Objective To explore the changes of the expression of islet amylodi polypeptide ( IAPP ) and somatostatin( SS) of islet in type 1 diabetes mice, and the mechanism of the expression changes .Methods We established the diabetes model in C57BL/6J mice by low-dose streptozotocin ( STZ) injection.The excisions of the pancreas tails removed on the 3th, 7th, 10th, 14th, 21th and 28th day.Tissues were assessed by immunohistochemical SABC, immunofluorescence in the study .Results 1.Numerical density on area ( NA ) of IAPP positive cells in experimental group (EG) decreased since the 3th day, but average absorbance increased since the 7th day.2.NA of SS-IR cells in EG increased since the 3th day, and average absorbance increased since the 7th day.3.The results of immunofluorescence double staining showed that , IAPP and SS could coexpression in part of cells in islets .Conclusion The number of IAPP positive cells in type 1 diabetes is decreased , but the immunoreaction increased .Immunoreaction and number of SS positive cells increase .Both of them are involved in the pathogenesis of type 1 diabetes.

AIM: To investigate the protective mechanism of extract of Ginkgo biloba (EGB) on apoptosis of hippocampus neuronal cells in type 1 diabetic encephalopathic rats. METHODS: Thirty-six male Sprague-Dauley rats were divided into 3 groups: control group, diabetic group and EGB-treated group. Streptozotocin was injected intraperitoneally to the animals in later two groups to induce diabetes. The rats in EGB-treated group were injected intraperitoneally with EGB, and the same volume of normal saline was injected to the rats in other groups. At the end of the 12th week, the spatial learning and memory abilities of rats in each group were examined by Morris water maze test. Blood glucose and serum insulin concentration were measured. The neuron densities in hippocampus were measured by Image-Pro Plus 6.0 software. The expressions of Bax, Bcl-2, caspase-3 were assayed by Western blotting and immunohistochemistry. RESULTS: Compared to control group, the level of blood glucose (P<0.01), the protein expression of Bax (P<0.01) and caspase-3 (P<0.01) in hippocampus neuronal cells, and the ratio of Bax/Bcl-2 (P<0.01) and the escape latency (P<0.01) in diabetic group, were significantly increased, while the serum insulin concentration (P<0.01), the neuronal density (P<0.05) in CA1,CA2 hippocampal regions and the platform searching score (P<0.01) were significantly deceased. After treated with EGB, the serum insulin concentration (P<0.05), the neuronal density (P<0.05) in CA1,CA2 hippocampal regions and the platform searching score (P<0.01) were significantly increased, while the level of blood glucose (P<0.01), the protein expression of Bax (P<0.05), caspase-3 (P<0.05) in hippocampus neuronal cells, the ratio of Bax/Bcl-2 (P<0.01) and the escape latency (P<0.05) were significantly deceased than those in diabetic group. The protein expression of Bcl-2 in hippocampus neuronal cells did not alter in any experimental rats. CONCLUSION: EGB improves the spatial learning and memory capacity in diabetic rats by decreasing the expression of Bax, Bax/Bcl-2 ratio and down-regulating caspase-3 to reduce neurocyte apoptosis and increase the neuron density in CA1, CA2 hippocampal regions, suggesting that effective regulation of neuron apoptosis associated genes may be one of the mechanisms for EGB to treat diabetic encephalopathy.

