ObjectiveTo study the effect of α7 ( α7 AChR) agonist nicotine on regulating sensitivity of regular chemotherapeutic agent in cholangiocarcinoma cells,and explore the possible target.MethodsThe effect of nicotine and α-BTX pretreatment on the survival ability of cholangiocarcinoma cells was investigated when applied with 5-FU by using MTT and Flat cloning formation experiment.ResultsApplied with 5-FU,in various con centrations nicotine stimulating group( 10-3 g/L,10-4 g/L,10-5 g/L ),the survive rate of QBC939 was 128％,124％,118％,while that in α-BTX stimulating group and combined stimulation group was 92％,94％,93％,92％,respectively.The cloning formation ability of nicotine- stimulating group (6.2 ± 0.40) was significantly higher than α- BTX stimulating group (3.2 ± 0.20 ),combined stimulation group ( 3.2 ± 0.20 ) and control group ( 3.4 ±0.33).ConclusionNicotine can prevent chemotherapy-induced apoptosis,and improve cholangiocarcinoma cell survival via α7 nicotine acetylcholine receptor in vitro.

Circulating tumor cells (CTCs) is the source of tumor metastasis and recurrence. Many studies have suggested that tissue factor (TF) can be used as a novel marker of CTCs. This review focuses on the effects of TF on promotion of formation of CTC, activation of tumor cell proliferation, tumor cell survival, tumor angiogenesis and immune escape of tumor as well as its mechanisms; it also discusses the current status of TF/CTCs-targeted strategy in tumor treatment and the challenges faced, in hope of providing the theoretical basis for the application of TF/CTCs-targeted strategy in this field.

Objective To study the assosiation of angiopoietin-like protein 2 (ANGPTL2) to lower extremity arterial disease in type 2 diabetes mellitus.Methods A total of 360 type 2 diabetic patients were divided into three groups:without (group A),with mild to moderate (group B),and severe (group C) lower extremity arterial disease according to the ankle brachial index.And,120 healthy subjects serveed as control group.The levels of ANGPTL2,high sensitivity C-reactive protein (hsCRP),free fatty acid,biochemical index,and fasting insulin were detected.And,waist-to-hip ratio (WHR),body mass index (BMI),insulin resistance index were calculated.Results The level of ANGPTL2 was getting higher and higher as lower extremity arterial disease progressing.The level of ANGPTL2 in group B was higher than that in group A [1.27 (1.09,1.51) vs 0.88 (0.66,1.07) μg/L,P＜0.05],and the level of ANGPTL2 in group C was higher than that in group B [1.70 (1.45,1.91) vs 1.27 (1.09,1.51)μg/L,P＜0.05].Pearson correlation analysis showed that the ANGPTL2 level in serum positively correlated with hsCRP,BMI,WHR,cholesterol,low-density lipoprotein-cholesterol,fasting insulin,and insulin resistance index.Logistic regression analysis showed that the levels of ANGPTL2,hsCRP,and glycosylated hemoglobin were significantly associated with lower extremity arterial disease in type 2 diabetes mellitus.Conclusion It is possible that ANGPTL2 is one of the risk factors of lower extremity arterial disease in type 2 diabetes mellitus,and which is likely to be of great significance in the early prediction,disease assessment,and prognosis evaluation for lower extremity arterial disease in type 2 diabetes mellitus.

Cannula ; Catheters ; Drainage ; Duodenal Diseases ; Humans ; Intestinal Fistula/surgery*

Carcinoma, Hepatocellular ; pathology ; Humans ; Liver Neoplasms ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Peptide Fragments ; pharmacology ; Peptides ; pharmacology ; Tumor Cells, Cultured

To block tumor cell adhesion, inhibit tumor metastasis and recurrence, the anti-adhesion peptide-trimeric beta peptide (DLYYLMDLSYSMKGGDLYYLMDLSYSMKGGDLYYLMDLSYSMK, beta3) was designed. The DNA fragment of beta3 was cloned into expression vector pET-His and the fusion protein His-beta3 was expressed in E. coli. BL21(DE3)plysS. After 1.5 hours' induction with IPTG, His-beta3 peptide was expressed significantly amounting to 10% of the insoluble proteins and 4% of the total proteins. 20mg of beta3 peptide was obtained from one litter culture medium after purification by using metal-chelating sepharose 6B FF. The purity of beta3 is 92.2% according to Gel-Pro analysis. The anti-adhesion effects of beta3 peptide, beta1 peptide (DLYYLMDLSYSMK) and GRGDS on the hepatocellular carcinoma cell line SMMC-7721 and the high metastasis hepatocellular carcinoma cell line HCCLM6 were studied. The result showed the beta3 blocked the adhesion of HCCLM6 cells and SMMC-7721 cells to fibronectin (FN) specifically. The inhibition effect was dose-dependent and time-dependent and the inhibition rate of beta3 was higher than three times concentration of beta1 and GRGDS. This suggested that pET-His-beta3/BL21(DE3)plysS was a suitable expression system for beta3, and the expressed beta3 specially inhibited the adhesion of cancer cells.
Amino Acid Sequence ; Antineoplastic Agents ; pharmacology ; Cell Adhesion ; drug effects ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Peptide Fragments ; pharmacology ; Peptides ; genetics ; metabolism ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; Tumor Cells, Cultured

