Animals ; Cell Culture Techniques ; methods ; Cell Separation ; methods ; Culture Media ; Epithelial Cells ; cytology ; Respiratory System ; cytology ; Swine

OBJECTIVETo observe interventional effects of anti-viral therapy and Compound Qin-gre Granule (CQG) on host cellular immune functions of acute virus infection patients.

METHODSThirty acute virus infection patients were recruited to detect peripheral lymphocyte subsets. They were randomly assigned to two groups, the Western medicine treatment group (treated with anti-virus Western medicine) and the integrative medicine treatment group (treated with anti-virus Western medicine plus CQG). T-cell subsets were re-examined 7 days later. Changes between before and after treatment were observed. Effect on host cellular immune functions and efficacy were compared between the Western medicine treatment and the integrative medicine treatment.

RESULTSCompared with the normal control group, the percentage of peripheral T cells increased, and the percentage of B/NK cells decreased in acute virus infection patients (P < 0.01). Meanwhile, in T cell subsets, the percentage of CD8+ T cells and CD8+ CD38+ T cells increased (P < 0.05, P < 0.01); and percentages of CD4+ T cells, CD4+ CD28 + T cells, and CD8+ CD28+ T cells decreased (P < 0.05, P < 0.01). After one-week treatment, percentages of CD4+ T cells, CD4+ CD28+ T cells, and CD8+ CD28+ T cells increased (P < 0.05, P < 0.01), while the percentage of CD8+ CD38+ T cells decreased (P < 0.01). More significantly, these changes were greater in the integrative medicine treatment group than in the Western medicine treatment group (P < 0.05).

CONCLUSIONSDisarranged cellular immune functions existed in acute virus infection patients. CQG could significantly improve viral infection induced immunologic derangement and immunologic injury.

Drugs, Chinese Herbal ; therapeutic use ; Humans ; Lymphocyte Count ; Lymphocyte Subsets ; T-Lymphocyte Subsets ; Virus Diseases ; drug therapy

The rate of corneal graft rejection is still high for high-risk keratoplasty although immune suppression drug is routinely used.The role of traditional Chinese medicine in corneal transplantation is concerned gradually.Heijingpaichitang on the prevention and treatment of rats with high-risk corneal allograft rejection needs further study.Objective This study was to investigate the inhibitory effect of heijingpaichitang on high-risk corneal transplantation immune rejection in rats.Methods Sixteen female SD rats were used as the donors and 32female Wistar rats were served as recipients.The high-risk corneal trasplantation models were established by corneal suture in 32 Wistar rats,and then homogeneity variant SD-Wistar corneal transplantation was performed.The recipients were randomized into model control group,cyclosporinc A (CsA)group,heijingpaichitang group and CsA +heijingpaichitang group.CsA,heijingpaichitang and CsA + heijingpaichitang was orally administered 4 days after operation once per day for 15 days,and normal saline solution was used at the same way in the model control group.Ocular anterior segment reaction was examined under the slit lamp and corneal opacification,edema and neovasculation were scored based on Larkin' s criteria.Rejection index of the corneal graft was recorded and the graft survival time was calculated.The pathological examination of the corneal graft was carried out in all rats,and the inflammatory cells in the corneas and CD4+ cells in the periphery blood were assayed using flow cytometry.The use of the animals complied with ARVO Statement.Results Corneal graft rejection occurred in 10 days after operation in the model control group,12-13 days in the CsA group and heijingpaichitang group and 22 days in the CsA +heijingpaichitang group.Compared with model control group,the scores of the corneal opacification,corneal edema and neovascularization were significantly lower in the CsA group,heijingpaichitang group and CsA+heijingpaichitang group (P＜0.05),and all the scores were declined in the CsA+ heijingpaichitang group compared with CsA group and heijingpaichitang group(P＜0.01),but no significant differences were seen in the scores between the CsA group and heijingpaichitang group(P＞0.05).The mean survival time of grafts was (10.38 ±1.69)days in the model control group,(22.50 ± 3.07) days in the CsA + heijingpaichitang group,with the significant difference (t =-9.790,P =0.000).The pathological examination of graft showed that the lymphocytes and new blood vessels were less in the CsA+heijingpaichitang group compared with CsA group and heijingpaichitang group 15 days after operation.Flow cytometry verified that the number of lymphocytes in graft,CD4+cells and CD4+/CD8+ in periphery blood were significantly lower in the heijingpaichitang group,CsA group and CsA+heijingpaichitang group compared with model control group (P＜0.05).Conclusions Heijingpaichitang can inhibit immune rejection to certain extent in high-risk corneal transplantation rat and has a similar effect to 0.1％ CsA.Heijingpaichitang and 0.1％ CsA have a synergistic effect.

