蝎皮蛋白质组分对正常小鼠免疫功能的影响

目的研究蝎皮蛋白质组分对正常小鼠免疫功能的影响,为蝎皮开发利用和发现新的活性物质提供实验依据。方法采用刃天青和M TT法检测蝎皮水溶性总蛋白(STP)、总角蛋白(STKP)及水溶性蛋白组分Ⅱ(STPⅡ)对正常小鼠T、B淋巴细胞转化、NK细胞杀伤活性的作用。结果STP和STKP ig给药后,小鼠脾T淋巴细胞增殖反应高于正常对照组,但只有STP小剂量组与对照组比较差异显著(P<0.001)。B淋巴细胞增殖反应结果显示STKP大、小剂量均可加强增殖反应,与对照组比较差异极显著(P<0.05、0.01)。STP和STKP大、小剂量组NK细胞杀伤活性与对照组相比有升高趋势。STPⅡ两个剂量组100、200 m g/kg ig给药均能明显抑制正常小鼠T、B淋巴细胞增殖反应,与对照组相比差异极显著。STPⅡ大剂量组NK细胞杀伤活性与对照组相比有降低趋势,但无统计学意义。结论STP和STKP具有一定的提高免疫功能的作用,STPⅡ则具有抑制免疫功能的活性。揭示蝎皮蛋白质组分具有免疫调节药理活性,值得深入研究。

双水相体系萃取分离葡萄酒下脚料中葡萄皮蛋白质

采用响应面法对双水相体系分离葡萄酒下脚料中葡萄皮蛋白质条件进行优化。在单因素实验基础上,选取pH、NaCl质量分数、样品加入量和温度为影响因子,利用Box-Behnken中心组合进行四因素三水平的实验设计,以蛋白质萃取率为响应值,进行响应面分析。结果表明,葡萄皮蛋白质的最佳萃取条件:质量分数为22%聚乙二醇(PEG600)-20%(NH4)2SO4体系,萃取pH7.1,NaCl质量分数1.2%,样品加入量2.41mL,萃取温度35.2℃。在此条件下,葡萄皮蛋白质萃取率的理论值为31.66%,实测值为31.08%。

反胶团萃取分离葡萄酒下脚料中葡萄皮蛋白质

采用阳离子表面活性剂十六烷基三甲基溴化铵(CTAB)反胶团萃取葡萄皮中的蛋白质。分别考察水相pH值、表面活性剂浓度、离子强度、蛋白质浓度、有机溶剂与助剂种类和比例、萃取温度对前萃取率的影响,并采用二次正交旋转组合的试验方法,考察前萃取过程中离子强度、表面活性剂浓度、水相pH值对前萃取率的影响。结果表明:32.68 mmol/L CTAB/三氯甲烷-正丁醇(体积比4∶1)的反胶团体系用于前萃取水相pH为6.4,包含0.4 mol/L NaCl的葡萄皮中蛋白质的浸提液,前萃取率可达56.54%。理想的前萃取条件是:pH 6.4、NaCl浓度0.4 mol/L、CTAB浓度32.68 mmol/L、蛋白质浓度1 mg/mL、萃取温度30℃。在该条件下,采用pH 3.8、后萃取温度20℃、0.8 mol/L NaCl水相进行后萃取,后萃取率可达71.29%。

