Background Studies showed that inflammatory process participates in the pathogenesis anddevelopment of diabetic retinopathy targeting retinal vascular endothelial cells (RVECs).A growing body of evidence revealed that metformin reduces the risk of micro-and macro-vascular complications by protecting blood-brain barrier,however,whether it plays a protective effect on human retinal vascular by similar mechanism is still unelucidated.Objective This study was to investigate the effects of metformin on the proliferation,migration and secreting monocyte chemotactic protein-1 (MCP-1) and interleukin-8 (IL-8) of human retinal vascular endothelial cells (RVECs) under the stimulation of tumor necrosis factor-alpha (TNF-α).Methods RVECs were cultured and divided into normal control group,metformin (5 mmol/L) group,TNF-α 2.5 ng/ml group,and TNF-α+metformin (5,10,20 and 40 mmol/L,respectively) groups.Corresponding drugs were added into medium according to grouping for 24 hours.Cell numbers were calculated before and after treatment.The metabolic activity (absorbancy) of RVECs was measured with MTS assay.Cell migration of RVECs was assessed with transwell migration assay.The MCP-1 and IL-8 concentrations in the cell supernatant were detected by ELISA assay.Results The number of the cells was significantly different among the normal control group,metformin group,TNF-α group,and TNF-α+metformin (5,10,20 and 40 mmol/L,respectively) groups (F =189.31,P ＜ 0.01).The metabolic activities of RVECs were 0.32 + 0.02,0.32±0.03,0.97 ± 0.02,0.90 ± 0.05,0.76 ± 0.15,0.74 ± 0.05 and 0.41 ± 0.03;migrated cell numbers were (1 214±49),(1 200±45),(1 648±43),(1 309±48),(1 279±73),(961±60) and (942±106)/field;the concentrations of MCP-1 were (0.385 ±0.050),(0.362±0.060),(2.285 ±0.200),(1.131 ±0.180),(0.622 ± 0.120),(0.537±0.090) and (0.492±0.130) μg/ml,and those of IL-8 were (0.385±0.080),(0.390±0.120),(1.123±0.130),(0.899±0.180),(0.680±0.060),(0.417±0.090) and (0.335±0.100) μg/ml in the normal control group,metformin group,TNF-α group,and TNF-α + metformin (5,10,20 and 40 mmol/L,respectively) groups,showing significant differences among the groups (F =73.31,103.89,150.92,268.32,all at P＜ 0.01).The cell number,cell metabolic activity,migrated cell number,and MCP-1 and IL-8 levels in the cell supernatant were evidently increased in the TNF-α group compared with the normal control group,and those in the TNF-α+10 mmol/L metformin group,TNF-e +20 mmol/L metformin group and TNF-α+40 mmol/L metformin group were significantly decreased in comparison with the TNF-α group (all at P＜0.05).Conclusions Metformin can inhibit TNF-α-induced proliferation,migration and MCP-1 and IL-8 secretion of the cells,and therefore plays a protective role on RVECs in the inflammatory environment.

Aim To research the effects of total flavones of rhododendra(TFR)on transient receptor potential vanilloid receptor 4(TRPV4)in cerebral basilar arteries(CBA)of rats subjected to ischemia/reperfusion(IR)injury.Methods The model of total brain IR was established by four-artery occlusion(4-VO)method in rats.Arterial pressure perfusion and cell membrane potential recording methods were used for surveying the dilatation and hyperpolarization of TFR and ruthenium red(RR,an inhibitor of TRPV4)in the KCl-preconstricted CBA ex vivo in rats subjected to IR.Quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot were utilized to investigate the TRPV4 mRNA and protein expressions of TFR and RR in cerebrovascular endothelial cells of CBA in vivo in rats subjected to IR.Results 11～2 700 mg·L-1 TFR significantly induced concentration-dependent hyperpolarization and dilatation in the KCl-preconstricted CBA in rats subjected to IR.TFR still produced degenerative hyperpolarization and dilatation by removal of endothelium in CBA,which was remarkably attenuated as compared with endothelium-intact group(P<0.01).After removal of NO and PGI2 vasodilatation,TFR obviously elicited the hyperpolarization and dilatation that were further decreased by RR(an inhibitor of TRPV4)in IR CBAs.TFR pretreatment apparently increased the level of TRPV4 mRNA and protein expressions in IR CBAs.These effects were restrained by RR,an inhibitor of TRPV4.Conclusions TFR could mediate endothelium-dependent and endothelium-independent effects.The endothelium-derived dilatation may be related to the increase of endothelium activity and endothelium-derived hyperpolarizing factor(EDHF)generation and release that have been promoted by TFR,and secondarily activating TRPV4,which results in Ca2+ inflow and subsequent hyperpolarization of vascular smooth muscle cell membrane and vasorelaxation.

