OBJECTIVE: To determine the expression pattern of EZH2 protein in the process of human fetal development and provide new and multi-organ information for understanding the possible roles of EZH2 during the whole fetal development. METHODS：33 human fetuses at 25 different gestational weeks and 1 neonatal sample were obtained through abortion or stillbirth. The organ or tissue including esophagus，stomach，small intestine，colon，liver， pancreas，parotid gland，thyroid，adrenal gland，lung，trachea，heart，large artery，kidney，bladder，uterus， ovary，testis，spleen，thymus，cerebrum were fixed，embedded，sectioned，then stained with hematoxylin and eosin to identify their morphological structures. Tissue microarrays were constructed and followed by immunohistochemical staining of EZH2. RESULTS：EZH2 was expressed in fetal organs investigated and exhibited different expression levels in different tissues and at various gestational ages. The positive staining of EZH2 protein was mainly located in the nucleus，as well as in the cytoplasm in some cells. CONCLUSION：EZH2 protein  was  generally expressed  in human fetal organs and tissues and its expression exhibited tissue specificity and dependence on the stages of fetal development.

OBJECTIVE: To explore the effects of curcumin on apoptosis-related gene expression in HaCaT cells. METHODS：Immunocytochemistry and Western blot were used to detect the effects of curcumin on apoptosis-related protein expression. RESULTS：Immunocytochemistry staining results showed that the expression levels of  P53，Bax，Fas were upregulated significatntly while the products of Bcl-2 proteins were downregulated significantly in cells treated with 7.5 μg/mL curcumin. Western blot analysis confirmed the same results. CONCLUSION：Curcumin induced apoptosis of HaCaT cells by inhibiting expression of  Bcl- 2 gene and promoting the apoptosis genes p53，Bax and Fas activation.  Curcumin could induce apoptosis by regulation of a variety of apoptosis-related genes to activate the apoptosis signaling pathway.

OBJECTIVE: To investigate whether Oct4 could activate miR-302 promoter in the F9 mouse embryonic carcinoma cells. METHODS：PGL3-P-miR-302 was created by cloning the PCR- amplified region of the miR-302 putative promoter.  PGL3-P-miR-302 promoter reporter plasmids (0.5  μg) were co-transfected with 0.5 μg or 1.0 μg Oct4 expression vector into 293T cells. PGL3-P-miR-302 promoter reporter plasmids (0.5 μg) were co-transfected with Oct4 siRNA(2.0 μg siRNA1 and 2.0 μg siRNA2) into F9 cells. Luciferase activity was assayed. RESULTS：PGL3-P-miR-302 was created，and the cell-specific activity of this promoter was identified by its  transfection into F9 cells. With 0.5 μg  and 1.0 μg Oct4 overexpression in 293T cells，miR-302 promoter activity increased 2.6-fold and 4.3-fold，respectively. After siRNA-mediated knockdown of Oct4，miR-302 luciferase activity fell to 68% compared with control siRNA. CONCLUSION：This study showed that the miR-302 cluster acted downstream of the Oct4 regulation network in F9 cells.

OBJECTIVE: To explore the effect of herbicide 2，4-D butyl ester on the expressions of peroxidase (POD) and esterase (EST) in Carassius auratus serum. METHODS：0.5，1.0，1.5，2.0 mL/L concentrations of 2，4-D butyl ester (according to its 24 h LC50) and 0.9% saline control group were prepared.  Carassius auratus received intraperitoneal injection of 0.1 mL/g dose of each concentration. Tail blood of Carassius auratus and the supernatant of centrifugation were collected after 24 h. SDS-polyacrylamide gel electrophoresis was used to test the toxicity on POD and EST.  The experimental data were analysed with ANOVA.  RESULTS：POD and EST enzyme activities were inhibited at low concentrations(0.5 mL/L)，induced at medium concentrations(1.0 mL/L) and decreased at high concentrations (1.5，2.0 mL/L). Compared with the control group. the effect of 2，4-d butyl ester on migration rate of EST isoenzyme was greater than POD.  CONCLUSION：2，4-D butyl ester affected the activities of POD and EST in Carassius auratus and  2，4-d butyl ester changed the EST isoenzyme mobility.