XPD和P53对HepG2.2.15增殖及乙型肝炎病毒X蛋白表达的影响

目的:探讨人着色性干皮病基因D(XPD)和P53对人肝癌细胞株HepG2.2.15增殖及乙型肝炎病毒X蛋白(HBx)、Bcl-2和Bax基因表达的影响。方法:用脂质体转染法将重组质粒pEGFP-N2/XPD和空载质粒pEGFP-N2转染HepG2.2.15细胞,转染后第2天给予20μmol/L的Pifithrin-α(P53抑制剂)孵育24h。分为空白对照组、pEGFP-N2组、pEGFP-N2/XPD组、pEGFP-N2/XPD+Pifithrin-α组及Pifithrin-α组。用RT-PCR和Westernblotting方法检测XPD、HBx、Bcl-2和Bax表达的变化;用MTT法观察细胞增殖活力;用流式细胞仪检测细胞周期。结果:(1)重组质粒pEGFP-N2/XPD的转染使得XPD表达增高(均P<0.001);XPD表达增高使得HBx和Bcl-2表达降低,同时使得Bax表达增高,而Pifithrin-α能抑制XPD这一作用(均P<0.01)。(2)XPD表达增高抑制了细胞增殖活力,而Pifithrin-α能抑制XPD降低细胞活力的作用(均P<0.001)。(3)XPD表达增高引起了细胞G1期增加、S期减少,而Pifithrin-α能抑制XPD这一作用(均P<0.001)。结论:XPD是通过P53途径抑制肝癌细胞增殖,下调HBx和Bcl-2的表达并上调Bax的表达;XPD、P53和HBx三者之间能相互作用、相互影响,共同调节肝癌的发生与发展。

晚期胃癌XPG及GSTP1基因多态性与奥沙利铂化疗效果的关系

目的探讨人着色性干皮病G组(XPG)及谷胱甘肽巯基转移酶P1(GSTP1)基因多态性与晚期胃癌以奥沙利铂为主的化疗敏感性的关系。方法 80例Ⅳ期胃癌病人化疗前采集外周静脉血,提取DNA,用实时荧光PCR技术检测XPG C46T及GSTP1 A105G基因的单核苷酸多态性(SNP)分型,比较不同基因型与化疗效果的关系。结果 80例病人XPG C46T基因C/C与T/T+C/T及GSTP1 A105G基因A/A与A/G+G/G基因型化疗有效率比较差异均有显著意义(χ2=5.459、5.291,P<0.05)。同时携带XPG C46T C/C和GSTP1 A105G A/G+G/G基因型病人化疗敏感性是同时携带XPG C46T C/T+T/T和GSTP1 A105G A/A基因型病人的4.886倍(OR=4.886,P<0.05)。结论 XPG C46T、GSTP1 A105G基因多态性可能与晚期胃癌病人接受以奥沙利铂为主一线化疗药物化疗后的效果有关。

人剪切修复基因XPD对人肝癌SMMC-7721细胞中Ets-1和Cdk6基因的调控作用

目的:观察野生型人剪切修复基因XPD转染入人肝癌细胞SMMC-7721后,细胞内Ets-1和Cdk6基因的表达变化及对SMMC-7721肝癌细胞增殖的影响。方法:将人工合成的pEGFP-N2-XPD重组质粒通过Lipofectamine 2000TM转染SMMC-7721细胞。设重组质粒转染细胞SMMC-7721-pEGFP-N2-XPD(XPD)组、空载质粒转染细胞SMMC-7721-pEGFP-N2(N2)组、脂质体组、无转染空白对照组。分别用逆转录-聚合酶链反应(RT-PCR)和蛋白印迹法(Western blot)检测细胞中XPD、Ets-1、Cdk6基因mRNA和蛋白质的表达量,并用流式细胞仪检测细胞周期变化,四甲基偶氮唑盐微量酶反应比色法(MTT)检测各组细胞的增殖活力。结果:XPD组中的XPD的mRNA和蛋白质表达较其他3组明显增高(P<0.001),而Ets-1、Cdk6 mRNA和蛋白质表达较其他3组明显减少(P<0.001)。转染pEGFP-N2-XPD重组质粒后细胞停滞在G1期,难于进入S期。转染了野生型XPD的SMMC-7721细胞增殖能力减弱。结论:XPD基因可能通过抑制Ets-1、Cdk6基因的表达影响肝癌细胞的生长。

Epithelial-mesenchymal transition mediated tumourigenesis in the gastrointestinal tract

Epithelial-mesenchymal transition (EMT) is a highly conserved process that has been well characterised in embryogenesis. Studies have shown that the aberrant activation of EMT in adult epithelia can promote tumour metastasis by repressing cell adhesion molecules,including epithelial (E)-cadherin. Reduced intracellular adhesion may allow tumour cells to disseminate and spread throughout the body. A number of transcription proteins of the Snail superfamily have been implicated in EMT. These proteins have been shown to be over-expressed in advanced gastrointestinal (GI) tumours including oesophageal adenocarcinomas,colorectal carcinomas,gastric and pancreatic cancers,with a concomitant reduction in the expression of E-cadherin. Regulators of EMT may provide novel clinical targets to detect GI cancers early,so that cancers previously associated with a poor prognosis such as pancreatic cancer can be diagnosed before they become inoperable. Furthermore,pharmacological therapies designed to inhibit these proteins will aim to prevent local and distant tumour invasion.

