Objective To analyze the hepatitis E virus (HEV) infection status and the molecular characteristics of HEV isolated from pregnant women in Zhejiang Province.Methods Totally 98 236 serum samples collected from pregnant women during the year 2013 to 2015 were tested for HEV IgM by enzyme-linked immunosorbent assay (ELISA) and samples positive for IgM were detected for nucleic acid of HEV by nested polymerase chain reaction (PCR).The whole gene of HEV open reading frame 2 (ORF2) was further amplified and the prevalence was analyzed in nucleic acid-positive samples.Results Three hundred and fifty-two out of 98 236 serum samples were tested positive for HEV IgM,with positive rate of 0.36％.All the samples were simple positive of HEV IgM except for two samples co-infected with hepatitis B virus.HEV specific nucleic acid fragments were detected positive from three serum samples.Further phylogenetic analysis revealed that all the three HEV isolates in this study belonged to HEV genotype 4.Three isolates did not cluster in one branch,although they shared high nucleic acid homology and more than 97 ％ of amino acid homology.Variations were found significantly between sample sequence and other published HEV 4 isolates,including two variable regions found in the ORF2 gene (1-460 nucleotide and 620-870 nucleotide).However,the synonymous and non-synonymous substitutions rates in the two regions were similar,and neutral selection was the main evolutionary pressure.Conclusions HEV infection rate in pregnant women of Zhejiang Province is similar with the published data.The HEV isolates obtained in this study belong to genotype 4 with high variation rate.

Objective To study the expressions of matrix metalloproteinase-1 3 (MMP-1 3)and p73 in gastric adenocarcinoma,and to explore the associations of the expressions of MMP-1 3 and p73 with the clinico-pathological features,and to evaluate their clinical significances for the prognosis of gastric adenocarcinoma metastasis.Methods The immunohistochemistry SP methods was used to evaluate the expressions of MMP-1 3 and p73 in 1 43 cases of gastric adenocarcinoma and 55 normal tissues adjacent to carcinoma,and their associa-tions to the clinicopathologic features were analyzed.Results The expression of MMP-1 3 in gastric adenocarci-noma was significantly higher than that in adjacent tissues of cancer (67.1 3% vs 1 6.35%),with a significant difference (χ2 =41 .1 0,P =0.000).The expression of p73 in gastric adenocarcinoma was significantly higher than that in adjacent tissues of cancer (58.74% vs 1 2.73%),with a significant difference (χ2 =33.86,P =0.000).In the gastric adenocarcinoma,the expression of MMP-1 3 was associated with peripheral lymph node metastasis (χ2 =1 1 .835,P =0.001 ),depth of invasion (χ2 =5.1 77,P =0.032)and TNM stage (χ2 =1 1 .1 07,P =0.001 ),but it was not correlated with the ages of patients (χ2 =0.1 1 3,P =0.853),tumor size (χ2 =0.338,P =0.591 )and tumor differentiation level (χ2 =3.628,P =0.072).In the gastric adenocarci-noma,the expression of p73 was associated with peripheral lymph node metastasis (χ2 =1 1 .440,P =0.001 ), tumor differentiation level (χ2 =5.407,P =0.025)and TNMstage (χ2 =9.497,P =0.003),but it was not correlated with the ages of patients (χ2 =1 .567,P =0.222),tumor size (χ2 =0.841 ,P =0.392)and depth of invasion (χ2 =0.554,P =0.498).The expression of MMP-1 3 was positively correlated with the expression
of p73 in gastric adenocarcinoma group (r =0.684,P =0.000).Conclusion Both MMP-1 3 and p73 may participate in the development of gastric adenocarcinoma,which can be used as an important index for the eval-uation of invasiveness and metastasis in gastric adenocarcinoma.

Aim To discuss the impact of important ac-tive ingredient of garlic diallyl sulfide———DATS on platelet activating factor (PAF ) mediated melanoma metastasis and its mechanisms.Methods ①MTT was used to test the effect of different concentrations of DATS on B16F10 and A375 melanoma cell growth number;②Scratch test and transwell were employed to test the effect of different concentrations of DATS on B16F10 and A375 melanoma cell migration;③ West-ern blot was used to test the effect of DATS on expres-sion of MMP-2,ERK,p38 induced by PFA;④Intrave-nous injection of tumor metastasis model was used to check the inhibition of DATS in PAF-mediated melano-ma metastasis.Results B16F10 cells relative growth rate fell to 73.21% and 48.78%,respectively,when DATS concentration reached 50 and 100 μmol·L-1 . DATS inhibited the levels of PAF-induced migration of melanoma cells B16F10 and vertical migration signifi-cantly,and inhibited B16F10 cells migration induced by PAF through inhibiting the expression of MMP-2, paxillin protein,FAK and other proteins.Conclusion DATS can significantly inhibit PAF-induced tumor metastasis, which is related to the inactivation of MAPKs.

