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.

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 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.

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 investigate the changes in the phosphorylation of N-methyl-D-aspartate (NMDA) receptors in spinal dorsal horns in a rat model of neuropathic pain induced by chronic compression of dorsal root ganglion (CCD).Methods Ninety-six male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups (n =32 each) using a random number table:control group (group C),sham operation group (group S) and group CCD.In CCD group,a small stainless steel needle (4 mm in length and 0.8 mm in diameter) was inserted into the L4,5 intervertebral foramen in pentobarbital sodium-anesthetized rats,developing intervertebral foramen stenosis and hence producing a chronic steady compression of the dorsal root ganglion.In group S,the intervertebral foramen was only exposed without inserting the needle.The paw withdrawal latency (PWL) was measured at 1 day before operation and 1,2 and 4 weeks after operation.The rats were then sacrificed after measurement of PWL and the lumbar enlargement segments of the spinal cord were removed for determination of the expression of interleukin-1β (IL-1β) and phosphorylation of NMDA receptor NR1 subunits at serine 896 (pNR1S896) on the injured side by immunohistochemistry and Western blot.Results Compared with group C,the PWL was significantly shortened at each time point after operation,and the expression of IL-1β and pNR1S896 was up-regulated in group CCD,and the PWL was shortened at 1 week after operation,the expression of IL-1β and pNR1S896 was up-regulated,and no significant change was found in PWL and expression of IL-1β and pNR1S896 at the other time points in group S.Compared with group S,the PWL was significantly shortened at each time point after operation,and the expression of IL-1β and pNR1S896 was up-regulated in group CCD,Compared with the baseline value at 1 day before operation,the PWL was slowly shortened starting from 1 week after operation,and decreased to the lowest level at 2 weeks after operation and maintained at this level for 2 weeks,the expression of IL-1β and pNR1S896 was slowly up-regulated starting from 1 week after operation,and increased to the highest level at 2 weeks after operation and maintained at this level for 2 weeks in group CCD,and the PWL was shortened at 1 week after operation,and the expression of IL-1β and pNR1 S896 was up-regulated in group S.Conclusion The development and maintenance of neuropathic pain induced by CCD are related to phosphorylation of NMDA receptors in spinal dorsal horns of rats.

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.

ObjectiveTo analyze the potential risk factors for relapse and development of epilepsy in patients with benign in-fantile convulsions associated with mild gastroenteritis (BICE).MethodsA total of 264 cases of BICE were recruited. Accord-ing to the frequency of convulsions, the patients were divided into single group (n=134, convulsion once), and multiple group (n=130, convulsions≥2 times). According to convulsion duration, the patients were divided into short-term group (n=186, con-vulsions duration <5 minutes) , and long-term group (n=78, convulsion duration≥5 minutes). The clinical data obtained during hospitalization and follow-up were analyzed.ResultsIn multiple group, 9.23% were relapsed and 6.15% developed epilepsy. In single group, 2.99% were relapsed and 0.75% developed epilepsy. There were signiifcantly different in the rate of relapses and development of epilepsy between two groups (P<0.05). In the long-term group, 12.82% were relapsed and 8.97% developed epi-lepsy. In the short-term group, 3.23% were relapsed and 1.08% developed epilepsy. There were signiifcantly different in the rate of relapses and development of epilepsy between the two groups (P<0.05).ConclusionsThere are the risks of relapse and development of epilepsy in BICE patients. Convulsions≥ 2 times and≥5 minutes may be the risk factors of relapse and devel-opment of epilepsy.

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:The use of three-dimensional cel culture techniques can better simulate the cel ular microenvironment, providing new tools for tissue engineering research.
OBJECTIVE:To analyze the biomaterial selection and application characteristics in three-dimensional cel culture as wel as applications in tumor tissue engineering.
METHODS:We searched Wanfang database and PubMed database 1998-2015 years for relevant literature using keywords of“three-dimensional cultures;scaffold;cel growth;cel differentiation;tumor tissue engineering”in Chinese and English, respectively.
RESULTS AND CONCLUSION:The selection and application of three-dimensional scaffold materials is one of the keys. So far, scaffold materials, such as col agen gels, gelatin sponge, agarose, chitosan, demineralized bone matrix, cannot provide the extracel ular matrix similar to the micro-environment in which seed cel growth and proliferation are not affected, and the ability to secrete type II col agen and glycosaminoglycan is decreased, although they can provide three-dimensional space for seed cel s. Biomimetic scaffold characterized as little trauma and strong plasticity gradual y shows its unique advantages. Three-dimensional culture conditions raise pro-angiogenic growth factor secretion from tumor cel s, and this feature is positively correlated with the occurrence of in vivo tumor angiogenesis.