The Mutagenicity of the Lyophilized Agkistrodon Haly （pallas） Venom

Objective To investigate the mutagenicity of Methods The mutagenicity of the Lyophilized Agkistrodon the Lyophilized Agkistrodon Haly （pallas） Venom. Haly （pallas） Venom was studied by the micronucleus test, the Ames test and the CHL test. Results No significant differences in the micronuclear frequences were found in any of the testing groups compared with negative control in mice （p〉0.05）; The mutagenic activity of the Lyophilized Agkistrodon Haly （pallas） Venom was not observed in TA97, TA98, TA100 and TA102 with and without S9 activation under the doses used; In CHL test, the effect of chromosomal damage of the Lyophilized Agkistrodon Haly （pallas） Venom was not found either in CHL cells. Conclusion The Lyophilized Agkistrodon Haly （pallas） Venom has no mutagenicity, and it might be useful in future.     

Study on the Mutagenicity of Diesel Exhaust Particles

The mutagenicity of diesel exhaust particles(DEP)was studied by using Salmonella typhimurium strains TA98 and TA100 in vitro and mice micronucleus in vivo test.DEP from six kinds of medium and heavy-duty diesel vehicles,which were made in China and imported,were tested.The vehicles wer eoperated under free accelerating condition.The results showed that the EP contained mutagenic activity.An increase in the number of the Salmonella TA98 was observed in the presence and especially in the absence of S9 Mmix.Positive results were also obtained from mice micronucleus assay.The frequency of mice bone marrow micronucleated polychromatic erythrocytes(M PCE)was increased and it showed a definite dose-response relationship.Comparing the different types of the vehicles,we found that the mutagenicity of DEP from domestic made vehicles was stronger than that from the imported ones.     

Mutagenicity and Induction of Drug—metabolizing Enzyme Activity by LPG Combustion Particulates in Rats

Methylene chloride extracts of particulates from liquefied petroleum gas(LPG)combustion appliance were studied by using Ames test,micronucleus test and inducibility of pulmonary and hepatic aryl hydrocarbon hydroxylase(AHH)and glutathione S-transferase(GST)in rats.The extracts showed mutagenicity for Salmonella typhimurium strain TA98 and its derivatives TA98NR and TA98/1,8-DNP6 with or without S9 mix.The revertants in strains TA98NR and TA98/1,8-DNP6 were less than 40%and 50% of than in strain TA98 without S9 mix,respectively.Positive results were obtained in mouse bone marrow micronucleus assay.Intratracheal instillation of the extracts led to increase in pulmonary(but not hepatic)AHH and GST activities in rats.In was seen that AHH was more sensitive than GST to induction by the extracts.     

Antioxidant and Antigenotoxic Activity of Bioactive Extracts from Corn Tassel简

This study is designed to evaluate antioxidant and antigenotoxic activities of corn tassel extracts（CTTs）. The major bioactive components of CTTs include flavonoid, saponin and polysaccharide. The antioxidant properties of the three bioactive components of CTTs were investigated by Ferric Reducing Antioxidant Property（FRAP） and 1, 1-diphenyl-2-picrylhydrazyl（DPPH） assays. The activities of the extracts were determined by assessing the inhibition of mutagenicity of the direct-acting mutagen fenaminosulf, sodium azide, and indirect-acting mutagen 2-aminofluorene using the Ames test（strains TA98 and TA100）. The results showed that the extraction rates of flavonoid, saponin, and polysaccharide from the dried corn tassels were 1.67%, 2.41% and 4.76% respectively. DPPH and FRAP assay strongly demonstrated that CTTs had antioxidant properties. CTTs at doses of 625, 1250 and 2500 μg per plate reduced 2-aminofluorene mutagenicity by 12.52%, 28.76% and 36.49% in Salmonella typhimurium TA98 strain assay respectively and by 10.98%, 25.27% and 37.83%, at the same doses in Salmonella typhimurium TA100 assay system, respectively. 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide（MTT） assay showed that the different concentrations of CTTs inhibited the proliferation of MGC80-3 cells in a dose-dependent manner（P0.01）. It is concluded that these integrated approaches to antioxidant and antigenotoxicity assessment may be useful to study corn tassel as a natural herbal material.     