AIM To observed the effect of procyanidins on DNA damage induced by carcinogen or hydroxyl radical (OH?),and inhibitory effect on topoisomerase. METHODS Using labeled TdR uptake assay and single cell gel electrophonesis. Topoisomerase was extracted from B16F10 cell lines, used agarose gel electrophoresis assay. RESULTS It was showed that hydroxyl radical (OH?) or different doses of MNNG can induce severe DNA damage of L 929 cell lines. Procyanidins can alleviate the damage induced by MNNG or OH?. It shows certain dose dependent effect relationship . But there is no significant inhibitory against Topo Ⅱ activity by procyanidins even at a dose of 50 ?mol?L -1 . CONCLUSION Procyanidins from grapes seeds can protect DNA damage,but not inhibit topoisomerase activity. This is one of cancer chemoprevention mechanism of Procyanidins.

Tumor metastasis is one of the important biological characteristics of malignant tumor,which is closely related with the prognosis of the cancer patients.High expression of ADAM8 in varieties of tumors was revealed in many recent studies,and such aberrant expression played a crucial role in regulating of tumor metastasis.Studies showed that overexpression of ADAM8 attenuated the intercellular adhesion effect,promoted tumor angiogenesis,and enhanced the degradation of ECM as well as the releasing of cytokines.Therefore,suppression of ADAM8 may lead to inhibition of tumor metastasis,which makes ADAM8 a particular attractive target as it can be used as a prognostic indicator and a potential therapeutic target of malignant tumor.A review about the relations between ADAM8 protein′s abnormal expression and tumor occurrence was discussed in this paper,also include discussion about the mechanisms of ADAM8 protein′s disorder-induced tumor formation,as well as therapeutic strategies based on ADAM8-targeted,which may provide references for follow-up research and clinical treatment.

Purpose To evaluate the value of real-time contrast-enhanced ultrasound (CEUS) combined with comparative observation in determining the initial time of peripheral lung lesion,and to provide a new method for the diagnosis of peripheral lung disease by CEUS.Materials and Methods Sixty patients with peripheral lung disease diagnosed by chest X-ray or CT examination were examined by CEUS,and the lesions and perfusion of surrounding lung tissue were observed in real time and comparatively.The time of initial enhancement of peripheral lung lesion was determined by real-time comparative observation method,that is,to compare the time of contrast agent to reach the peripheral lung lesion with the time to reach the adjacent gas-containing lung tissue.All lesion specimens were obtained by surgery or ultrasound-guided percutaneous biopsy,and the pathological diagnosis was performed.According to the pathological results,the peripheral lung lesions were classified into the malignant group,the chronic inflammation group and the pneumonia group.Results Sixty cases of peripheral lung lesions were identified as 38 cases of malignant tumors (including 15 cases ofsquamous cell carcinomas,9 cases of adenocarcinoma cell carcinomas,2 cases of large cell carcinomas,9 cases of small cell carcinomas and 3 cases of metastatic carcinomas),14 cases of chronic inflammation lesions (including 7 cases of tuberculomas,3 cases of inflammatory pseudotumors and 4 cases of lung abscess) and 8 cases of pneumonia.The detection rates of pulmonary arterial phase and bronchial artery phase using the real-time comparative observation method were 100％ (60/60) and 85％ (51/60),respectively.The initial time of pulmonary arterial phase and bronchial artery phase were (6.1 ± 0.9) s and (10.5 ± 1.6) s,respectively.The initial time of pulmonary arterial phase was sooner than bronchial artery phase,the difference was statistical significance (P＜0.05).Conclusion The real-time CEUS combined with comparative observation method has important diagnostic value in judging the initial enhancement time of peripheral lung lesion and can provide a new method for the accurate diagnosis of peripheral lung disease,which is worth popularizing.

