A Correlational Study Between Executive Functions and Conditional Discriminations in Children with Autism / Um Estudo Correlacional Entre Funções Executivas e Discriminações Condicionais em Crianças com Autismo

Abstract Neuropsychology is a science that allows tracing the profile of cognitive impairments and preserved skills to design appropriate treatments and educational practices aiming at a better quality of life for the individual. This is basic correlational research, which objective was to verify if the results found in the executive functions in children with autism spectrum disorder (ASD) are predictive of or have some correlation with the performance in conditional discrimination through choice tasks according to the identity matching-to-sample (MTS) model. Correlations revealed significant associations between neuropsychological tests and MTS tasks. Future research may further explore MTS tasks for the assessment and intervention of individuals with ASD.

Resumo A Neuropsicologia é uma ciência que permite traçar o perfil dos comprometimentos cognitivos e habilidades preservadas a fim de delinear tratamentos e práticas educativas adequadas, almejando melhor qualidade de vida do indivíduo. Trata-se de uma pesquisa básica correlacional, cujo objetivo foi verificar se os resultados encontrados nas funções executivas em crianças com Transtorno do Espectro do Autismo (TEA) são preditivas de ou tem alguma correlação com o desempenho em discriminação condicional por meio de tarefas de escolha de acordo com o modelo MTS de identidade. As correlações revelaram associações significativas entre os testes neuropsicológicos e as tarefas de MTS. Pesquisas futuras poderão explorar melhor as tarefas de MTS para avaliação e intervenção de indivíduos com TEA.

Avaliação da atividade citotóxica dos extratos etanólicos da casca e das folhas da Terminalia fagifolia Mart. sobre células normais e tumorais / Assessment of the cytotoxic activity of ethanol extracts from the bark and leaves of terminalia fagifolia mart. on normal and tumor cells

Introdução: A procura por novas alternativas terapêuticas, como as que utilizam as plantas medicinais, tem despertado grande interesse da comunidade científica na busca por tratamentos mais eficientes para as doenças, incluindo o câncer. Terminalia fagifolia Mart. é uma planta medicinal encontrada no Cerrado brasileiro, usada popularmente no tratamento de aftas e tumores. Objetivos: Avaliar a atividade citotóxica dos extratos etanólicos da casca e das folhas da Terminalia fagifolia em linhagens celulares NIH 3T3 e L929 e tumorais PC3 e B16F10. Métodos: Foi realizada a metodologia de determinação da viabilidade celular em ensaio com monocamada de células utilizando o ensaio MTS. As linhagens NIH 3T3, L929, PC3 e B16F10 foram expostas por 24 horas a diferentes concentrações dos extratos etanólicos da casca e folhas da Terminalia fagifolia. Resultados: Os resultados adquiridos mostraram que os extratos apresentaram viabilidade celular, sendo considerada de moderada a alta, para as células normais NIH 3T3 e L929 e citotoxicidade severa para as células tumorais PC3 e B16F10. Dessa forma, torna-se necessária a continuidade dos estudos com essa planta, pois os extratos da casca e das folhas apresentaram atividades antitumorais muito promissoras. Conclusões: Os extratos da casca e das folhas demonstraram viabilidade celular ≥ 50% nas linhagens celulares normais NIH 3T3 e L929 e demonstraram atividade citotóxica para as linhagens tumorais PC3 e B16F10, apresentando redução da viabilidade celular em torno de 60% e 70%, respectivamente. (AU)

Introduction: The search for new therapeutic alternatives, as the ones that use medicinal plants, has awaken a huge interest from the scientific community in seeking through more efficient treatments for diseases, including cancer. Terminalia fagifolia Mart. is a medicinal plant found in Brazilian "Cerrado", popularly used in aphthas and tumor treatment. Objectives: To evaluate the cytotoxic activity of ethanolic extracts of the bark and leaves of the Terminalia fagifolia in cell lines NIH 3T3 and L929 and tumor cells PC3 and B16F10. Methods: The determination methodology in cellular viability was held in an assay with cells monolayer's using the MTS assay. The NIH 3T3, L929, PC3 and B16F10 lines was exposed for 24 hours in differents ethanolic extracts concentrations. Results: The acquired results showed that the extracts had cellular variability is considered moderate to high for the normal cells NIH 3T3 and L929 and severe cytotoxicity to tumor cells PC3 and B16F10. This way, it is necessary to continue studying this plant, since both the bark and leaves extracts have great antitumor activity. Conclusion: The bark and leaves extracts showed cellular variability ≥ 50 % in normal cell lines NIH 3T3 and L929 and demonstrated cytotoxic activity for tumor cells PC3 and B16F10, presenting reduction of cell variability around 60% and 80%, respectively. (AU)

