《癌变·畸变·突变》是中国科学技术协会主管、中国环境诱变剂学会主办、汕头大学医学院承办、科学出版社出版的国家级学术期刊。系“中国科技论文统计源期刊”(中国科技核心期刊)。根据中国学术期刊综合引证报告(2003版)的统计,本刊影响因子为0.379。在肿瘤学类期刊中排名第4。 2. 办刊宗旨 通过介绍环境因子致癌、致畸变和致突变领域的新理论、新技术、新方法以及国内外研究动态,进行学术交流,促进本学科的繁荣与发展。 3. 栏目 设有“专家述评”、“论著”、“肿瘤防治”、“检测研究”、“相关医学基础与临床”、“技术与方法”及“综述”等栏目。 4. 稿件内容 主要报道环境因子与肿瘤发生、胎儿畸形 ...更多
Current Issue
30 March 2024, Volume 36 Issue 2
Evaluating the clinical application of computational methods for identifying splice variants in cardiomyopathy-associated genes
WANG Ziyi, LIU Hao, WU Dongyang, ZHAO Chunxia
2024, 36(2):  85-93.  doi:10.3969/j.issn.1004-616x.2024.02.001
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OBJECTIVE:Non-canonical splice variants are increasingly recognized as key contributors to inherited cardiomyopathies. Several computational methods have been developed to prioritize the effects of variants on splicing. However,their application in the genetic diagnosis of cardiomyopathies remains unclear. Thus, the performance of computational methods for cardiomyopathy-associated genes and of their clinical applications in hypertrophic cardiomyopathy (HCM) patients were evaluated. METHODS: Whole-exome sequencing was performed on 1 212 unrelated patients with HCM,and splice-site variants in cardiomyopathyassociated genes were collected from scientific publications. A splicing-region-specific strategy was employed to evaluate performance of the computational methods and to used optimal methods for prioritizing candidate splice variants. Splicing effects of these variants were assessed using in vitro Minigene assays. RESULTS: The performance evaluation showed that the optimal methods were Pangolin (deep exonic,core donor,and extended donor regions), MLCsplice (extended donor, core acceptor, and extended acceptor regions), MMSplice, and SpliceAI (extended donor region). Using these optimal methods,53.5% (23/43) of the variants were prioritized and confirmed to cause aberrant splicing. A total of 23 splice variants were found in 4.5% (54/1212) of the unrelated HCM cases. CONCLUSION: Based on the splicing-region- specificity, the prediction strategy effectively identified splice variants, thereby improving the diagnostic value of genetic testing for inherited cardiomyopathies.
Effect of low-dose ionizing radiation on oxidative stress and DNA damage repair in human lymphocytoid cells
SUN Xin, LI Shuang, LU Xue, CAI Tianjing, LIU Ya, LIU Qingjie, ZHANG Wei
2024, 36(2):  94-99.  doi:10.3969/j.issn.1004-616x.2024.02.002
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OBJECTIVE: To investigate effect of low-dose 137Cs γ-irradiation on oxidative stress, DNA damage repair and indued DNA damage in normal human lymphocytoid cells (AHH-1). METHODS: AHH-1 cells were irradiated with 137Cs γ-rays at a dose rate of 8.32 mGy/min,with doses of 0 (unirradiated),0.01, 0.02,0.05,0.075,0.1,and 0.2 Gy,and cultured for 1,24,48 and 72 h after irradiation,respectively. Cell viability was examined by a CCK-8 kits. The level of oxidative damage was detected by malondialdehyde (MDA), superoxide dismutase (SOD) and reactive oxygen species (ROS) kits. γH2AX and 53BP1 foci were performed by immunofluorescence. Changes in the expression levels of DNA damage repair-related genes CDKN1A, DDB2 and POLH mRNA were detected by real-time fluorescence quantitative PCR. RESULTS: Compared with the unirradiated group,cell viability were significantly increased at 24 and 48 h after irradiation (P<0.05). The differences in MDA levels and SOD activities were changed significantly in the 0.2 Gy (P<0.05); the relative fluorescence intensity of ROS was significantly increased in the doses of 0.02,0.05,0.075 Gy and 0.2 Gy at 48 h after irradiation. The number of γH2AX and 53BP1 focus increased with the doses,and there were good dose-response relationships at 1 h following radiation exposure (P<0.01). Compared with the unirradiated group,the relative mRNA expression level of DDB2 and POLH genes were significantly up-regulated at 48 h after irradiation (P<0.05). CONCLUSION:Exposure to low-doses of ionizing radiation induced changes in oxidative stress,DNA damage and expression of DNA damage repair-related genes in AHH-1 cells.
