Background Researches showed that the mitochondrial-mediated apoptosis and autophagy play an important role in the survival of retinal ganglion cells (RGCs) in glaucoma.However,whether high pressure will lead to mitochondrial-mediated apoptosis and autophagy is not elucidated.Objective This study was conducted to explore whether elevated pressure can directly induce the mitochondrial-mediated apoptosis and autophagy in cultured RGC-5 cells in vitro.Methods RGC-5 cells were exposed to self-made pressure device and treated by 0,20,40 and 60 mmHg pressure for four hours in pressurized bottles.RGC-5 cells were incubated simultaneously in sealed incubator bottles and served as normal control.The morphological changes of the cells were examined under the inverted phase-contrast microscope.The apoptosis percentages of the cells were detected by flow cytometry.Mitochondrial membrane potentials of the cells were assessed using the JC-1,a fluorescent dye.And the expressions of cytochrome C (Cyt-c),microtubule associated protein light chain 3 (LC3) and Beclin-1 protein in the cells was detected by Western blot.Results Cultured cells showed fusiform shape in the normal control group and 0 mmHg group with more dendritic process.The cell density was obviously reduced and the number of dead cells was increased in the 20,40 and 60 mmHg groups.The apoptotic percentage was (15.69 ± 0.77)％,(15.77 ± 1.14)％,(18.30± 1.07) ％,(23.28 ± 1.33)％ and (34.47± 1.17)％ in the normal control group and 0,20,40 and 60 mmHg groups,respectively,showing a significant intergroup difference (F =150.90,P＜0.001),and the apoptotic percentage in the 40 mmHg group and 60 mmHg group was significantly increased in comparison with the normal control group (both at P＜0.01).The mitochondrial membrane potential was high in the normal control group and 0 mmHg group,with the reddish fluorescence in the cells,and the reddish fluorescence was weakened in the 20 mmHg group and 40 mmHg group.The lowing of mitochondrial membrane potential was seen in the 60 mmHg group,with the green fluorescence.Compared with the normal control group,the expression of Cyt-c,LC3-Ⅱ and Beclin-1 proteins in pressured groups was correspondingly increased (all at P＜0.05).Conclusions Elevated pressure induces the morphologic change of RGC-5 cells,results in mitochondrial membrane potential reduction and mitochondrial-mediated apoptosis and autophagy.

Objective To evaluate the diagnostic value of the direct MR arthrography in the meniscus degeneration and tear of knee.Methods Thirty subjects undefined by MR imaging having meniscus degeneration and tear of knee underwent MR arthrography.The diagnostic accurate rates in meniscus degeneration and tear by using MR imaging and MR arthrography were evaluated according to the arthroscopy results.Results The diagnostic accurate rates of meniscus degeneration and tear were 80% and 96.7% by conventional MRI scan and MR arthrography respectively.Conclusion The direct low-feild MR arthrography can improve the diagnostic accuracy of meniscus degeneration and tear of knee.

Objective Receptor was screened to find out the interaction between outer membrane protein 25 of Brucella(OMP25) and Brucella infected macrophages.Methods Recombinant OMP25 protein was expressed and purified.The 7 peptide was screened out with OMP25 protein by phage display technology.OMP25-7 peptide interactions were further confirmed by enzyme linked immunosorbent assay(ELISA) after candidate peptides were predicted with bioinformatics software.Four potential 7 peptides were synthetized.Their functions were estimated after Brucella abortus 2308 infected murine macrophages RAW264.7.Results Forty-two cyclic 7 peptides were screened out based on purified OMP25 by the M13 Phage Library Kit.Four peptides,named as ST-7,MT-7,TF-7 and SN-7,were confirmed with ELISA.Colony forming unit(CFU) result demonstrated ST-7,MT-7,TF-7 and SN-7 showed some function for inhibition of intracellular parasite.ST-7 inhibition ratio was 14.4％-51.8％ for Brucella abortus 2308; MT-7 inhibition ratio was 49.6％-69.5％ for Brucella abortus 2308; TF-7 inhibition ratio was 43.2％-74.4％ for Brucella abortus 2308; SN-7 inhibition ratio was 57.5％-82.2％ for 2308.ELISA result showed,that compared to control group,tumor necrosis factor α(TNF-α)increased no statistical significance in each time point of ST-7,MT-7 experiment groups (all P ＞ 0.05).There was no statistical significance in 8 h ago of TF-7,SN-7 experiment groups (all P ＞ 0.05),but there was statistical significance in 12,24 h (all P ＜ 0.05).Conclusions We have obtained four effective OMP25 receptors through phage display technology; TF-7 and SN-7 have inhibited Brucella abortus 2308 invasion and intracellular survival in murine macrophage cells,while ST-7 and MT-7 have only prevented the invasion against Brucella abortus 2308.