Difference in susceptibility of carbapenem-resistant Klebsiella pneumoniae to tigecycline detected by MTS and instrumental MIC assay / 中国感染控制杂志

Objective To explore clinical distribution characteristics of carbapenem-resistant Klebsiella pneumoniae (CRKP), analyze enzyme production of strains and verify the in vitro antimicrobial activity of tigecycline.Methods Antimicrobial susceptibility testing results of 53 strains of CRKP isolated from clinical specimens of patients in a hospital from January to December 2015 were analyzed, carbapenemase production of target strains was detected by modified Hodge test, metallo-β-lactamase was detected by EDTA synergy test, minimum inhibitory concentration(MIC) of tigecycline susceptibility testing result detected by instrument was confirmed by MIC test strip(MTS method).Results 53 CRKP strains were mainly isolated from patients in intensive care unit (n=14, 26.42％) and burn unit(n=13, 24.53％);sputum(n=23, 43.40％) and wound secretion(n=15, 28.30％) were the main specimen sources;isolation rate of CRKB was highest in the elderly≥60 years old, 35 strains(66.04％)of CRKP were isolated.CRKP was most sensitive to tigecycline(96.2％).The modified Hodge test showed that 48 strains(90.6％) produced carbapenemases and 15 strains produced metallo-β-lactamase.MICs of tigecycline-resistant strains detected by instrument were all confirmed as susceptibility by MTS.Conclusion CRKP mainly produce carbapenems in this hospital, some strains can produce two types of different β-lactamases;antimicrobial susceptibility testing showed that tigecycline has good antimicrobial activity against CRKP, tigecycline-resistant strains detected by instrument must be confirmed by MTS method.

Efficacy of Human Platelet Rich Fibrin Exudate vs Fetal Bovine Serum on Proliferation and Differentiation of Dental Pulp Stem Cells

BACKGROUND AND OBJECTIVES: The imperative role of dental pulp stem cells (DPSCs) in regenerative therapy demands an in-vitro expansion which must deal with the safety and ethical problems associated with fetal bovine serum (FBS). The primary aim of this study was to compare the effects of human platelet rich fibrin (hPRF) exudate Vs FBS on proliferation and osteodifferentiation of human dental pulp stem cells (hDPSCs). The secondary one was to determine the optimum concentration of hPRF exudate inducing hDPSCs proliferation and osteodifferentiation. METHODS: The direct method was used to prepare hPRF exudate. hDPSCs were isolated from impacted mandibular third molars of twelve donors by the outgrowth method. For cell viability and proliferation rate testing, 96 well plates were used and the assay was done in duplicate and the trial repeated four times under the same conditions. Six wells were used to contain 10% FBS, serum free media, 1%, 5%, 10% and 20% concentrations of hPRF exudates, respectively. The proliferation assay was carried out by MTS tetrazolium cell proliferation assay kit and Elisa reader. The study design for osteodifferentiation protocol was exactly as the proliferation one and instead the assay was carried out by alizarin red with Elisa reader. RESULTS: Compared to 10% FBS, 10% hPRF exudate was the optimum concentration for hDPSCs proliferation, while 1% hPRF exudate was the optimum concentration for osteodifferentiation of hDPSCs. CONCLUSIONS: Avoiding the risk of zoonosis which may be occurred with FBS, it is recommended to use 10% hPRF exudate for proliferation and 1% for osteodifferentiation.