Objective To investigate the effects of advanced glycation end-products(AGEPs)on the function of normal keratinocytes in vitro so as to explore the role of AGEPs in impaired wound healing. Methods Normal rat keratinocytes were incubated with different concentrations of AGEPs.After 48 hours of culturing,the cell proliferation rates were measured by MTT colorimetric determination.The cell cycle distributions and apoptosis were analyzed with flow cytometry,and the migration was investigated by 24-well fluorimetric cell migration assay kit by exposing to 100?g/ml AGEPs.Nuclear extracts from these cells were examined for binding of nucleotides containing NF-?B consensus by immunocytochemistry and EMSA in vitro.Results The proliferations of normal keratinocytes were significantly arrested and many cells were induced to early apoptosis compared with control ones(P＜0.05)by exposing to AGEPs for 48 hours. Meanwhile AGEPs also irritated keratinocytes migration compared with control ones(P＜0.05).Inhibiting the activation of NF-?B could partly recover the proliferation of keratinocytes,reverse apoptosis and attenu- ate migration.Conclusion AGEPs are correlated with the migration,proliferation and apoptosis of kera- tinocytes by NF-?B.

OBJECTIVETo evaluate the impact of nicotine- and tar-free cigarette smoke extract (fCSE) on the serum testosterone (T) level and erectile function of male rats.

METHODSWe randomized 30 male SD rats to three groups of equal number to receive subcutaneous injection of PBS (1.0 ml / 300 g body weight per day), fCSE (1.0 ml/300 g body weight per day), and reduced glutathione hormone (GSH, 200 mg per kg body weight per day) in addition to fCSE (fCSE + GSH), respectively, all for 8 weeks. Then we evaluated the erectile function of the rats by measuring the maximal intracavernous pressure (MICP), mean arterial pressure (MAP), ICP/MAP ratio, time of stimulation to MICP (Tmax), and cavernosal filling fate (CFR). We determined the serum T level, the activities of superoxide dismutase (SOD) , malondialdehyde (MDA), and nitric oxide synthase (NOS) in the cavernosal tissue, and also observed the morphological changes of the corpus cavernosum.

RESULTSCompared with the controls, the rats of the fCSE group showed obvious decreases in the levels of serum T ([5.37 ± 1.43] vs [3.22 ± 1.11] μg/L), NOS ([2.90 ± 0.27] vs [1.67 ± 0.18] U/mg) , and SOD ([18.41 ± 1.09] vs [13.36 ± 1.18] U/mg prot) and erectile function-related indexes MICP ([85.92 ± 6.36] vs [58.99 ± 10.76] mmHg), MICP/MAP (0.86 ± 0.09 vs [0.56 ± 0.08]), and CFR (2.14 ± 0.44 vs 0.89 ± 0.44), but markedly increased Tmax ([29.90 ± 5.78] vs [42.90 ± 8.56]s), with a positive correlation between the serum T level and CFR (r = 0. 364, P < 0.05). Masson staining revealed a lower ratio of the corpus cavernosum smooth muscle tissue to collagen fiber in the fCSE group (0.27 ± 0.04) than in the control (0.98 ± 0.12). Compared with the fCSE group, the fCSE + GSH group exhibited significantly improved MICP ([58.99 ± 10.76 ] vs [77.95 ± 7.71] mmHg), MICP/MAP (0.56 ± 0.08 vs 0.77 ± 0.09), and CFR (0.89 ± 0.44] vs 1.76 ± 0.42) and shortened Tmax ([42.90 ± 8.56 ] vs [32.10 ± 5.84 ] s). The ratio of the corpus cavernosum smooth muscle tissue to collagen fiber was higher in the fCSE + GSH than in the fCSE group (0.77 ± 0.09 vs 0.27 ± 0.04) but still lower than in the control (0.98 ± 0.12).

