AIM To develop a murine model for investigating the effects of prenatal cocaine exposure on the development of fetal nerve system. METHODS A nutritionally paired control group of dams injected with saline and pair fed with the COC dams were set up. Another two groups were COC groups injected with cocaine HCl and SAL group administrated with saline. After injection twice daily during gestation days 8～17,mice were decapitated on E17 and blood and brain samples were collected for pharmacological analysis and neurotransmitter analysis by HPLC.RESULTS Pharmacological analysis revealed that cocaine was found in maternal and fetal plasma at 15 min following ip administration to embryonic day E17 pregnant mice. Though COC dams and SPF dams had the same feeding condition, compared with the latter, the former had higher maternal concentrations of DA and 5 HT, lower fetal weight, brain weight, striatum weight and higher concentrations of DA and 5 HT in striatum, P

AIM To investigate the effects of in utero cocaine exposure on the fetal development, when fetuses were exposed to equal total dose but different dose and timing. METHODS Pregnant dams were randomly separated into three groups: SAL, COC20 and COC40. On E17, recorded body weight, brain weight and striatum weight of all groups, and examined the concentrations of DA and 5 HT in fetal striatum by HPLC. RESULTS Body weight of SAL, COC40, COC20 groups decreased progressively in turns. Brain weight of COC20 group and COC40 group was lower than that of SAL. Only the brain/body ratio of COC40 was decreased ( P

Objective:To investigate the effect of c-maf gene on the MM cells' proliferation and invasion activity. Methods:Lipo-fectin Reagent was used to transfect c-maf siRNA into multiple myeloma cell of RPMI8226. The mRNA expression level of c-maf was detected by RT-PCR.Cell growth curve was measured by MTT assay. Transwell chamber test was used to measure MM cells' in vitro in-vasion activity. The cell cycle distribution were assessed by flow cytomentry. The protein expression levels of survivin,MMP-2, MMP-9, ARK5 and cyclin B1 were detected by Western blot. We also detected the activity of Caspase-3/7. Results:The c-maf siRNA was effectively transfected into cells and the mRNA expression of the c-maf gene was inhibited.MTT test and Transwell chamber test showed that the proliferation and in vitro invasion activity of transfected cells were significantly lower than those of other two groups (P<0.05). Cell cycle of c-maf siRNA transfected group cells was arrested in G2/M phase. The expression levels of survivin,MMP-2, MMP-9,ARK5,cyclin B1 and the activity of Caspase-3/7 between c-maf siRNA transfected group and the other two groups were sta-tistically different (P<0.05). Conclusion:c-maf gene by c-maf siRNA can remarkably inhibit proliferation and invasion of multiple my-eloma cell lines of RPMI8226. C-maf gene may be used as the target for multiple myeloma gene therapy.

Objective To investigate the interaction of human testis sperm-associated antigen 4 like(SPAG4L) protein containing Sad,UNC-84(SUN) domain with nuclear envelop spectrin repeat proteins 3 (Nesprin-3) containing Klarsicht,ANC-1 and Syne homology (KASH) domain.Methods SPAC-4L protein domains were analyzed with the bioinformatics method.After the SPAG4L plasmid was transfected into Ntera-2 cells,the subcellular localization of SPAG4L was observed.The interaction of SPAG4L with Nesprin-3 was determined by immunofluorescence,co-immunoprecipitation(co-IP) and bimolecular fluorescence complementation (BiFC) technology,respectively.Results Bioinformatics analysis results showed that there was a transmembrane structure in SPAG4L protein.Subcellular localization results demonstrated that SPAG4L protein located in the nuclear membrane and cytoplasm.Immunofluorescence,Co-IP,and BiFC results showed that there was an interaction between SPAG4L and Nesprin-3,and that a LINC(linkers of the nucleoskeleton to the cytoskeletan) complex was formed.Conclusion SPAG4L may interact with Nesprin-3 to form a LINC complex,which is important for understanding the function of SPAG4L protein in spermatogenesis.

