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.

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.

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 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.

The effect of the autonomic nerves on the transmural dispersion of ventricular repolarization (TDR) under acute myocardial ischemia in intact canine was investigated. Using the monophasic action potential (MAP) recording technique, MAPs of the epicardium (Epi), mid-myocardium (Mid) and endocardium (Endo) were recorded simultaneously by specially designed plunge-needle electrodes at the left ventricular free wall under acute myocardial ischemia in 12 open-chest dogs. MAPD90 and TDR among three myocardial layers as well as the incidence of the early afterdepolarization (EAD) before autonomic nervous stimulation and during autonomic nervous stimulation were compared. It was found that 10 min after acute myocardial ischemia, TDR was increased from 55 +/- 8 ms to 86 +/- 15 ms during sympathetic stimulation (P < 0.01). The TDR (53 +/- 9 ms) during parasympathetic stimulation was not significantly different from that of the control (55 +/- 8 ms) (P > 0.05). The EAD was elicited in the Mid of 2 dogs (16%) 10 min after acute myocardial ischemia, but the EAD were elicited in the Mid of 7 dogs (58%) during sympathetic stimulation (P < 0.01). It was concluded that: (1) Sympathetic stimulation can increase the transmural dispersion of repolarization and induce early afterdepolarizations in the Mid under acute myocardial ischemia, which provide the opportunity for the ventricular arrhythmia developing; (2) Parasympathetic stimulation has no significant effect on the transmural dispersion of repolarization under myocardial ischemia.
Action Potentials/physiology ; Autonomic Nervous System/*physiopathology ; Electric Stimulation ; Heart Ventricles/innervation ; Heart Ventricles/*physiology ; Myocardial Ischemia/*physiopathology ; Neuromuscular Junction

The effect of the autonomic nerves on the transmural dispersion of ventricular repolarization in intact canine was investigated. By using the monophasic action potential (MAP) recording technique, monophasic action potentials (MAPs) of the epicardium (Epi), midmyocardium (Mid) and endocardium (Endo) were recorded simultaneously by specially designed plunge-needle electrodes at the left ventricular free wall in 12 open-chest dogs. MAPD90 and transmural dispersion of repolarization among three myocardial layers as well as the incidence of the EAD before autonomic nervous stimulation and during autonomic nervous stimulation were compared. The results showed that the MAPD90 of Epi, Mid and Endo before autonomic nervous stimulation were 278 +/- 11 ms, 316 +/- 16 ms and 270 +/- 12 ms respectively, the MAPD90 of Mid was significantly longer than that of Epi or Endo (P<0.01). MAPD90 of Epi, Mid and Endo were shortened by 19 +/- 4 ms, 45 +/- 6 ms, 18 +/- 3 ms respectively during sympathetic stimulation. Compared with that of the control, the transmural dispersion of repolarization during sympathetic stimulation was shortened from 44 +/- 4 ms to 15 +/- 3 ms (P<0.01), but early afterdepolarizations were elicited in the Mid of 5 dogs (41%) during sympathetic stimulation. Parasympathetic stimulation did not significantly affect the MAPD90 in the three layers. It is concluded that there is the transmural dispersion of ventricular repolarization in intact canine. Sympathetic stimulation can reduce transmural dispersion of repolarization, but it can produce early afterdepolarizations in the Mid. Parasympathetic stimulation does not significantly affect the transmural dispersion of ventricular repolarization.
Action Potentials/physiology ; Autonomic Nervous System/*physiology ; Electric Stimulation ; Electrocardiography ; Electrodes ; Endocardium/innervation ; Endocardium/physiology ; Heart Ventricles/innervation ; Heart Ventricles/*physiology ; Myocardium/cytology ; Neuromuscular Junction ; Pericardium/innervation ; Pericardium/physiology

To elucidate the mechanism of arrhythmia in healed myocardial infarction (HMI), the changes of action potential duration (APD), transient outward potassium current (Ito), delayed rectifier potassium current (IK) and inward rectifier potassium current (IK1) of left ventricular myocytes in non-infarcted zone of HMI were investigated. Rabbits were randomly assigned into two groups: HMI group, in which animals were subjected to thoracotomy and ligation of the circumflex coronary and sham-operated group, in which rabbits underwent thoracotomy but no conorary ligation. 3 months after the operation, the whole myocyte patch clamp technique was used to record APD, Ito, IK, and IK1 of ventricular myocytes in non-infarcted zone. Our results showed that the membrane capacitance was larger in HMI group than in sham-operated group. Action potential duration was significantly lengthened in HMI group and early afterdepolarization (EAD) appeared in HMI group. The densities of Ito, I(K, tail), and IK1 were reduced significantly in HMI group, from 6.72 +/- 0.42 pA/pF, 1.54 +/- 0.13 pA/pF and 25.6 +/- 2.6 pA/pF in sham-operated group to 4.03 +/- 0.33 pA/pF, 1.14 +/- 0.11 pA/pF and 17.6 +/- 2.3 pA/pF, respectively. It is concluded that the reduced densities of Ito, I(K, tail) and IK1 in ventricular myocytes of non-infarcted zone in HMI were responsible for the prolongation of APD and the presentation of EAD which played important roles in the development of malignant arrhythmia in HMI.
Action Potentials ; Arrhythmia/*etiology ; Heart Ventricles/metabolism ; Myocardial Infarction/complications ; Myocardial Infarction/metabolism ; Myocardial Infarction/*pathology ; Myocytes, Cardiac/*cytology ; Patch-Clamp Techniques ; Potassium Channels/*metabolism