Abstract:Objective: To establish porcine naive-like induced pluripotent stem (iPS) cell line followed by labelling with the red fluorescent protein via nucleofection. The labelled porcine naive-like iPS cells could be used for tracing their development and differentiation. Methods: Firstly, bama miniature pig embryonic fibroblasts (PEF) were nucleofected with TetO-FUW-OSKM (OSKM: mouse derived transcriptional factors OCT4, SOX2, KLF4 and c-MYC) and FUW-M2rtTA vectors. Then, these cells were incubated in the medium with both leukemia inhibitory factor (LIF) and basic fibroblast growth factor (bFGF), supplemented with doxycycline hyclate. The established iPS cell lines were identified for their pluripotency. Secondly, the iPS cells were nucleofected with pEF1alpha-DsRed-Express Vector which could express the red fluorescent protein, then the maintainence of pluripotent characters of these labeled cells were analyzed. Results: The established iPS showed domed morphology and could survive long-term single-cell passaging (30 passages). They were alkaline-phosphatase positive, maintained a normal karyotype, expressed various pluripotency markers and required LIF/STAT3 signaling pathway for self renewal. The iPS cells could form embryonic bodies(EB), which expressed markers of three germ layers after culture in LIF free medium. The iPS cells were transfected with red fluorescent protein vector successfullyand kept showing red fluorescence for 10 passages. The results of alkaline phosphatase staining and immunofluorescence staining suggested that the pluripotency of these cells had not been compromised after red fluorescent protein labelling. Conclusion: The red fluorescent protein labeled porcine naive-like iPS cell lines were established successfully.

Abstract:Objective: To investigate the effects of MDM2 inhibitor SP-141 on proliferation, apoptosis and migration in MGC803 and BGC823 human gastric cancer cells. Methods: The cell viability, proliferation, cell cycle distribution, apoptosis and migration were detected by CCK-8 method, colony formation assay, flow cytometry analysis, Hoechst staining assay and wound-healing assay, respectively. Western Blot was performed to determine the levels of MDM2 and relative biomarkers in SP-141 treated MGC803 and BGC823 cells. Results: The mRNA expressions of MDM2 was significantly increased in gastric cancer tissues compared with adjacent non-cancerous tissues (P< 0.01) in TCGA gastric cancer database. MDM2 expressions did not show obvious differences among five different human gastric cancer cell lines. SP-141 inhibited the cell viability and colony numbers in MGC803 and BGC823, and induced cell cycle arrest at G2/M phase. SP-141 induced cellular apoptosis with down-regulating Bcl-2 as well as up-regulating Bax, cleaved-Caspose-3 and cleaved-PARP1 expressions. SP-141 also suppressed the wound closure in MGC803 cells, and accompanied by the decreased FAK protein expression. Conclusion: The MDM2 specific inhibitor SP-141 effectively suppresses cell proliferation, migration, and promotes apoptosis in human gastric cancer cells.

Abstract:Objective: To investigate the expression of PARP14 and its biological functions in pancreatic ductal adenocarcinoma (PDAC). Methods: The public gene chip data of PDAC obtained from the Gene Expression Omnibus (GEO) database and the Cancer Genome Atlas (TCGA) database were used to analyze the expression of PARP14 between pancreatic tumors and its adjacent tissues, The Kaplan-Meier method and Log rank test were used to assess survival analysis according to PARP14 expression in PDAC from TCGA. The biological role of PARP14 in regulating the proliferation of PDAC cells was evaluated by CCK-8 assay and colony formation experiment in BXPC-3 and CFPAC-1 cells transfected with PARP14 siRNAs. Results: The expression level of PARP14 in PDAC tissues were higher than that in the adjacent tissues or healthy controls according to the GEO databases (P<0.001). TCGA database further revealed that higher expression of PAPR14 was significantly associated with poorer survival of PDAC patients (P<0.05). Besides, PARP14 siRNAs were successfully transfected into PDAC cells BXPC-3 and CFPAC-1, and transfection with PARP14 siRNAs significantly suppressed proliferation of PDAC cells (P<0.05). Conclusion: The expression of PARP14 is up-regulated in PDAC tissues and high expression of PARP14 predicts poor survival, suggesting its role as a potential oncogene in PDAC.