Plasmodesmata: Dynamic Channels for Symplastic Transport

Plasmodesmata (PDs) are cytoplasmic structures that link adjacent cells to form the symplast of a plant. PDs are involved extensively in a plant's life by mediating symplastic transport of a wide range of ions and molecules. Major components of a plasmodesma (PD) include a plasma membrane, a desmotubule, and a cytoplasmic annulus, all of which are readily detectable by electron microscopy. Both the plasma membrane and the desmotubule contain proteinaceous particles, thought to be involved in altering the size of the cytoplasmic annulus. Cytoskeleton elements (actin and myosin) are essential for maintaining the integrity of PDs. Together with these elements, calcium_binding proteins probably play a significant role in regulating PD function. Symplastic transport occurs through the cytoplasmic annulus for the great majority of solutes, while other substances may traverse through the desmotubule internal compartment, the desmotubule shell, or the plasma membrane. The symplast is subdivided into several domains with varying molecular size exclusion limits (ranging from <1 kD to >10 kD). Plasmodesmata can be either primary or secondary; the former are developed during new wall formation and the latter are made in existing walls. The dynamic nature of plasmodesmata is also reflected by their changing frequencies, which, in turn, depend on the developmental and physiological status of the tissue or the entire plant. While diffusion is the major mechanism of symplastic transport, plasmodesmata are selective for certain ions and molecules. Upon viral infection, viral movement proteins interact with PD receptor proteins and, as a result of yet unknown mechanisms, the plasmodesmata are remarkably dilated to allow viral movement proteins and the bound viral genome to enter healthy cells. Some proteins of plant origin are also able to traverse plasmodesmata, presumably in ways similar to viral movement proteins. Some of these plant proteins are probably signal molecules contributing to cell differentiation and other activities. Other proteins move cell_to_cell in a non_specific manner.

Hyperlipidemia intensifies cerulein-induced acute pancreatitis associated with activation of protein kinase C in rats

AIM： To investigate the effects of hyperlipidemia on acute pancreatitis （AP） and the possible mechanisms. METHODS： Rat models of hyperlipidemia and AP were established by Triton WR1339 and cerulein respectively. Human albumin was used to treat AP complicated by hyperlipidemia. In each group, we compared the histological score, volume of ascites, ratio of pancreatic wet/dry weight, serum amylase （AMY） and pancreatic acinar cell apoptosis. The level of protein kinase C （PKC） membrane translocation in pancreatic tissue was detected by Western blot.RESULTS： In the hyperlipidemia model established by Triton WR1339, triglyceride （TG） increased remarkably and reached its peak 6 h after injection, and most rats developed mild acute pancreatitis. Histological score, volume of ascites, ratio of wet/dry weight and serum AMY in AP animals with hyperlipidemia were obviously higher than those in AP animals （P 〈 0.05） and decreased after albumin therapy but not significantly （P 〉 0.05）. Apoptotic cells detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling （TUNEL） increased in AP animals with hyperlipidemia and did not change distinctly after albumin therapy. PKC membrane translocation level increased in AP animals with hyperlipidemia and decreased remarkably after albumin therapy （P 〈 0.05）.CONCLUSION： Hyperlipidemia may induce AP or intensify pancreatic injury. Albumin therapy can not alleviate pancreatic lesion effectively. PKC activation may be one mechanism by which AP is intensified by hyperlipidemia.

