial acetic acid-concentrated sulphuric acid as the color-developing agent and gitogenin as the control sample. The content of the total saponins in the samples was calculated by using standard curve. Results Gitogenin showed a good linear relationship at range of 14～70 μg/ml (r=0.999 2) with the average recovery of 96.8% (RSD=2.97%, n=6). Conclusion The method is easy, accurate and sensitive with a higher repeatability, which can be used in the quantitative determination of the total saponins from Flos Hostae Plantagineae.

OBJECTIVETo analyze the influence of optimal codon usage on the expression levels and immunogenicity of DNA vaccines, encoding the human papillomavirus type 6b (HPV 6b) E7 gene.

METHODSThe full length E7 gene of HPV 6b was modified to substitute human preferred codon for rarely used codon, and three mutations were introduced into the pRB binding site of HPV 6b E7 to eliminate its transformation potential. The codon optimized and mutated E7 gene (hu-mE7) were cloned into the Kpn I and EcoR I site of the pcDNA3 mammalian expression vector, the in vitro expression of the hu-mE7 gene and the immunogenicity of hu-mE7 DNA vaccine were compared with the wt-E7gene.

RESULTSThe in vitro expression of pcDNA3-hu-mE7 was much higher than the classical wt-E7 plasmid in monkey COS-1 cell line. Mice immunized intramuscularly with the pcDNA3-hu-mE7 showed that the codon modified E7 gene induced a stronger IFN-gamma ratios than the wt-E7 gene.

CONCLUSIONSThese results suggest that the optimized codon usage contributes to the enhancement of gene expression and immunogenicity of HPV 6b E7 gene.

Animals ; Cell Line ; Codon ; genetics ; Female ; Gene Expression ; Genes, Viral ; genetics ; Genetic Vectors ; Mice ; Papillomaviridae ; genetics ; Papillomavirus Vaccines ; Transfection ; Vaccines, DNA ; immunology ; Viral Proteins ; biosynthesis ; genetics ; Viral Vaccines ; immunology

Objective To understand the prevalence of older people with mild cognitive impairment (MCI) in Zhejiang province and to provide the basis for elderly early detection and diagnosis of Alzheimer' s disease (AD).Methods 1211 more than 60-year-old elderly populations were selected in Zhejiang province,and were given screening questionnaire by general information,the montreal cognitive assessment (MoCA) and mini-mental state examination (MMSE).Results MCI prevalence of elderly populations in Zhejiang was 20.7％ and the AD prevalence was 4.5％.The patient' s gender,age,education level,nature of work,sleep status,marital status,whether or not participating in physical exercise,having smoking and drinking habits,whether combined with hypertension and diabetes for MCI prevalence of the elderly were statistically significant (P＜0.05).Conclusion We should pay attention to take appropriate measures in preventing the cognitive decline for populations as elderly,especially for women,senior,no spouse,engaged in manual labor,low education level,poor quality of sleep and no physical exercise,with smoking and drinking,combined with hypertension and diabetes.

Animals ; Interleukin-1 ; biosynthesis ; genetics ; Liver ; blood supply ; Lung ; metabolism ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics