BACKGROUND ＆ AIM：To study the expression of heat shock protein 47(HSP47) in normal forelimbs and short forelimb malformations，normal palates and cleft palates during mouse embryogenesis. MATERIAL AND METHODS: At GD10, gestational mice of the treatment and the control groups were administered with 80 mg/kg all-trans retinoic acid and the same volume vegetable oil separately. The forelimbs of embryos were harvested in GD11-GD18，and the normal palates and cleft palates were obtained in GD15-GD17. The relative abundance of HSP47 of all samples was measured by reverse transcript polymerase chain reaction (RT-PCR). RESULTS: HSP47 was expressed in all samples. To the normal forelimbs and short forelimb malformations, with the embryos aging, there was the increasing tendency of the expressional abundance of HSP47 in GD11-GD18，their peaks of expression were especially at GD16 and GD17, and kept stable in the following age. The expressional abundance of HSP47 of the normal palates was stable in GD15-GD17, and the peak of that of the cleft palates was at GD16. The expression of HSP47 of the abnormal limbs was stronger than that of the normal limbs in GD11-GD18. The expression of HSP47 of the cleft palates has no difference with that of the normal palates in GD15, but it was stronger than that of the cleft palates in GD16-GD17. CONLUSION: The expressional abundance of HSP47 in normal limbs and short limb malformations，normal palates and cleft palates induced by all-trans retinoic acid during mouse embryogenesis was increased by stress action.

BACKGROUND ＆ AIM： To investigate the mutation frequency and molecular mutation of spontaneous mutations on the tk gene of mouse lymphoma cells. MATERTAL AND METHODS：Performing four times parallel test. Mutation frequency of tk locus were analyzed by the mouse lymphoma assay(microitier procedure), and Loss of heterozygosity(LOH) analysis withallic special PCR. Subsequently, we determined the DNA sequence of negative LOH's clones. RESULTS：Loss of heterozygosity analysis of the tk locus revealed that the most spontaneous mutants lost the functional tk allele (average LOH 79.8％). LOH of spontaneous mutations showed there were no significant difference between large clone(LC) and small clone(SC).Spontaneous mutations were substitutions, and most of them were transversions. CONCLUSION： Functional tk allic gene lose was major mutational event in spontaneous mutations and point mutations were substitutions.

BACKGROUND ＆ AIM: Compare the anti-tumor effects of Cortex Periplocae and its analogues extracted in vitro. MATERIAL AND METHODS: We investigated the suppressive effect of Cortex Periplocae and its analogues extract on the proliferation of different tumor cells from human and mouse by using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide(MTT) method. RESULTS: Cortex Periplocae extract markedly inhibited the proliferation of tumor cells such as K562,BGC-823,TE-13 with dose-dependent manner.There exist markedly difference among Cortex Periplocae and its analogues extracts of their affect on the proliferation of different tumor cells from human and mouse(P<0.01). CONCLUSION： Cortex Periplocae and its analogues extracts markedly inhibited the proliferation of many tumor cells in vitro, with dose-dependent manner; The Cortex Periplocae worked better than its analogues; Cortex Periplocae as anti-tumor substance is hoped advanced development.

BACKROUND＆AIM: To discuss the protective effects of ammonia against phosgene on mice , in order to find a new therapy for lung injury induced by phosgene. MATERIAL AND METHODS: During phosgene exposure, the treated groups were protected by different dose of ammonia. After 3 h, all mice were anatomized to measure malondialdehyde(MDA),reduced glutathione(GSH),oxidized glutathione(GSSG)of lung and glutathione peroxidase(GSH_Px)of blood by fluorescence_metry. RESULTS: MDA,GSH,GSSG were different significantly between control and positive group. With compared with positive group, MDA and GSSG of trial group was lower(P<0.05). CONCLUSION: There were some oxidative injury occurred in lung during phosgene exposure and ammonia may have protective effect on it.

