BACKGROUND ＆ AIM: For clinical diagnosis of malignant tumors and research the function of PAK4 protein, we prepared the McAbs of PAK4 protein. MATERIALS AND METHODS: The Balb/C mice were immunized with the recombinant plasmid of PAK4(the vector is P3XFLAG_CMV_10).By cell fusion and cell cloning, the McAbs of PAK4 were prepared. RESULTS: Two strains of hybridoma cells, producing anti_PAK4 antibody, named 2D1(PAK4) a subclass of IgG and 3H4(PAK4) a subclass of IgM, were obtained after fusion followed by three or four screening. The ascites of McAbs were prepared by injecting the hybridoma cells into the mice abdomen. Indirect ELISA results showed that the ascites of McAbs revealed high affinity with the PAK4 protein at the titer of 1∶1.0×104. Immunohistochemistry showed that the McAbs of PAK4 reacted specifically with the cells of breast cancer. CONCLUSION:These results demonstrated that the McAbs of PAK4 could be a specific marker for diagnosis of cancer.

BACKGROUND ＆ AIM: To study the relationship between CR1 genomic density polymorphism of erythrocyte with the quantitative expression of ECR1 and immune functions in patients with Non_Hodgkin Lymphoma. MATERIALS AND METHODS: ECR1 gene in erythrocytes was measured with PCR_RFLP method.Expression level of ECR1 was assessed by the quantitative assay. Soluble CD4, CD8 levels in serum were detected by ELISA. The RBC_C3bRR, RBC_ICR and TRR levels in erythrocyte were detected by the method established by Guo Feng. RESULTS: Expression of ECR1 gene in erythrocytes of Non_Hodgkin Lymphoma patients was markedly lower than that in normal persons(P<0.01), but HL type and LL type were markedly higher than that of normal people(P<0.01). The expression level of ECR1 of Non_Hodgkin Lymphoma patient was markedly lower than normal persons(P<0.01). There was positive correlation between CR1 genomic density polymorphism and expression level of ECR1 in erythrocytes and lymphocytic immune function. CONCLUSION：There was positive correlation between CR1 genomic density polymorphism with expression level of ECR1 in erythrocyte and immune function in the body. That could affect the prognosis of patients with Non_Hodgkin.

BACKGROUND ＆ AIM:To investigate the in vitro effects of DNA damage and chromosome aberration induced by sodium pentachlorophenol. MATERIALS AND METHODS: Different concentrations of sodium pentachlorophenol were added to the culture medium for cells of hamster ovary (CHO).DNA damage and chromosome aberration thus induced were evaluated by using single cell gel electrophoresis(SCGE, comet assay)and chromosome aberration test. RESULTS: The frequencies of DNA damage caused by sodium pentachlorophenol in the treatment groups with dosages of 80 and 160 μg/ml were significantly higher than the control group, and showed a dose_response relationship. Sodium pentachlorophenol also induced a significantly increased chromosome aberration rate both with and without adding S9 mix. CONCLUSION: Sodium pentachlorophenol could induce DNA damage and chromosome aberration of CHO cells.

BACKGROUND ＆ AIM： To study the different serum protein spectra of patients with laryngeal carcinoma and vocal cord polyps controls,and establish a serum biomarker pattern for screening laryngeal arcinoma. MATERIALS AND METHODS： Serum samples from 33 laryngeal carcinoma patients and 30 age_ and sex_ matched patients with vocal cord polyps were analyzed by SELDI_TOF on a ProteinChip reader, PBSII_C. Protein profiles were generated using WCX2 protein chips. Protein peak clustering and classification analyses were performed utilizing the Biomarker Wizard and Biomarker Pattern software packages, respectively. RESULTS： The results showed that 27 peaks showed significant differences between laryngeal cancer patients and vocal cord polyps patients, twelve of which were up_regulated in the cancer patient samples, and the others were down_regulated. Three protein peaks, 2 279 Da,4 626 Da and 5 663 Da, were automatically chosen for system training and development of a classification tree.We divided all patients by using this classification tree,the corresponding sensitivity was 93.94％ and the specificity was 100％. CONCLUSION： Our results suggested that serum could be a useful source for the detection of specific biomarkers for laryngeal carcinoma. Proteinchip Array System was a useful tool for a high throughput screening of large_sized serum samples to identify potential biomarkers for carcinoma.

