Our studies tried to demonstrate Eha (Et haemolysin activator) could regulate the resistance of the bacterium against acidification to survive in the macrophage and explain its underlying molecular mechanism.When the bacteria infected the macrophages at time intervals,intracellular survival rate in bafilomycin-treated macrophages was higher than that with untreated cells,and the rate of wild type ET 13 was higher than that of its eha mutant,respectively (P＜0.05).The survival rate of the wild type was higher than that of the mutant under acid treatment (P＜0.05).To determine the conditions that induced the highest eha expression,we constructed a pMP220-Peha LacZ plasmid and determined the lacZ expression under different conditions.After exposure of pH6.3 medium for 2 h time,we performed the whole transcriptomic profiles of the wild type and mutant by RNA-sequencing.We identified 147 differentially-expressed genes ([log2 ratio| ≥1),113 and 34 of which were significantly up-and down-regulated,respectively in the mutant,comparing with the wild type.These findings were validated by qRT-PCR.GO functional analysis revealed that these genes were divided into 25 categories,including the bacterial catalysis,cellular composition,combination,localization,metabolism,processing,and transportation.Based on the KEGG database,these genes were distributed in 55 pathways,such as two-component system,ABC transporters,and microbial metabolism in diverse environments.Overall,Eha is an important regulator to affect all kinds of target genes and pathways for E.tarda to adapt to an acid environment.These results could be helpful for further investigations of the mechanisms by which E.tarda survives in macrophages.

Mesenchymal stem cells (MSCs) have good immune regulatory function,can inhibit many immune cell proliferation,direct effects on activation and proliferation of T cell,play a role in immune regulation by Treg cells or by the secretion of soluble factors regulating Thl/Th2 secretion and reaction equilibrium,inhibit the inflammation through the anti-inflammatory,regulation of cytokines expression at the same time,so reduce the expression levels of matrix metalloproteinase-2 and-9,which may promote lacrimal gland tissue damage,and thus play a role in immune regulation.MSCs can reduce the autoimmune dry eye clinical index,recovery secretion function of part lacrimal gland.This article reviews the research advances in inmmmne regulation of MSCs on autoimmune dry eye.

Mesenchymal stem cells (MSCs) are multipotent stromal cells derived from the mesoderm that can differentiate into a variety of cell types.Recently,as the study of MSCs in the treatment of various autoimmune diseases is maturing,more and more researchers have advocated the immune function of MSCs and the treatment for dry eye.The rate of dry eye is higher.The duration of therapy in dry eye,especially immune related dry eye,is long,and the therapeutic effect is poor.Dry eye has affected people's quality of life,so it is important to find new methods to treat it.This article reviews MSCs function in dry eye.

Objective:To investigate the mechanism and effects of autophagy on cisplatin(DDP)-induced apoptosis in human gas-tric cancer cell line SGC7901. Methods:Cell proliferation was determined by an MTT assay after the SGC7901 cells were treated with DDP and/or chloroquine. Cell apoptosis was determined by flow cytometry. Autophagy and related protein expressions were detected by Western blot. Autophagy was quantitatively analyzed by fluorescence microscopy after monodansylcadaverine staining was per-formed. Results:The cells were treated with 5 mg/L of DDP for 24 h, the rate of cell apoptosis was (21.07 ± 2.12)%. Autophagy, char-acterized by an increase in the number of autophagic vesicles and LC3-II protein level, was observed in DDP-treated cells. After autoph-agy was inhibited by chloroquine, the rate of cell apoptosis was increased to (30.16 ± 3.54)%. In addition, caspase-3 and P53 protein levels were increased, but Bcl-2 protein was decreased. Conclusion:Autophagy protected human gastric cancer cell line SGC7901 from DDP-induced apoptosis. In addition, the inhibition of autophagy could promote apoptosis. The combined therapy of DDP and chlo-roquine may be a promising therapeutic strategy for gastric cancer.