Abstract:Objective: To explore the regulatory mechanism of RNPC1 on the expression of Snail family transcriptional repressor 1 (Snail) in BT474 breast cancer cells and the possible role of RNPC1 in breast cancer lymph node metastasis. Methods: Immunohistochemical (IHC) staining was used to detect and compare the expression of RNPC1 in 42 cases of breast cancer tissues with lymph node metastasis and 39 cases without lymphatic metastasis. The difference of RNPC1 gene level in breast cancer tissues with or without lymph node metastasis was calculated via utilizing TCGA database. The expression of RNPC1 in BT474 cells was knocked down or induced by lentiviral vector, and the effect was identified by qRT-PCR and Western blot. Dual-luciferase activity assay was applied to test and verify the target gene of RNPC1. Results: Compared with breast cancer tissues with lymph node metastasis, expression of RNPC1 was significantly higher in those without lymph node metastasis. The qRT-PCR and Western blot results showed that knockdown of RNPC1 in BT474 cells up-regulated Snail levels, whereas overexpression of RNPC1 resulted in down-regulation of Snail expressions. The dual-luciferase activity assay confirmed that 3′-untranslated region (3′-UTR) was required for RNPC1 to decrease the expression of Snail. Conclusion: Our findings suggested that RNPC1 might inhibit breast cancer metastasis via targeting Snail.

Abstract:Objective: To investigate the effect of nicorandil, an adenosine triphosphate-sensitive potassium channel opener, on pulmonary vascular remodeling in monocrotaline-induced mice model of pulmonary artery hypertension. Methods: CD1 male mice were randomly divided into control group, monocrotaline group, and monocrotaline+nicorandil group(n=6 for each group). The latter 2 groups were subcutaneously injected with 400 mg/kg monocrotaline weekly for 8 weeks to induce pulmonary artery hypertension model. The mice in the monocrotaline+nicorandil group were treated with 10.5 mg/kg nicorandil daily by intragastric administration. After 8 weeks, the right ventricular systolic pressure was evaluated by right ventricular puncture through the diaphragm; the index of right ventricular hypertrophy was calculated; the morphologic changes of right ventricle cardialmyocytes were detected by HE staining; the thickness of pulmonary arterial media wall was measured by elastin staining; α-smooth muscle actin (α-SMA) immunohistochemitry was used to assay pulmonary arterioles muscularization. Results: Daily treated with nicorandil (10.5 mg/kg) for 8 weeks significantly decreased RVSP (P<0.01), attenuated the index of right ventricular hypertrophy (P<0.05), alleviated the thickness of pulmonary arterial media wall (P<0.01), declined the ratio of fully-muscularized pulmonary arterioles (P<0.01), and prevented the injury of right ventricle cardialmyocytes. Conclusion: Nicorandil could relieve monocrotaline-induced pulmonary artery hypertension by inhibiting pulmonary vascular remodeling. It could be a potential drug for preventing and treating pulmonary artery hypertension.

Abstract:Objective: To investigate the beneficial effects of astragaloside IV (AS-Ⅳ) on oxidative stress injury induced by hydrogen peroxide (H2O2) in cultured human aortic endothelial cells (HAECs). Methods: HAECs were cultured in vitro and randomly divided into Control group, H2O2 treated group，and AS-Ⅳ treated group. The cells were treated with H2O2（400 μmol/L) for 36 hours as H2O2 group. For AS-Ⅳ treatment, the cells were first treated with H2O2 for 12 hours, then AS-Ⅳ(50 μg/mL) were added into culture medium for 24 h. After 36 h, all the cells were examined for oxidative stress injury and mitochondrial antioxidative function. Results: Compared with the Control cells, the addition of H2O2 significantly increased the MDA content, NADPH oxidase activity, and mitochondrial ROS content of the cells, whereas the SOD activity, Mn-SOD activity and mitochondrial transmembrane potential level were significantly decreased. The administration of AS-Ⅳ significantly decreased NADPH oxidase activity, MDA content and mitochondrial ROS content, and increased SOD activity, Mn-SOD activity and mitochondrial transmembrane potential level, compared with those of H2O2 treated injured cells. Conclusion: In the present study, AS-Ⅳ attenuates H2O2 induced oxidative stress in HAECs. The enhancement of mitochondrial antioxidant function by AS-Ⅳ may be one of the possible pathways of AS-Ⅳ to protect HAECs from oxidative stress.