BACKGROUND ＆ AIM： To explore the significance of mutation of the exon 9 domain of Kai_1/CD82 gene in the progression and metastasis of colorectal cancer (CRC). MATERIALS AND METHODS： The mutation of the domain of exon 9 was detected by using PCR and judged by sequencing in 40 CRC. The deletion of exon 9 was identified by using RT_PCR and validated by sequencing. RESULTS： The mutation in the Kai_1/CD82 gene was found in 18 of 40 colorectal cases (45％).The frequency of mutation was significantly higher in the colorectal tumors with lymphatic metastasis than those without(P<0.05), and in the advanced stage(Dukes C and D) than in the earlier stage(Dukes A and B) of the cancer (P<0.05). However, there was no significant difference of the mutation frequency when stratified by age and sex of the patients, histological type as well as the degree of differentiation of the CRC (P>0.05). CONCLUSION： The mutation of the Kai_1/CD82 gene was associated with the progression and metastasis of CRC. This can be used for assessing the progression and metatasis of colorectal cancer and for guiding clinical therapy.

BACKGROUND ＆ AIM: To investigate the protein expressions of PTEN, NF_κBp65 and CyclinD1 in gastric adenocarcinoma and the relationship between their expressions and clinicolpathological features. MATERIALS AND METHODS: Protein expressions of PTEN, NF_κBp6 and CyclinD1 in paraffin embedded tissues from 73 cases of gastric adenocarcinoma and 20 normal gastric mucosa tissues were analyzed by immunohistochemical method. RESULTS: ①The positive rate of PTEN protein expression was 42.5％(31/73)in gastric adenocarcinoma, lower than that in normal gastric mucosa tissues 90％(18/20)(P<0.01); The positive rates of NF_κBp65 and CyclinD1 protein expression were 58.9％(43/73)and 60.3％(44/73),respectively in gastric adenocarcinoma, higher than those in normal gastric mucosa tissues 20％(4/20)and 25％(5/20),respectively(both P<0.01); ②The expression of PTEN in well_differentiated adenocarcinoma was higher than that in poorly_differentiated adenocarcinoma (P<0.01). Also, the loss or decreased expression of PTEN significantly correlated with infiltrative depth, lymph node metastasis and clinicolpathological stage (P<0.01 or P<0.05). Contrary to PTEN, the expression level of NF_κBp65 in poorly_differentiated adenocarcinoma was higher than that in well_differentiated ones(P<0.05), and the increased expression of NF_κBp65 significantly correlated with infiltrative depth, lymph node metastasis and clinicolpathological stage(P<0.05 or P<0.01).The expression level of CyclinD1 only correlated with infiltrative depth of gastric adenocarcinoma(P<0.05); ③Significant relationships were found between PTEN and NF_κBp65 or CyclinD1(P<0.05, P<0.01), while a positive correlation could also be found between NF_κBp65 and CyclinD1(P<0.05). CONCLUSION: This expreiment suggested that PTEN, NF_κBp65 and CyclinD1 may play certain roles in the oncogenesis and progression of gastric adenocarcinoma, but to different extents. Combined evaluation of PTEN, NF_κBp65 and CyclinD1 may be helpful to assess the malignant degree,treatment and prognosis of gastric adenocarcinoma.