Comparison of several common methods for measuring the biological activity of recombinant human granulocyte colony stimulating factor / 中国药学杂志

OBJECTIVE: To compare several common staining and detection methods using NFS-60 cells for biological activity test of recombinant human granulocyte colony stimulating factor (rhG-CSF). METHODS: The biological activity of rhG-CSF was detected using some common methods, named NFS-60 cells/MTT staining, NFS-60 cells/MTS staining, NFS-60 cells/CCK-8 staining, and NFS-60 cells/fluorescence staining. The biological activity was detected using the NFS-60 cells/MTT method using dual wavelength (570 nm detection, 630 nm reference) and single wavelength (570nm detection and 630nm detection). The biological activity was detected using NFS-60 cells/MTS dynamic detection method and NFS-60 cell/CCK-8 dynamic method. Then, the results were analyzed and compared. RESULTS: The different methods were not significantly different (P>0.05); the difference between the dual wavelength detection and single wavelength detection of NFS-60 cells/MTT was not significant (P>0.05). The results from NFS-60 cells/MTS dynamic detection method, NFS-60 cells/CCK-8 dynamic method and NFS-60 cells/MTT method had not significant difference (P>0.05). CONCLUSION: The biological activity determination results of the tested methods using NFS-60 cells are consistent. This study provides basis for utilization of results from different laboratories using different methods, support for expansion of the biological activity detection method of rhG-CSF in Chinese Pharmacopoeia, and reference for expansion of the biological activity detection method of other cytokines.

Inhibition effects of fungal immunomodulatory proteins from Ganoderma spp .on different gastric cancer cell lines in vit ro / 药学实践杂志

Objective To study the cell proliferative effects of fungal immunomodulatory proteins from Ganoderma spp . on 26 gastric cancer cell lines in vitro .Method 26 human gastric cancer cell lines were treated with FIPs by MTS assay .The average optical density (OD) in 490 nm and inhibition rate (GI50 )was counted by Universal Microplate Spectrophotometer . Results Three FIPs showed similar profiling in 26 human gastric cancer cell lines after 72 h treatment in cell proliferation as-say ,which except for NUGC-4 and OCUM-1 did not showed obvious anti-proliferative effect ,the other 24 human gastric cell lines showed some anti-proliferative effects ,especially for 7 cell lines(NUGC-3 ,GTL-16 ,HGC-27 ,IM95m ,SNU-638 ,SNU-216 and SNU-5) showing strong potency ,with their GI50 less than 50 μg/ml .Conclusion FIPs showed strong anti-prolifera-tive effects in some human gastric cancer cell lines in vitro ,which had potential to be further developed as anti-gastric cancer drugs .

Diferenças metabólicas no hipocampo contralateral entre pacientes concordantes e discordantes na esclerose mesial temporal (EMT): avaliação com espectroscopia de protóns por ressonância magnética (1H-ERM) / Metabolic differences in the contralateral hippocampus of concordant and discordant mesial temporal sclerosis (MTS) patients: evaluation with proton magnetic resonance spectroscopy (1H-MRS)

OBJETIVO: Pacientes com esclerose mesial temporal (EMT) são clinicamente classificados como concordantes ou discordantes de acordo com a extensão da atividade epileptogênica. O objetivo desse estudo é verificar se as alterações metabólicas no hipocampo são diferentes nos dois grupos. MATERIAIS E MÉTODOS: Foram avaliados 33 pacientes diagnosticados com EMT, 23 concordantes (33±9 anos) e 10 discordantes (33±10 anos), e 28 controles (33±10 anos). Foi obtida espectroscopia por ressonância magnética de ambos os hipocampos (ipsi e contralateral) com aparelho de 3T e com a sequência PRESS de vóxel único com TE/TR=135/1500ms. Os principais metabólitos foram quantificados com o LCModel. Para a comparação entre os grupos foram realizados testes estatísticos com nível de significância de 0,05. RESULTADOS: Para todos os pacientes foi observada redução de NAAipsi e Cr ipsi quando comparado com o lado contralateral, porém esta diferença foi significativa apenas para o grupo de pacientes concordantes. Quando comparado com o grupo controles houve uma redução significativa de Cr ipsi e NAAipsi para os dois grupos e da relação NAA/Cr ipsi para o grupo de pacientes concordantes enquanto que a relação NAA/Cr contra estava diminuída apenas no grupo de pacientes discordantes. CONCLUSÃO: Nossos achados sugerem uma maior diminuição do NAA/Cr ipsi no grupo de pacientes concordantes e da relação NAA/Cr contra no grupo de pacientes discordantes apontando para um padrão diferente de alteração metabólica para os dois grupos. Porém é preciso aumentar o tamanho da amostra para confirmar estes resultados.