Bioinformatics analysis of key genes and immune cell infiltration in cervical squamous cell carcinoma
GAO Han, ZHANG Yi, ZHENG Zhanyue, TAN Qitao, YANG Minli, SUN Yan
2024, 36(2):  100-106.  doi:10.3969/j.issn.1004-616x.2024.02.003
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OBJECTIVE: To use chip data of datasets to identify key pathogenic genes for cervical squamous cell carcinoma, and to investigate mechanisms of action regarding the level of immune cell infiltration. METHODS: The GSE63514 dataset was downloaded from the GEO database. Using the dataset and the limma package in R language,d differentially expressed genes of cervical squamous cell carcinoma. The clusterProfiler package in R language was used to conduct functional enrichment analysis (GO term and KEGG pathway). Then, Protein interaction network maps were constructed, and key genes were screened by using the String and Cytoscape software. The Cancer Genome Atlas (TCGA) database was used to obtain the gene expression profile of cervical squamous cell carcinoma for identification of hub genes. Disease-critical genes were screened using the random forest analysis, followed by the analysis of immune cell infiltration patterns based on the CIBERSORT deconvolution algorithm. RESULTS:569 differentially expressed genes were identified,of which 262 were up-regulated and 307 were down-regulated. The GO analysis showed that the carcinomas were mainly involved with cell cycle process and mitosis. The KEGG analysis showed that the carcinomas were mainly highly correlated with cell cycle and motor proteins. The random forest analysis based on hub genes screened:5 key genes:TTK,CDK1,KIF23,KIF2C,NUSAP1. The results of immune infiltration showed that in naïve CD4+ T cell expression was significantly upregulated in the carcinomas. CONCLUSION: The genes TTK, CDK1, KIF23, KIF2C and NUSAP1 were closely related to the development of cervical squamous cell carcinoma, and immune cells such as naive CD4+ T cells had an important role in the development of the carcinomas.
Relationships between SPP1, DEC1, C1QTNF6 and clinicopathologic outcomes of oral squamous cell carcinomas
FU Yongqing, XU Sanhui, ZHAO Yan, WANG Lili
2024, 36(2):  107-111,117.  doi:10.3969/j.issn.1004-616x.2024.02.004
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OBJECTIVE: To investigate correlations of serum secreted phosphoprotein 1 (SPP1), differentiated embryo-chonrocyte expressed gene 1 (DEC1) and the complement C1q/tumor necrosis factor associated protein 6 (C1QTNF6) levels with clinicopathologic outcomes of oral squamous cell carcinomas. METHODS: Immunohistochemical staining and electrochemiluminescence immunoassay were used to detect protein expression levels of SPP1,DEC1 and C1QTNF6 in tumor tissues and serum samples from 88 patients with oral squamous cell carcinoma. Pearson correlation and Kaplan-Meier survival analyses were used to analyze correlations between protein expression levels of serum SPP1, DEC1 and C1QTNF6 and clinicopathological outcomes. Multivariate Cox regression analysis was used to analyze the risk factors affecting the prognosis of patients with oral squamous cell carcinoma. RESULTS:Results from immunohistochemical staining showed that positive expression levels of SPP1, DEC1 and C1QTNF6 proteins in carcinoma tissues were 1.94, 2.98 and 2.35 times higher than those in adjacent normal tissues (P<0.05 or P<0.01). Results from the electrochemiluminescence immunoassay showed that expression levels of serum SPP1,DEC1 and C1QTNF6 in patients were up-regulated by 8.61,6.20 and 4.03 times (P<0.05 or P<0.01) compared with normal controls. Pearson correlation analysis results showed that expression levels of serum SPP1,DEC1 and C1QTNF6 were positively correlated with the occurrence of multifocal neurotropic invasion, degree of tumor invasion, lymph node metastasis,and higher TNM stage (P<0.05). Kaplan-Meier survival analysis showed that patients with high serum SPP1, DEC1 and C1QTNF6 protein expression levels had a low overall survival rate. Multivariate Cox regression analysis showed that multifocal neurotropic invasion,high tumor invasion,lymph node metastasis and high TNM stage were the risk factors for prognosis of patients (P<0.05). CONCLUSION:Abnormally elevated serum SPP1, DEC1 and C1QTNF6 proteins in patients were positively correlated with multifocal perineural invasion, tumor invasion, lymph node metastasis and higher TNM stage, but negatively correlated with the prognosis. Systematic detection of expression levels of SPP1, DEC1 and C1QTNF6 in serum of patients have important value and significance for mastering patient's conditions and for guiding clinical practices.