BACKGROUND ＆ AIM: To investigate the regulation of histone H3 acetylation and the expression of p21WAF1/CIP1 by NS_398 in HepG2 cells. MATERIALS AND METHODS: The effect of NS_398 on the proliferation of HepG2 cells was evaluated by MTT. The apoptotic cells were determined by flow cytometric(FCM)analysis.Total proteins and mRNA were extracted from HepG2 cells treated with or without NS_398 with IC50 200 μmol/L at various time points (4,8,12,24,48 h).By using Western blot, the levels of acetylated histone H3 and p21WAF1/CIP1 protein expression were assayed. RT_PCR was used to detect the expression level of p21WAF1/CIP1 mRNA. RESULTS: NS_398 inhibited cell proliferation and induced apoptosis in HepG2 in a concentration_dependent manner. DNA ploidy analysis showed that S phase cells were significantly decreased with increasing NS_398 concentration.Histone H3 acetylation was obviously increased by NS_398 in a time_dependent manner. The expression levels of p21WAF1/CIP1 mRNA and protein were up_regulated in a time_and dose_dependent manner. CONCLUSION: The level of acetylated histone H3 was up_regulated by NS_398, and the expression of p21WAF1/CIP1 was increased at the same time.

BACKGROUND ＆ AIM: To study the effects of GFW on tumor apoptosis and to provide basis for the development of GFW. MATERIALS AND METHODS： Each of the 30 mice received 0.2 ml S180 tumor cell suspension subcutaneously in the right axilla . The mice were randomly divided into model group,GFW group and CP group and with 10 mice in each group.Another 10 mice without cancer were used as control. The model group received intragastric normal saline,0.02 ml/(g·d); the GFW group was given intragastric administration,0.02 ml/(g·d) whilst the CP group received intraperitoneal injection of CP at 20 mg/(kg·d),The medicines were given to the mice in each group after they were inoculated, once a day for 10 days .The mice were sacrificed and the tumor removed. The S180 mice sarcoma model was employed to detect the inhibiting effects of GFW, the flow cytometer was used for determination of apoptosis and the ultrastructural changes assessed by electron microscope.The expression of Survivin mRNA was evaluated by situ hybridization. RESULTS: For S180 sarcoma ,the tumor inhibition by GFW was 38.93％. The apoptosis in GFW group was 17.79％ by flow cytometer. Some typical apoptotic cells and apoptotic bodies were identified through electron microscope. Chromatin gathered on the side of the cell, while the nucleus was condensed. Survivin mRNA was markedly suppressed in the GFW group when compared with the model group. CONCLUSION: GFW induced tumor cell apoptosis, and its mechanism might be closely related with down_regulating the expression of the survivin gene.

BACKGROUND ＆ AIM: To investigate the differentiation of mice B16 melanoma induced by proanthocyanidins from lotus seedpod (LSPC)in vivo. MATERIALS AND METHODS: B16 melanoma_bearing mice were used to study the inhibitory effect of LSPC on B16 melanoma, the effect of LSPC on tyrosinase activity, and the ability of melanin formation, and the morphology change of B16 melonoma and the expression of S_100 protein. RESULTS: LSPC(60－120 mg/kg)could dose_dependently inhibit the growth of B16 melanoma,The effects in the group with dosage of 120 mg/kg was the most marked. The inhibition rate of B16 melonoma was 55.3％, while the activity of tyrosinase, the content of melanin increased 28.5％ and 23.8％,respectively. There were significant differences compared with control group(P<0.05). The expression of S_100 protein was down_regulated. CONCLUSION: LSPC could significantly inhibit the growth of mice B16 melanoma cells, and induce the differentiation of B16 melanoma cell in morphology and function.

BACKGROUND ＆ AIM: To determine the acute oral toxicity and genetic toxicity of ethanol extracts from Momordica charantia(MC), so as to provide experimental basis for its exploitation. MATERIALS AND METHODS: The tests were performed according to GB159193_2003 Procedures and Methods for Toxicological Assessment of Food. RESULTS: The acute oral LD50 of ethanol extracts from Momordica charantia was greater than 21.5 g/kg. The genetic toxicity indicated by Ames test, micronucleus test of polychromatic erythrocytes in bone marrow and sperm shape abnormality test in mice was negative. CONCLUSION: In our experimental context, the acute toxicity of ethanol extracts from Momordica charantia was classified as non_toxic. Genetic toxicity was not found.