PURPOSE: Patients with mesial temporal sclerosis (MTS) are clinically classified as concordant or discordant according to the extent of epileptogenic activity. The aim of this study is to determine whether the metabolic changes in the hippocampus are different in the two groups. MATERIALS AND METHODS: 33 patients diagnosed with MTS, 23 concordant (33±9 years old) and 10 discordant (33±10 years old) and 28 controls (33±10 years old) were evaluated. We obtained magnetic resonance spectroscopy of both hippocampi (ipsilateral and contralateral) on a 3T scanner with single voxel PRESS sequence with TE/TR=135/1500ms. The main metabolites were quantified with LC Model. For comparison between groups statistical tests were performed with a significance level of 0.05. RESULTS: In all patients a reduction of NAAipsi and Cr ipsi was observed, when compared to the contralateral side, but this difference was only significant for the group of concordant patients. In comparison with controls a significant reduction of Cr ipsi and NAAipsi was observed for both groups and for Naa/Cr ipsi in the group of concordant patients while NAA/Cr contra was reduced only in the group of discordant patients. CONCLUSION: Our findings suggest a stronger decline of NAA/Cr ipsi in the group of concordant patients and of NAA/Cr contra in discordant patients, suggesting a slightly different metabolic pattern for both groups. However, we need to increase the sample size to confirm these findings.

Comparison of Experimental Conditions of CCK-8 and MTS for Human Amniotic Epithelial Cells Proliferation Assay / 中国康复理论与实践

@# Objective To explore the optimal experiment conditions of CCK-8 and MTS for cell proliferation assays in human amniotic epithelial cells and to evaluate the cytotoxicity of these reagents. Methods Human amniotic epithelial cells (hAECs) in logarithm growth stages were prepared in different cell concentrations with DMEM/F12 and 10% FBS. The sensitivity and optimal wavelengths was determined based on the optical density (OD) measured at 450 nm and 492 nm. The optimal time was determined under the conditions of the same cell concentration and defined OD values. HAECs were treated with DMSO, CCK-8 and MTS for 1 h, 2 h, 3 h, and 4 h, respectively. 24 h later, cytotoxicity of the CCK-8 and MTS was evaluated by determination of cell proliferation and Trypan Blue staining. Results The optimal detection wavelength was 450 nm for CCK-8, and 492 nm for MTS. The sensitivity of CCK-8 was slightly lower then that of MTS. The optimal time for incubation hAECs with CCK-8 was 4 h within 1~4 h. The inhibitory on cell proliferation and cytotoxicity of CCK-8 were weaker then those of MTS. Conclusion CCK-8 is a convenient reagent with low cytotoxicity for detection of the proliferation of hAECs.

Strain of implants depending on occlusion types in mandibular implant-supported fixed prostheses

PURPOSE: This study investigated the strain of implants using a chewing simulator with strain gauges in mandibular implant-supported fixed prostheses under various dynamic loads. MATERIALS AND METHODS: Three implant-supported 5-unit fixed prostheses were fabricated with three different occlusion types (Group I: Canine protected occlusion, Group II: Unilaterally balanced occlusion, Group III: Bilaterally balanced occlusion). Two strain gauges were attached to each implant abutment. The programmed dynamic loads (0 - 300 N) were applied using a chewing simulator (MTS 858 Mini Bionix II systems, MTS systems corp., Minn, USA) and the strains were monitored. The statistical analyses were performed using the paired t-test and the ANOVA. RESULTS: The mean strain values (MSV) for the working sides were 151.83 microepsilon, 176.23 microepsilon, and 131.07 microepsilon for Group I, Group II, and Group III, respectively. There was a significant difference between Group II and Group III (P < .05). Also, the MSV for non-working side were 58.29 microepsilon, 72.64 microepsilon, and 98.93 microepsilon for Group I, Group II, and Group III, respectively. One was significantly different from the others with a 95% confidence interval (P < .05). CONCLUSION: The MSV for the working side of Groups I and II were significantly different from that for the non-working side (Group I: t = 7.58, Group II: t = 6.25). The MSV for the working side of Group II showed significantly larger than that of Group III (P < .01). Lastly, the MSV for the non-working side of Group III showed significantly larger than those of Group I or Group II (P < .01).