BACKGROUND ＆ AIM: To investigate the effects of different stress factors on HeLa cell proliferation,apoptosis and heat shock protein 70(HSP70) expression. MATERIAL AND METHODS:Cell culture technique,3_(4,5_dimethylthiazol_2_yl)_2,5_diphenyl tetrazolium(MTT) flow cytometry,immunocytochemistry,western dot. RESULTS:The present study showed that cell apoptosis appeared in all experiment group;there was a tendence to increase HSP70 expression treated with Ethylenediamine tetra_acetic acid (EDTA) and Ca2＋; The protein level of HSP70 was elevated significantly in the groups of H2O2,heating and aminophylline (P<0.01) than control group; the protein level of HSP70 was higher (P<0.05) than control group in the groups of dexamthasone and tween_80. CONCLUSION: The results suggested that the expression of HSP70 in HeLa cells could be elevated and apoptosis could be induced by stress factors.

BACKGROUND ＆ AIM: To observe the toxicity effect and the extent on rats via tail venous continuously injection dicycloplatin, carboplatin and carboplatin added cyclobutane dicarbonylic acid, and compare the degree of toxicity in the three kinds of platinum. MATERIAL AND METHODS: The rats were divided into two dosage groups (5 mg/kg,15 mg/kg)for every tested drug and one negative control group. There are three disposed periods. The drugs were injected via tail venous for five days continuously and quitted the drugs for nine days in every period. RESULTS: There is different extent toxicity effect on rats of the three kinds of tested drugs at same dosage. The major target organ are blood system, digestive system and reproductive system. The toxicity effect of carboplatin stresses on restraining the system of marrow hemopoiesis and damaging the function of kidney, and the toxicity effect of dicycloplatin stresses on damaging the germ cell, but carboplatin added cyclobutane dicarbonylic acid between them. CONCLUSION: Summarizing the toxicity effect of the three tested drugs, the sequence size of the toxicity effect is carboplatin, carboplatin added cyclobutane dicarbonylic acid and dicycloplatin.

BACKGROUND ＆ AIM: To study the detection of micrometastasis in peripheral blood of patients with cardial carcinoma and its clinical implication. MATERIAL AND METHODS: Cytokeratin 20(CK20) mRNA in peripheral blood of 58 patients with cardial carcinoma was detected by using nested reverse transcriptase-polymerase chain reaction (RT-PCR) technique. RESULTS: The positive rate of CK20 mRNA in peripheral blood of the 58 patients was 36.2％. The positive rate of CK20 mRNA in patients with lymph node metastasis was 51.5％, while 16％ in patients without lymph node metastasis, the two rates were significantly different. CONCLUSION: Using nested RT-PCR technique to detect CK20 mRNA in peripheral blood of patients with cardial carcinoma is helpful for early diagnosis of micrometastasis, the positive expression is related to metastasis of lymph node.

BACKGROUND ＆ AIM: To analyze the genotype frequency of alcohol dehydrogenase-2 (ADH2), and evaluate the relationship between ADH2 polymorphism and drinking behavior in Chinese. MATERIAL AND METHODS: Epidemiological data were collected from 207 healthy persons in Taixing city of Jiangsu province, China, and ADH2 genotypes were determined by polymerase chain reattion restriction-fragment length polymorphism,(PCR -RFLP) method. RESULTS: The frequencies of , and genotypes were 12.6％, 46.8％ and 40.6％ respectively. Significant differences were found in the frequency of ADH2 genotypes between drinkers and no-drinkers. Cumularivy amounts of alcohol consumption were found statistically different among drinkers with different ADH2 genotypes. CONCLUSION: Drinking behavior was influenced by the individual genotypes of ADH2.