Objective To observe the effect of Hydroxycamptothecin (HCPT) on proliferation of human lung cancer cell line A549 in vitro. Method Using cell calture and MTT assay to observe the effect of HCPT on proliferation of human lung cancer cell line A549. Result Lower concentration of HCPT had no evident inhibitory effect on the human lung cancer cell line A549 after 24 h. The effect was evident when the concentration of HCPT was up to 50 ?g/mL, and the inhibitory rate was 44.17%. The inhibitory rate was 50.28% when the concentration of HCPT was up to 100 ?g/mL. The inhibitory effect of HCPT became more significant with the stimulation of the time, and the inhibitory rate of 100 ?g/mL concentration of HCPT was 70.98% after 48 h. Conclusions HCPT can inhibit the proliferation of human lung cancer cell A549 in vitro. The effect is dose and time dependent.

Objective To investigate the expression and significance of CXCL 9 in systemic lupus erythematosus (SLE ) . Methods The level of CXCL9 in peripheral blood from 20 patients with SLE and 20 normal controls were measured by enzyme linked immunosorbent assay .Compare the difference of the expression level of CXCL 9 in peripheral blood between two groups and analyzed their correlation with ages ,duration ,SLE diseases activity index (SLEDAI) .Results The level of CXCL9 in peripheral blood in patients with systemic lupus erythematosus was (1 549 .14 ± 362 .74)pg/L ,but in normal controls was(602 .54 ± 83 .70)pg/L .The level of CXCL9 in peripheral blood in patients with systemic lupus erythematosus was obviously higher than that in controls by statistical analysis ,there was significant difference between the two groups (P<0 .01) .The expression level of CXCL9 in pe‐ripheral blood of patients with systemic lupus erythematosus was positively correlated with SLEDAI (r=0 .892 ,P<0 .01) .Conclu‐sion CXCL9 may be involved in the pathogenesis of systemic lupus erythematosus .

This study aimed at exploring the effects of Griffipavixanthone,a natural small molecule extracted from Garcinia esculenta,on the autophagy and the mechanism behind it.Cervical cancer HeLa cells were selected as the model in vitro.Confocal microscopy,flow cytometry and immunoblotting were performed to know the impacts of Griffipavixanthone on signaling pathway over autophagy flux and calcium level;while nude mouse model was established to explore its anti-cancer action in vivo.It was found that Griffipavixanthone significantly induced GFP-LC3 puncta formation and the accumulation of LC3-Ⅱ/LC3-Ⅰ,and increased the protein expression of SQSTM1 in HeLa cells in a time and concentration dependent manner.MTT assay results showed that Griffipavixanthone inhibited the growth of HeLa cells in a dosage dependent manner,and eliminated the tolerance of tumor cells to nutrient starvation,which fastened the death of cells.Griffipavixanthone also down-regulated the concentration of calcium in the cytoplasm.Furthermore,the in vivo study indicated that intratumoral injection of Griffipavixanthone presented the anti-cancer effect to a certain extent.In conclusion,it was suggested that Griffipavixanthone inhibit autophagy with the potential to be developed into a clinical anti-cancer agent.Griffipavixanthone sensitized HeLa cells to nutrient stress-induced cell death through inhibiting the autophagy flux and the level of calcium in the cytoplasm.