The study on tumor metabolism has been gradually be-come a hot spot in recent years .A lot of proteins involved in the regulation of tumor metabolism especially the glucose transporter protein 1(GLUT1).As a key regulatory factor mediating energy metabolism within tumor cells , GLUT1 can regulate the glucose intake and maintain the basic level of metabolism in tumor cells . More importantly, the abnormal expression of GLUT1 was asso-ciated with many kinds of tumors , of which GLUT1 was used to meet the energy requirement for the fast growth of tumor .Thus GLUT1 also played a crucial role in growth , differentiation and metastasis of tumor cells and prognosis of tumors .Meanwhile , as three-dimensional crystal structure of GLUT 1 was determined , it is possible to design the small molecular inhibitors of GLUT 1, which can realize “starve to death” tumor cells.GLUT1 can be a particularly attractive target for tumor treatment and interfer-ence.The relationship between abnormal expression of GLUT 1 protein and tumor metabolism was reviewed . Moreover , the mechanism of tumor metabolism regulated by GLUT 1 protein ex-pression and treatment of cancers were discussed , which may provide references for future research and clinical treatment .

Tumor metastasis is one of the most important biologi-cal characteristics of malignant tumor, and it is also the main factor resulting in poor prognosis and leading to failure of treat-ment. In recent years, studies have shown that the extracellular matrix ( ECM) of tumor microenvironment plays a critical role in tumor metastasis. Tumor ECM fibrogenesis could form the cross-linked network structure, which not only provides nutrition and support to tumor, also it is necessary to tumor growth and inva-sion. These research results indicate that blocking ECM fibro-genesis may exert an inhibitory effect on tumor metastasis. Therefore, targeting ECM fibrogenesis has become a particularly attractive strategy as it can be used in the treatment of metasta-sis-related diseases. The ECM fibrogenesis in tumor is reviewed in this paper as well as the treatment strategies on tumor metas-tasis by targeting ECM fibrogenesis, which may provide refer-ences for follow-up research and clinical treatment.

Aim To screen the potential inhibitors of post-transcriptional activity of pro-inflammatory media-tor TNF-α from the lipophilic constituents in Chinese Medicine Salvia miltiorrhiza Bunge ( Danshen) , we es-tablished dual luciferase reporter gene system pGL3-TNF-α3′UTR ( 3′untranslated region ) co-transfected with Renilla control gene. Methods Complementary DNA ( cDNA) template was obtained from human um-bilical vein endothelial cells ( HUVECs ) . The full length DNA of TNF-α 3′-UTR was amplified through PCR, and then connected the luciferase reporter vector pGL3-control after enzyme digestion. pGL3-TNF-α 3′UTR constructs were co-transfected with pSVRenilla into the mononuclear macrophages RAW264. 7 cells. The relative activity of reporter genes was measured by dual luciferase reporter ( DLR ) assay system after the stimulus of lipopolysaccharide ( LPS ) in presence or absence of tanshinones compounds. Results The pGL3-TNF-α3′UTR luciferase reporter gene was suc-cessfully constructed. The cloning DNA fragment and sequence were both consistent with the GENBANK da-tabase. LPS significantly induced the relative reporter activityof RAW264 . 7 cells transfected with pGL3-TNF-α 3′UTR. Among four tanshinones compounds, we found only cryptotanshinone could significantly de-crease LPS-induced relative reporter activity. Conclu-sion The pGL3-TNF-α 3′UTR construct combined with DLR assay system was successfully established, which can be used to discover the agents such as cryp-totanshinone that regulate the post-transcription of TNF-α in treatment of inflammatory and malignant dis-eases.

Tumor metastasis is one of the most important biologi-cal characteristics of malignant tumor, and it is also the main factors that cause treatment failure and poor prognosis. Clinical studies have shown that the number of platelets in patients with malignant tumor increased more significantly than that in benign tumor patients and healthy people, which indicate that platelet might be involved in the development process of tumor. Further study found that in the process of cancer spreading to blood, platelet could interact with tumor cells to form tumor emboli, helped tumor cells escape from immune surveillance, thus pro-moted the tumor metastasis. In recent years, related mechanisms on platelets in promoting tumor metastasis were revealed gradual-ly, and several targeted therapies based on platelets were also carried out. This paper reviews the role of platelet in mediating tumor metastasis by hematogenous spread and its mechanisms and discusses the therapy strategies that target platelet, which may provide references for follow-up research and clinical treat-ment.