Studies on the Mutagenicity of Hair Dyes Made in China

A total of 13 commercial hair dye products made in China were tested for mutagenicity in 2 short-term bioassays, the histidine-requiring mutants of Sahnonella typhhnurium (strains TA98 and TA100) and the micronucleus test with mouse bone-marrow polychromatic erythrocyte cells in vivo. The results showed that the 13 hair dyes were not mutagenic in strains TA98 and TA100 with and without S-9. In the micronucleus test, no mutagenic effect was observed.     

Mutagenicity of Urine From Individuals Exposed to LPG Combustion Products

The mutagenicity of urine from individuals exposed to the combustion products of liquefied petroleum gas(LPG)was detected with Salmonella typhimurium TA98 and its newly developed derivatives YG1021(nitroreductase overproducing)and YG1024(O-acetyltransferase overproducing).The detection showed significantly increased mutagenicity for two YG strains and increased positive rates for all three strains in the presence of both rat liver S9 and β－glucuronidase.Further analysis demonstrated that urine samples taken from smoking and non-smoking exposed individuals exhibited significatly higher mutagenic potency(revertants/10μl urine concentrate)than their corresponding controls.These results indicate that the increased urine mutagenicity is caused by the exposure to LPG combustion products or smoking.The mutagenic potency of urine samples of all exposed individuals tested with YG 1024 was found to be about 7 times higher than with TA98.The difference in mutagenic potency was smaller for the same samples when comparison was made betweenTG1021 and TA98.This suggests that the mutagenic compounds present in the urine samples contain mainly armatic compounds as glucuronide conjugates.Our results demonstrate that YG1024 is more sensitive than TA98 in detecting the mutagenicity of these samples.In addition ,no significant difference in the mutagenic potency between the‘pure’ exposed(non-smokers‘) and the ‘pure’ smokers’(unexposed) samples was found in all three tester strains.This might mean that the exposure extent of mutagens/carcinogens in LPG combustion products for exposed individuals roughly corresponds to the smoking level of smokers who smoke 20-40 cigarettes per day.Furthermore,the results also suggest that synergism might exist in the mutagenic effects of exposure to LPG combustion products and cigarette smoking.     

A Study on Genotoxicity of Cooking Fume from Rapeseed Oil

The present article reports the genotoxic potential of rapeseed oil cooking fume investigated by a battery of short-term tests (Ames test, SCE/V79 in vitro and mice micronucleus in vivo test). The results showed that the cooking fume contained mutagenic activity. In the presence of S9 mix, an increase in the number of the Salmonella TA98 was observed at doses ranging from 1.0 to 5.0 mg/plate, and the SCE frequencies of V79 cell were markedly raised at doses ranging from 0.05 to 0.5 mg·ml~(-1). The positive result was also obtained in mice micronucleus assay, the mice had inhaled the cooking fume a week earlier. The frequency of mice bone marrow MN-PCE was increased and it showed a remarkable time-dose-response relationship during the 4 weeks exposure. The results suggested that this cooking fume exposure may be a risk factor of lung cancer in Chinese women.     

Mutagenic and Estrogenic Effects of Organic Compounds in Water Treated by Different Processes:A Pilot Study

Objective In this study, a pilot-scale investigation was conducted to examine and compare the biotoxicity of the organic compounds in effluents from five treatment processes (P1-P5) where each process was combination of preoxidation (O3), coagulation, sedimentation, sand filtration, ozonation, granular activated carbon, biological activated carbon and chlorination (NaClO).
Methods Organic compounds were extracted by XAD-2 resins and eluted with acetone and dichlormethane (DCM). The eluents were evaporated and redissolved with DMSO or DCM. The mutagenicity and estrogenicity of the extracts were assayed with the Ames test and yeast estrogen screen (YES assay), respectively. The organic compounds were detected by GC-MS.
Results The results indicated that the mutation ratio (MR) of organic compounds in source water was higher than that for treated water. GC-MS showed that more than 48 organic compounds were identified in all samples and that treated water had significantly fewer types and concentrations of organic compounds than source water.
Conclusion To different extents, all water treatment processes could reduce both the mutagenicity and estrogenicity, relative to source water. P2, P3, and P5 reduced mutagenicity more effectively, while P1 reduced estrogenicity, most effectively. Water treatment processes in this pilot plant had weak abilities to remove Di-n-butyl phthalate or 1, 2-Benzene dicarboxylic acid.     

