Objective To study changes of AMPA receptors expression in nucleus accumbens and hypothalamus of methamphetamine dependent rats,and the therapeutical effect of rhynchophylline.Methods SPF male rata were randomly divided into normal control group,model group of methamphetamine,low dose of rhynchophylline group and high dose of rhynchophylline group(n=8 in each group).Experiment of conditioned place preference(CPP)was used to build the model of methamphetamine dependent rata.Western blotting was used to examine the changes of GluR2/3 subunits expression.The time of staying in drug-paired compartment of rats was used independent-samples t test to gather statistics,and the photodensity of proteinum strap was used One-Way ANOVA to gather statistics.Results Compare with rats in normal control group(the time of staying in drug-paired compartment of rats was(383.00±38.20)s),the rats produced CPP after treated with methamphetamine(the time of staying in drug-paired compartment of rats was(536.20±57.49)s),and low(30mg/kg) and high (60 ms/kg)dose of rhynchophylline(the time of staying in drug-paired compartment of rats were(299.80±15.96)s and(189.40±59.02)s)both could eliminate CPP effect.Compare with rats in normal control group (the ratio of value of average gray scale were(0.54±0.04)INT·mm~2 and (0.70±0.04)INT·mm~2),GluR2/3 subunits expression in nucleus aecumbens increased significantly in model group(the ratio of value of average gray seale was(0.89±0.03)INT·mm~2)and low dose of rhynchophylline group(the ratio of value of average gray seale was (0.93±0.03)INT·mm~2,P<0.01),which decreased significantly in hypothalamus(the ratio of value of average gray scale were (0.53±0.03)INT·mm~2 and (0.52±0.02)INT·mm~2,P<0.01).But GluR2/3 subunits expression in nucleus accumbens and hypothalamus of rats in high dose of rhynchophylline group(the ratio of value of average gray scale were (0.57±0.06)INT·mm~2 and (0.65±0.01)INT·mm~2) just liked the expression of normal control group(P>0.05).Conclusion GluR2/3 subunits expression of methamphetamine-induced CPP rats increased in nucleus accumbens but decreased in hypothalamus.High dose of rhynchophylline can reverse such changes and rebound the expression to normal level.

This study was aimed to explore the influence of breast cancer associated fibroblasts (CAFs) in migration and invasion of breast cancer cell line MCF-7, and investigate whether hepatocyte growth factor (HGF) is involved in this process. Primary breast CAFs and their corresponding normal breast fibroblasts (NFs) were obtained by collagenase digestion. On the basis of the co-culture, the migration and invasion capacity of MCF-7 cells was compared between CAFs and NFs by Transwell. The difference in the HGF expression between them was detected by ELISA. The secretion of HGF was knocked down by using RNA interference technology in CAFs. Then the changes of migration and invasion capacity of MCF-7 cells were investigated by Transwell. Eventually, we isolated high-purity CAFs and NFs, and the CAFs had a stronger ability in promoting MCF-7 migration and invasion than the NFs. ELISA results demonstrated that CAFs secreted higher HGF, and the capacity of MCF-7 migration and invasion was declined after knocking down the secretion of HGF in CAFs by RNA interference. It is suggested that CAFs can promote MCF-7 migration and invasion through HGF in vitro.

AIM:To investigate the role of peripheral blood leukocytes and polymorphonuclear neutrophils (PMNs) on intestinal injury following mesenteric ischemia/reperfusion (IR) in rats. METHODS: Twenty adult, male Sprague-Dawley rats, weighing 200-230 g, were randomly divided into two groups. The control group (CON) consisting of 10 rats was subjected to laparotomy and separation of superior mesenteric artery (SMA) only. The ischemia/reperfusion (IR) group consisting of 10 rats, was subjected to laparotomy, followed by occlusion of the superior mesenteric artery (SMA) by an atraumatic microvascular clamp for 30 min. At the end of ischemic period in IR, the microvascular clamp was removed and the intestinal segment was reperfused for 60 min. The pathological changes of the ileal mucosal tissue were evaluated. The apoptosis of intestinal mucosal epithelial cells was examined by terminal deoxylnucleotidy-l transferase mediated-dUTP nick end-labeling (TUNEL). The enzymatic activity of casapse-3 in mucosal cells was determined using a colorimetric assay. The percentages of apoptotic peripheral blood leukocytes and PMNs were measured by flow cytometry using Annexin-V/PI double staining assay. The numbers of peripheral blood leukocytes in each animal was measured at baseline, 30 min of ischemia, and 30 min and 60 min of reperfusion. RESULTS: (1) Compared to CON group animals, the most severe mucosal injury was observed in IR group under optical microscope. (2) The number of apoptotic mucosal epithelia cells and enzymatic activity of caspase-3 were significantly higher in IR than those in CON group (P

