Objective To study the evolutionary and mutant characteristics of Ebora virus envelope glycoprotein.Methods A total of 100 Ebora virus envelope glycoproteins amino acid sequences isolated during 1976 and 2014 were collected from National Center for Biotechnology Information (NCBI).Multiple sequence alignment and phylogenetic tree analysis were performed to investigate the evolutionary and mutant characteristics of Ebora virus envelope glycoprotein.Results Glycoprotein amino acid sequences of Ebora virus isolated during 1976 and 2014 showed only 54.00％-65.00％ homology among different subtypes,while 95.00％-100.00％ homology in same subtypes.Ebola virus isolated from different regions in 2014 showed a 99.70％-100.00％ homology of glycoprotein amino acid sequences in the same subtype.The homology of glycoprotein amino acid sequences of Ebola-Zaire virus isolated from Sierra Leone in 2014 was 100.00％,but three strains of Ebola-Zaire virus isolated from Guinea showed diversity in glycoprotein amino acid sequences.Glycoprotein amino acid sequences of Ebola virus with different subtypes were on different branches of phylogenetic tree.Glycoprotein amino acid sequences of Ebola-Zaire virus isolated from Sierra Leone in 2014 were on one branch,and those of Ebola-Zaire virus isolated from other countries during 1976 and 2014 were on the another branch.Conclusions Glycoprotein amino acid sequences of Ebora virus vary with time and region.Ebola-Zaire virus isolated from different regions in 2014 may be two variants with the same origin,and hybrid phenomenon is not observed among virus of different subtypes.

AIM: To investigate the behavior of depression in chronic alcoholism and withdrawal model of mice, and to explore the co-mechanism of alcoholism and depression.METHODS: A novel model of chronic alcoholism was constructed in this study.The animals were divided into normal control group, and alcohol 7 d, 14 d, 21 d and 28 d groups.The mice were given alcohol preference test on the 6th, 13th, 20th and 27th days.After the test, alcohol were withdrawn for 1 d, then the next day the mice were given behavior test of depression.After the test, the mice were sacri-ficed.The contents of 5-hydroxytryptamine (5-HT) and norepinephrine (NE) were detected by HPLC.The expression of cAMP response element-binding protein ( CREB) and brain-derived neurotrophic factor ( BDNF) was detected by Western blot.RESULTS:The mice showed an obvious drinking phenomenon, and the immobility time of forced swimming test and tail suspension test was significantly increased, with increasing drinking days and withdrawal times.5-HT level in 7 d group mice only increased in frontal cortex (P<0.05).However, compared with control group, 5-HT levels in hippocampus and cortex were decreased on the 21th and 28th days (P<0.01).NE levels in 21 d and 28 d groups were decreased in hippo-campus and frontal cortex (P<0.05), and no significant change was observed in 7 d and 14 d groups.The protein levels of p-CREB and BDNF were significantly decreased in hippocampus and frontal cortex of 12 d and 28 d groups (P<0.05), and no significant change was observed in 7 d group and 14 d group.CONCLUSION:The co-mechanism of alcoholism, withdrawal and depression is related to 5-HT.5-HT-cAMP-CREB-BDNF signaling pathway may be a common mechanism for alcoholism and depression.

Object To study the inhibitory effect and mechanism of extract from cultured cell of Taxus chinensis (Pilg.) Rehd. on hepatocarcinoma cell line SMMC 7721. Methods Trypan blue stain assay was used to determine tumor cell growth curve, flow cytometry and Giemsa staining were used to analyze cell cycle and cell apoptosis. Results Inhibitory effect happened on tumor cell line SMMC 7721, G 2/M stage of tumor cells increased with the addition of T. chinensis cell extract by carrene and apoptosis happened. Conclusion The extracts from cultured cell of T. chinensis have inhibitory effect on SMMC 7721 cells and can further induce cell apoptosis.

Objective:To optimize the overall desirability of orthogonal design sith multi index evaluations. Methods:Four multi index evaluations were applied to optimize the overall desirability of an orthogonal design,and their results wer analyzed. Results:According to the optimum conditions,there was no difference among the four multi index evaluations. Conclusion:multi index test can be used to represent the most desirable variables in experimental design.

OBJECTIVE: To compare dispersibility between two kinds of plasdone used as the hydrophilic adhesive in nimesulide dispersible tablets.METHODS: Nimesulide dispersible tablets were prepared with Plasdone S630 and Plasdone K30 as the hydrophilic adhesive respectively which were dissolved in 5% alcohol solution or 5% water solution.By friability,hardness and dispersibility tests,the influence of S630 and K30 in quality of nimesulide dispersible tablets were observed.RESULTS: Plasdone S630 alcohol solution or aqueous solution showed same or better results in friability,hardness and dispersibility compared with Plasdone K30 alcohol solution or aqueous solution.CONCLUSION: Plasdone S630 could be used as a alternate of K30 for hydrophilic adhesive in dispersible tablets.