BACKGROUND ＆ AIM: To study the influence of arsenic trioxide (As2O3)on the telomerase activity and the growth inhibition in melanoma B16 cells,aim to provide new theoretical and experimental bases for the treatment of melanoma by As2O3. MATERIAL AND METHODS: The growth inhibition was analyzed by methyl thiazolyl tetrazolium(MTT);the telomerase activity in melanoma B16 cells was detected by telomeric repeat amplification protocol,enzyme-linked immunosorbant assay(TRAP-ELISA) and telomeric repeat amplification protocol,poly acryl amide gel electrophoresis silver-staining （TRAP-PAGE-silver staining）. RESULTS: As2O3 could inhibit the growth and telomerase activity of B16 cells remarkably, the rates of inhibition had both dose-dependent and time-dependent manner. CONCLUSION: The inhibition of B16 cells by As2O3 may be enhanced with the increase of As2O3 concentration and prolong of co-culture time.

BACKGRAND ＆ AIM： To investigate the relationship among the expression of connexin43(Cx43)，connexin26(Cx26)，vascular endothelial growth factor-C(VEGF-C)，Estrogen receptor(ER)，Progesterone receptor(PR)； the clinicopathological characteristics in breast carcinoma. MATERIAL AND METHODS： A total of 125 paraffin-embedded breast carcinoma and non-carcinomatous breast tissues were stained using S-P method, and the patients' prognosis significance was analyzed by Multivariable Cox proportional hazards model according to the clinical follow-up material. RESULTS： The expression levels of Cx43 and Cx26 were significantly lower in the tumors (38.74％ and 43.24％ respectively) than that in the non-carcinomatous breast tissues (P<0.05 and P<0.01, respectively). And the expression level of VEGF-C was significantly higher in the tumors (47.75％) than that in the non-carcinomatous breast tissues (P<0.05). Out of 53 VEGF-C positive staining, the positive rate of lymph node metastasis was 37(69.81％), which was significantly higher than that in the no lymph node metastasis group (P<0.05). By the Cox regression model, VEGF-C, Cx26, and lymph node metastasis were closely associated with prognosis. VEGF-C and lymph node metastasis were risk factors. The hazard ratios of VEGF-C expression and lymph node status were 11.118 and 10.126,respectively . P=0.007 and 0.042, respectively. There was significant association with prognosis. CONCLUSION： The lack of Cx43, Cx26 gap junctions is a main feature of human breast cancer tissues. Breast tumor malignant degree increased with Cx43 and Cx26 positive expression decreased. It was significantly higher in VEGF-C positive expression and ER, PR negative expression in lymph node metastasis group than that in the no lymph node metastasis group. VEGF-C, Cx26 and Lymph node metastasis status may be indicators of prognosis of breast carcinoma, which would be helpful in assessment prognosis and selecting the higher risk cases for further treatment.

BACKGROUND ＆ AIM: To study the effect of photodynamic therapy (PDT)on ultrastructure of esophageal carcinoma cells. MATERIAL AND METHODS: After action of photosensitizer for 6,12,and 24 hours respectively, cultured esophageal carcinoma cells were irradiated with Ultraviolet B(UVB) for half an hour. The ultrastructure of esophageal carcinoma cells in experimental groups and control groups were observed under transmission electron microscope. RESULTS: PDT resulted in condensation of mitochondria,reduction of rough endoplasmic reticulum, dilation of smooth endoplasmic reticulum,breakup of nuclear membrane and pyknosis of nucleolus. The severity of injury of membranous structures increased with the time prolonging of photosensitizer action. CONCLUSION: PDT can do damage to membranous structures of esophageal carcinoma cells,which suggests that PDT may have clinical efficacy for treatment of esophageal carcinoma.