AIM: To investigate the effects of antagonistic peptide specifically binding to the second extracellular loop of CC chemokine receptor 5 (CCR5) on inflammatory cell infiltration and TNF-α expression in lung tissues of asthmatic mice.METHODS: The asthmatic model of BALB/c mice was induced by ovalbumin (OVA) and the optimal sensitization concentration of OVA was screened.After modeling, the mice were intervened by gradual concentrations of antagonistic peptide via tail-vein injection.The pathocytological analysis and grading were performed in the lung tissues with HE staining.The expression of TNF-α at mRNA and protein levels in the lung tissues was determined by real-time PCR and Western blot.RESULTS: The optimal concentration of OVA was 500 mg/L (0.1 mL) as this concentration of OVA stably induced moderate degree of inflammation in the BALB/c mice.Treatment with different concentrations (1.5 g/L, 2.5 g/L and 3.5 g/L) of antagonistic peptide at 0.2 mL through tail-vein injection inhibited the expression of TNF-α, and markedly reduced the degree of inflammation in the lung tissues.The optimal concentration of antagonistic peptide was 2.5 g/L as the lung inflammation degree in 2.5 g/L group alleviated by 2 grades, and the number of inflammatory cells was also significantly reduced.Moreover, the mRNA expression abundance of TNF-α was nearly decreased by 90%, and the protein expression of TNF-α was decreased by 70% compared with model group.Meanwhile, the use of antagonistic peptide at 2.5 g/L before OVA stimulation confirmed the preventive function to some degree.In this group, the lung inflammation degree alleviated by 1 grade, and the expression of TNF-α at both mRNA and protein levels decreased by nearly 50%.CONCLUSION: The antagonistic peptide of CCR5 effectively inhibits the expression of TNF-α and relieves the inflammation in the asthmatic mouse lung tissues in a concentration-dependent manner.

Purpose A RP-HPLC method was used to determine genioside and breviscapine in plasma and to study its pharmacokinetics in rat, respectively.Methods Rat plasma samples were collected after a single dose of Zhideng injection and pharmacokinetic parameters of genioside and breviscapine were estimated,respectively.Results A good linear relationship was obtained between 0.2-40.0 μg/mL for breviscapine, and 0.5-200.0 μg/mL for genioside.The recoveries from plasma were larger than 85%,and RSDs of inter-day asaay and intra-day assay were below 10%. The pharmacokinetic results showed that genioside and breviscapine were rapidly eliminated from plasma after iv administration of three doses of Zhideng injection.The mean half-life was 72.6 min and 21.6 min,respectively.Conclusion The established HPLC method was suitable for the pharmacokinetic study of genioside and breviscapine.

AIM:To study the effect of gossypol on the cognitive function of type 2 diabetic rats,and to explore its mechanism. METHODS:Thirty male Sprague-Dawley rats were divided into three groups randomly:normal group,type 2 diabetic group and gossypol treated group. After fed with high-fat diet for 4 weeks,the later two groups were injected with streptozotocin intraperitoneally to establish type 2 diabetic rat model. The animals in gossypol treated group were given gossypol at dosage of 15 mg/kg once per day for 4 weeks by gavage. Since 5th week,the times of gavages were changed into once per week at the same dosage and lasted to 12th week. Learning and memory abilities of rats were assayed with Morris water maze test. The concentration of blood glucose was measured by biochemical method. The levels of serum corticosterone and insulin were detected by enzyme-linked immunosorbent assay and radioimmunoassay,respectively. The protein expressions of 11?-HSD1 and GR in cerebral cortex and hippocampus were determined by Western blotting. The morphological changes of cerebral cortex and hippocampus were observed under light microscope and transmission electronic microscope,respectively. RESULTS:Compared to normal group,the karyopyknosis,dilation of golgiosome and mitochondria swelling of neuron from cerebral cortex and hippocampus were prominent in diabetic group. The concentrations of blood glucose,serum corticosterone and insulin increased significantly (P