Muire-Torre syndrome: A case of sebaceous epithelioma with thyroid cancer / 대한내과학회지

Muir-Torre syndrome (MTS) is an autosomal dominant genodermatosis characterized by at least one rare sebaceous neoplasm occurring in association with at least one internal malignancy. The visceral neoplasms most frequently associated with MTS are colorectal and genitourinary cancer, accounting for approximately 50 and 25% of cases, respectively. MTS rarely occurs in association with head and neck cancers. We report a rare case of MTS involving follicular thyroid carcinoma in an 84-year-old female.

Muire-Torre syndrome: A case of sebaceous epithelioma with thyroid cancer / 대한내과학회지

Muir-Torre syndrome (MTS) is an autosomal dominant genodermatosis characterized by at least one rare sebaceous neoplasm occurring in association with at least one internal malignancy. The visceral neoplasms most frequently associated with MTS are colorectal and genitourinary cancer, accounting for approximately 50 and 25% of cases, respectively. MTS rarely occurs in association with head and neck cancers. We report a rare case of MTS involving follicular thyroid carcinoma in an 84-year-old female.

Homozygous deletion of p15、p16 genes and its co-deletion of p15/16 genes in serum DNA of the epithelial ovarian cancer / 中国癌症杂志

Background and purpose:It has been confirmed that homozygous deletion of p16/p15 gene and its co-deletion of p16/p15 genes were related to the occurrence, progress and prognosis of epithelial ovarian cancer. However, the mono-deletion and co-deletion of the genes has been detected with tissue but not in serum DNA of the epithelial ovarian cancer. In this article, we studied the relationship between homozygous deletion of p16/p15 gene and its co-deletion of p16/p15 genes in serum DNA of the epithelial ovarian cancer.Methods:Primers were used to amplify exon 2 of p16 and exon 2 of p15 gene by polymerase chain reaction. Homozygous deletions of the p16, p15 and co-deletion of p16/p15 genes were studied in either serum DNA of 165 patients with epithelial ovarian cancer, their counterpart lymphocytes DNA, serum DNA of 25 benign ovarian cyst or of 15 health donors.Results:The homozygous deletion rates of either p15 or p16 gene were 27.9%(46/165)and 27.3%(45/165)serum DNA in the patients with epithelial ovarian cancer respectively, while the co-deletion rate of p16/p15 genes was 24.2% (40/165). However, the deletions of p15/p16 genes and its co-deletion were not found in serum DNA of the counterpart lymphocytes,25 benign ovarian cyst and 15 health donors (The P values were 0.000、0.000 and 0.000 respectively). The deletions of either p15 or p16 gene for the patients with stage Ⅰ~Ⅱ were 14.3%(5/35) and 11.4%(4/35), 33.3%(25/75) and 32.0%(24/75) for the patients with stage Ⅲ, 29.1%(16/55) and 30.9% (17/55) for stage Ⅳ, respectively. Although there was no significant differences among the groups, the deletion of p15 and p16 genes in the patients with advanced stage were higher than that with early stage. The deletion was not found to be associated with histopathology of epithelial ovarian cancer.Conclusions:Homozygous deletions of the p16, p15 genes and its co-deletion of p15/p16 genes were commonly found in the serum DNA of epithelial ovarian cancer and might be associated with clinical stage of the disease. It was suggested that detection with serum DNA may be used as a micro-invasive approach and the deletion of genes might served as biological markers for the development and prognosis of the patients with epithelial ovarian cancer.

A Case of Muir-Torre Syndrome / 대한피부과학회지

Muir-Torre syndrome (MTS) is an autosomal dominant genodermatosis first described in 1967 by Muir and in 1968 by Torre. It is characterized by the presence of at least one sebaceous gland tumor and the presence of a low-grade internal malignancy. The sebaceous neoplasia are typically adenomas, sebaceomas/sebaceous epitheliomas, or carcinomas. Most common internal malignancy is colorectal adenocarcinoma, but also neoplasia of the uterus, ovary and kidney may occur. Recent studies about genetic defect revealed MTS was caused by germline mutations in DNA mismatch repair genes and microsatellite instability. We report a case of MTS, multiple sebaceous adenomas with colorectal cancer in a 67-year-old male.