Objective:To observe the clinical effect of tuina plus foot bath with Chinese medicine for patients with diabetic foot (DF) in early stage.Methods:A total of 70 patients with early-stage DF were randomly allocated by the random number table into two groups,with 35 cases in each group.Patients in the control group received conventional medication,while patients in the observation group received tuina plus foot bath with Chinese medicine on the basis of conventional medication.The clinical efficacy was compared after 2 courses of treatment.Results:After treatment,intra-group comparisons of ankle-brachial index (ABI) showed statistical significance in both groups (both P＜0.05).The curative rate was 83.3％ in the observation group,with the total effective rate of 96.7％,versus 29.4％ and 76.5％ in the control group,respectively,and the between-group comparisons showed statistical significance (both P＜0.05),indicating a better effect in the observation group.Conclusion:Tuina plus foot bath with Chinese medicine has a good therapeutic effect for DF patients in early stage.

Objective To investigate the anti-apoptotic function of clusterin in LNCaP cell line and the role of clusterin antisense oligodcoxynucleotide(AS-ODN)in TNF-?-induced death of LNCaP cell. Methods The wild type LNCaP(group L),LNCaP transfected with the control vector(group M),LNCaP transfected with full-length clusterin expression vector(group A,ie,study group)were cultured.For the de- tection of cytotoxic effect of TNF-?,MTT and ELISA methods were used to determine the cell proliferation and apoptosis of the 3 clones,and the changes of proliferation and apoptosis in A cell after transfection of clusterin AS-ODN were also assessed.Results MTT method showed that the cell proliferation activity(A value)of groups L,M,and A were 0.84?0.03,0.85?0.04,0.95?0.03,respectively;the difference be- tween groups L and M was not significant(P＞0.05);but compared with group A the cell proliferation activ- ity was significantly lower in groups L and M(P＜0.01 for both).ELISA resuhs showed that the A values of groups L,M,and A were 0.59?0.04,0.62?0.03,0.33?0.04,respectively;the difference between groups L and M was not significant(P＞0.05);but compared with group A,the apoptosis rates were significantly higher in groups L and M(P＜0.01 for both).In group A,A values of cell proliferation activity in subgroups control,AS-ODN,TNF-?,TNF-?+AS-ODN were 1.30?0.03,1.25?0.03,0.99?0.03,0.80?0.03, respectively;the differences between each group were significant(P＜0.05 for all).And the A values of cell apoptosis in the above 4 groups were 0.02?0.00,0.21?0.02,0.63?0.07,1.16?0.04,respectively,the differences between each group were significant(P＜0.01 for all).Conclusions Stable transfection and subsequent expression of clusterin result in resistance to the cytotoxic effect of TNF-?.Transfection with clus- terin AS-ODN enhances cytotoxic effect of TNF-?in A cells.These results suggest that clusterin plays an im- portant role in anti-apoptotic function in LNCaP cell line.

Objective To study the pelvic fascia related to pelvic autonomic nerve and detect the anatomical localization of pelvic autonomic nerve by marker in adult male.Methods Twelve pelvises of adult male harvested from cadavers were studied by dissection.Results Hypogastric nerve was embedded in the posterior leaf of the visceral pelvic fascia.Pelvic plexus was situated between vesicohypogastric fascia and visceral fascia.Pelvic nerve branch of seminal vesicle and prostate was located at the anterolateral part of Denonvilliers fascia.Sacral promontory,ureter,junction of Denonvilliers fascia,visceral fascia and seminal vesicle could be regarded as anatomical markers for pelvic autonomic nerve.Conclusion The anatomical characteristics of pelvic autonomic nerve can be used for protecting and isolating pelvic autonomic nerve in total mesorectal excision of adult male.