BACKGROUND ＆ AIM: To investigate the effects of quercetin on proliferation, cell cycle and apoptosis of colon carcinoma cell line SW480 and to study its probable molecular mechanisms. MATERIALS AND METHODS: SW480 cells were treated with different concentrations of quercetin(10, 20, 40, 80, 160 μmol/L)for 24 h, 48 h and 72 h. Then the cell proliferation, cell cycle, apoptosis rate and cyclin B1 protein expression of SW480 cells were analyzed using MTT assay, flow cytometry and immunoblot methods, respectively. RESULTS: Proliferation of SW480 cells was promoted by quercetein at a concentration of 20 μmol/L(P<0.05), while it was significantly inhibited by quercetin at concentrations of 40, 80, 160 μmol/L(P<0.01), in a dose_and time_dependent manner. After treatment with 20, 40, 60, 80 μmol/L quercetin for 48 h, the percentages of SW480 cells at G0/G1 phase were decreased, and those at G2/M phase were increased markedly (P<0.01). when SW480 cells were incubated with 80 μmol/L quercetin for 48 h, the apoptosis rate increased from baseline of 2.68％±1.04％ to 13.32％±4.62％(P<0.01). The expression of cyclin B1 protein could be down_regulated by quercetin at concentrations of 40, 60, 80 μmol/L. CONCLUSION: Quercetin at lower concentrations could promote proliferation of colon carcinoma cell line SW480, while it could inhibit proliferation and induce apoptosis of these cells at higher concentrations. Quercetin might exert its anti_tumor effect by blocking the cell cycle, inducing apoptosis and down_regulation of cyclin B1 protein.

BACKGROUND ＆ AIM: To study the in vitro anti_tumor effect of 6_epi_ ophiobolin K、ophiobolin K and ophiobolin G from microorganisms. MATERIALS AND METHODS: The effects of the three compounds on the proliferation of six human tumor cell lines were investigated by MTT method with IC50, and the inhibitory activities of the three compounds against cathepsin B were assayed for evaluation of anti_tumor invasiveness and metastasis in vitro. RESULTS: Compared with negative control, 6_epi_ophiobolin K,ophiobolin K and ophiobolin G showed significant inhibitory activities to Hct_15,Hct_116,MDA_MB_231,MCF_7,A549 and K562 cell lines. 6_epi_ophiobolin K to the above tumor cell lines demons trated IC50 of 5.70,3.60,28.00,8.40,2.00 and 3.10 μg/ml, respectively. Ophiobolin K showed IC50 of 1.40、0.81、4.26、3.70,0.91 and 1.10 μg/ml,respectively,while Ophiobolin G revealed IC50 of 4.10,4.60,26.10,17.10,6.70 and 6.10 μg/ml,respecfively.All three compounds have moderate inhibitory activities to athepsin B with the IC50 of 46,26 and 23 μg/ml,respectively. CONCLUSION: The three compounds all had anti_proliferative effects on tumor cell lines.

BACKGROUND ＆ AIM: To establish a model of mouse hepatoma H22 and to study the effects of Pingtiaoyin on the tumor. MATERIALS AND METHODS: The model of transplanted tumor was made by vaccination of hepatoma H22 in the right axilla.The model mice were divided into 4 groups:0.9％ NaCl,Pingtiaoyin,Pingtiaoyin＋5_Fu and 5_Fu.The diameter of transplanted tumor, inhibition rates,thymus and spleen indexes,the rates of life prolongation and the expression of VEGF in carcinoma and peri_carcinoma tissues were analyzed. RESULTS: The diameter of the tumor and the expression of VEGF in the carcinoma were reduced, thymus and spleen indexes,the inhibition rates and the life prolongation rates were increased after treatment with Pingtiaoyin. CONCLUSION: Pingtiaoyin could improve the thymus index,reduce the negative effect of 5_Fu on the immune function, and prolong the survival of the model mouse. Pingtiaoyin also could inhibit the growth of transplanted tumor in mouse through increasing the differentiation of carcinoma cells,reducing their proliferation and downregulating the expression of VEGF in carcinoma and pre_carcinoma tissues.