Ultrasonic microbubbles were used to open blood-brain barriers (BBB) with a reversed and limited behavior feature in the study, which could improve the brain-targeted delivery of anti-tumor drugs. The glioma rat model was prepared. Low-frequency ultrasound was combined with microbubbles to affect the permeability of BBB compared with the permeability of independently administered Evans blue (EB) crossing BBB. Time point and length of ultrasound were investigated whether they affect the permeability of BBB and the damage of brain tissue. The effect of the growth time of glioma on BBB permeability was explored. Only glioma had a very little impact on BBB permeability. However, ultrasonic microbubbles opened the BBB with the features of temporary, limited and reversed behavior and improved EB and magnetic resonance imaging contrast agent penetrating BBB. A length of 30 s ultrasound is appropriate for opening BBB and no damage of brain tissue. Drugs should be injected before ultrasound so that they enter into brain as BBB opening. Ultrasonic microbubbles can open BBB effectively and safely, which improve drugs penetrating BBB under proper time point and length.

AIM:To investigate the effect of phosphodiesterase 4 (PDE4) inhibitor rolipram on the levels of cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), cAMP response element-binding protein (CREB), phosphorylated CREB (p-CREB) and brain-derived neurotrophic factor (BDNF) in the hippocampus and the prefrontal cortex (PFC) of alcoholism model mice.METHODS:The mice (n=60) were randomly divided into control group , con-trol+rolipram group, alcoholism model group, and alcohol +rolipram (0.1, 0.5 and 1 mg/kg) groups.The mice were given alcohol preference test on days 6, 13, 20 and 27.After the test, the mice received withdrawal of alcohol for 1 d.On day 28, the mice were given behavior test of depression , and after the test, the mice were sacrificed.The cAMP levels in the hippocampus and PFC were detected by ELISA , and the protein levels of PKA , CREB, p-CREB and BDNF were detec-ted by Western blot.RESULTS:The mice showed an obvious drinking phenomenon (P<0.01), and the immobility time of forced swimming test and tail suspension test was significantly increased (P<0.01), with increasing drinking days and withdrawal times .However , chronic treatment with rolipram for 28 d reversed this phenomenon .Moreover , the cAMP lev-els in the hippocampus and PFC were significantly decreased after 28 d alcohol treatment ( P<0.01 ) , and pretreatment with rolipram (1 mg/kg) obviously reversed this decrease (P<0.01).Parallel to these changes of cAMP , the protein lev-els of PKA, p-CREB and BDNF were also decreased in the hippocampus and PFC (P<0.01), and 28 d rolipram adminis-tration inhibited the decreased cAMP , PKA, p-CREB and BDNF levels in the hippocampus .Moreover, 28 d rolipram ad-ministration also reversed decreased cAMP , PKA and p-CREB in the PFC.CONCLUSION:Rolipram treatment protects against alcohol-induced depression-like behaviors , and also reduces alcohol drinking .These effects may be related to PDE4-cAMP-PKA-CREB-BDNF pathway .

Aim To investigate the effect of ASX (trans-astaxanthin)on the expressions of NF-κB p65 , iNOS and TNF-αin the hippocampus and the prefron-tal cortex of chronic alcohol mice.Methods 40 mice were randomly divided into control group,7 d,14 d, 21 d,28 d alcohol-treated group,the mice were given alcohol preference testing on day of 6,13,20,27. Mice were subjected to alcohol withdrawal for one day after testing.In order to determine the exact time point of cognitive memory impairment in mice after alcohol consumption,they were given morris water maze test after alcohol preference testing. The other 40 mice were randomly divided into control group, alcohol group and ASX group (20,40,80 mg·kg-1 ).After chronic ASX administration, mice were given one probe trial of 60 s in which the platform was removed from the pool to evaluate escape latency,the number of times the animal crossed the previous location of the platform,time spent in the target quadrant,and swim-ming speed.The expressions of NF-κB p65 ,iNOS and TNF-αwere detected by western blotting after behav-ioral testing.Results The mice showed an obvious al-cohol-related phenomenon on 2 1 and 28 days after al-cohol treatment,and escape latency significantly in-creased,entries in target quadrant and duration in tar-get quadrant significantly decreased with increasing drinking days and withdrawal times.The results also suggested that 2 1 days chronic ASX treatment reversed this learning deficit.Moreover,the expression of NF-κB p65 ,iNOS and TNF-αin the hippocampus were significantly increased after 2 1 d alcohol treatment (P<0.001),and pretreatment with ASX (40,80 mg· kg-1 ) could obviously inhibit these changes (P <0.001);Parallel to these changes in the hippocam-pus,the level of NF-κB p65 ,iNOS and TNF-αwere also increased in the prefrontal cortex (P<0.001 ), however,only ASX (80 mg · kg-1 ) administration could inhibit the increase (P<0.05 ).Conclusion These results indicate that ASX pretreatment can pro-tect against alcohol-induced memory impairment via the inhibition of NF- κB p65 ,iNOS and TNF-αexpres-sions in hippocampus and prefrontal cortex.