Abstract:Objective: To develop and validate a sensitive high performance liquid chromatography-tandem mass spectrometry method for the simultaneous determination of clopidogrel and its metabolites and subsequently to use it to investigate pharmacokinetic properties of clopidogrel in mice. Methods: Clopidogrel and its metabolites were determined at an electron spray ionization (ESI) source using multiple reaction monitoring (MRM) in positive ion mode after protein precipitation by acetonitrile，in which piroxicam was added as an internal standard. Chromatographic separation was achieved on a Poroshell 120 SB-C18 column (100×2.1 mm, 2.7 μm, Agilent Technologies) using a mobile phase that consisted of both solvent A (0.1% formic acid in water) and solvent B (0.1% formic acid in acetonitrile) in a gradient elution manner. The pharmacokinetics of clopidogrel was investigated in mice receiving a single dose of clopidogrel (10 mg/kg) by gavage administration. Results: The simultaneous assay of clopidogrel and its metabolites achieved good linearity (r> 0.99) within the range of the spiked plasma concentrations. The recoveries were greater than 80%, and no significant matrix effect was found. The both intra- and inter-day RSDs were less than 15%, and the accuracy ranged from -2.40% to 5.00%. The stability of the method was good. In addition, a series of pharmacokinetics parameters were obtained in mice after receiving a single oral dose of clopidogrel through this analytical method. Conclusion: The modified methodology was highly sensitive, accurate, and reproducible for the simultaneous determination of clopidogrel and its three major metabolites in mouse plasma and therefore, it was successfully applied in pharmacokinetic studies of clopidogrel in mice.

Abstract:Objective:To investigate effects and underlying me chanisms of miR-199a on prolifer ation of HepG2. Methods:The miR-199a gene was cloned to lentiviral expression vectorpLV-GFP-puro with EGFP by recombining DNA technology,and positive clones were i dentified by DNA sequencing. The 293T cells were cotransfected with lentiviral packaged systems and miR-199a gene plasmid by lipofectinregeant to package the lentiviral partic les.The HepG2 cells were infected with purified recombinant lentivirus. The transfec tion efficiency was assessed under fluores cent micros cope;MTT assay was used to detect cell proliferation;Western Blot was used to analyzae expressions of miR-199A and NF-kB related anti-apoptotic genes(TRAF2,cIAP2,Bfl-1 and cFLIP)in transfected HepG2 cells.Results:The recombinant lentiviral vectors of miR-199a gene were successfully constructed. The virus reached a titer of 1.0.8×108TU/mL after being packaging,purification and concentration. Interestingly,the transfected cells held high expression of miR-199a,which inhibited the pression of miR-199a,cells.ln addition,the transfencted HepG2 cells stablely expressed IkBa,effectively inhibited NF-kB activity,and further suppressed expressions of NF-kB related anti-apoptotic genes(TRAF2.c IAP2,Bfl-1 and cFLIP).Conclusion:The miR-199a recombinant lentiviral vector had been successfully constructed and effectively transfected HepG2 cells, which inhited the proliferation of HepG2 cells . The underlying me chanisms may beinhibit NF-kB activity,and further inhibitexpressions of NF-kB related anti-apoptotic genes by miR-199a.

Abstract:Objective：To investigate the proliferation and migration effect of cortisol on hormone-related breast cancer MCF-7 cells. Methods：MCF-7 cells, originated from a hormone-related breast cancer woman, were treated with different concentration of cortisol. The effect of cortisol on cell proliferation was determined by MTS assay and the rate of cell survival was calculated. The effect of cortisol on cell migration was examined by wound-healing assay.Immunoblot analysis detected the protein expression of phospho mTOR (p-mTOR)， which is involved in cell proliferation，and MMP-9, MMP-2，which are related to cell migration. Results: Cortisol at concentration of 10 nmol/L significantly increased cell proliferation（P<0.05）and cell migration（P<0.01）compared with the vehicle group. Furthermore, cortisol at concentration of 10 nmol/L obviously increased the protein expression involved in cell proliferation (pmTOR，P<0.05) and cell migration (MMP-9, P<0.05； MMP-2，P<0.01). Conclusion: Cortisol at concentration of 10 nmol/L effectively induced cell proliferation and cell migration by activating mTOR pathway and upregulating MMP-9 and MMP-2 protein expression.

Abstract:Objective: To screen the differently expressed miRNAs study the expression andfunction of let-7f-5p in unexplained recurrent spontaneous abortion (URSA) and investigate the roles of miRNAs played in URSA. Methods: The miRNA-sequencing was applied to screen miRNAs that differently expressed in URSA placental villous; The qRT-PCR was used to validate the miRNA’s expression of screened miRNAs; The DAVID was applied to analyze pathway; The qRT-PCR was conducted to measure the expression of target genes in p53 pathway. Results: The expression of let-7f-5p was markedly up-regulatedhighly expressed in URSA placental villous; The targets of let-7f-5p were abundant in p53 pathway, and the expression of MDM-X and MDM2 was significantly down-regulated, while the expression of p53 was significantly up-regulated. Conclusion: let-7f-5p might promote cell apoptosis in URSA by regulating the expression of MDM-X/MDM2/p53 in p53 pathway.