BACKGROUND ＆ AIM: R. chingii Hu is a newly identified herbal source in Hubei Province, although it has been used to prevent and help treat hyperglycemia and hyperlipidemia for hundreds of years by local people. Rubusoside is thought to be its special ingredient. In order to develop and utilize this new resource,toxicology tests of this new herb were conducted. MATERIALS AND METHODS: The safety of Hubei R. chingii Hu was evaluated systematically by a series of acute, mutagenic and sub_chronic toxicological tests. RESULTS: The acute toxicity test showed that the maximum tolerated oral dose of R. chingii Hu was more than 20.0 g/kg body weight in mice. No mutagenicity was found, as judged by mouse bone marrow cell micronucleus test，Ames test and mouse sperm abnormality test. In the subchronic study, no deaths or clinical signs, or abnormal hematological, biochemical and histopathological changes were found in rats after gavaging 2.5, 5.0 and 10.0 g/(kg·d) of Hubei R. chingii Hu for 90 days. CONCLUSION: The results indicated that Hubei R. chingii Hu may be safely used as a source of health food.

BACKGROUND ＆ AIM： To study isolatel and combined effects of the organic oxidant cumene hydroperoxide (CHP), the inorganic oxidant H2O2 and several antioxidants, on nitric oxide (NO) contents, total carbonyl and malondialdehyde (MDA) of hepatoma cells in different phases of cell cycle. MATERIALS AND METHODS：Hepatoma cells H299(1×106/ml) were synchronized,and then divided into eight groups:the control group(DMSO),CHP (50 μmol/ml), H2O2(50 μmol/ml), CHP＋H2O2 (oxidant componds: H2O2 of 50 μmol/ml plus CHP of 50 μmol/ml), Vit A(10 μmol/ml), Vit E (10 μmol/ml),Na2SeO3 (50 nmol/ml),AO(antioxidant compounds of Vit A＋Vit E＋Na2SeO3 , among them: Vit A and Vit E were 5 μmol/ml each, Na2SeO3 was 25 nmol/ml). They were separately added to cultured cells at G1, S, G2 and M phase of the cell cycle. The cell contents of NO, total carbonyl (T_carbonyl) and MDA were determined. RESULTS： The cell cycle showed better synchronization. The synchronized cells in G1 phase was 86％, in S phase was 78％,and in G2/M phase was 61％. We found that NO of cells stimulated by oxidant componds (CHP and H2O2) at G1 phase were significantly increased (P<0.05) while they were inhibited by Vit A (P<0.05). The oxidant componds (CHP and H2O2) had stimulatory function, and Vit A、Vit E、Na2SeO3 had obvious inhibitory effects on T_carbony contents throughout the whole cell cycle (P<0.05). The levels of MDA in synchronized cultured cells were elevated by H2O2 at G2/M phase, and were decreased by antioxidant compounds Vit A＋Vit E＋Na2SeO3 at G2 phase (P<0.05). CONCLUSION： After synchronization, the actions of organic oxidant, inorganic oxidant, and the tested antioxidants varied according to the different phases in the tumor cell cycles. Hepatoma cells in G1 phase were sensitive to oxidant compounds (CHP and H2O2) and Vit A on the oxidative injury NO level. The cellular oxidative injury marker MDA contents in hepatoma cells in G2 phase were significantly altered by H2O2 and antioxidant compounds (Vit A＋Vit E＋Na2SeO3).