Objective: To investigate the clinicopathologic characteristics of intractable epilepsy. Methods: Based on the classification criteria proposed by the International League Against Epilepsy (ILAE), a retrospective analysis of the pathological characteristics was done in 822 patients who underwent epilepsy surgery in Sanbo Brain Hospital, Capital Medical University, from June 2008 to December 2012. Results: The mean age of epilepsy onset was 9.9 years, mean duration of epilepsy was 11.9 years. Complex partial seizures were the main presenting features. Histopathological study showed 33 cases (4.01%) with mild forms of cortical malformations, 690 cases (83.94%) with focal cortical dysplasia (FCD) and 99 cases with others (including 39 pure hippocampal sclerosis, 20 cystosclerosis, 19 Sturge-Weber syndrome, 8 tuberous sclerosis complex, 6 without significant pathological changes, 5 gyral malformations and 2 hamartoma). Among the 690 FCD cases, 106 were FCD typeⅠ, 91 were FCD typeⅡ and 493 were FCDⅢ(Ⅲa: 160, Ⅲb: 106, Ⅲc: 26 and Ⅲd: 201). Conclusions FCDⅢd is the most common histopathological subtype causing intractable epilepsy, mainly due to focal hypoxia/ischemia in the perinatal period, which results in scarring of local brain tissue; this is followed by other isolated forms of FCD (FCDⅠand FCDⅡ), and then FCD Ⅲa and FCD Ⅲb. The reason to distinguish isolated forms of FCD (types Ⅰ and Ⅱ) from FCD Ⅲ and to subclassify FCD Ⅲ is to allow better definition of cortical dyslamination. Therefore, the pathogenic factors of intractable epilepsy can be grouped in greater details, and facilitate the diagnosis and potential curative treatment of intractable epilepsy.

OBJECTIVE: To explore the effects of cluster needling at the scalp points on the expression of choline acetyl transferase (ChAT) and choline cholinesterase (AchE). METHODS: A total of 60 Wistar rats were randomized into a sham-operation group, a model group, a medication group and a cluster needling group, 15 rats in each one. In the model group, the medication group and the cluster needling group, the models of Alzheimer's disease (AD) were established by the orienteering injection with Aβ1-42 in the bilateral hippocampal CA1 in the rats. In the sham-operation group, the distilled water was injected in bilateral hippocampus of rats. In the medication group, the lavage with aricept was adopted for the basic treatment, once a day, for 4 weeks consecutively. In the cluster needling group, on the base of the treatment as the medication group, the cluster needling at the scalp points was adopted, once a day, 6 times a week, for 4 weeks totally. In the sham-operation group and the model group, the normal feeding was provided. After intervention, the learning and memory ability was measured with Morris water maze in the rats of each group. The changes in the hippocampal gross structure were observed with HE staining. The changes in the positive expressions of hippocampal ChAT and AchE were determined with the immunohistochemical method. RESULTS: Compared with the sham-operation group, the escape latency was prolonged and the percentage of the second quadrant and the frequency of platform leaping were reduced in the rats of the model group (all <0.01). Compared with the model group, the escape latency was shortened and the percentage of the second quadrant and the frequency of platform leaping were increased in the rats of the cluster needling group and the medication group (<0.05, <0.01). Compared with the medication group, the escape latency was shortened and the percentage of the second quadrant and the frequency of platform leaping were increased in the rats of the cluster needling group (all <0.05). Compared with the sham-operation group, the expression of ChAT was decreased and that of AchE increased in the model group (both <0.01). Compared with the model group, the difference was not significant in ChAT expression (>0.05) and the expression of AchE was reduced (<0.05) in the medication group; the expression of ChAT was increased (<0.05) and that of AchE decreased (<0.01) in the cluster needling group. Compared with the medication group, the expression of ChAT was increased and that of AchE decreased in the cluster needling group (both <0.05). CONCLUSION The effect mechanism of cluster needling at the scalp points on AD could be related to the up-regulation of ChAT expression and down-regulation of AchE expression in the hippocampus. The combined treatment with the cluster needling and aricept achieves the better therapeutic effect on AD.
Alzheimer Disease ; Animals ; Choline O-Acetyltransferase ; Hippocampus ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Scalp

Our previous studies have shown that puerarin, an active component of the traditional Chinese medicine-Pueraria Lobata, can improve glycometabolism in high-fat diet (HFD) mice with diabetes by activating the glucagon-like peptide-1 receptor (GLP-1R) pathway. This study intends to further evaluate the effect of puerarin on depressive symptoms in HFD mice. Long-term HFD induces type 2 diabetes and depressive-like symptoms in mice. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of the Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Traditional Chinese Medicine (approval No. AEWC-025). The experiment was divided into: control group, model group, model/puerarin (150 mg·kg^-1·day^-1) group, and model/fluoxetine (15 mg·kg^-1·day^-1) group. The oral glucose tolerance test (OGTT) and behavioral experimental analysis were performed after 6 weeks of continuous administration. Afterwards, enzyme-linked immunosorbent assay (ELISA) was used to detect interleukin-1β (IL-1β), interleukin-6 (IL-6), 5-hydroxytryptamine (5-HT), and corticosterone (CORT) in serum of mice for each group. Western blot assays were used to detect the level of activation and expression of proteins related to neuroplasticity and depressive disorder in the hippocampus. Moreover, HT-22 cell line was used to investigate the protective effect of puerarin on cell morphology and survival. The results show that puerarin can effectively maintain the survival of HT22 in an environment with high glucose and corticosterone. Meantime, the glycemic regulation of diabetic mice was improved after treatment of puerarin, the depressive symptoms were alleviated, the 5-HT increased, and the corticosterone, IL-1β, and IL-6 decreased in the serum. The up-regulation of related proteins in GLP-1R/Wnt/mTOR (mammalian target of rapamycin) signaling in hippocampus suggests that its effect on ameliorating depression in diabetic mice may be related to the activation of GLP-1R/Wnt/mTOR signaling pathway. This study shows that puerarin can significantly ameliorate the depressive symptoms of HFD induced diabetic mice which might be achieved through activating the GLP-1R/Wnt/mTOR signaling pathway and improving hippocampal neuroplasticity.