Objective To observe the effects of blood pressure by intravenous infusion of different doses ofoxytocin in cesarean section.Methods Sixty full-term pregnant women undergoing cesarean section with continuous epidural anesthesia were divided into three groups by random digits table method with 20 cases each:group A,B and C.Three groups were injected 10 U oxytocin in uterine muscle after infant delivery.Group A,B and C received 5,10 and 20 U oxytocin (sodium lactate ringer,500 ml) continuous intravenous infusion at the speed of 10 ml/min.If happened uterine contractions bad,they were sublingual administering 0.2 mg misoprostol.If happened severe hypotension,they were intravenous injected 5 mg ephedrine.The change of mean arterial pressure (MAP) and heat rate before anesthesia (T0),after fetal childbirth (T1),5 m in (T2),10 min (T3),30 min (T4) after infusion of oxytocin and the dosage of ephedrine and misoprostol were recorded.Results There were no significant differences in MAP and heart rate at every time point between group A and B (P＞ 0.05).MAP decreased and heart rate increased in group C at T2,T3 compared with those in group A and B,and there were significant differences (P＜ 0.05).The number of cases of sublingual misoprostol were increased in group A (7 cases) compared with that in group B (2 cases) and group C(1 case).The 8 patients injected ephedrine in group C were more than group A(1 case) and group B (3 cases).Conclusion Cesarean section after the delivery of the fetus in the uterus muscle injection of oxytocin 10 U,after 10 U of oxytocin added 500 ml sodium lactate ringer injection at the speed of 10 ml/min intravenous infusion has little effect on the blood pressure and heart rate,and has good uterine contractions.

Objective To evaluate the effect of anticoagulant therapy in the patients with non-small cell lung cancer (NSCLC).Methods One hundred and fifty-nine patients with NSCLC without venous throm-boembolism (VTE)were divided into anticoagulant therapy group (81 cases)and control group (78 cases)by random number table method.The 81 cases in anticoagulant therapy group were treated with anti-tumor therapy and anticoagulant therapy,using low molecular heparin calcium 5 000 U subcutaneous injected for 1 0-30 days, once every 1 2 hours.The 78 cases in control group were merely treated with anti-tumor therapy.Results After treated with anticoagulation therapy,patients in anticoagulant therapy group had prolonged prothrombin time [(1 3.56 ±4.30)s vs (1 5.1 6 ±2.1 2 )s;t =3.1 95,P =0.001 ],active partial thromboplastin time [(28.24 ±5.28)s vs (30.26 ±3.28)s;t =2.71 2,P =0.007)],and a lower FIB [(3.85 ±0.75)g/l vs (4.25 ±2.65)g/l;t =2.971 ,P =0.003]compared with the patients in control group.The incidence of thrombosis rates of the two groups were 2.47% and 1 6.67% respectively,with statistical significance (χ2 =9.901 ,P =0.002).Both the 1 ,2 years overall survival rates of patients in anticoagulant therapy group were longer than those in control group,with statistical significances (χ2 =5.496,P =0.026;χ2 =4.540,P =0.046),while the 1 ,2 years progression-free survival rates of patients in the two groups were no statistical sig-nificances (χ2 =2.034,P =0.1 82;χ2 =0.091 ,P =0.395 ).Adverse reactions such as hemorrhage (4.94% vs 6.41 %),thrombocytopenia (9.88% vs 8.98%),skin necrosis incidence (3.70% vs 1 .28%) in the anticoagulant therapy group and control group were no statistical significances (χ2 =0.51 6,P =0.685;χ2 =0.008,P =1 .000;χ2 =0.847,P =0.632).Conclusion For patients with NSCLC,prophylactic antico-agulant therapy can improve coagulation status,reduce the incidence of thrombosis,prolong OS,and no obvi-ous adverse reactions.