CONCLUSIONNicotine- and tar-free cigarette smoke extract reduces the serum T level and erectile function of rats, which is related to oxidative stress. Antioxidant therapy can improve erectile function but has a limited value for morphological protection of the penile tissue.

Animals ; Erectile Dysfunction ; chemically induced ; Male ; Malondialdehyde ; metabolism ; Muscle, Smooth ; pathology ; Nicotine ; Nitric Oxide Synthase ; metabolism ; Penile Erection ; drug effects ; Penis ; pathology ; Rats ; Rats, Sprague-Dawley ; Smoke ; adverse effects ; Superoxide Dismutase ; metabolism ; Tars ; Tobacco ; adverse effects

OBJECTIVETo study the utility of fractional exhaled nitric oxide (FeNO) in young children at different stages of asthma.

METHODSFifty-eight children with newly diagnosed asthma (aged 1-3 years) at the acute exacerbation stage between April and June, 2014 were recruited. After 3 months' treatment, the children switched into the chronic persistent stage (n=34) or remission stage (n=24). Thirty aged-matched healthy children served as controls. FeNO levels and lung function were measured for all subjects. The best cut-off value of FeNO for the diagnosis of asthma was evaluated by receiver operating characteristic (ROC) curve.

RESULTSThe FeNO levels in children with asthma at various stages were higher than controls (P<0.05). The FeNO levels in the acute exacerbation stage were highest, followed by the chronic persistent stage (P<0.05). FeNO level was correlated to the stages of asthma (r=-0.382, P<0.001). The cut-off value of FeNO for the diagnosis of asthma was 22.75 ppb by ROC curve, with the sensitivity of 0.933 and the specificity of 0.388.

CONCLUSIONSThe children with asthma at different stages have different FeNO levels. Measurement of FeNO is useful in the diagnosis of asthma in young children.

Asthma ; diagnosis ; metabolism ; Breath Tests ; Humans ; Nitric Oxide ; metabolism ; ROC Curve

OBJECTIVETo examine fractional exhaled nitric oxide (FeNO) values in 1-3-year-old children with asthma and analyze the correlation of FeNO with peripheral blood eosinophils (EOS) and lung function in these children.

METHODSA total of 111 children aged 1-3 years with asthma were enrolled. The children were classified into acute exacerbation (n=62) and remission groups (n=49) according to their symptoms. FeNO values, lung function, and peripheral blood EOS count were measured in these children. Sixty age-matched healthy children were enrolled as the control group.

RESULTSFeNO values were significantly higher in the acute exacerbation group (24.4 ppb) than in the remission group (18.0 ppb) and the control group (13.7 ppb) (P<0.05). The FeNO values in the remission group were significantly higher than in the control group (P<0.05). FeNO values were not significantly correlated with peripheral blood EOS count and lung function parameters (PEF, TEF25, TEF50, and TEF75).

CONCLUSIONSMeasurement of FeNO is useful to evaluate the disease activity in children with asthma aged 1 to 3 years, but the FeNO values are not correlated with peripheral blood EOS count and lung function.