Expression levels of the serum KRT17 protein in patients with locally advanced cervical cancer and its impact on response to neoadjuvant chemotherapy
ZHANG Xiaofeng, SONG Fang, ZHU Linzhong
2024, 36(2):  112-117.  doi:10.3969/j.issn.1004-616x.2024.02.005
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OBJECTIVE: To determine expression levels of the serum keratin 17 (KRT17) protein in patients with locally advanced cervical cancer (LACC) and its impact on response to neoadjuvant chemotherapy (NACT). METHODS: From January 2017 to June 2018, 70 cervical cancer patients who visited our hospitalwere selected and they were treated with arterial interventional neoadjuvant chemotherapy combined with surgery. Before treatment, their serum KRT17 protein expression levels were measured by enzyme-linked immunosorbent assay (ELISA). According to the median level,they were divided into low expression and high expression groups. The progression free survival (PFS), total survival (OS), pelvic progression free survival (PPFS),and pelvic progression free survival (PPFS) of the two groups of patients were compared to evaluate changes in the expression levels and in clinical significance with chemotherapy. There was a difference in distant metastasis free survival (DMFS). RESULTS: The median serum KRT17 level in LACC patients was 0.65 μg/L,and the range was 0.49-1.26 μg/L. There were 33 patients in the low expression group and 36 in the high expression group. The area under the subject operating characteristic (ROC) curve (AUC) of serum KRT17 for predicting the efficacy of NACT in the patients was 0.897 [95%CI (0.816,0.978)],with sensitivity and specificity of 90.6% and 81.6%,respectively. The median PFS of patients with low and high expression of serum KRT17 was 57.23 and 48.75 months, respectively; The median OS was 58.1 and 55.5 months, respectively; The median PPFS was 56.9 and 49.9 months, respectively; The median DMFS was 57.1 and 50.1 months, respectively. The Kaplan Meier survival curve shows a positive correlation between increased serum KRT17 expression and shortened PFS, OS, PPFS, and DMFS in LACC patients. CONCLUSION: Expression of KRT17 protein in the serum of LACC patients was elevated,and the elevated levels can be a useful predictor for poor prognosis in LACC patients who were treated with NACT.
Network pharmacology-based prediction of Pueraria lobata for treating liver injury
LIU Shibu, ZHENG Jifan, ZHANG Li, MA Libo, DUO Hui, LIU Mengqi
2024, 36(2):  118-123.  doi:10.3969/j.issn.1004-616x.2024.02.006
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OBJECTIVE:To investigate the active ingredients,targets, action pathways and mechanisms of actions of Pueraria lobata for treating liver injury. METHODS:According to the public databases such as TCMID, TCMIP, TCMSP, GeneCards, and basic network pharmacological methods such as KEGG pathway analysis, potential pathways of pueraria for treating liver injury were predicted. RESULTS: The main active components of puerariae showed intersection with liver injury. Functional analysis, enrichment analysis and molecular docking showed that formononetin,β-sitosterol and ESR1,coumestrol,3′-methoxydaidzein and PPARG have good affinity for docking. The interaction between formononetin and ESR1 was the strongest (-8.04 kcal/ mol). It was concluded that the mechanism of Pueraria lobata treating liver injury was related to ESR1 and PPARg-mediated pathway. CONCLUSION:The use of Pueraria lobata for treatment of liver injury may be mainly due to regulating the growth,differentiation and physiological functions of hepatocytes through estrogen receptor signaling pathway and PPAR-γ activation and regulation of target gene transcription expression pathway,and alleviating the mechanism of oxidative stress induced liver injury and hepatocyte apoptosis.