AIDS Vaccines ; HIV Infections ; prevention & control ; Humans ; Nigeria

Objective To establish the model of autoimmune dry eye and analyze the level of cytokines and the pathological changes of autoimmune dry eye rabbits treated with human umbilical cord mesenchymal stem cells (MSCs).Methods Twentyfour female New Zealand rabbits were randomly divided into three groups:Normal control group,dry eye group and MSCs treatment group,8 cases in each group.After treated with MSCs for six weeks,the pathological changes were observed,and the expression of Th1 and Th17 cell differentiation related cytokines mRNA in lacrimal gland tissue was detected by real-time quantitative PCR.The percentage of regulatory T cells (CD4 +Foxp3 + cells) in the lacrimal gland and spleen tissues of the dry eye model group and MSCs treatment group was measured by flow cytometry.Results After treatment with MSCs for 6 weeks,HE staining from each rabbit lacrimal gland showed that,the normal control group showed no or only a few lymphocytes;The lacrimal gland cells were atrophied in the dry eye group,the distribution of lymphocyte scattered around the gland and small blood vessels;Compared with dry eye group,the lacrimal gland lymphocyte infiltration reduced in MSCs treatment group,and the cell morphology were better.Compared with dry eye group,lymphocyte infiltration and aggregation in the conjunctival conjunctiva in MSCs treatment group were significantly reduced,the epithelial structure was intact,and the degeneration and atrophy cells were rare.The expression of inflammatory factors including interferon-γ(IFN-γ),Interleukin-17 (IL-17) and transcription factor T-bet mRNA were decreased in MSCs treatment group,there were significant difference compared with dry eye group (all P ＜ 0.05);Thl7 cell associated cytokines IL-17 expression levels were decreased,but no significant difference was found between two groups (P ＞ 0.05);The expression of transcription factor RORrt mRNA in dry eye group was significantly decreased,the difference was statistically significant (P ＜ 0.05).The expression of inflammatory factors TNF-α in the lacrimal gland tissue of MSCs treatment group was significantly lower than that in the dry eye model group,while the expression of the anti-inflammatory factor TGF-β was significantly higher than that in the dry eye model group,and the differences were statistically significant (all P＜0.05).The regulatory T cells (CD4 + Foxp3 + cells) in the lacrimal gland of dry eye model group accounted for 10％ of lymphocytes was,while the treatment group accounted for 27.8％,MSCs group was significantly higher than dry eye group,the difference was statistically significant (P ＜ 0.05).Conclusion MSCs can reduce the histopathological changes of the immune dry eye,and they may have immunoregulatory effects on autoimmune dry eye.The mechanisms of immunomodulatory effects may be related to the balance of inflammatory cytokines and anti-inflammatory cytokines.

Objective To observe and analyze the effect of B. Adolescentis and L. Acidophilus on the proportion of Th cell Th1/Th2 in peripheral blood of UC-mice in acute stage and recovery stage. Method 40 BABL/c mice were induced by 3% DSS water for 7 clays free drinking and then with distilled water for 10 days. They were randomly allocated in 4 groups: NS group, SASP group, BF0624 group and LT0637 group, also the fifth group-10 normal animals. The blood of mice were collected by removing their eyes at day 8 and day 18, and then the mononuclear cells were iso]atecl. The proportion of Th1/Th2 was analyze through flow eytometry,hy labeling the specific antibody ot Th cellular membrane with the CD4 antibody, and the cytoplastie antigen of Thl or Th2 with IIA antibody or IF'N-γ antibody. Result The proportion of Th1/Th2 in normal mouse was 0. 84 -0. 94,and it raised up in DSS-mice at both acute stage and recovery stage. It decreased unequally after 7 or 17 days'B, adoleseentis, L. Acidophilus and SASP treatment, but that of all three groups were lower than NS group (2.21±0. 83). Even the proportion got close to the normal animals after 17 days'L. Acidophilus -treated. Conclusion The proportion of Thl/Th2 increased at the acute stage and recovery stage of DSS-mice. Both B. Adoleseentis and L. Acidophilus had more effective than SASP on decreasing the proportion of Th1/Th2 at two stages,ospecially L. Acidophilus.