BACKGROUND ＆ AIM: In order to understand the mechanism of methotrexate (MTX) further and to investigate the genotoxic target organs.MATERIAL AND METHODS： DNA damage and the correlation with dosage treated with MTX were studied by using the alkaline single cell gel electrophoresis assay(SCGE). Liver, spleen, bone marrow, thymus, kidney, testicle, stomach and peripheral lymphocytes of mice were isolated at 1、3、6、12、24 h after MTX intraperitoneal injection. RESULTS: Significant increase in DNA migration and comet frequency in the spleen, thymus, bone marrow and peripheral lymphocytes were induced after intraperitoneal treatment of MTX at a dose of 1.25～5 mg/kg. The migration of nuclear DNA and comet frequency of spleen, thymus, bone marrow and peripheral lymphocytes in the dose-response study showed a dose-dependent increase. CONCLUSION: The results indicate that DNA SSBs could be induced by MTX in some cells of mice. There are difference in sensitivity of various organs in the mice and cells of spleen, thymus, bone marrow and peripheral lymphocytes may be the important target cells of MTX.

BACKGROUND ＆ AIM： The current study investigates the effects of rotary strong magnetic field(RSMF) exposure on rats reproductive toxicity, in order to evaluate the safety of RSMF. MATERIAL AND METHODS：In these tests we adopted three dosage groups, that were treated with MF 0.5, 1, 2 h everyday, a negative group and positive group. Then sperm malformation test, traition malformation test and micronucleus test were performed. The growth of rat offspring, G band of chromosome and percent of micronucleus was compared. RESULTS： 0.4 T MF have no toxicity to rat sperm, gravid rat, growth of rat embryo and chromosome. CONCLUSION：0.4 T MF is safe to the reproduction of rat.

BACKGROUND ＆ AIM: To further study the antimutation effect of grape seed extraction which contains plenty of phenol chemical compound . MATERIAL AND METHODS: The grape seecl extraction was investigated by using mice bone marrow cell micronucleus test and Ames test. RESULTS: The result showed that grape seed extraction inhibited the effect of the positive substance inducing mutation on TA98、TA100 in the condition of whether or not adding S9.Feeding mice with grape seed extract in different doses 30 d,it showed that cyclophosphamide(CPA) induced mice bone marrow cell micronucleus rate were inhibited obviously. CONCLUSION: The experiments suggested that grape seed extraction has the antimutation effect.

BACKGROUND ＆ AIM: To evaluate the value and potential clinical implications of microsatellite instability(MSI) in diagnosis of bladder cancer recurrence. MATERIAL AND METHODS: The analysis by PCR for detecting MSI was done in 20 cases of nomal samples and 60 cases of urine samples from 42 patients with bladder carcinoma in postoperative follow-up. The analytic parameters included hematuresis ,tumor size, number of tumor, tumor grade and 5 microsatellite bio-markers. RESULTS: There were 17 cases of MSI＋ in 20 recurrent cases, of which 15 cases got hematuresis samples. The sensitivity, specificity and positive predictive value was 85.00％, 82.50％ and 83.33％, respectively, with the analysis by the results of MSI in diagnosis of bladder cancer recurrence. No correlation was found between MSI and tumor grading. CONCLUSION: The results indicate that MSI is a good bio-marker in diagnosis of bladder cancer recurrence for patients with bladder carcinoma in postoperative follow-up, especially for which with hematuresis.

BACKROUND ＆ ATM： To investigate the relationship between glioma malignancy grading and P21WAF1/CIP1, P16 gene expressing levels. MATERIAL AND METHODS： The expression levels of P21WAF1/CIP1 and P16 proteins were detected in 98 cases of brain glioma using SABC immunohistochemical staining. Eight normal brain tissues were used as control. RESULTS： The positive expression rates of P21WAF1/CIP1 and P16 protein in glioma were 58.16％ and 42.86％respectively, anda significant difference was observed between gliomas and control.The expression intensity of P21WAF1/CIP1 and P16 decreased with the gliomas grade increasing. Coexpression of P21WAF1/CIP1 and P16 was found in glioma and normal brain tissues. CONCLUSION： The positive expression rate and coexpression rate of P21WAF1/CIP1 and P16 protein may reflex the malignant grade of glioma in some extent,and they can be as a sensitive index for the gliomas grading.