Expression and significance of miR-544a in patients with colorectal cancer
TONG Jing, WANG Dan
2024, 36(2):  124-128,137.  doi:10.3969/j.issn.1004-616x.2024.02.007
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OBJECTIVE: To explore expression level and clinical prognostic value of microRNA-544a (miR-544a) in tissues of colorectal cancer patients. METHODS:From January 2016 to February 2019, 150 patients with colorectal cancer were selected. Real time fluorescence quantitative PCR was used to detect expression levels of miR-544a in tissues, and immunohistochemistry was used to detect the expression of Ki67,CD4,and CD8 in tissues. Mann Whitney U-test,chi square test,and Fisher′s exact test were used to analyze relationships between miR-544a expression level in cancer tissue and clinical pathological indicators of patients. Univariate and multivariate Cox regressions were used to analyze factors affecting the prognosis of colorectal cancer. The Kaplan-Meier method was used to analyze relationships between expression levels of miR-544a in tissues and disease-free survival (DFS) and overall survival (OS) of patients. RESULTS: The relative expression level of miR-544a in colorectal cancer tissue (1.85±0.15) was significantly higher than that in the adjacent normal tissues (1.06 ± 0.21) (P<0.01). The expression level of miR-544a was related to the incidence of peripheral nerve invasion,T staging,and N staging (all P<0.01),while the high expression group of miR-544a had a higher incidence of nerve invasion and a later T and N staging. The immunohistochemical results showed that the Ki67 score of the miR-544a high expression group was 120.2±33.7,while the Ki67 score of the low expression group was 112.4±40.8,with no significant difference between the two groups (P> 0.05). The CD4 and CD8 scores of patients in the miR-544a high expression group were 15.5±5.3 and 24.8± 7.6, respectively, lower than those in the miR-544a low expression group (21.3 ± 7.2 and 29.6 ± 8.9, respectively), with statistically significant differences (all P<0.05). Univariate and multivariate COX regression showed significant correlations between tumor N,M staging,and miR-544a expression levels and postoperative DFS and OS in CRC patients (all P<0.01). CONCLUSION: Upregulation of miR-544a expression in colorectal cancer tissue may be associated with delayed N and M stagings in colorectal cancer patients,and miR-544a may be a potential biomarker for immunotherapy efficacy.
Diagnostic value of sperm-associated antigen 9 combined with PSAD and fPSA/tPSA ratio for prostate cancer in the PSA grey zone
SHEN Hongmei, LIU Peilong, GU Ping, BEN Liangliang, XU Zhenyu
2024, 36(2):  129-132,163.  doi:10.3969/j.issn.1004-616x.2024.02.008
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OBJECTIVE:To study the diagnostic value of sperm-associated antigen 9 (SPAG9) in urine combined with prostate-specific antigen density (PSAD),free prostate-specific antigen (fPSA)/total PSA (tPSA) ratio in blood for the diagnosis of prostate cancer in the PSA grey zone. METHODS:From January 2018 to December 2022, 309 patients with serum PSA in the gray zone (4-10 ng/mL) who were admitted to the Second People's Hospital of Nantong were selected as the study subjects. They were divided into case and control groups: 58 prostate cancer case group and 251 benign cancer control group according to the pathological results of prostatocentesis. Before surgery, urine and blood samples from patients were collected and SPAG9,PSAD and fPSA/tPSA were determined. Differences in clinical data between the two groups were identified, and the related factors of prostate cancer were analyzed by logistic regression. The receiver operating characteristic curve (ROC) was drawn, and the area under the curve (AUC) was calculated. The diagnostic value of each indicator for prostate cancer in PSA gray zone was analyzed. RESULTS:Among the two groups of patients,there were no significant differences in body weight,tPSA,white blood cell count, hemoglobin,platelet count,total bilirubin,albumin,creatinine,uric acid and prostate volume between them (P>0.05). However,the age,urinary SPAG9,fPSA/tPSA and PSAD of the case group were significantly higher than those of the control group (P<0.05). Multivariate logistic regression analysis showed that SPAG9, fPSA/ tPSA and PSAD were related factors for prostate cancer. ROC curve analysis showed that SPAG9,fPSA/tPSA and PSAD alone and in combination could diagnose prostate cancer. The combined efficacy of the 3 indexes was ideal,and the sensitivity and specificity were 80.00% and 80.88% respectively. CONCLUSION:Urine SPAG9 can be used as a marker for the diagnosis of prostate cancer in PSA grey zone,and the combination of SPAG9,serum fPSA/tPSA and PSAD can provide good diagnostic efficacy.