objective To investigate the recurrence of stroke,clinical prognosis and vascular changes in patients with ischemic stroke due to middle cerebral artery stenosis. Methods The ischemic stroke patients with symptomatic middle cerebral artery stenosis were enrolled continuously and followed up prospectively for six months. The recurrence of ipsilateral stroke,clinical prognosis and dynamic changes of vessels were analyzed. Results Eighty patients were included,and 20.0% of the patients(16 cases)presented with recurrence of ipsilateral ischemic stroke and 56 cases (70.0%)with a good outcome(modified Rankin scale[mRS]≤1)during the 6 months follow-up;38.6% patients (27 cases) presented with significant vascular changes with progression in 12 cases (17.1%)and regression in 15 cases (21.4%). Conclusion The patients with simple symptomatic middle cerebral artery stenosis have an high rate recurrence of ipsilateral stroke but have good prognosis;Lesioned artery of the majority of patients in the short period after stroke was stable,but vascular stenosis in some patients could appear progression or remission.

Objective To observe the effects of electroacupuncture at Ganshu (BL18) and Zusanli (ST36) on depression state and gastrointestinal motility and relevant protein expressions in brain of rats with stress-induced depressive functional dyspepsia. Methods Totally 48 Wistar rats were randomly divided into normal, model, electroacupuncture (EA) and sham-operation group, with 12 rats in each group. Depressive functional dyspepsia model was established by chronic unpredictable mild stress protocols. From day 15 to day 28, the EA group was given electroacupuncture at BL18 and ST36, and the sham-operation group was stimulated by electroacupuncture at 1/3 and 2/3 of the tail. Body weight test, open field test and sucrose preference test were measured on day 0, day 14 and day 28. After 14-day administration, gastric residual volume and intestinal propulsion rate of semisolid paste were detected. The expression of βCaMK inⅡ lateral habenulas and BDNF in hippocampus were measured by Western blot. Results Compared with the normal group, the weight, the sucrose preference rate, the behavior score, the small intestinal propulsion rate of semisolid paste and the expression of BDNF in hippocampus decreased significantly both in the model group and sham-operation group (P<0.01), and the gastric residual volume of semisolid paste and the expression of βCaMK inⅡ lateral habenulas increased significantly (P<0.01). Compared with the model group, the weight, the sucrose preference rate, the behavior score, the small intestinal propulsion rate of semisolid paste and the expression of BDNF in hippocampus increased significantly in EA group (P<0.01), and the gastric residual volume of semisolid paste and the expression of βCaMK inⅡ lateral habenulas decreased significantly (P<0.01). Compared with the sham-operation group, the expression of βCaMKⅡ in lateral habenulas decreased significantly, the expression of BDNF in hippocampus increased significantly in EA group (P<0.01). Conclusion Electroacupuncture at Ganshu (BL18) and Zusanli (ST36) can improve depression and gastrointestinal motility of rats with depressive functional dyspepsia, which may be related to the inhibition of βCaMK inⅡ lateral habenulas and the activation of BDNF in hippocampus dyspepsia.

Objective To study the effects of prenatal cocaine exposure on the development of offspring's brains by building a murine model. Methods We weighted the body weight and brain weight of offspring on P10 from COC and SAL groups and observed the development of neuron and astrocyte in cerebral cortex by toluidine blue staining and immunohistochemistry. Results The brain weight and body weight from COC were both reduced on P10 compared with SAL group.We discovered prenatal cocaine exposure induced polarity disorder and dysplasia of neuron in cerebral cortex;the number of the astrocytes in corpus callosum and hippocampus regions decreased.Conclusion\ Pregnatal cocaine exposure can result in abnormal development of cerebral cortex of offsprings which may play an important role on cocaine induced abnormal behavior.\;[