Objective To investigate the characteristics and relationship of cognitive function and resting state functional magnetic resonance imaging(fMRI) amplitude of low-frequency fluctuations (ALFF)in childhood and adolescence-first-onset obsessive-compulsive disorder(OCD).Methods Twenty-four childhood and adolescence-first-onset OCD patients (patient group) and thirty-three age/gender/education-matched healthy controls (healthy control group) were enrolled in this study.The participants' cognitive function was tested with trail making test (TMT),symbol coding,Hopkins verbal learning test-revised (HVLT-R),brief visuospatial memory test-revised (BVMT-R),verbal fluency test (VF),digital span test,maze test and Stroop test.Then a resting-state fMRI scan was conducted for each participant.Amplitude of low-frequency fluctuation approach was used to explore the differences of resting-state brain function between patients and controls,and analyze the significant brain regions by adopting t test.The cognitive functions of patients and healthy control group in the significant brain regions were compared by using Pearson correlation analysis.Results The patient group had lower scores than healthy control group in symbol coding,BVMT-R and digital span test,and the differences were statistically significant [(45.24 ± 10.32)scores vs.(56.85 ± 9.57) scores,(21.94 ± 6.99) scores vs.(30.58 ± 3.92) scores,(64.06 ± 20.00) scores vs.(78.03 ±18.08) scores;t =3.96,4.90,2.50;all P ＜ 0.05].Corrected by Alphasim,patient group showed significantly increased ALFF in the left and fight temporal lobe,left and fight posterior lobe of cerebellum,right middle temporal gyrus,but ALFF decreased in the area of left and right lenticula,left putamen compared with the healthy control group (all P ＜ 0.05).The patients group showed that TMT scores were positively correlated with the value of ALFF in the right temporal lobe (r =0.67),and color scores were positively correlated with the value of ALFF in the right middle temporal gyrus (r =0.53),but colour scores were negatively correlated with the value of ALFF in the right tcmporal lobe (r =-0.54);the healthy control group showed that BVMT-R and digital span test scores were positively correlated with the value of ALFF in the left temporal lobe (r =0.47,0.41),but digital span test scores were negatively correlated with the value of ALFF in the right temporal lobe (r =-0.49),and colour scores were negatively correlated with the value of ALFF in the left temporal lobe (r =-0.35),while word and color-word scores were negatively correlated with the value of ALFF in the right middle temporal gyrus (r =-0.39,-0.36);the correlation coefficient above had statistical significance (all P ＜ 0.05).Conclusions The childhood and adolescence-first-onset OCD patients had cognitive impairment,including speed of processing,visual learning and memory,working memory;and the patients showed abnormal spontaneous neural activity at some brain regions.The cognitive dysfunction may be associated with abnormal spontaneous neuronal activity in childhood and adolescence-first-onset OCD.

Objective To investigate the expression of monocyte-macrophage-related factors and interstitial fibrosis in kidney tissues of rats with ureter obstruction and recanalization .Methods Forty-eight male Spragur-Dawley rats were divided randomly into the obstructive group:sham (n=6), unilateral ureteral obstruction(UUO)3 days (n=6), UUO 7 days (n=6), and UUO 14 days (n=6) and recanalization group:bilateral ureteral obstruction(RBUO)0 day (n=6), 3 days after RBUO (n=6), 7 days after RBUO (n=6), and 14 days after RBUO (n=6).The kidneys were excised on day 3, 7, and 14, and the deposition of collagen fibers in kidney was detected with HE and Masson staining . Immunohistochemical analysis was performed to evaluate the protein expressions of monocyte chemoattractant protein -1 (MCP-1), macrophage colony-stimulating factor (M-CSF) and activated-macrophage marker CD68.Real-time PCR was used to detect the mRNA expressions of MCP-1 and M-CSF.TGF-β1 levels were determined by ELISA .Results Fibrosis observed with HE and Masson staining was obviously increased in kidney tissue of UUO rats , and aggravated as time prolonged, but alleviated in rats with recanalization .TGF-β1 levels were increased obviously in the UUO group , but decreased in rats with recanalization compared with those in BUO rats .In UUO rats, mRNA and protein expression levels of MCP-1 and M-CSF were increased .MCP-1 and M-CSF expression was gradually decreased in rats with recanalization compared with those in BUO rats .The dynamic change in expression of MCP-1 and M-CSF in both UUO rats and recanalization rats was consistent with the change in expression of CD 68. Conclusion Dynamic change in expression of MCP-1 and M-CSF in kidney tissues reflects change of activated and accumulated monocyte -macrophages , which may be one of the major mechanisms contributing to fibrosis induced by ureter obstruction .Renal fibrosis is alleviated by down-regulated expression of monocyte-macrophages factors with recanalization operation .