Evaluation of the in vitro and in vivo genotoxicity of almond skins

Objective It aims to study potential genotoxicity of almond skins.Methods A bacterial reverse mutation assay was performed on S.typhimurium strains TA97,TA98,TA100,TA102,and TA1535 in the absence or presence of S-9 mixture at a dose range of 312.5 to 5 000 μg/plate.A micronucleus test and a mammalian bone marrow chromosome aberration tests were performed in Swiss Albino (CD-1) mice at doses of 625,1 250,and 2 500 mg/kg bw used.Results Almond skins exerted no mutagenic activity in various bacterial strains o...     

Mutagenicities of 22 Kinds of Foods After Nitrite Treatment

The mutagenicities of 22 kinds of foods,including soy sauce,fish sauce,shrimp paste,sausages,sundried fishes,rice cracker,deep fried pork skin,salted vegetable and spices after nitrite treatment,were detected by meansof Ames test with preincubation.Mutagenicity assay was employed on Sal-monella typhimurium TA100,and meanwhile,the validity of each experi-ment was checked by using the known mutagen,AF-2,as the positivecontrol.16 kinds of foods showed marked direct-acting mutagenicity towardSalmonella typhimurium TA100 after nitrite treatment.Shrimp paste prod-uced in Bankok was the strongest one among these samples.Its specificmutagenicity was 37000 revertants/g.The amount of precursor,tyramine,in shrimp paste was estimated fromthe area of tyramine peak on HPLC by using the authentic tyramine as thestandard.Results showed that one gram shrimp paste contained 439μgtyramine.The mutagenicities of foods suggested that nitrosatable precursors,suchas amine or amide,were present in foods,and they could be converted intoendogenous carcinogen,nitrosamine,in vivo.Therefore,detection of muta-genicities of foods has significance in preventing cancer.     

Retrieval and Amplification of DNA from Unstained Histopathological Sections

Testing of compounds for carcinogenic potential in vivo involves various experimental designs.A few of these techniques are directed to demonstrate the genotoxicity and mutagenicity of the compound by histopathology.These changes shown by histochemical means include monoclonal antibody directed cellular markers.Development of the polymerase chain reaction technique(PCR)for amplification of DNA has facilitated the investigation of molecular events related to the formation of malignant neoplasms.We describe here a method for screening tissues for mutations of the H-ras gene using monoclonal antibodies directed toward normal and mutant p21 proteins.Formalin-fixed,paraffinembedded tissue sections are used to subsequently confirm the gene mutation by PCR amplification of the H-ras gene.The results indicated a successful application of this technique to demonstrate the presence of p21 oncoprotein in the tissues tested.     

Management of the occlusion and stenosis of the aortic arch branches in the course of Takayasu’s arteritis

Takayasu＇s arteritis （TA） is a chronic inflammatory disease of the aorta and its thoracic and abdominalparts, most commonly involving the aortic arch and the arteries that arise from it. TA is an autoimmune disease, although certain HLA-linked genetic predispositions have been observed. TA affects females 2 to 8 times more fi＇equently than males. The onset age is between 10 to 40 years old.1 TA is extremely uncommon in Poland. Corticosteroids are the choice of treatment for TA,     

A SAS macro for testing differences among three or more independent groups using Kruskal-Wallis and Nemenyi tests

As a nonparametric method,the Kruskal-Wallis test is widely used to compare three or more independent groups when an ordinal or interval level of data is available,especially when the assump-tions of analysis of variance (ANOVA) are not met.If the Kruskal-Wallis statistic is statistically signifi-cant,Nemenyi test is an alternative method for further pairwise multiple comparisons to locate the source of significance.Unfortunately,most popular statistical packages do not integrate the Nemenyi test,which is not easy to be calculated by hand.We described the theory and applications of the Kruskal-Wallis and Nemenyi tests,and presented a flexible SAS macro to implement the two tests.The SAS macro was demonstrated by two examples from our cohort study in occupational epidemiology.It provides a useful tool for SAS users to test the differences among three or more independent groups using a nonparametric method.     