OBJECTIVE: To study the effects of the heated A375 melanoma cells on natural killer (NK) cells and dendritic cells (DC) generated from human cultured peripheral blood mononuclear cells (PBMCs) in vitro，and to explore the mechanism. METHODS：NK cells and DCs cultured from PBNCs in vitro and  reacted with A375 melanoma cells in  a  water-bath at 43℃. Using CCK-8 kit to test the killing rate of NK cells and DCs against A375 melanoma cells at the effector∶ ratio of efficacy to target(E/T) (NK∶DC∶A375) of 1∶2∶1，3∶6∶1，and 6∶12∶1.  ELISA was used to measure concentrations of γ-INF released by NK cells at the effector∶E/T (NK∶DC∶A375) of 1∶2∶1，3∶6∶1 and 6∶12∶1. RESULTS：At each E/T ，the killing rate of the heated group against A375 melanoma cells were stronger than the unheated group (P<0.05)，and the killing rate increased as the concentrations of NK and DC increased (P<0.05). The concentration of γ-INF released by the heated group was also more than the unheated group(P<0.05). CONCLUSION：Heated A375 melanoma cells could induce NK to release more γ-INF and significantly increased the activity of NK cells ，in which DC played an important role.

OBJECTIVE: This study aimed at evaluating the survival of patients older than 70 with lung cancer and explore the independent prognostic factors in this group of patients. METHODS：The modified cumulative illness rating scale-geriatric(MCIRS-G) was scored for a cohort of elderly patients with lung cancer. Total score (TSC)，severity index(SV) and comorbidity index(CM) were obtained. Clinical features were also used. All patients underwent a follow-up for mortality. Univariate analysis and multivariate analysis were used to identify factors associated with prognosis in the enrolled patients. RESULTS：The overall median survival was 30.52 months and the incidence of complications was 82.26%. TSC，SV and CM were positively correlated with age(r were 0.656，0.739 and 0.677，respectively，P<0.05). By univariate analysis，age，pathological type，clinical stages，American Eastern Cooperative Oncology Group(ECOG) performance status (PS)，differentiation degree，surgery，TSC，SV and CM were significantly related to prognosis and survival in geriatric lung cancer patients(P<0.05). By multivariable analysis，clinical stages，ECOG PS，differentiation degree，surgery，TSC，SV and CM were independent prognostic factors. CONCLUSION：Clinical stages，differentiation degree，comorbidity，ECOG PS and surgery may be independent prognostic factors in the elderly with lung cancer. The MCIRS-G could effectively assess the influence of comorbidities on the prognosis of lung cancer in this patient group.

OBJECTIVE: To investigate the promoter methylation status of N-myc downstream-regulated gene 2 (NDRG2) and its correlation with mRNA and protein expressions of the gene in gastric cardia adenocarcinoma (GCA). METHODS：MSP method was used to measured the promoter methylation status of NDRG2 in 97 GCA tumor tissues and corresponding normal tissues. RT-PCR and immunohistochemistry were used to examine mRNA and protein expression，respectively，of NDRG2 in 97 GCA tumor tissues and corresponding normal tissues. RESULTS：The promoter methylation frequency of NDRG2 in GCA tumor tissues (49.5%) was significantly increased in comparison to that in corresponding normal tissues (5.1%) (P<0.05). The methylation frequency of NDRG2 in stages Ⅲ and Ⅳ tumor tissues was significantly higher than that in stages Ⅰ and Ⅱ tumor tissues. The methylation frequency of NDRG2 in poorly differentiated group was significantly higher than that in well and moderately differentiated group (P<0.05). NDRG2 mRNA expression in tumor tissues was significantly lower than that in corresponding normal tissues (P<0.05). Positive protein expression of NDRG2 in tumor tissues (44.3%，43/97) was significantly lower than that in corresponding normal tissues (94.8%，92/97) (P<0.01). The protein expression of NDRG2 was inversely correlated with its promoter methylation status in GCA tissues. CONCLUSION：Promoter hypermethylation of NDRG2 may be one of the mechanisms that lead to gene inactivation in gastric cardia adenocarcinoma.