Effect of Native and Modified Lipoproteins, and Albumin on Human Mesangial Cell Proliferation / 대한신장학회잡지

BACKGROUND: Modified lipoproteins may be involved in nephro- and glomerulosclerosis. Diabetic nephropathy-like lesions have also been induced in a rat model by glycated and glycoxidized albumin. In cultured rat or human mesangial cells, enhanced cell proliferation and production of mesangial matrix in response to lipoproteins and their modified forms have been demonstrated by [3H]-thymidine incorporation and cell counting assays. But these methods are relatively complex and most of them have used only one or two of the lipoprotein, albumin and their modified forms. METHODS: We investigated the effects of native and modifed lipoproteins, and albumin on cultured human mesangial cell proliferation using non-radioactive colorimetric method by MTS/PMS assay. Lipoproteins added were low density lipoprotein(LDL), high density lipoprotein(HDL), very low density lipoprotein(VLDL), oxidized LDL(oxidation with copper sulfate in vitro) and glycated LDL and we also used albumin, glycated albumin, and interleukin-1beta as a positive control. RESULTS: Interleukin-1beta promoted the proliferation of cultured human mesangial cells up to concentration 20 ng/mL. LDL induced the proliferation of mesangial cells in a concentration-dependent manner up to concentration 100 microgram/mL. HDL and VLDL had no significant proliferative effect. Oxidized LDL caused the proliferation of mesangial cells at low concentration up to concentration 25 microgram/mL. Addition of glycated LDL resulted in a concentration- dependent inhibition of mesangial cells. Albumin and glycated albumin inhibited the proliferation of mesangial cells at low concentration of 100 microgram/mL, but cell growth was increased at higher concentrations. CONCLUSION: We demonstrated the effects of the single and modified proteins on the proliferation of cultured human mesangial cell by relatively simple colorimetric method. Results were almostly identical to those of previous studies obtained by radioactive method or cell counting assay.

Expression and significance of the mRNA and protein of Multiple tumor suppressor 1(MTS_1) in gastric cancer and its precancerous lesions / 中国癌症杂志

0.05). But there were significant differences between them and the stronger positive rate cases. There were significant differences between the positive and stronger positive rate of MTS 1 protein comparing normal gastric mucosa to small intestinal type(84.6%,61.5%), colonic type(85.7%, 50.0%) of IM and gastric carcinoma (83.0%,60.4%)( P

DETECTION OF MTS1/p16 GENE EXPRESSION IN COLORECTAL CANCER BY RT-PCR / 解放军医学杂志

To investigate the deletion of MTS1/p16 gene in human colorectal carcinoma,the homozygous deletion of MTS1/p16 gene was checked by reverse transcriptase polymerase chain reaction from 60 patients with colorectal carcinoma.The specimens consisted of 35 colorectal carcinoma,15 tumor-adjacent tissues and 10 normal colorectal mucosa. It was found that 17 14% (6/35) colorectal carcinoma specimens showed homozygous deletion of MTS1/p16 gene.No deletion was detected in tumor-adjacent tissue and normal colorectal mucosa.Deletion of MTS/p16 gene may play a role in the progression of colorectal carcinoma.

Toxicity of Bile Acids on Colon Cancer Cell Lines / 대한암학회지

PURPOSE: Cytotoxicity of the bile acids on colon cancer cell lines was studied to know which bile acid was most cytotoxic to colonic mucosal epithelium. We performed agarose gel electrophoresis whether this toxicity was caused by detergent effect of the bile acids or by apoptotic pathway. MATERIALS AND METHODS: HT29, LoVo, SW620 colon cancer cell lines were exposed to lithocholate, cholate, deoxycholate and chenodeoxycholate with 50, 100, 150, 200, 250, 300 pM as final concentration in DMEM culture media for short time (for 2 hours) and for long time (for 5 days). Agarose gel electrophoresis was performed on each colon cancer cell lines (HT29, LoVo, SW620, SW480) after 1, 2, 3, 4, 5 days exposure to deoxycholate with 150 pM concentration to detect intemucleosomal fragmentation. RESULTS: There was no toxicity after short time exposure in all bile acids concentration and in all colon cancer cell lines. Of the bile acids, deoxycholate was most toxic for all colon cancer cell lines. And DNA fragmentation was noticed after 2 days exposure with deoxycholate. Only LoVo cell line showed apoptotic DNA pattern after 4 days of exposure with deoxycholate. CONCLUSION: Bile acids (especially deoxycholate) are suggested to be possible agents to cause apoptosis in colonic mucosal epithelium.