OBJECTIVE To investigate the effect of ferulic acid(FA)on motor function of rats with a spinal cord injury model(SCI)and its possible effects on expression of microtubule-associated protein light chain 3(LC3),Beclin1,Bcl-2 and Bax. METHODS SD rats were randomly divided into 3 groups:sham group,SCI group,FA(100 mg·kg-1,po)group. Rats were subjected to moderate contusion inju?ries using a vascular clip for 2 min to establish an SCI animal model beforfe they were given BBB scores and inclined plate scoring function test on 6 h,1,3,7 and 14 d after SCI. After the test,rats were sacrificed. Spinal cords were observed by hematoxylin eosin(HE)staining. Furthermore,the expressions of LC3,Beclin1,Bcl-2 and Bax were detected by Western blotting. RESULTS Compared with the sham group,BBB scores and inclined plate function scores significantly decreased in model group. The BBB scores decreased from 21 in sham group to(0.5±0.5)in SCI group,and inclined plate function scores decreased from 70° in sham group to(5.8±2.0)° in SCI group. However,this was reversed by FA treatment. BBB scores and inclined plate function scores increased from 3.0 ± 1.7 to 6.2 ± 3.6(P<0.05)and from (13.3 ± 4.1)° to(26.7 ± 8.7)°(P<0.05)after FA was po given for 7 d,respectively. HE staining showed the gradual emergence of internal spinal cord edema,while the structural changes associated with spinal cord injury could be significantly reversed by administration of FA. Moreover,the expression of LC3-Ⅱ/LC3-Ⅰand Beclin1 was significantly increased in SCI 1 d group(P<0.01),but was decreased in 14 d group when compared with SCI 1 d group(P<0.05). The expression of Bcl-2 was increased in SCI 14 d group,and the ratio of Bcl-2/Bax was increased on 14 d after SCI(P<0.05). Compared with the SCI group,LC3-Ⅱ/LC3-Ⅰand Beclin1 expression was increased in FA-treated 1 d group(P<0.05),Bcl-2 expression was increased in FA-treated 14 d group and the ratio of Bcl-2/Bax was significantly increased on 14 d after SCI(P<0.05). CONCLUSION FA has a therapeutic effect on SCI,which may be related to the impact of neuronal autophagy and apoptosis. Meanwhile,autophagy of SCI may be a process of gradual enhancement followed by weakening,and the anti-apoptosis effect may gradually increase with autophagy.

Objective To investigate the protective effects of left ventricular myocardial perfusion after delivery of acidic fibroblast growth factor (aFGF) in rats with diabetic cardiomyopathy (DCM) by using ultrasound‐targeted microbubble destruction ( UTMD ) with real‐time myocardial contrast echocardiography( RT‐MCE) . Methods Among 64 male SD rats ,52 rats were randomly selected and were induced DCM by streptozotocin through intraperitoneal injecting ,the other rats as normal control group . DCM rats were randomly divided into the DCM model group ,aFGF only group ,SonoVue‐aFGF group and the SonoVue‐aFGF+ UTMD group in this study . The aFGF only group rats were injected with aFGF solution through tail vein ,the SonoVue‐aFGF group were injected with SonoVue‐aFGF solution through tail vein and SonoVue‐aFGF+ UTMD group rats were injected with SonoVue‐aFGF solution through tail vein and using UTMD simultaneously . All rats underwent conventional echocardiography and RT‐MCE exams before and 4 weeks after intervention . Left ventricular ejection fraction ( LVEF) and left ventricular fraction shortening( LVFS ) were measured by conventional echocardiography . The plateau intensity ( A ) ,initial slope of the curve (β) and myocardial blood flow ( A ×β) of left ventricular anterior wall at the papillary muscle level were measured in left ventricular short‐axis view by RT‐MCE . Results Before intervention , LVEF and LVFS in the DCM model group ,aFGF only group ,SonoVue‐aFGF group and the SonoVue‐aFGF+UTMD group were significantly lower than in the normal control group ( P <0 .01) .During 4 weeks after intervention ,LVEF and LVFS in the aFGF only group ,SonoVue‐aFGF group and the SonoVue‐aFGF+UTMD group were slightly increased than those in the DCM model group ,but no statistically significant differences were found ( P > 0 .05) ,however A and A × β in the SonoVue‐aFGF+ UTMD group were significantly increased than those in the DCM model group( P < 0 .01) . Compared with the same group before intervention ,A and A ×βin the SonoVue‐aFGF+UTMD group were higher ( P <0 .05) and these in the DCM model group were lower during four weeks after intervention ( P < 0 .05) . Conclusions Acidic fibroblast growth factor delivered by using UTMD can improve the left ventricular myocardial perfusion in diabetic cardiomyopathy rats .