AIM:To observe the effect of the extract of Ginkgo biloba(EGB)on pituitary-testicular axis and the mRNA expressions of luteinizing hormone receptor(LHR)and steroidogenic acute regulatory protein(StAR).METHODS:Thirty male Sprague-Dauley rats were divided into three groups randomly:normal control group,type II diabetic group and EGB treatment group.After fed with high-fat diet for 4 weeks,the later two groups were injected with strepozotocin intraperitoneally to induce type II diabetes mellitus.The EGB treatment group was given EGB at the dose of 50 mg/kg once a day for 12 weeks by intragastric administration.The normal control and diabetic group were given normal saline of equal volume per day for 12 weeks.The indices of blood glucose,insulin and low-density lipoprotein-cholesterol(LDL-c)were measured.The morphologic change of testicular tissue was observed under light microscopy(LM)and transmission electron microscopy(TEM)respectively.The concentrations of blood luteinizing hormone(LH)and testosterone(T)were assayed by the technique of enzyme linked immunosorbent assay(ELISA).The mRNA expressions of LHR and StAR from Leydig cells were detected by RT-PCR.RESULTS:The concentrations of blood glucose,insulin and LDL-c increased obviously,and the testis weights lessened obviously in type II diabetic groups compared to those in normal control groups.Rare spermatogenic cells of seminiferous tubule and germinal arrest were observed in diabetic group under LM.Ultrastructural analysis of testicular tissue by TEM showed dilation of the endoplasmic reticulum and mitochondrial swelling in Leydig cell and sertoli cell in diabetic group.The level of blood LH and T decreased in typeⅡ diabetic groups in comparison with that in the normal control group.Compared to normal groups,the mRNA expression of StAR in type Ⅱ diabetic groups decreased,while the mRNA expression of LHR increased.After the treatment of EGB,the pathological change of testis was relieved,the concentrations of blood glucose,insulin and LDL-c were decreased,the level of blood LH and T,and the mRNA expression of StAR were increased,and the mRNA expression of LHR descended compared to type Ⅱ diabetic groups.CONCLUSION:EGB may increase the LH-induced testosterone production by correcting metabolic disorder of glucose and lipid,improving the function of pituitary-testicular axis and regulating the expression of LHR and StAR mRNA.

Objective To explore the expression of long noncoding RNA ( lncRNA) RN7SK and its screening value in clinical diagno-sis of gastric cancer. Methods The expression of lncRNA RN7SK in gastric cancer cells and paired gastric cancer tissues were meas-ured by qRT-PCR. The screening efficacy was detected by the receiver operating characteristic ( ROC) curve. Results The expression levels of lncRNA RN7SK in gastric cancer lines SGC-7901(3.91±0.53),MGC-803(3.44±0.29),HGC-27(4.04±0.87)and BGC-823 (4.30±1.13) were markedly higher than that in human gastric mucosal epithelial cell line GES-1, and the difference had statistical sig-nificance(1.02±0.27, t=12.33, 7.48, 7.20 and 4.90, P<0.05). The expression of RN7SK was up-regulated in 85.5％ (47/55) of gastric cancer tissues while down-regulated in the the other 8 tissues ( 14. 5％) . The area under ROC curve ( AUCROC ) of lncRNA RN7SK expression in gastric cancer patients was 0.827 and the 95％ confidence interval (CI) was from 0.746 to 0.907. When the cut-off value was 0.618, the sensitivity and specificity were 0.836 and 0.782 respectively. Conclusion The level of lncRNA RN7SK should be overexpressed in gastric cancer tissues so that it may have high screening efficacy and could be used as a potential molecular marker for the diagnosis of gastric cancer.