OBJECTIVE: To investigate the effects of imperatorin on Rab27a and tyrosinase in cultured human epidermal melanocytes. METHODS：Human epidermal melanocytes were separated，purified and cultured in vitro. Melanocytes were randomly divided into control group and imperatorin group. After 48 h incubation with culture medium M254 or imperatorin(50 or 25 μmol/L)，the morphology of melanocytes was examined by inverted phase contrast microscopy. The effects of imperatorin on Rab27a and tyrosinase in melanocytes were investigated by immunofluoresent assay. The ultrastructures of melanocytes and melanosomes were studied by transmission electron microscopy. RESULTS：The melanocytes cultured with imperatorin for 48 h had shorter dendrites. Immunofluorescence microscopy showed that the location of Rab27a and tyrosinase in melanocytes were not obviously changed，but the fluorescence intensity of Rab27a was enhanced (P<0.05) and tyrosinase attenuated (P<0.05). Fewer melanosomes found in the cultured human epidermal melanocytes by transmission electron microscopy. CONCLUSION：Imperatorin promoted the expression of Rab27a and inhibited tyrosinase activity.

OBJECTIVE: To study the function of Yu-mu-li-sheng-ru-ling capsule in milk secretion in rats. METHODS：SD rats with overload lactation model were used. The pregnant rats were randomly divided into four groups，including test groups with different dosage (1 800，900，450 mg/kg) and negative control group. The mother rats  were gavaged for 25 d continuously during lactation. Body weight of mother and baby rats，survival ratio of baby rats and breast tissue pathology examination of mother rats were evaluated. RESULTS：No significant differences were observed in body weight of  treated  mother rats. For 450 and 900 mg/kg groups，baby rat survival ratios were 89%-94%，average body weight was insignificantly increased compared with that in negative control group(P>0.05). For 1 800 mg/kg group，baby rat survival ratio was 100%，average body weight increased significantly during early and late stage of lactation (P<0.05). Increased milk in breast tissue，filled acinus cavities and expanded mammary gland tubes were obviously observed. CONCLUSION： At 10 times  higher  than clinical dosage (1 800 mg/kg)，Sheng-ru-ling capsule could facilitate milk secretion in lactating rats，increase baby rat body weight and reduce malnutrition death.

OBJECTIVE: To explore the relationship between the protein expressions of MMP-2 and E-cadherin and the development and metastasis of esophageal carcinoma. METHODS：The expressions of E-cadherin protein and MMP-2 protein were examined by immunohistochemistry (IHC) in 45 cases of esophageal carcinoma，21 cases of atypical hyperplasia and 24 cases of normal esophageal mucosa tissues. RESULTS：The positve rates of MMP-2 protein were 40.00% (18/45) in esophageal carcinoma，38.1% (8/21) in esophageal atypical hyperplasia and 4.17% (1/24) in normal tissues. There were significant differences among three groups (P<0.05). Additionally，the expression of MMP-2 protein had a correlation with lymph node metastasis (P<0.05). The positve rate of MMP-2 protein in esophageal cancer tissues with lymph node metastasis was 55.56% (15/27) ， significantly higher than that in esophageal cancer tissues without lymph node metastasis (16.67%，3/18). The positve rate of MMP-2 protein in esophageal cancer tissues with fibrous membrane infiltration was higher than that in esophageal cancer tissues without fibrous membrane infiltration(48.57%，17/35；10.00%，1/10). The positive rate of E-cadherin protein in normal tissues (100%) was much higher than that in esophageal carcinoma (28.89%) and esophageal atypical hyperplasia (71.43%). According to these results，the expression of E-cadherin protein had a negative correlation with the expression of MMP-2 protein (χ2=4.615，r=-0.793，P<0.05). CONCLUSION：There were significant relationships between the high expression of MMP-2 protein and low expression of E-cadherin protein with the development and metastasis of esophageal carcinoma. Furthermore，the expression of E-cadherin protein had a negative correlation with the expression of MMP-2 protein in the invasion and metastasis of esophageal carcinoma.