Targeting of RhoE inhibits epithelial-mesenchymal transition during colorectal cancer cell migration

Objective Despite microRNA （miR-200b） being proved to promote the proliferation of colorectal cancer （CRC） cells, the relationship between miR-200b and epithelial-mesenchymal transition （EMT） of CRC cells remains poorly understood. The aim of the study was to investigate the relationship between miR-200b and EMT during CRC cell migration. Methods The effect of miR-200b on EMT-associated markers E-cadherin and vimentin was evaluated by western blot in CRC cells （SW620 and HT-29） by treatment with miR-200b mimics and inhibitors. A lucifer- ase reporter assay was employed to detect downstream targets of miR-200b. Transwell migration assays were used to detect CRC cell migration. Results Westem blots revealed that treatment with miR-200b mimics led to up-regulation of E-cadherin and down-regulation of vimentin, metalloproteinase （MMP）-9, and MMP-2, whereas treatment with miR- 200b inhibitor exhibited opposite effects on expression of E-cadherin and vimentin. Luciferase reporter assays demonstrated that RhoE （RND3） was targeted by miR-200b. Two predicted target sites of miR-200b were present in the 3＇-UTR of RhoE. Predicted target site 1 was from nucleotides 1584 to 1591, and site 2 was from nucleotides 1729 to 1735. RhoE knockdown cell lines were also established to investigate the impact of RhoE and miR-200b on EMT and cell migration. RhoE knockdown enhanced the effect of miR- 200b mimics, up-regulating E-cadherin and down-regulating vimentin. RhoE knockdown also inhibited cell migration. Furthermore, miR-200b mimic treatment further promoted the inhibitory effect of RhoE knock- down on cell migration.

α-fetoprotein involvement during glucocorticoid-induced precocious maturation in rat colon

AIM： TO investigate the role of a-fetoprotein （AFP）, a cancer-associated fetal glycoprotein, in glucocorticoidinduced precocious maturation in rat colon. METHODS： Colons from suckling Sprague-Dawley rats were used in this study. Corticosterone acetate at a dose of 100 μg/g body weight was given to normal pups on days 7, 9 and 11 after birth to induce hypercorticoidism. Control animals were injected with identical volumes of normal saline. Some rats receiving corticosterone 7 d after birth were also treated with mifepristone （RU38486）, a glucocorticoid cytoplasm receptor antagonist to investigate the effects of glucocorticoids （GCs）. The morphological changes of the crypt depth and villous height of the villous zone in colon were observed as indicesof colon maturation. Expression levels of AFP in colons were detected by reverse transcriptase polymerase chain reaction and Western blotting. To identify the cellular lo- calization of AFP in developing rat colons, double-immu- nofluorescent staining was performed using antibodies to specific mesenchymal cell marker and AFP. RESULTS： Corticosterone increased the crypt depth and villous height in the colon of 8- and 10-d-old rats with hypercorticoidism compared with that in the control ani- mals （120% in 8-d-old rats and 118% in 10-d-old rats in villous height, P = 0.021; 145% in 8-d-old rats and 124% in 10-d-old rats in crypt depth, P = 0.017）. These increases were accompanied by an increase of AFP ex- pression in both mRNA and protein （2.5-folds in 8-d- old and 2.5-folds in 10-d-old rats higher than in control animals, P = 0.035; 1.8-folds in 8-d-old and 1.3-folds in 10-d-old rats higher than in control animals, P = 0.023）. Increased crypt depth and villous height and increased expression of AFP in the colon of rats with hypercorti- coidism were blocked by mifepristone. Both had positive staining for AFP or vimentin, and overlapped in mesen- chymal cells at each tested colon. CONCLUSION： GCs promote the development of rat colon. AFP appears to be involved, in part, in mediating the effects of GCs in the developmental colon.