Objective:To investigate the effects of Mor-platin, a novel mitochondrial platinum complex, on proliferation and migration of human hepatoma carcinoma HepG2 cells. Methods:Cell counting kit-8 (CCK-8) assay was used to analyze cell proliferation of Mor-platin and classic anticancer drugs, particularly cisplatin, in HepG2 cells. A laser confocal microscope was used to observe whether Mor-platin can target mitochondria. The morphological changes in cellular mitochondria after treatment with Mor-platin were ob-served on a transmission electron microscope. Cell apoptosis was measured by flow cytometry, and cell invasion was evaluated by three-dimensional tumor spheroid model. Results:Mor-platin can inhibit cell proliferation and is dose dependent. The half inhibitory concentration (IC50) of Mor-platin is lower than that of cisplatin. Laser confocal images showed that Mor-platin can target cell mito-chondria and enrich cell mitochondria. Transmission electron microscopy images showed that cell mitochondrial morphology changed after Mor-platin treatment. Furthermore, cell mitochondrial membrane is incomplete and mitochondrial cristae are reduced. Cell apoptosis caused by Mor-platin is dose dependent. The three-dimensional tumor spheroid model showed that the cell areas of the group subjected to Mor-platin treatment are smaller than those of the control group. Conclusion:Mor-platin can target cell mitochon-dria, change the cell mitochondrial morphology, inhibit cell proliferation, and thus promote cell apoptosis. It also showed better anti-cancer effects than cisplatin. Furthermore, Mor-platin can inhibit three-dimensional tumor spheroid invasion. These results suggest that Mor-platin is a potential antitumor drug.

Cerebral cysticercosis is the most common worldwide parasitic infection of the central nervous system. Intraventricular involvement is apparent in 15% to 28.8% of cases with neural compartment infestation.' Although different forms of the disease (parenchymatous, subarachnoid, and mixed form ) have been treated successfully with chemotherapy, direct surgical excision of simple cystic lesions appears to be an adequate primary therapeutic strategy in the majority of intraventricular forms. In recent years, however, some authors have advocated the use of anthelmintic treatment in all cases of intraventricular cysts so that surgical procedures of the posterior fossa and their potential complications can be avoided. The strict definition for managing the spectrum of intraventricular infestation remains controversial. We present our experience in the treatment of a patient with primary isolated intraventricular cysticercosis.
Albendazole ; therapeutic use ; Anthelmintics ; therapeutic use ; Child ; Female ; Humans ; Neurocysticercosis ; therapy ; Ventriculoperitoneal Shunt

BACKGROUNDPromoter analysis is currently applied to detect the expression of the targeted gene in studies of signal transduction and transcriptional regulation. As a reporter gene, luciferase plays an important role and has been used widely in the promoter assay.

METHODSHuman embryonic lung fibroblast cells (2BS), HeLa cells and MCF-7 cells were transfected with various genes embedded by lipofectamine. This study determined various factors that affect promoter activity determination, such as the selection of the reporter genes and internal references, the dose and the type of the vectors carrying the transcription factors, the host cells and the instruments.

RESULTSThe sensitivity of the luciferase assay was much higher than that of enhanced green fluorescence protein (EGFP). Moreover, promoter activity is increased in a dose-related manner only in certain ranges outside of which the results may be reversed and the promoter activity is related to the expression vector which is carrying the cDNA. Otherwise, the length of the promoter, internal references and the host cell can also influence the promoter activity.

CONCLUSIONSTo detect the promoter activity accurately, a few factors including dose, vector, length and host cell which influence reporter gene assay aforementioned should be considered.

Cells, Cultured ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Luciferases ; genetics ; Plasmids ; Promoter Regions, Genetic ; Transfection

Adult ; Albendazole ; therapeutic use ; Cerebellar Diseases ; diagnosis ; drug therapy ; Humans ; Magnetic Resonance Imaging ; Male ; Neurocysticercosis ; diagnosis ; drug therapy