ObjectiveTo detect the Alu expression in the stool of patients with pancreatic cancer and investigate its value in the diagnosis of pancreatic cancer.MethodsStool samples were obtained from patients with pancreatic cancer (PC) ( n =41 ),chronic pancreatitis (CP) ( n =27 ) and healthy subjects ( n =23 ),the DNA was extracted from the stool and the expression of Alu repetitive sequences was subjected to quantitative analysis by the real-time PCR.ResultsThe expressions of Alu repetitive sequences in PC,CP,and healthy subjects were (5.17 ± 0.99 ),( 3.79 ± 0.94),(0.28 ± 0.35 ) rig/g,and the difference among the three groups was statistically significant (P ＜0.05).The AUC of PC was 74.8％ with the 95％ CI 0.661 ～0.835,and the sensitivity,specificity was 75.6％ and 67.1％,respectively.ConclusionsAlu repetitive sequences are highly expressed in the stool of patients with pancreatic cancer,and it is of value in the diagnosis of pancreatic cancer.

Objective To detect the level of plasma miR-155 in patients with pancreatic cancer and evaluate its diagnostic value for pancreatic cancer. Methods Sixty-two cases of pancreatic cancer patients, 61 patients of chronic pancreatitis and 36 normal controls were included in the study. miR-155 in the total RNA extracted from the plasma was measured using real time PCR. The diagnostic parameters and relationship of the miR-155 with clinical characteristics in pancreatic cancer patients were analyzed. Results The relative abundance of plasma miR-155 in pancreatic cancer, chronic pancreatitis and normal controls were 5.41 ±3.14,2.59 ±2.49 and 0.77 ± 1.17, and the value was significantly higher in pancreatic cancer group compared with those of chronic pancreatitis and normal group (P ＜ 0. 01 ). No significant correlation between the level of plasma miR-155 and age, sex, tumor size in pancreatic cancer patients was found. But there was a negative correlation between the level of plasma miR-155 and TNM staging (r = -0. 323, P = 0.01 ). For discriminating pancreatic cancer from normal control, the AUC ROC of miR-155 was 0. 943 (95% CI 0.902-0.985), and the sensitivity and specificity were 87.1% and 83.3%, respectively. For discriminating pancreatic cancer from chronic pancreatitis, the AUC ROC of miR-155 was 0.762 (95% CI0. 678-0. 846),and the sensitivity and specificity were 64.5% and 73.8%, respectively. For discriminating pancreatic cancer from chronic pancreatitis and normal, the AUC-ROC of miR-155 was 0. 829(95% CI0. 767-0.892), and the sensitivity and specificity were 62.9% and 84.5%, respectively. Conclusions Plasma miR-155 was significantly increased in patients with pancreatic cancer, and can be used for diagnosis of pancreatic cancer.

Objective To investigate the expression of miRNAs in endoscopic ultrasound (EUS)-guided fine needle pancreatic aspirates (FNA) and its clinical significance. Methods The expression of miRNA-210, miRNA-21, miRNA-196a, miRNA-181a, miRNA-181b, miRNA-155 and miRNA-16 in EUS-FNA specimen of 23 pancreatic cancer and 13 benign pancreatic masses was detected with quantitative real-time polymerase chain reaction (qRT-PCR) assays and its clinical significance was analyzed. Results The relatively expression quantity of miRNA-210, miRNA-21,miRNA-196a, miRNA-181a and miRNA-181b (0.86±1.10, 0.69±0.64, 0.32±2.50, 0.16±0.83and 0.56 ±0.88, respectively) was significant higher in pancreatic cancer than those in the benign pancreatic masses (-0.11±0.98,-0.03±0.97,-1.50±1.40,-0.53±1.10 and -0.28±1.10,respectively) (P value was 0. 012, 0. 011, 0. 024, 0. 036 and 0.015, respectively). The relatively expression quantity of miRNA-16 and miRNA-155 in pancreatic cancer and benign pancreatic masses was - 0.11 ± 0.69, 0.08 ± 1.04, - 0.73 ± 1.26 and - 0.19 ± 1.19 respectively, no statistic difference (P value was 0. 067 and 0. 467). The high expression of miRNA196a was positively correlated with pancreatic cancer lymphatic metastasis and clinical TNM staging. Conclusion There were multi miRNAs with abnormal high expression in the pancreatic cancer. Of those, miRNA-196a was associated with the clinical malignant features of pancreatic cancer.