Objective To examine the association between the polymorphisms of brain-derived neurotrophic factor (BDNF)gene with heroin dependence.Methods Genomic DNA was isolated from the venous blood leukocytes of 308 unrelated patients with heroin dependence and 31 7 healthy individuals.Seven single nucleotide polymorphisms (SNPs)were genotyped using MassARRAY system.Data were analyzed using HaploView 4.0 and SPSS 20.0 software.Results There was a significant difference in the genotype frequency of rs6265 between heroin dependence group and healthy control group (χ2 =1 5.1 5 1,P =0.001).The rs6265 G allele was significantly higher than in controls (χ2 =9.864,P =0.002,OR =1.429,95% CI =1.143 -1.786).Furthermore,there was also a significant difference in the genotype frequency of rs13306221 between heroin dependence group and control group (χ2 =7.699,P =0.006).The rs13306221 G allele was significantly higher than in controls (χ2 =7.137,P =0.008,OR =0.539,95% CI =0.340-0.853).Strong linkage disequilibrium (LD)was observed in one block (D’> 0.9;r 2 >0.8),and significantly less G-G haplotype frequency of block 1 (χ2 =4.546;P =0.033)was found in heroin dependence group. Conclusion Our findings support the role of BDNF rs6265 and rs13306221 polymorphisms in heroin dependence and may guide future studies to identify other genetic risk factors for heroin dependence.

Objective To investigate the protective effects and mechanisms of granulocyte-colony-stimulating factor (G-CSF)on a rabbit model of chronic myocardial ischemia.Methods Myocardial ischemia models were created by partial ligation of the left anterior descending coronary artery in Japanese white male rabbits.Rabbits were subcutaneously injected with G-CSF (G-CSF group)or saline (control group)for 6 days after myocardial ischemia.The percentage of CD34-positive cells in the peripheral blood was evaluated by flow cytometry,and CD34-positive cells homing and vWF expression in the ischemic myocardium were determined by immunohistochemistry.Results Rabbits in G-CSF group had a higher survival rate than those in control group (P <0.05).Immunohistochemistry of the ischemic myocardium showed that compared with control group,G-CSF group had increased homing of CD34-positive cells on day 7 post-surgery,and more vessels on day 28 post-surgery by anti-von Willebrand factor staining.In addition,we observed an increase in the percentage of CD34-positive cells in the peripheral blood in G-CSF group.Conclusion G-CSF produces an obvious protective effect against chronic myocardial ischemia in rabbits by increasing stem cell mobilization,homing to ischemic myocardium and accelerating neovascularization.

Objective To study the effects of endoplasmic reticulum stress on liver damage in young rats fed with high-fat diet.Methods We divided 48 male weaned young rats randomly into high-fat diet group and control group,which were separately fed with high-fat diet and normal diet.After feeding 8,12 and 1 6 weeks,the body weight and visceral fat of the rats were measured.The serum liver function was measured.The morphology of livers was observed by HE and transmission electron microscopy. The mRNA expressions of ATF6 and GRP78 in hepatocytes were measured with RT-PCR.Results ① The body weight and visceral fat weight of rats in high-fat diet group increased compared with those in control group (P <0.05).② Alanine aminotransferase and aspartate aminotransferase in high-fat diet group increased slightly over time (P >0.05);alanine aminotransferase at week 1 6 was increased significantly compared with that in controls (P < 0.05 ).③ Liver cells in high-fat diet group had steatosis at week 8 and the steatosis became more serious between week 12 and week 1 6.④ In high-fat diet group at week 8 there were a large number of lipid droplets in the cytoplasm,and the cell structure was close to that of normal cells;rough endoplasmic reticulum was nearly normal and the ribosome was visible.At week 12 and week 1 6,besides a large number of lipid droplets,we could also see that some substances with line-like structure deposited in rough endoplasmic reticulum pool.⑤ The expressions of ATF6 and GRP78 mRNA in hepatocytes in high-fat group at weeks 8, 12 and 1 6 were significantly increased compared with those in control group (P <0.05). Conclusion High-fat diet in infants can cause visceral fat accumulation,fatty degeneration of hepatocytes and liver injury.ATF6-mediated endoplasmic reticulum may be closely related to the liver injury which results from highfat diet.