Abstract:Objective :To investigate the effects of leptin on aortic valvular interstitial cells (VICs) transdifferentiation and reveal the role of leptin in the development of calcific aortic valve disease (CAVD). Method Porcine aortic VICs were separated and cultured via collagenase digestion. The VICs were incubated with leptin in different concentrations for 24 hours and the best stimulation concentration was determined by detecting ALP activity in the cell lysate. After incubated into optimal stimulation concentration for 24 hours, mRNA and protein expressions of α-SMA and OPN were detected by RT-PCR and Western blot， respectively. Results: Treatment with leptin enhanced the ALP expression of VICs and the optimal concentration was 100 ng/ml. Leptin treatment significantly promoted the mRNA and protein expression of OPN while α-SMA was not changed. Conclusion: Leptin can promote VICs calcification through inducing VICs transdifferent into osteoblast-like cells.

Abstract:Objective：To investigate the effect of a small molecule deubiquitinating enzyme inhibitor，PR-619 on Herpes simplex virus(HSV) replication. Methods：HSV replication and viral protein expression were detected via In-cell Western and Western blot. Viral gene expression on mRNA level was evaluated using Real-time PCR. We studied the inhibitory effect of PR-619 on viral replication，including viral mRNAs transcription and proteins expression. Results：PR-619 down-regulated HSV immediate-early and late gene expression(ICPO，ICP4 and gD) to inhibit viral replication effectively； PR-619 increased amount of ubiquitin-conjugates to suppress intracellular ubiquitin cycle，which may cause inhibitory effect on viral replication.and block viral induced NF-κB pathway activation. Conclusion：HSV effective replication is dependent on functional ubiquitin. Deubiquitinating enzymes is vital for intracellular ubiquitin cycle，which is associated to viral replication. This study could give deep insight into finding novel anti-HSV agents or genital microbicides.

Abstract:Objective: To investigate the regulatory role of IQGAP1 in cytoskeletal rearrangement of podocytes. Methods: Conditionally immortalized mouse podocytes were used to perform cell culture and stimulated by puromycin aminonucleoside (PAN). RT-PCR and Western blot analysis were used to measure the level of IQGAP1 mRNA and protein，respectively. The cytoskeleton of F-actin was evaluated by FITC-conjugated phalloidin staining under laser confocal scanning microscope and semi-quantitative system with cortical-F-actin score (CFS) was used to analyze the degree of actin cytoskeleton arrangement. We further explored the changes of cytoskeletal rearrangement after up- and down-regulation of IQGAP1. Results:①when compared with the control group, the expression of IQGAP1 mRNA and protein was significantly decreased after PAN stimulation in mouse podocytes(P<0.05). PAN stimulation induced cytoskeletal rearrangement including cortical F-actin ring formation, stress fiber attenuation and CFS up-regulation; ②the above rearrangement was exacerbated by IQGAP1 siRNA transfection (P<0.05); ③ PAN induced cortical F-actin ring formation and CFS up-regulation was abated after IQGAP1 plasmid transfection (P<0.05). Conclusion: IQGAP1 can inhibit PAN induced cytoskeleton rearrangement in podocytes and plays an important role in maintaining normal podocyte cytoskeleton.

Abstract:Objective：To verify changes of NF-κB in hypercoagulable state of rats and find new targets to treat thrombotic diseases. Methods：Eighty rats were splited into model group and control group. The model group was injected with adrenaline hydrochloride at the dosage determined in pre-experiment，while the control group was injected with normal saline. After 12，18，24，48 and 96 h，eight rats of each group were anaesthetized to collect blood sample. Preparing plasma samples were prepared to test PT，APTT， and the whole blood viscosity， and serum were collected to test the level of NF-κB. Results：In the pre-experiment，death rate of the model group of 0.7 mg/kg was lower，and compared with the control group，PT and APTT were shorter(P＜0.05) and the whole blood viscosity was higher(P＜0.05)；The model group showed distinct differences on PT and APTT compared with the control group at time of 12 h and 18 h(P＜0.05)，the whole blood viscosity of the model group is higher than that of the control group within 24 h，and the level of NF-κB of the model group is higher than that of the control group within 48 h. Conclusion：The best dosage of adrenaline is 0.7 mg/kg. Level of NF-κB in rats with hypercoagulable state is higher than that of the control group，which decreases with time. The hypercoagulable state in rats may be related to the increasing expression of NF-κB pathway.