Asthma ; blood ; physiopathology ; Breath Tests ; Child, Preschool ; Eosinophils ; physiology ; Female ; Humans ; Infant ; Lung ; physiopathology ; Male ; Nitric Oxide ; metabolism

Objective To examine the concentrations of hepatocyte growth factor(HGF)and vascular endothelial growth factor (VEGF)in the patients with diabetic neovascular glaucoma(NVG),proliferative retinopathy without neovascularization of the iris(PDR) and idiopathic macular role(IMH),and to determine the relationship between HGF and VEGF.Design Prospective case series. Participants 14 patients(14 eyes)with NVG,22 patients(22 eyes)with non-NVI PDR,19 patients(19 eyes)with IMH.Methods The double-antibody sandwich enzyme-linked immunosorbent assay(ELISA)was used to measure the intravitreous level of HGF and VEGF.Main Outcome Measures The concentrations of HGF and VEGF in the same specimens.Results The concentrations of HGF (mean?SD)in vitreous of NVG group,PDR group and IMH group were(12908.42?2946.46)、(9770.86?3802.99)、(4160.54?2044.80)pg/ ml,respectively(Kruskal-Walls test,X~2=32.36,P=0.000).In addition,significant differences were observed between groups of the concentrations of HGF(P＜0.01,respectively).The intravitreous concentrations of VEGF(mean?SD)in three groups were(823.50?718.58)、(821.82?786.27)、(22.73?3.20)pg/ml(X~2=28.30,P=0.O00),respectively(Kruskal-Walls test,X~2=32.36,P=0.000).There was no significant correlation between the concentrations of HGF and those of VEGF in the same specimens from each group(P＞0.05, respectively).Conclusions The intravitreous concentrations of HGF in the patients with diabetic NVG are obviously higher than those with IMH and non-NVI PDR,suggesting that HGF appears to be associated with mediating the neovascularization of retina and iris in NVG.However,it is not found there is any relationship between the intravitreous levels of HGF and VEGF.

425 halophilic bacteria strains were isolated by using six different media from thirty salt-soil samples collected from Heijing ancient salt mine,Yunnan. By the growth on NaCl grandient concentration and other screening methods,79 strains of extremely halophilic microorganisms were chosen for further research. Based on colonies color,shape and size,15 stains were selected to be sequenced. The sequencing results revealed that 11 of them were haloarchaea. According to the phylogenetic analysis,they belong to four genera of the family Halobacteriaceae. The blast results showed that the similarities of six sequences were higher than 97 % with the validly described species of the following four genera: Halorubrum,Natronococcus,Natrialba,Halalkalicoccus. The similarities of other five sequences with any validly described species were less than 97 %,therefore,the taxonomic positions of the following five strains YIM-ARC 0032,YIM-ARC 0036,YIM-ARC 0037,and YIM-ARC 0050 could be determined according to further polyphasic taxonomy data.The results indicated that there was a considerable diversity of haloarcheaea in salt-mine environment of Heijing ancient salt mine,Yunnan and it was worth continuing to research on this area.

Background:X-linked inhibitor of apoptosis (XIAP) is a vital factor in the anti-apoptosis mechanism of tumors and is highly expressed in renal cell carcinoma (RCC).However,the mechanism through which XIAP regulates DNA damage repair is unknown.This study investigated the regulatory mechanism of XIAP in etoposide-induced apoptosis in two Caki-1 cell lines with high or low XIAP expression.Methods:The two cell lines were established using RNA interference technology.The differentially expressed proteins in the two cell lines were globally analyzed through an isobaric tags for relative and absolute quantitation-based quantitative proteomics approach.Proteomic analysis revealed 255,375,362,and 5 differentially expressed proteins after 0,0.5,3,and 12 h of drug stimulation,respectively,between the two cell lines.The identified differentially expressed proteins were involved in numerous biological processes.In addition,the expression of histone proteins (H1.4,H2AX,H3.1,H3.2,and H3.3) was drastically altered,and the effects of XIAP silencing were accompanied by the marked downregulation of H2AX.Protein-protein interactions were assessed and confirmed through immunofluorescence and Western blot analyses.Results:The results suggested that XIAP may act as a vital cell signal regulator that regulates the expression of DNA repair-related proteins,such as H2AX,and influences the DNA repair process.Conclusions:Given these functions,XIAP may be the decisive factor in determining the sensitivity of RCC cell apoptosis induction in response to chemotherapeutic agents.