The gut-brain axis (GBA) is a nerve-endocrine mediated bidirectional communication system between the gut and brain, which links the cognition and emotion in brain to peripheral intestinal function. In recent years, many researches have showed that colonized intestinal microbiota plays an important role in the communication between gut and brain. On one hand, microbiota can influence the development and function of brain via GBA. On the other hand, brain can also change the composition of gut microbiota. These findings gradually become a novel medical research highlight, i.e. the microbiota-gut-brain axis. This paper reviews the interaction between gut microbiota and brain via GBA in order to provide supports for studying functions of gastrointestinal tract and brain, as well as the treatment of related diseases.

Objective To investigate the effect of atorvastatin on the expressions of long non-coding RNA (LncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1)and inflammation factors in human umbilical vein endothelial cells (HUVECs) stimulated by high glucose. Methods The expression of MALAT1 in HUVECs incubated with high glucose(30 mmol/L) for different time periods were detected by real-time PCR. Under high glucose condition, the expressions of MALAT1, interleukin-6(IL-6), and interleukin-8 (IL-8) in HUVECs were detected after MALAT1 was silenced by siRNA or atorvastatin was added. Results (1) After HUVECs were incubated with high glucose for different time periods, the expressions of MALAT1 were increased to some extent(P<0.05), with the peak at 12h (P<0.01). The levels of IL-6 and IL-8 expression and secretion were increased after HUVECs were stimulated by high glucose for 12h (P<0.05). (2)The silence of MALAT1 markedly suppressed high glucose-stimulated expression and secretion of IL-6 and IL-8 (P<0.05). (3) Atorvastatin significantly inhibited high glucose-stimulated expressions of MALAT1, IL-6, and IL-8(all P<0.05). Conclusions High glucose induces the secretion of inflammatory factors by stimulating MALAT1 expression in endothelial cells. Atorvastatin significantly inhibits high glucose-stimulated MALAT1 expression and decreases inflammatory reaction.

ObjectiveTo evaluate the efficacy of stereotactic body radiotherapy combined with coinstan taneous gemcitabine,and gemcitabine alone for advanced pancreatic cancer.Methods56 advanced pancreatic cancer patients were assigned into observation group,which accepted stereotactic body radiotherapy combined with coinstantaneous gemcitabine 500 mg/m2,d1,d8.Other 50 patients were assigned into the control group which only accepted gemcitabine 1 000 mg/m2,d1,d8,d15.Stereotactic body radiotherapy was delivered with a total dose of 4 000-4 500 cGy in 10 fractions.ResultsCT examinations were carried out 2 months after treatment.The response rate of the observation group and control group was 82％ and 16％ respectively,and the pain relief rate was 67％ and 17％ respectively.The time to progression of the observation group was 14 months,and was better than that of the control group(7.5 months,x2 =7.31,P =0.032).The median survival time of the observation group and control group was 15.8 months and 13.2 months,and the difference had no statistical significance(x2 =3.28,P =0.082).ConcolusionStereotactic body radiotherapy combined with gemcitabine has a better overall response rate and a pain relief rate.It can prolong the time to progression,but can't improve the overall survival.