Objective To study the effectiveness of intravenous Lidocaine on subjective tinnitus in the elderly, and analyze its correlation with the effectiveness of subsequently oral Carbamazepine.Methods A prospective case-control study was conducted in 120 elderly patients with subjective tinnitus, who were divided into two groups.The control group (n=60) was treated with oral Carbamazepine 0.1 g Bid for one month, and the treatment group (n=60) was treated with 2％ lidocaine by intravenous injection.Patients in the treatment group were observed on responses in tinnitus during the period with Lidocaine injections.The treatment group was further divided into the improvement subgroup and the no-improvement subgroup, according to responses to intravenous lidocaine.All the patients in the treatment group received oral carbamazepine 0.1 g Bid for one month following the intravenous lidocaine injection phase.Results In the control group, 5 patients showed marked improvement, 20 patients showed partial improvement, 33 patients showed no improvement, and 2 patients showed symptom deterioration, with an effectiveness rate of 41.7％ (25/60).In the treatment group, 6 patients showed marked improvement, 26 patients showed partial improvement, 24 patients showed no improvement, and 4 patients showed symptom deterioration, with an effectiveness rate of 53.3％ (32/60).Of the 32 patients in the improvement subgroup, 6 cases showed marked improvement, 16 cases showed partial improvement, 8 cases showed no improvement, and 2 cases showed symptom deterioration after subsequent oral Carbamazepine treatment, and the effectiveness rate was 68.8％ (22/32).Of the 28 patients in the no-improvement subgroup, 1 case showed marked improvement, 9 cases showed partial improvement, 16 cases showed no improvement, and 2 cases showed symptom deterioration, and the effectiveness rate was 35.7％ (10/28).The effectiveness rate in the improvement subgroup was higher than in the no-improvement subgroup (68.8％ vs.35.7％, x2 =6.55, P＜0.05).The effectiveness of lidocaine followed by carbamazepine was better than that of carbamazepine alone (68.8％ vs.53.3％, x2 =6.13, P＜0.05).Conclusions Oral Carbamazepine treatment following intravenous Lidocaine injection in elderly patients with subjective tinnitus has better therapeutic outcomes than treatment with oral Carbamazepine alone, but is not as effective in patients with negative responses to Lidocaine as in patients with positive responses.

Objective To explore the impact of TIMP 3 regulated by miR-181b as a target gene on invasion and migration of hepatocellular carcinoma (HCC) in vitro.Methods The expressions of miR-181b were detected using SYBR Green real-time fluorescence quantitative polymerase chain reaction on liver cancer specimens and on HCC cell lines.The protein expression of TIMP 3 in HCC was detected using westen blot,and SKHep-1 as a cell line expressing high miR-181b was chosen through reporter gene experiment.TIMP-3 as a target gene regulated by miR-181b and its effect on invasion and migration treated by anti-miR-181 b were studied using transwell and cell scarification test,respectively.Results The expression of miR-181b in HCC was higher than cancer-adjacent tissues and normal liver tissues.The differences among them were significant.There was a correlation between the high expression of miR-181b and invasiveness and metastasis in HCC.The protein expression of TIMP-3 in HCC was significantly lower than normal liver tissues and cancer-adjacent tissues.Expression of miR-181b mRNA was detected in various HCC cell lines such as Hep3B,HepG2,Huh 7,SKHep-1,SNU182,SNU449 and hepatocyte,with the expression of miR-181b in SKHep-1 being the highest (P＜0.01).TIMP3-3UTR was low when the expression of miR-181b was high (P＜0.05).The invasion and migration abilities of SKHep-1 were significantly inhibited by anti-miR-181b (P＜0.05).Conclusion The data suggested that miR-181b promoted invasion and migration of SKHep-1 by down-regulating TIMP-3 in HCC.

Functional gastrointestinal disease is a group of clinical syndrome of non-organic disease. Its various clinical symptoms have a certain specificity and overlap phenomenon, and the mechanism is not clear. TCM believes thatphychological factorsare an important cause. Emotion failurecan effect spleen and stomach functionthrough liver and heart directly or indirectly.At present, the phychological factors and the relationship between functional gastrointestinal disease are getting attentiongradually. It is recognized that the mechanism of phychological factorsmay be related to brain axis dysfunction, mast cell activation, intestinal flora and so on. This article expounded the above-mentioned mechanism and reviewed the detailed TCM intervention measures to functional gastrointestinal disease in recent years.

Irritable bowel syndrome( IBS)is a common functional gastrointestinal disease,relevant investigations on pathogenesis of IBS mainly focus on genetic susceptibility, social psychological stress, visceral hypersensitivity, dysregulation of brain-gut axis,dysbiosis of intestinal flora,and dysimmunity of intestinal mucosa. This article reviewed the correlation between dysbiosis of intestinal microbiota and dysregulation of brain-gut axis in IBS.