OBJECTIVE: To study the association between XRCC1 rs25487 and CCNH rs2234942 polymorphism with the susceptibility to breast cancer and benign breast tumor in Han women in Gansu area. METHODS：Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was used for the polymorphism of XRCC1，CCNH in 101 cases of breast cancer，101 benign breast tumors and 101 disease-free controls collected in Gansu. Logistic regression analysis was used for comparing the genotypes or clinical pathological characteristics with the risk of breast cancer. Chi-square test was used to compare menarche age and disease onset age with the risk of breast cancer or benign tumor. RESULTS：Logistic regression analysis  showed that XRCC1 rs25487，GG genotype increased the risk of breast cancer (P=0.001，OR=6.39，95%CI：2.18－18.65). Regarding the clinicopathological immunohistochemical characteristics，the distribution of AA/AG genotype was significantly different between PR+ and PR- at XRCC1 rs25487(P=0.04，OR=0.29)，while the distribution of AG/GG genotype was significantly different in Her-2+ cases and Her-2- (P=0.008，OR=0.45). Chi-square test found that the menarche age of GG/AG genotype in XRCC1 rs25487 and GG genotype in CCNH rs2234942 was significantly different in breast cancer and benign tumor (P= 0.001， 0.043，0.049). The disease onset age of GG genotype in both XRCC1 rs25487 and CCNH rs2234942 was significantly different in breast cancer and benign tumor(P=0.019，0.048). CONCLUSION： GG genotype in XRCC1 rs25487 increased the risk of breast cancer. AA/AG genotype in XRCC1 rs25487 with PR+ and AG/GG genotype in CCNH rs2234942 with Her-2+ reduced the risk of breast cancer.

OBJECTIVE: To study Survivin shRNA interference on CDK4 mRNA and protein levels of esophageal cells ECA109. METHODS：Survivin shRNA interference technology was applied to liposomes transfection esophageal ECA109 cells，RT-PCR and Western blot methods was applied to  evaluate genes expression of Survivin and CDK4. Flow cytometry was used to detect the proliferation of cell lines index changes after ECA109 esophageal cell transfection. RESULTS：After Survivin shRNA transfection of esophageal ECA109 cell，Survivin mRNA and protein expressions were obviously down-regulated. CDK4 mRNA expression was also down-regulated. Cell ratio was increased in G2 phase but decrease in S phase，with the cell cycle blocked. CONCLUSION：Survivin was effective on adjusting CDK4 gene expression at mRNA level. The role of silencing Survivin down-regulated expression of CDK4 mRNA，suggesting  that Survivin could be a new gene regulatory factor in the tumor signal control network.

OBJECTIVE: This study aimed to analyze the expression，clinical significance of epithelial membrane protein-1(EMP1) in nasopharyngeal carcinoma. METHODS：Immunohistochemistry and western blot were used to analyze EMP1 protein expression in 75 cases of nasopharyngeal carcinoma and normal tissues to study the relationship between EMP1 expression and clinical factors. RESULTS：Immunohistochemistry：The level of EMP1 protein expression was found to be significantly lower in nasopharyngeal carcinoma tissue than normal tissues (P<0.05). Western blot：The relative amount of EMP1 protein in nasopharyngeal carcinoma tissue was found to be significantly lower than in normal tissues (P<0.05). The level of EMP1 protein expression  correlated with lymph node metastasis，clinical stage and histological grades (P<0.05). CONCLUSION：Decreased expression of EMP1 may play important roles in the pathogenesis and progression of nasopharyngeal carcinoma.