Preoperative preparation and surgical separation of conjoined pygopagus twins

Conjoined twins are very rare with an estimated ,incidence of about 1 in 200 000 births with a male-female ratio of 1：3 The separation ot conjolned twins presents a challenge to surgeons and also a test for comprehensive efficiency of a hospital. Recently, a rare pair of pygopagus twins were admitted to our hospital and a successful surgical separation was carded out.     

Expression of CⅡTA Gene in Five Human Malignant Hematological Cell Lines and Its Significance

The role of MHC class Ⅱ transactivator (CⅡTA) in constitutive or IFN-γ inducib|e expression of HLA molecules in human malignant hematological cell lines was investigated. The expression of HLA molecules and CⅡTA protein was detected by Western blot, immunohistochemistry and flow cytometry. The expression of CⅡTA gene was determined by RT-PCR. The capability of peripheral blood T cell reaction stimulated by tumor cells was monitored by mixed lymphocyte reaction. It was found that the HLA Ⅱ-positive tumor cells expressed the CⅡTA quite well, andthe expression of HLA Ⅰ＋Ⅱ was increased in the tumor cells with constitutive or inducible expression of CⅡTA after induced by IFN-γ. The tumor cells which did not express CⅡTA after in-duced by IFN-γ were not response to the expression of HLA Ⅱ promoted by IFN-γ. It suggests a correlation between the inability of some malignant hematological cell lines in response to IFN-γ for HLA expression and the deficiency in the inducible expression of CⅡTA, indicating CⅡTA might take part in the regu|ation of HLA Ⅰ＋Ⅱ expression in the tumor cells, which might p|ay an important role in tumor immunologic escape.     

Genotoxic and nongenotoxic effects of glycidyl methacrylate on human lung fibroblast cells

Objective To evaluate the genotoxic and nongenotoxic effects of short-term exposure to glycidyl mathacrylate (GMA) on human lung fibroblast cells (2BS cells) in vitro. Methods DNA strand breakage was determined by single cell gel electrophoresis, and DNA ladder formation assay and flow cytometric analysis were carded out to detect apoptic responses of cells to GMA exposure.The HPRT gene mutation assay was used to evaluate the mutagenicity, and the effect of GMA on gap junctional intercellular communication (GJIC) in the exposed cells was examined with the scrape loading/dye transfer technique. The ability of GMA to transform 2BS cells was also tested by an in vitro cell transformation assay. Results Exposure to GMA resulted in a dose-dependent increase in DNA strand breaks but not apoptic responses. GMA was also shown to significantly induce HPRT gene mutations and morphological transformation in 2BS ceils in vitro. In contrast, GMA produced a concentration-dependent inhibition of GJIC. Conclusions GMA elicits both genotoxic and nongenotoxic effects on 2BS cells in vitro. The induction of DNA damage and gene mutations and inhibition of GJIC by GMA may casually contribute to GMA-induced cell transformation.     

In vitro safety evaluation and anticlastogenic effect of BacoMind on human lymphocytes