ObjectiveTo investigate the relationship of lymphotoxin β receptor (LTβR) and classical nuclear factor-κB (NF-κB) activation pathway in the pathogenesis and progress of cystitis and bladder cancer.MethodsThe LTβR and P65 mRNA expression were detected by Real-time quantitative PCR in 108 cases of fresh bladder tissue specimens (75 cases of bladder cancer,10 cases of inflammation and 23 normal bladder mucosa cases grouped by the tissue classification ),and protein expression were analyzed by immunohistochemistry assay in 118 cases of paraffin-embedded biopsy specimens (73 cases of bladder cancer,30 cases of cysitis and 15 normal bladder mucosa cases).The correlation analysis between the expressions of LTβR and P65 with clinical pathological data was then performed.Differences between LTβR and P65 mRNA and protein expression level were compared in different groups of bladder tissues using Kruskal-Wallis H test and the Chi-square test.Results( 1 )The mRNA expressions of LTβR and NF-κB/P65were higher in bladder cancer than those in normal group ( LTβR:29.4 ( 14.2 - 46.7 ) × 10 - 3/1.2 ( 0.3 -7.0) ×10-3,Z=-5.508; P65:9.7 (2.7 -21.1) ×10-3/1.0(0.8 ～1.8) ×10-3,Z=-5.030,P＜0.05 ).There were significantly differences between bladder cancer with different histological grades ( LTβR:18.2(2.1-31.3) × 10-3/ 28.4(16.6-36.2) × 10-3/47.9(34.3 -70.5) ×10-3,x2K-W=20.378;P65:4.9(1.3 - 12.0) × 10-3/7.4(3.0-21.9) × 10-3/17.0(10.0 ～28.3)× 10-3 ,x2K-W2 =15.219,P all ＜0.05) and lymph node metastasis (LTβR:27.2(9.7-40.1) ×10-3/39.4(26.7 -52.6) ×10-3,Z=-2.552; P65:7.4(2.3-15.6) ×10-3/13.4(6.7-23.3) ×10-3,Z=-2.026,P＜0.05).(2)The positive rates of LTβR and phosphorylated P65 ( p-P65 ) protein in cancer were higber than those of normal group (LTβR:69.8％/13.3％,x2 =16.600 ; p-P65:56.2％/6.7％,x2 =12.220,P ＜ 0.05 ).Upregulation of LTβR and p-P65 were associated with the histological grade (LTβR:56.3％/70.0％/90.4％,x2 =7.055; p-P65:40.6％ /60.0％/76.2％,x2 =6.679,P ＜0.05) and with lymph node metastasis (LTβR:58.3％/92.0％,x2 =8.849; p-P65:52.1％/64.0％,x2 =5.088,P ＜0.05).(3)There was a positive correlation between LTβR and P65 expression ( mRNA:r =0.654,P ＜ 0.05,protein:r =0.399,P ＜ 0.05 )in the bladder cancer and cystitis (r =0.521,P＜0.05).ConclusionsThe activation of LTβR and P65 was associated with progression and metastasis of bladder cancer.The activation of classical NF-κB pathway by LTβR may be achieved by P65.

This study was aimed to explore the influence of breast cancer associated fibroblasts (CAFs) in migration and invasion of breast cancer cell line MCF-7,and investigate whether hepatocyte growth factor (HGF) is involved in this process.Primary breast CAFs and their corresponding normal breast fibroblasts (NFs) were obtained by collagenase digestion.On the basis of the co-culture,the migration and invasion capacity of MCF-7 cells was compared between CAFs and NFs by Transwell.The difference in the HGF expression between them was detected by ELISA.The secretion of HGF was knocked down by using RNA interference technology in CAFs.Then the changes of migration and invasion capacity of MCF-7 cells were investigated by Transwell.Eventually,we isolated high-purity CAFs and NFs,and the CAFs had a stronger ability in promoting MCF-7 migration and invasion than the NFs.ELISA results demonstrated that CAFs secreted higher HGF,and the capacity of MCF-7 migration and invasion was declined after knocking down the secretion of HGF in CAFs by RNA interference.It is suggested that CAFs can promote MCF-7 migration and invasion through HGF in vitro.

Objective To investigate the abnormal expression of Golgi protein 73(GP73) in CD4+ T lymphocytes of the pa-tients with hepatocellular carcinoma (HCC) and its influence of Th1/Th2/Th17 subtype differentiation .Methods Fifty cases of HCC hospitalized in this hospital from May 2015 to February 2016 and 50 healthy volunteers as controls were selected .Peripheral blood was collected and CD4+ T lymphocytes were isolated ;then the expression levels of GP73 and nuclear factor kappa-light-chain-enhancer (NF-κB) in CD4+ T lymphocytes were determined by using RT-qPCR and Western blotting methods ;furthermore ,the se-cretion levels of IL-4 ,IL-17 and IFN-γin the supernatants were examined by using ELISA method .Results GP73 mRNA expres-sion in peripheral blood CD4+ T lymphocytes in the HCC patients were significantly up-regulated compared with the healthy volun-teers ,the difference was statistical difference (P<0 .05) .The expression level of in GP73 overexpression group was significantly in-creased(P<0 .05) ,while which in the GP73 interference group was significantly decreased (P<0 .05) .Over-expression of GP73 in-duced significant increase of IL-4 and IL-17 levels and significant decrease of IFN-γ(P<0 .05);silencing GP73 induced marked de-crease in the expression of IL-4 and IL-17 in CD4+ cells and obvious increase of IFN-γ(P<0 .05) .Conclusion GP73 is over-ex-pressed in peripheral blood CD4+ T cells of HCC patients ,moreover GP73 is very likely to participate in the inflammatory reaction by activating NF-κB to cause the unbalance of Th1/Th2/T17 and promote the occurrence and development of HCC .