Subacute toxicity of a dietary supplement containing Cyclocarya paliurus and Fagopyrum tataricum in rats
ZHOU Qinghong, HE Ning, ZHANG Shujing, LIU Hongbo, WEI Jiemin, QIAN Zhiyong
2024, 36(2):  133-137.  doi:10.3969/j.issn.1004-616x.2024.02.009
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OBJECTIVE: To investigate the subacute toxicity of a dietary supplement containing Cyclocarya paliurus and Fagopyrum tataricum in rats. METHODS: Wistar rats were randomly divided into four groups according to body weight,including three dosage groups of the dietary supplement (1.67,3.33 and 6.67 g/kg) and one control group fed with a conventional basal diet. The corresponding feed was given for 28 days with free access to water. Body weight, food utilization rate, blood routine, blood biochemistry, urine parameters, organ coefficients and histopathological examination of the major organs were observed after anesthesia and dissection. RESULTS:The experimental rats had normal body weight growth and no obvious toxic reactions were observed. There was no statistical difference in hematological, biochemical, urine parameters and organ coefficients between the experimental and the control groups (P>0.05). No abnormality was found in gross observation during animal dissection. Some experimental groups showed statistically significant differences (P<0.05) in heart and kidney weight compared with the control group. Histopathological examination revealed no typical pathological histological changes or specific damage caused by the dietary supplement containing Cyclocarya paliurus and Fagopyrum tataricum. CONCLUSION:Under the conditions of this experiment,the dietary supplement containing Cyclocarya paliurus and Fagopyrum tataricum did not induce obvious toxicity in rats after oral exposure for 28 days.
Study on perinatal toxicity of serum thymic factor in rats
YANG Yu, LI Rongguang, WANG Dan, HUANG Yali, SHAN Chun, LIU Shibu, LIN Fei
2024, 36(2):  138-142.  doi:10.3969/j.issn.1004-616x.2024.02.010
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OBJECTIVE: To investigate effects of serum thymic factors on the growth and development, learning ability and reproductive ability of rats during the late pregnancy,delivery,lactation;as well as during postnatal periods of embryos and fetuses. METHODS:After successful mating,female SD rats were into 20 in each group. From the 15th day of pregnancy to the end of the lactation period, serum thymus factors were administered by continuous subcutaneous injection at 0.88,1.75 and 3.50 mg/kg respectively,and 0.9% chloride was provided. Sodium injection was the solvent control group. The body weight and food intake of the F0 generation female mice were regularly weighed;the birth survival rate and lactation survival rate of F1 generation offspring mice were determined,and the physiological development and neonatal reflexes reach the standard age, and the learning and memory abilities were trained. The reproductive behavior of the offspring after mating in the 12th week after birth was observed, and the various indicators of embryotoxicity was determined. RESULTS: After injection of serum thymic factors,the average body weight and food intake of the F0 generation pregnant and lactating mice in the solvent control group and the test drug groups were normal at each week,and the total number of offspring,sex ratio and birth survival rate (96.3%-97.9%) and the survival rate during lactation (97.7%-98.9%) were basically consistent. The average body weight and food intake of F1 generation mice during lactation and after weaning were basically normal. The physiological development and neonatal reflex time point examination items of the offspring in the solvent control group were all normal at the standard age,swimming adaptation time (28.5±15.8) s,spatial positioning establishment time (134.9±44.1) s,and darkroom electric shock reflex establishment time (4.9±3.1) s,the number of electric shocks to establish memory is 1.7±0.9;the cage mating rate of F1 generation sexually mature rats in the solvent control group was 100%,the pregnancy rate was 90%,the average number of implantations was 10.6±2.1,and the average number of live fetuses was 10.2±1.9, the number of stillbirths was 3,and the number of malformed fetuses was 1. The above indicators did not show significant changes in each dose group of the tested drug. Compared with the solvent control group, the differences were not statistically significant. CONCLUSION:Under the conditions of this experiment,no obvious abnormalities were found in the growth and development,neurobehavior,and reproductive ability of rats in late pregnancy and F1 generation embryos and fetuses in the three dose groups of the solvent control group and serum thymus factor,and no perinatal toxicity was found.