Effects of fasting and preoperative feeding in children

AIM： To investigate whether children should undergo surgery without a long period of fasting after feeding. METHODS： Eighty children with inguinoscrotal disorders （aged 1-10 years） were studied prospectively. They were divided into eight groups that each contained 10 children who were fed normal liquid food （NLF） and a high-calorie diet （HCD） 2, 3, 4 and 5 h before surgery, in two doses at 6-h intervals. NLF was given to four groups and HCD to the other four. In all groups, glucose, prealbumin and cortisol levels in the blood were measured twice： just after oral feeding and just before the operation. After the establishment of adequate anesthesia, gastric residue liquid was measured with a syringe. RESULTS： Blood glucose levels in all patients fed NLF and HCD were high, except in patients inthe HCD-4 group. There was no significant difference in the blood prealbumin levels. There was a significant increase in the blood cortisol levels in the NLF-2 （14.4± 5.7）, HCD-2 （13.2 ± 6.0）, NLF-3 （10.9± 6.4）, and HCD-5 （6.8 ± 5.7） groups （P 〈 0.05）. CONCLUSION： The stress of surgery may be tolerated by children when they are fed up to 2 h before elective surgery.Key words： Cortisol; Diet; Fasting; Food; Glucose; Liquids; Prealbumin

Proteomic analysis of pancreatic intraepithelial neoplasia and pancreatic carcinoma in rat models

AIM:To detect the proteomic variabilities of pancreatic intraepithelial neoplasia(PanIN)and pancreatic carcinoma(PC)induced by 7,12-dimethylbenzanthracene(DMBA) in rat models and to identify potential biomarkers.METHODS:Sixty adult male Sprague Dawley rats were randomized into three groups.The rats had DMBA implanted into their pancreas for one(n=20)or two months(n=20)or assigned to the normal group(n =20).The rats were killed after one or two months,and were evaluated histopathologically.Three tissue samples from each group of rats with either normal pancreas,PanIN(PanIN-2)or PC were examined by 2D-DIGE.The different expression spot features were analyzed by matrix-assisted laser desorption/ionizationtime of flight/time of flight(MALDI-TOF/TOF)tandem mass spectrometry.The expression of enolase 1,a differentially expressed protein,was identified by immu-nohistochemistry.RESULTS:There was significant difference in the proportions of neoplastic changes between the 1-and 2-mogroups(P=0.0488).There was an increase in the frequency of adenocarcinomas in the 2-mo group compared with the 1-mo group(P=0.0309).No neoplastic changes were observed in any of the animals in the normal group.Enolase 1,pancreatic ELA3B,necdin,Hbp23,CHD3,hnRNP A2/B1,Rap80,and Gnb2l1 were up-regulated in the PanIN and PC tissues,and CEL,TPT1,NME2,PCK2,an unnamed protein product,and glycine C-acetyltransferase were down-regulated in the PanIN and PC tissues.The immunohistochemical results showed that enolase 1 expression was up-regulated in the pancreatic cancer tissues of rats and humans.CONCLUSION:The pancreatic protein expression changes induced by DMBA suggest potential molecular targets for the early diagnosis and treatment of PC.

Substitution of Silkworm Pupae （Bombyx mori） for Fish Meal in Broodstock Diets for Snakeskin Gourami （ Trichogaster pectoralis）

Substitution of silkworm pupae （Bombyx mori） for fish meal in broodstock diets for snakeskin gourami （Trichogaster pectoralis） was conducted in order to evaluate the effect of silkworm pupae on feed digestibility and broodstock performance. Snakeskin gourami broodfish were fed with isocaloric diets （DE 3,000 a： 50 Kcal/kg） contained 32% crude protein. The levels of substitution were 0%, 50%, 75% and 100% of protein from fish meal （equal to 0%, 14.57%, 21.75% and 29.03% silk worm by weight）. The study was assigned in CRD with 4 treatments and 3 replications. Six hundred of snakeskin gourami with five-month old and average weight of 78.67 ± 2.15 g in female and average weight of 73.00 ± 2.49 g in male were stocked in 12 cages. Fifty fishes of 25 females and 25 males were reared in each cage and fed experimental diet for five months. The result indicated that protein digestibility of test diets were significantly decrease （P 〈 0.05） according to the level of silkworm pupae in diets where as the dry matter digestibility showed no significantly different （P 〉 0.05）. The growth performance and egg fecundity were significantly decreased （P 〈 0.05） according to the levels of silkworm pupae. In contrast, egg quality in term of fry number on first spawned, fingering number on first spawned were not significantly different （P 〉 0.05）. The survival rate of one month nursing fish on first spawned were significantly different （P 〈 0.05）. Hence, silkworm pupae could substitution for 50% of protein from fish meal （14.57% by weight） in snakeskin gourami broodstock diets without any adverse effect on egg quality.