Objective To investigate the most sensitive methods for diagnosing spontaneous internal carotid artery dissection (sICAD) and spontaneous vertebral artery dissection (sVAD) respectively,for the sake of earlier and more accurate diagnosis.Methods Consecutive patients with sICAD and sVAD who visited the Department of Neurology and Radiology,Huashan Hospital Affiliated to Fudan University during 2008-2013 were retrospectively reviewed and the sensitivity of CT angiography (CTA),magnetic resonance T1-weighted fat-suppressed images (MR T1-FS) and digital subtraction angiography (DSA) for the diagnosis of sICAD and sVAD was compared.Results Eighty patients (62 male,18 female; mean age (45.7 ± 11.9) years) were included in the study.There were 99 arterial dissections in total,45 cases of sICAD,52 cases of sVAD and 2 cases of spontaneous middle cerebral artery dissections.The sensitivity of CTA,DSA and MR T1-FS for diagnosing sICAD was 97.5％ (39/40),90.0％ (36/40) and 69.6％ (16/23) respectively,while for sVAD was 89.8％ (44/49),84.6％ (44/52) and 100.0％ (27/27) respectively.Conclusions sICAD and sVAD have significant differences in many aspects including diagnostic strategies.CTA and MR T1-FS seem to be the most sensitive methods for the diagnosis of sICAD and sVAD respectively.Although DSA has been considered as the gold standard for the diagnosis of artery dissection,this imaging technique does not allow analysis of artery wall thickness,thus also has limitations.It is likely that the diagnostic sensitivity will be improved by combining CTA and MR T1-FS.

The treatment for severe pancreatitis includes surgical and non-surgical methods,and the key points of treatment include surgical timing,surgical method selection and the management of postoperative complications.Hepatic artery thrombosis after surgery for severe pancreatitis is rarely seen,and few experiences in the diagnosis and treatment for this disease have been summarized.A patient with the course of severe pancreatitis of 10 years and suffered from 3 different kinds of diseases including thrombosis of right hepatic artery was cured by open surgery for 2 times and intervention therapy in the Affiliated Hospital of Hainan Medical College in October 2011.The treatment experience was summarized based on the clinical data of this patient.

BACKGROUND:Specific down-regulation of interleukin 1 beta (IL-1β) may al eviate the pain behaviors effectively after peripheral nervous injury. Compared with smal interference RNA (siRNA), short hairpin RNA (shRNA) could inhibit the expression of target gene more stably and efficiently. However, simple shRNA could not enter target cel s to down-regulate target gene efficiently. Adenovirus vectors have wide host range, high infection efficiency and stable expression in host cel s.
OBJECTIVE:To construct recombinant adenovirus vector expressing shRNA targeting IL-1βgene and detect its effect on the expression of target gene.
METHODS:Three siRNAs were designed on the basis of the nucleotide sequence of IL-1βobtained from NCBI and then three shRNAs (shRNA1, shRNA2 and shRNA3) were synthesized. The annealed shRNA product and adenovirus vector pHBAd/U6/GFP digested by BamH I and EcoR I were connected to construct the recombinant adenovirus vector shuttle plasmid expressing shRNA targeting IL-1β. After sequencing, HEK 293 cel s were co-transfected by the shuttle plasmid and skeleton vector, and three recombinant adenovirus vector expressing shRNA targeting IL-1β(rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3) were packaged and amplified. Rats H9C2 cel s were infected by recombinant adenovirus vector expressing shRNA targeting IL-1βand fluorescence microscope was used to observe the infection efficiency. The effect of recombinant adenovirus vector expressing shRNA targeting IL-1βon the expression of target gene was detected by western blot assay.
RESULTS AND CONCLUSION:The sequencing results showed that the sequences of three shRNAs adenovirus vector shuttle plasmid were consistent with the sequences of three designed shRNAs. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 were constructed successful y. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 could down-regulate the expression of IL-1βin rat H9C2 cel s and the down-regulation effect of rAd/shRNA2 was the most significant.