ABSTRACT:Precision medicine is deliberate orchestrated by Obama’s advisers,and it is based on DNA and human genome project.Double helix structure discovery and the human genome project completed are the first and the second revolution of life science.DNA sequencing and genome technology which drive precision medicine have a far-reaching influence.

Objective To investigate the role of RNA-binding protein La protein in the invasion and migration of cervical cancer cells.Methods RNAi technology was used to silence the La protein in HeLa cell,a cell line of cervical cancer,and then screened by G418.Finally the stably expressed HeLa-shLa cell line was constructed and then wound healing,Transwell,Western blot and gelatin zymography assay were performed. Results After La protein HeLa was silenced by RNAi,the invasion and migration capabilities of HeLa cells were decreased significantly compared with those of the controls.Meanwhile,SiRNA-mediated depletion of La reduced the expression of MMP-2 and increased the expression of TIMP-2.Meanwhile the activity of MMP-2 was reduced too.Conclusion RNA-binding protein La promotes the invasion and migration of cervical cancer cells,which may be related to regulating its matrix metalloproteinases and inhibitors.

Objective To investigate the effect of allicin on the development of atherosclerosis in apoE-/-mice and explore its underlying mechanism from the perspective of protein S-nitrosylation.Methods Thirty male apoE-/- mice were randomly divided into 3 groups:control group (saline,ig),low-dose group (allicin,9 mg/kg·d, ig)and high-dose group (allicin,18 mg/kg·d,ig).They were fed with high cholesterol diet for 12 weeks.The levels of plasma lipids,oxidized-LDL (ox-LDL),malondialdehyde,tumor necrosis factor-alpha and nitric oxide (NO)were measured.The atherosclerotic lesions in aortic root were evaluated after hematoxylin and eosin staining and elastica van Gieson and immunohistochemical staining,respectively.Furthermore,in vitro experiments were performed using human umbilical vein endothelial cells (HUVECs).The HUVECs were treated with allicin (10μmol/L or 20 μmol/L)for 24 hours in the presence of ox-LDL (50 μg/mL).The level of NO in supernatant was measured by a nitrate/nitrite assay. The protein S-nitrosylation of the HUVECs was detected through immunofluorescence.Results The histological analysis revealed that allicin treatment not only significantly decreased the areas of the atherosclerotic lesion (all P <0.05)but also suppressed the macrophage accumulation and smooth muscle cell proliferation in the lesion.There was no significant difference in the levels of plasma lipids between control and treated groups.However,allicin exerted obvious anti-oxidative and anti-inflammatory effects. Interestingly,the allicin treatment led to marked increase of the plasma NO level (P <0.05)and aortic protein S-nitrosylation.The experiments in vitro further proved that the allicin up-regulated the levels of NO and protein S-nitrosylation in HUVECs treated with ox-LDL (P < 0.01 ).Conclusion Allicin can inhibit the development of atherosclerosis.The mechanism is associated with the up-regulation of protein S-nitrosylation in endothelial cells, which plays an important role in anti-oxidization and anti-inflammation.

Objective To evaluate the hot and cold characteristics of 10 antibiotics.Methods MTT assay was used to investigate the effects of 10 antibiotics on the growth and proliferation of cultured SMMC7721 cells and MFC-7 cells in vitro .Morphological changes were observed under the inverted microscope.Results The 10 antibiotics,namely,ampicillin,cefixime,cefpodoxime proxetil,cefaclor,cefalexin,azithromycin,clarithromycin, roxithromycin,doxycycline and oxytetracycline showed cold or cool characteristics.Morphological observation showed that cells treated with these drugs presented decreased cell density and turned round.Conclusion The results of this study demonstrated that the cytological method can be used to evaluate the hot and cold characteristics of western drugs.This simple and reliable method will promote research on Chinese medicalization of western drugs.