Objective BacoMind^TM （BM） is a standardized extract of Bacopa monnieri, which belongs to the family Scrophulariaceae and is a creeping annual plant found throughout the Indian subcontinent. It has been used by Ayurvedic medicinal practitioners in India for almost 3000 years and is classified as a medharasayana, a substance which improves memory and intellect. With the widespread traditional use as well as scientific validation of Bacopa monnieri for nootropic activity, a bioactive-rich unique phytochemical composition-BacoMind^TM was developed from B. monnieri for use as a cognition and memory enhancing agent. The present study aimed to investigate the in vitro toxicity of this formulation of BacoMind^TM on human lymphocytes and to rule out its possible contribution to mutagenicity. Methods In the present investigation the active ingredients present in BM were identified and quantified by high performance liquid chromatography （HPLC） and high performance thin-layer chromatography （HPTLC）. Antioxidant and anticlastogenic properties of BM were studied in vitro with and without metabolic activation. Doses of BM were chosen on the basis of mitotic index （MI） and cytokinesis-block proliferation index （CBPI）. Clastogenicity assays were performed at 31.2 μg/mL, 62.5 μg/mL, and 125 μg/mL, while the Salmonella reverse mutation assay （Ames test） was performed at doses of 61.72, 185.18, 555.55, 1666.67, and 5000.00 μg/plate. Results HPLC and HPTLC analysis of BM revealed the presence of bacoside A3. bacopaside Ⅰ, bacopaside Ⅱ, jujubogenin isomer of bacopasaponin C, bacosine, luteolin, apigenin, bacosine, and β-sitosterol D glucoside. BM demonstrated significant antioxidant activity. The number of chromosomal aberrations and the frequency of micronuclei induced by BM were not statistically significant up to a dose of 62.5 μg/mL. A subsequent dose of 125 μg/mL prior to metabolic activation induced mild clastogenicity, but it was found to be biologically insignificant as this effect was not     

磷酸羟基哌喹致突性的实验研究

应用Ames试验、体外染色体畸变试验、微核试验和显性致死试验对磷酸羟基哌喹(HPQP)的致突性进行了实验研究。结果表明,HPQP对TA97a、TA98、TA100和TA102无致突性。各剂量组的染色体畸变率均低于3％。以1/2、1/10和1/25LD_(50)的剂量诱发小鼠骨髓多染红细胞的微核率分别为5.7±2.55、5.3±1.70和4.3±2.11,与阴性对照无区别。根据活胎率,平均早死数等指标表明HPQP无显性致死突变。综合本组试验结果,可认为HPQP未显示有致突性。     

一个中国Gilbert综合征家系的临床和遗传学特征分析

Objective To perform clinical and genetic pedigree analyses of a Chinese male patient with Gilbert's syndrome and his relatives. Methods Blood sample were collected from the proband and his relatives by liver function test, etiological examination and genetic analysis to exclude other related diseases. The phenobarbital-responsive enhancer module (PBREM) , TATA box and common mutation sites in exons of UGT1A1 gene were amplified by polymerase chain reaction (PCR) and the products screened by direct DNA sequencing. Results c. -3279T ＞ G in PBREM, TA insertion in TATA box and Gly71Arg were observed in this family. A linkage disequilibrium is also noted between c. -3279T ＞ G and TA insertion. In four affected members, three are heterozygotes and one is homozygote. The correlation between genotype and phenotype with a high serum level of unconjugated bilirubin was confirmed. Conclusion c. -3279T ＞ G in PBREM, TA insertion in TATA box and Gly71 Arg are essential for the pathogenesis of Gilbert's syndrome in this Chinese family. Gilbert's syndrome in this family is inherited in an autosomal recessive manner.     

一个中国Gilbert综合征家系的临床和遗传学特征分析

Objective To perform clinical and genetic pedigree analyses of a Chinese male patient with Gilbert's syndrome and his relatives. Methods Blood sample were collected from the proband and his relatives by liver function test, etiological examination and genetic analysis to exclude other related diseases. The phenobarbital-responsive enhancer module (PBREM) , TATA box and common mutation sites in exons of UGT1A1 gene were amplified by polymerase chain reaction (PCR) and the products screened by direct DNA sequencing. Results c. -3279T ＞ G in PBREM, TA insertion in TATA box and Gly71Arg were observed in this family. A linkage disequilibrium is also noted between c. -3279T ＞ G and TA insertion. In four affected members, three are heterozygotes and one is homozygote. The correlation between genotype and phenotype with a high serum level of unconjugated bilirubin was confirmed. Conclusion c. -3279T ＞ G in PBREM, TA insertion in TATA box and Gly71 Arg are essential for the pathogenesis of Gilbert's syndrome in this Chinese family. Gilbert's syndrome in this family is inherited in an autosomal recessive manner.