hVEGF165 Expression in Escherichia coli Conserves Its Biological Function

The paper describes the expression of human protein VEGF165 in Escherichia coli and its purification. This growth factor isoform contains exon 7, which is essential for binding to extracellular domain of VEGF receptor 2, located on endothelial cells lining the surface of blood vessels. This binding stimulates the cascade of downstream signalling events leading to process known as angiogenesis, hVEGF165 overexpressed with His-tag in BL21 E. coli cells forms inclusion bodies （insoluble protein）, so the research found the procedure for its solubilization and purification on a Nickel based affinity chromatography. Although this eukaryotic signal protein needs posttranslational processing for its full function as a homodimer, author verified the biological activity of our hVEGF165 protein, obtained as monomer, by wound healing test.

The effect of alphastatin peptide suppressing the hypoxia-induced vasculogenic mimicry formation of glioma

Objective:To investigate the vasculogenic mimicry formation induced by hypoxia in Ⅱ-Ⅲ human glioma cell and the effect of alphastatin peptide suppressing the hypoxia-induced vasculogenic mimicry formation and the mechanism.Methods:MTT,Transwell and three-dimentional culture were used to detect the proliferation,migration and tubule formation of SHG44.The expression of vascular endothelial growth factor-α(VEGF-α),erythropoietin-producing hepatocellular carcinoma-A2 (EphA2) and matrix metalloproteinases 2 (MMP2) was detected by RT-PCR and Western blotting analysis.Results:The OD 490 in hypoxia group was 0.60±0.06 and in control group was 0.46±0.05.The number of cell migration was 178.71±18.81 in hypoxia group and 85.86±17.92 in control group.The tubule formation was 56.80±12.21 in hypoxia group and 4.20±2.62 in control group.The proliferation,migration and tubule formation in hypoxia group were significantly higher than that in control group.The expression of VEGF-α,EphA2 and MMP2 was upregulated in hypoxia.When various concentrations of alphastatin (100,1 000,10 000 nmol/L) were added to hypoxia group,the numbers of cell migration were 142.57±12.12,92.71±17.68,30.00±7.72 and the tubule formation were 47.71±10.58,18.86±8.40,8.43±5.62.The cell migration and tubule formation were significantly suppressed by alphastatin in a dose-dependent manner.In alphastatin group,the phosphorylation of EphA2 protein (P=0.037,F=4.629) and activation of MMP2 protein (P=0.005,F=9.331) were significantly suppressed but there was no change in VEGF-α protein.Conclusion:Ⅱ-Ⅲ human glioma cell is able to form vasculogenic mimicry induced by hypoxia and alphastatin peptide can suppress the hypoxia-induced vasculogenic mimicry.VEGF-α induced EphA2 phospharilation and MMP2 activation maybe the key pathway to form vasculogenic mimicry.

Effect of quercetin on expression of matrix metallo - proteinases and tissue inhibi tor of matalloproteinase -1 in cultured rat hepatic stellate cells

Objective: To study the effects of quercetin (QU) on matrix metallo-proteinases (MMPs) , the tissue inhibitor of matalloproteinase-1 (TIMP-1) , procollagen I and 2 proteoglycans (decorin and biglycan) mRNA expression in cultured rat hepatic stellate cell line HSC-T6 cells. Methods: Cells were treated with different concentrations of QU (12. 5, 25, 50 μmol/L) or drug solvent (0. 1 % Me2SO) for 24 h. mRNA expression was determined by reverse transcription polymerase chain reaction (RT-PCR). Results: QU (12.5 - 50 μmol/L) enhanced collagenase (rat MMP-13) and membrane typel-MMP (MMP-14) mRNA expression, decreased procollagen I mRNA expression in a concentration-dependent manner, but did not affect gelatinase-A (MMP-2) , TIMP-1, decorin and biglycan expression. Conclusion: QU may decrease matrix deposition and increase matrix degradation, which might be beneficial to liver fibrosis.