Objective To investigate the apoptotic effect of petasin on myeloma RPMI 8226 cells and the mechanisms.Methods The inhibition of petasin on the proliferation of myeloma RPMI 8226 cells was tested by trypan blue assay.Apoptosis of RPMI 8226 cells was measured by terminal-deoxynueleotidyl transferase mediated dUTP nick end labeling (TUNEL)assay and Hoechst 33258 staining assay.Effects of petasin on caspase-3,8 and 9 expressions,phosphorylation of ERK1/2,MEK(p-ERK1/2 ;p-MEK)and p38MAPK(p-p38MAPK)protein were analyzed by Western blot.Results Incubation by petasin for 24 h,48 h or 72 h could significantly inhibit the pro-liferation of myeloma RPMI 8226 cells (P <0.01,P <0.01,P <0.01).Petasin induced the apoptosis of myeloma RPMI 8226 cells in time-and concentration-dependent manners (P <0.05,P <0.05).Caspase inhibitor pretreat-ment could significantly inhibit the apoptosis of myeloma cells.After cultured with petasin for 72 h,the expressions of caspase-3,8 and 9 were obviously enhanced (P <0.05,P <0.01,P <0.05)and phosphorylation of p-p38MAPK of RPMI8226 cells was significantly increased (P <0.01).However,phosphorylation of p-ERK1/2 and p-MEK was decreased significantly (P <0.01,P <0.05).Conclusion Petasin can inhibit the proliferation of myeloma RPMI 8226 cells and induce apoptosis.The mechanism may be related to the activation of caspase-3,8 and 9 proteins and the changes in phosphorylation of p38MAPK,ERK1/2 and MEK.

Objective To investigate the mechanism of unbalanced expressions of endothelin receptors (ETA/ETB )in cerebral vasospasm (CVS)after subarachnoid hemorrhage (SAH).Methods The rat CVS models were established by injecting autologous blood into the cisterna magna the second time.Basilar artery morphology was observed under light microscope and immunofluorescence staining was conducted to dynamically detect ETA/ETB receptor expression.Results The cross-sectional area of the basilar artery in the SAH model group decreased at 2 d to 3 d,and then gradually returned to normal.ETA receptor expression in endothelial cells of the basilar artery increased at 2 d after SAH,peaked at 3 d and remained increased till 14 d.ETB receptor expression increased significantly in endothelial cells at 3 d,peaked at 7 d and remained the same level till 14 d.Conclusion The results suggest that ETA/ETB receptors play an important role in cerebral vasospasm after SAH.The specific expression differences of ETB receptor subtypes in the brain vascular layers need further study.

Objective To study the effects of different temporal resolution of spiral computed tomography (CT)on perfusion parameters and perfusion curve of cervical cervical cancer.Methods Ten cases of cervical cancer were clinically confirmed with CT perfusion scanning.The original data were acquired using temporal resolution of 0.75 s.Then the original data were grouped according to different temporal resolution,namely,1.5 s group,2.25 s group,3 s group,3.75 s group,4.5 s group,5.25 s group,and 6 s group (experiment group). According to the same mathematical model and ROI of the same part,perfusion parameters (BF,BV,MTT,and PS)in each group were calculated respectively and compared with the original data.Results BF and MTT were relatively sensitive to the change of temporal resolution.When the temporal resolution was 3 s,it had a significant impact.PS and BV were not so sensitive to the change of temporal resolution.Temporal resolution of 4.5 s had a significant effect on PS. There was a significant effect on BV until the temporal resolution was 5.25 s. Conclusion Changing the temporal resolution will lead to corresponding changes of perfusion curve and perfusion parameters.Under the premise that it does not affect the diagnosis,properly decreasing temporal resolution (circu-lar scanning temporal ≤2.25 s)of CT perfusion scanning of cervical cancer can reduce the radiation dose effectively.