[ ABSTRACT] AIM:To investigate the effect of silencing cell division cycle 25a ( CDC25a) gene on the prolifera-tion of human hepatoma HepG2 cells.METHODS:CDC25a gene in human hepatoma HepG2 cells was silenced by RNA interference.Real-time PCR was applied to detect the expression of CDC25a, cyclin E and CDK2 at mRNA levels in the HepG2 cells.Western blotting was applied to detect the expression of CDC25a at protein level.In addition, MTT assay, Giemsa staining and flow cytometry were used to measure the proliferation of human hepatoma HepG2 cells.RESULTS:The expression of CDC25a at mRNA and protein levels in RNA silence group was lower than those in negative control group and normal control group (P<0.05).The mRNA expression of cyclin E and CDK2 in silence group was lower than that in negative control group and normal control group (P<0.05).The cell proliferation in silence group was lower than that in negative control group and normal control group ( P<0.05) .The results of flow cytometry revealed that the cells in silence group were blocked in G1 phase.CONCLUSION:Infection of LV-CDC25a-RNAi recombinant to the HepG2 cells effec-tively inhibits the CDC25a gene expression and the proliferation of human hepatoma cells, and arrests the cells in G1 phase, suggesting that CDC25a gene may be a key target for the treatment of liver cancer.

Objective To explore the basic ingredients of the tree shrew’ s( Tupaia belangeri) milk and compare with the dairy ingredients of other milks.Methods We select ten seed tree shrews after delivery ( 1 ~21 ) d with lactation mother tree shrews, and use artificial passive breastfeeding method let the young tree shrews suck breast milk,we took the milk from the young tree shrews in the stomach, directly using aseptic operation with a syringe immediately, once every two days, for consecutive three to five times, and a total of 18 mL milk was taken from each seed tree shrew.Then the milk was detected according to the national standard method for component testing.Results The total solid content of the tree shrew’ s milk was 43.63%, including 26.01%of fat, 10.41%of protein, 0.45% of lactose and 0.99%of ash content.Compared with cow's milk, the tree shrew’ s milk contained 3.36 times of total solid contents, 1.24 times of ash, 2.74 times of protein, 6.67 times of fat, and 0.09 times of lactose.Compare with baby formula milk, the tree shrew’ s milk contained 1.44 times of total solid contents, 0.20 times of ash, 0.58 times of protein, 1.53 times of fat, and 0.06 times of lactose.The trace mineral composition of the tree shrew’ s milk showed that the calcium, phosphorus, potassium, sodium, magnesium, and iron contents were 1.83 times, 2.73 times, 1.25 times, 1.93 times, 1.28 times, and 1.48 times higher than those in the cow's milk, and were 0.66 times, 0.85 times, 0.34 times, 0.26 times, 0.85 times, 0.24 times lower than those in baby formula milk.Conclusions The main nutrients of tree shrew’ s milk is of high fat, high protein and low sugar, and it can provide a basis for tree shrews artificial brood and breeding work.

Objective: To investigate the clinicopathological features and immunophenotype of inflammatory myofibroblastic tumor (IMT) and their relationship with IMT diagnosis and prognosis. Methods:A total of 11 IMT cases with follow-up were analyzed morpho-logically and immunohistochemically. Results:The patients included 6 men and 5 women aged 13-66 years. The tumors were found in various anatomical sites, including lung, mediastinum, liver, intra-abdominal, and bladder. Histologically, the majority of the cases com-prised spindled fibroblastic and myofibrobalstic cells accompanied by chronic inflammatory cells in a myxoid or hyalinized stroma;the rest were individual cases of abscess formation. Prognosis mala was indicated for cases with features including atypia tumor cells with two cases demonstrating epithelioid morphology and nucleoli. Immunohistochemical study showed that vimentin, ALK, SMA, S-100, CD117, and CD34 were expressed in 91%(10/11), 55%(6/11), 100%(11/11), 27%(3/11), 18%(2/11), and 9%(1/11) of IMT, respective-ly. Ki-67 was expressed from 3%-40%respectively. CK, H-caldesmon, and DOG1 were negative in all cases. Follow-up data were avail-able for 11 patients and ranged from 4 to 22 months. Data showed that 7 patients were alive with no evidence of disease;4 patients were alive with tumor, whereas 3 showed aggressive biological behavior. Conclusion:IMTs had intermediate behavior or malignant po-tential. Most IMTs with aggressive behavior showed a minority of tumor cells with atypia, epithelioid morphology, and nucleoli. High proliferation index expression, ALK, SMA, and H-caldesmon can aid in IMT diagnosis.

AIM:To investigate the effect of recombinant lentiviral vector for RNA interference (RNAi) on the expression of fatty acid-binding protein 5 (FABP5) gene in hepatocellular carcinoma HepG2 cells and tumor formation in nude mice.METHODS:RNAi lentiviral vector was used in the experiment.Human hepatocellular carcinoma HepG2 cells were divided into 3 groups:the HepG2 cells in experimental group were transfected with the recombinant lentivirirus vector LV-shRNA-FABP5, the cells in negative control group were transfected with a control lentiviral vector LV-shRNA-NC, and the cells in normal control group were without any treatment.The nude mice were randomly divided into 3 groups.The growth of the transplanted tumor cells in the nude mice was observed.The tumor growth curve, volume and weight were de-termined 4 weeks after the cell inoculation.The expression of FABP5 was detected by real-time PCR, Western blot and im-munohistochemical staining.RESULTS:Transfection of the lentiviral vector FABP5-shRNA obviously reduced FABP5 ex-pression in the HepG2 cells.Tumor formation was all positive in the 3 groups of the nude mice inoculated with the tumor cells.Compared with normal control group and negative control group, the tumor growth slowed significantly in experimental group with smaller volume and weight.FABP5 expression in the transplanted tumor tissues was significantly down-regulated at mRNA and protein levels in experimental group as compared with normal control group and negative control group. CONCLUSION:RNAi-induced down-regulation of FABP5 effectively inhibits the growth of transplanted hepatocellular carcinoma, suggesting that FABP5 gene may be an effective target for gene therapy in treating liver cancer.

AIM:To explore the changes and significance of Kupffer cells in the process of tree shrew chroni -cally infected with hepatitis B virus (HBV).METHODS:The animals were divided into 3 groups.Group A consists of 6 tree shrews that were identified as persistently infected with HBV;group B consists of 3 tree shrews that were suspected as persistently infected with HBV;group C consists of 4 tree shrews that were not inoculated with HBV and were applied as normal controls.Liver biopsies were collected regularly from all animals , and the Kupffer cells were isolated , purified and primarily cultured.The techniques of flow cytometry , immunohistochemistry, lysosomal fluorescent probe staining and real-time RT-PCR were applied to determine the number and function of these Kupffer cells .RESULTS: The result showed that the count and proportion of CD 163+cells in group A were significantly higher than those in group B and group C ( P<0.05).Meanwhile, the fluorescence intensity levels of lysosomal , the number of lysozyme-positive cells and the mRNA ex-pression level of TNF-αin the Kupffer cells in group A were significantly lower than those in group B and group C ( P<0.05).CONCLUSION:Kupffer cells may play a regulatory role during host’s chronic HBV infection.