Some Agronomic and Chemical Traits of Blue Aleurone and Purple Pericarp Wheat （Triticum L.）

Thirteen diverse anthocyanin pigmented wheat genotypes originating from different countries were investigated for agronomic and chemical traits. The results showed significant variation among wheat genotypes in yield and grain physical characteristics, Released cultivars were superior in grain yield, but other genetic resources exceeded the cultivars in regard to test weight, grain mass, or seed plumpness. In case of phytochemical content both genotypic and environmental effects were important for the observed variations. The total phenolic content varied from 120 to 177 mg ferulic acid equivalent per 100 g dry weight; total anthocyanin content from 3.4 to 75.2 ppm cyanidin glucoside equivalent; yellow pigment content from 2.6 and 7.6 ppm beta-carotene equivalent; protein content from 11.3 to 19.1%. The study demonstrated that Ethiopian wheats are a source of high levels of anthocyanins and protein content.

Relationship between epithelial to mesenchymaltransition and chemoresistance of lung cancer

Objective： The aim of this study was to explore the correlation between epithelial to mesenchymal transition （EMT） and chemoresistance of non-small-cell lung cancer （NSCLC）. Methods： In vitro, the drug resistance index of cisplatin resistant lung adenocarcinoma cell line （A549/DDP） was detected by CCK-8 assay; the morphological change between A549/ DDP cells and lung adenocarcinoma cells （A549） was observed by phase contrast microscope; expression of EMT markers （including E-cadherin and vimentin） and resistance protein, excision repair cross-complementing 1 （ERCC1） was detected by immunocytochemistry. The expression of E-cadherin, vimentin and ERCC1 was investigated by immunohistochemistry in 120 cases of NSCLC, half of that were treated with pre-operative neoadjuvant chemotherapy （neoadjuvant chemotherapy group）, and the other underwent surgery alone （simple surgery group）. Results： There was a significant difference between the ICso （half maximal inhibitory concentration） of A549/DDP cells （5.20） and A549 cells （1.88） （P 〈 0.05）, and the drug resistance index of A549/DDP cells was 2.77. Compared with A549 cells, A549/DDP cells increased expression of ERCC1 （P 〈 0.05）. Moreover, A549/DDP cells showed morphological and phenotypic changes consistent with EMT： with spindle-shaped morphology, and decreased expression of E-cadherin and increased expression of vimentin. Immunohistochemistry showed significant positive correlation between the expression of ERCCI and vimentin （r = 0.496, 0.332, P 〈 0.05）, and significant negative correlation between the ERCCI and E-cadherin （r = -0.403, -0.295, P 〈 0.05） in neoadjuvant chemotherapy group and simple surgery group. In addition, compared with simple surgery group, the expression of ERCC1 （P = 0.003） and vimentin （P = 0.004） was significantly increased, and the expression of E-cadherin was decreased in neoadjuvant chemotherapy group （P = 0.032）. Cenclusion： A549/DDP cells acquired cisplatin-resistance and occurred EMT simultaneously; the phenomenon of chemoresistance and EMT was caused more easily in neoadjuvant chemotherapy group. As such, we further confirmed the close correlation between chemoresistance and EMT of NSCLC, and provided theoretical basis for the targeting therapy with EMT regulatory factor for chemoresistant NSCLC patients.

Population Bursts of Parvalbumin-Positive Interneurons Inhibit Spiking Pyramidal Cells in Spontaneously Active Cortical in Vitro Networks

Author present the interplay between different neuron types in the spontaneous electrical activity of low density cortical in vitro networks grown on MEA （multielectrode arrays） of glass neurochips. In 10% of the networks, the continuously spiking activity of some neurons was inhibited by synchronous bursts or superbursts of the majority of the other neurons. Immunohistochemical staining subsequent to MEA recordings suggest that the synchronously bursting neurons are parvalbumin-positive interneurons with abundant axonal ramifications. Blocking chemical synaptic transmission by Ca2＋-free medium revealed that the curbed spiking neurons are intrinsically active. It is assumed that these neurons are pyramidal cells which may be inhibited by groups of synchronously bursting interneurons. It is propose that the observed burst-induced inhibition is an important principle in the temporal organization of neuronal activity as well as in the restriction of excitation, and thus essential for information processing in the cerebral cortex.