Protective effect of aqueous extracts from Phyllanthus emblica on longevity and climbing ability of Drosophila melanogaster
LI Lingzhi, HU Xueqin, HUANG Yanan, JIA Yizhen, WANG Xu, GUO Xihan
2024, 36(2):  143-149.  doi:10.3969/j.issn.1004-616x.2024.02.011
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OBJECTIVE: phyllanthus emblica (PE) is a medicinal and food plant rich in a variety of phenolic substances and organic acids. In this study,we investigated the effect of PE aqueous extract on the lifespan and climbing ability of Drosophila melanogaster (D.melanogaster) under normal physiological and Cadmium (Cd) stress conditions. METHODS: PE aqueous extracts (1.6, 8 and 16 mg/mL) were applied to wild-type male and female D.melanogaster to find the optimal concentrations and treatment times to investigate their effects on lifespan and climbing ability,and on the SOD,CAT,and MDA. Then 5,10,20,40 mg/L CdCl2 was applied to both male and female D.melanogaster to find the optimal concentration that effectively reduced the lifespan of D.melanogaster. Then,the optimal concentration of CdCl2 and PE extracts were mixed and used to treat both male and female D.melanogaster and to explore the effects on D.melanogaster under Cd stress. RESULTS:Compared with the control,16 mg/mL PE aqueous extract extended the lifespan of male and female D.melanogaster under normal physiological conditions (P<0.05). Under the optimal concentrations of PE (1.6 mg/mL for females and 8 mg/mL for males), the climbing abilities of both were improved and showed gender differences,with the optimal time for females of 22 d and males of 10 d,but the optimal concentration of PE aqueous extracts did not increase the antioxidant enzyme activities in males and females for 10,22,33and 45 d,respectively. From treatment with 40 mg/L CdCl2,the average lifespans for both were reduced (P<0.01) compared with the control. Even with treatment from 40 mg/L CdCl2,8 and 16 mg/mL PE helped to prolong the lifespan of males but their climbing ability was significantly reduced after 18 d of treatment (P<0.05), while the treatments of 1.6, 8 and 16 mg/mL PE did not generate any significant effects on the lifespan and climbing ability of females (P>0.05). CONCLUSION:PE aqueous extract prolonged the lifespan and climbing ability of both male and female D.melanogaster under normal physiological conditions,and prolonged the lifespan of males under Cd stress,but reduced their climbing ability. In addition,there was a gender difference in the protective effect of PE on D.melanogaster.
The subchronic toxicity and teratogenicity of Radix Asparagi in rats
XIA Qiyue, JIANG Yong, WANG Dongxia, LI Bin
2024, 36(2):  150-154,158.  doi:10.3969/j.issn.1004-616x.2024.02.012
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OBJECTIVE: To investigate the subchronic toxicity and teratogenicity of Radix Asparagi by repeated oral administration to rats orally. METHODS:The 90-day oral toxicity of the Radix Asparagi was detected by eighty Sprague-Dawley rats (half male and half female,65.4-94.0 g) and rats were administered with the doses of 2 000,4 000 and 8 000 mg/kg and negative control for consecutive 90 days to observe their general conditions, weight changes and food intake of animals, food utilization rates, and to conduct hematology,biochemical and pathological tests. In the traditional teratogenic test,22 pregnant female SD rats were used in each group,and the doses of 1 000,2 000 and 4 000 mg/kg body weight and negative control group (distilled water) were administered orally. The pregnant rats were sacrificed on the 20th day of pregnancy to examine the pregnancy status and fetal development, and to observe the appearance, the bone and the internal organs of the fetus for teratogenicity evaluation. RESULTS:No toxic symptoms and death occurred in the 90-day oral toxicity test,and no toxic damage was observed in the histopathological examination in the experimental group. In the teratogenicity test,the incidence of sternum hypoplasia was 21.49% and 26.32% in the low and medium dose group,respectively,which was significantly higher than that in the control group (6.25% ) (P<0.01), and no other abnormalities were found. CONCLUSION: The no observed adverse effect level (NOAEL) in 90-day oral toxicity test was 8 000 mg/kg bw. In the teratogenicity study,there were no treatment-related effects observed and Radix Asparagi did not show teratogenic effect on SD rats.