BACKGROUND ＆ AIM: We explored the amino acid substitution in ErbB3 tyrosine kinase domain by bioformatics. MATERIALS AND METHODS: ErbB receptor family and tyrosine receptor family sequence alignments were performed to find out the important amino acid substitution sites in ErbB3 kinase domain, these sites was then located on EGFR crystal structure. RESULTS: A total of 54 substitution sites which might be meaningful were identified, in which 17 sites could be more important, R726E，L738T，E740H，D815N，K830Q，L836V，R838D might be the key substitution sites related to tyrosine kinase activity decrease. The tyrosine site near DFG motif in ErbB3 appeared to be ineffective. CONCLUSION: These substitution sites are related to the decrease of tyrosine kinase activity.

BACKGROUND ＆ AIM: Follow up cytogenetic tests was performed on the 6 victims exposed to 60Co radiation accident in Henan. MATERIALS AND METHODS: The samples of chromosome and CB micronucleus were prepared by using cultured whole blood, and analysis of chromosomal aberrations and CB micronuclei were made in the 6 victims exposed to 60Co radiation accident occurred in Xinxiang of Henan 1_2 years after irradiation. RESULTS： The frequencies of chromosomal aberrations in the victims had obviously declined in 1_2 years after irradiation. The proportion of unstable chromosomal aberration was higher in 1 year, and the rate of stable chromosomal aberrations was greater than that of unstable 2 year after irradiation. Frequencies of CB micronuclei in the victims had mostly declined into the background range in 1_2 years after irradiation. CONCLUSION： The results suggested that unstable chromosomal aberrations had gradually resolved, and stable chromosomal aberrations was still the majority as time progressed after irradiation. Late genetic effects of individuals exposed to high_dose irradiation can not be evaluated by analysis of CB micronucleus.

BACKGROUND ＆ AIM： To investigate umbilical cord blood chrome level of newborns in Guiyu town,an e_waste recycling area,and to discuss the effects of e_waste on newborn's health. MATERIALS AND METHODS： 152 full_term newborns belonged to two groups, one group from Guiyu (Guiyu group,n=100) and another group from other neighboring towns (control group,n=52). The chrome level of umbilical cord blood was determined with graphite atomizer absorption spectrophotometer. Questionnaires for mothers were used for analysis of related factors. RESULTS： Compared with the control group (20.30 μg/L), the geometric mean of chrome level 303.38 μg/L in the Guiyu group was significantly different (P<0.01). The Guiyu group of newborns with relatively high chrome levels in umbilical cord blood were from pregnant women who were working with e_waste recycling. CONCLUSION： There were close relationships between the Guiyu group of newborns with relatively high chrome levels in umbilical cord blood and local e_waste recycling activities as well chrome pollution locally. Environmental chrome pollution could threaten the children's health around the recycling site.

BACKGROUND ＆ AIM: To investigate the role of MT_ 3 in transitional cell carcinomas of bladder and the possible relation with MDR_1. MATERIALS AND METHODS: Using RT_PCR technique, the expressions of MT_3 and MDR_1 mRNA in 11 specimens from normal bladder mucosa and 23 specimens from bladder transitional cell cancer(BTCC) were measured. RESULTS: MT_3 and MDR_1_positive expression rates were found in 0 and 9.1％ in normal bladder mucosa ,respectively .But they were 78.3％ and 56.5％ in 23 BTCC specimens ，respectively. The positive rate was significantly higher in BTCC specimens than in normal tissue (P<0.05) .The expressions of MT_3 and MDR_1 were not related with the grade ,stage and recurrence after postoperative intravesical chemotherapy. CONCLUSION: MT_3 mRNA could not be identified in normal bladder mucosa, but highly expressed in bladder neoplasm，MT_3 might be used as a tumor marker.

BACKGROUND ＆ AIM: To find the median lethal dose of sodium formaldehyde sulfoxylate(SFS). MATERIALS AND METHODS: 50 NIH male mice were fed normal food and water before test and were randomly dividecl into five groups treated with SFS at different doses by gavage. The indices included toxic syndrome and death rate in each group. RESULTS: The lethal dose of SFS was 8 711 mg/kg. CONCLUSION: SFS actually belonged to the non_toxic grade by national criteria of toxins.