Mutations of CDKN2 (MTS1/p16I(NK4A)) and MTS2/p15(INK4B) genes in human stomach, hepatocellular, and cholangio-carcinomas

The CDKN2 (MTS1/p16(INK4A)) and MTS2/p15(INK4B) genes, encoding cyclin dependent kinase inhibitors, were found to be homozygously deleted at high frequency in cell lines from many different types of cancer and some primary cancers. To determine the frequency of CDKN2 and MTS2 mutations in human stomach, liver, and cholangio-cancers, molecular analyses of CDKN2 and MTS2 were performed on 4 stomach cancer cell lines, 14 primary stomach adenocarcinomas, 11 hepatocellular carcinomas, and 5 cholangiocarcinomas. Two (50%) of the four stomach cancer cell lines (SNU1, SNU5, SNU16 and Kato III) had mutations of the CDKN2 and MTS2 gene: SNU16, a homozygous deletion; SNU5, a nonsense mutation, CGA to TGA (Arg to stop) at codon 72 of the CDKN2 gene. No mutations were observed in the 14 primary stomach cancer tissues. In contrast to the mutations of CDKN2 and MTS2, Northern blot analysis showed that expression of CDKN2 was absent or decreased in all the remaining four stomach cancer cell lines and 11: of the 14 (79%) primary stomach adenocarcinomas. Five of the 11 (45%) hepatocellular carcinomas and one of the 5 (20%) cholangiocarcinomas have possible mutations in CDKN2 exon 2 and MTS2. One of hepatocellular carcinoma was expressed mobility shift on PCR-SSCP analysis and a missense mutation, GAC to GAA (Asp to Glu) at codon 105 of CDKN2 gene. These results suggest that mutations or inactivation of the CDKN2 gene may be a critical genetic change in the formation of stomach, hepatocellular, and cholangiocarcinomas.

Screening of antimicrobial peptide from marine clamworm Marphysa sanguinea and preliminary research on its antitumor effect / 中国海洋药物

Active fractions were obtained from the c(?)amworm homogenate by reverse-phase concentration, gel filtration and affinity column chromatography. The MTS/PMS assay was used in the screening of antimicrobial peptide in vitro. The MTS/PMS assay and ELISAAFP method were combined to evaluate the effects of antimicrobial peptide on the proliferation and ?-fetoprotein secretion of HepG2 cells as well as on the proliferation of normal mouse osteoblast MC3T3-E1. The results showed that the purified antimicrobial peptide from clamworm Murphysa sanguinea was a constitutive basic protein with MW 8.1 kDa and pI 8.6, which showed inhibition effects on the proliferation and ?-fetoprotein secretion of HepG2 cells at different levels. These effects have a positive correlation with the concentrations of antimicrobial peptide. No obvious inhibition and damage effects were observed on the normal osteoblast.

Study of non-linearity relation of MTS cytotoxicity assay / 中国药理学通报

AIM We found in experiment that the relation of cell number-OD in MTS cytotoxicity assay is non-linearity. In this paper we probe into the relations of cell number-OD and drug dose-inhibition rate. METHODS Cells HL-60 and Raji were used in this experiment. Assay on the relation of cell number-OD: Cells of different concentrations were seeded into 96 well plates. OD was read at 490 nm after incubating with MTS for 1～5 hour. The regression curves were estimated from the data with the software SPSS 11.0. Cytotoxicity assay: Cells were seeded into 96 well plate and incubated with drug of different doses for 72 hours. OD was read at 490 nm after incubating with MTS for 3 hour. The regression curves were estimated from the data with the software SPSS 11.0. RESULTS The cubic curves fit well the relations of cell number-OD of the 2 strains of cells at 5 time points and the coefficient R2 of these cubic curves is 0.997～1.000, greater than the R2 0.938～0.993 of linear model. For the relations of dose-inhibition rate, the cubic curves also have best fitness and their R2 is 0.998～1.000 greater than the R2 0.948～0.987 of linear model of logarithm dose-probit. CONCLUSION For the relations of cell number-OD in MTS cytotoxicity assay, the fitness of cubic curves are better than the linear model. A more accurate IC 50 would be obtained by the cubic curve equation than by probit regression that is in common use.