Abstract:Objective：To investigate the correlation between mean platelet volume(MPV) and abnormal drainage induced by postoperative anticoagulant therapy in patients undergoing coronary artery bypass grafting with valve replacement，to provide evidences for formulating postoperative anticoagulant treatment regimens. Methods：Retrospective analysis was performed on 72 patients received coronary artery bypass grafting with concomitant valve replacement.The patients were divided into 2 groups by P75 of the total drainage volume and the anticoagulation-related bleeding. Results：Preoperative MPV presented a negative correlation with preoperative platelet count(r=-0.511，P＜0.001)，and a positive correlation with postoperative drainage volume(r=0.300，P=0.013).In Group 2，patient’s age，proportion of patients with history of alcohol intake，using biovalve and postoperative platelet transfusion volume were all higher than those in Group 1，the preoperative MPVe was higher than that in Group 1，and the postoperative platelet count was lower than that in Group 1(all P＜0.05).Age，history of alcohol intake，and preoperative MPV were shown to be independent risk factors for postoperative abnormal drainage.When plotting the receiver operating characteristic(ROC) curve for MPV，the area under the curve(AUC) was 0.691(95%CI 0.553~0.829，P=0.012)，the optimal cut-off point was 10.75 fl，the sensitivity was 0.714，and the specificity was 0.617. Conclusion：In patients undergoing coronary artery bypass grafting with valve replacement，preoperative MPV presented significant correlation with abnormal drainage induced by postoperative anticoagulant therapy，indicating its good predictive value.For those patients with the preoperative MPV＞10.75 fl，older age，history of alcohol intake，and received biovalve replacement， exercise in postoperative anticoagulation should be careful，and attention should be paid to monitoring of coagulation function.

Abstract:Objective：The aim of our study is to investigate the alterations in frequency of regulatory T cells (Treg cells) and Thelper 17 (Th17) cells in peripheral blood in patients with Crohn’s disease (CD) and the relationship between the proportion of Treg cells and clinical features of CD. Methods: Forty-six patients with CD (31 quiescent and 15 active CD patients) were enrolled in our study． Eight cases who had gastrointestinal polyps treated and will have gastrointestinal endoscopy to check as normal controls. The disease activity of CD was evaluated by Crohn’s disease activity index (CDAI). Concurrently, blood platelet (PLT), mean platelet volume (MPV), platelet distribution width (PDW), erythrocyte sedimentation rate (ESR) and serum C-reactive protein (CRP) level were measured. The percentage of Treg and Th17 cells in peripheral blood from CD patients and control group were detected by flow cytometry. We also analyzed the relationship between the proportion of Treg cells and these inflammatory biomarkers. Results: In peripheral blood, the propotion of Treg cells in active CD patients was significantly decreased compared with patients in remission and control group (P < 0.05). Compared with quiescent CD patients, active CD patients had higher percentage of Th17 cells in peripheral blood (P< 0.05). ESR and serum CRP levels increased and MPV obviously decreased in active CD patients compared to quiescent patients. Negative correlation was found between proportion of Treg cells and ESR, serum CRP levels as well as CDAI (P < 0.05). In addition, there was a positive correlation between the proportion of Treg cells and MPV (P < 0.05)． Conclusion: The change of proportion of Treg and Th17 cells in peripheral blood and the relationship between the proportion of peripheral Treg and serum inflammatory biomarkers reflecting CD activities (e.g., CRP, ESR and MPV) indicate that the differentiation imbalance of Th17/Treg may be involved in the pathogenesis of CD.

Abstract:Objective：This study compared oncologic outcomes between thoracoscopic lobectomy and segmentectomy,as well as detected the relevant independent risk factors in patients with p-stage IA(T1aN0M0) pulmonary adenocarcinoma. Methods：The clinical, pathological, and survival data of 159 cases in stage IA pulmonary adenocarcinoma who underwent pulmonary resection in the First Affiliated Hospital of NJMU from December 2012 to January 2014 were retrospectively analyzed．Kaplan-Meier method was used for analysis of survival，and the Cox Regression analysis was used to examine independent predictors for prognosis. Results: The 3-year overall survival rates and progression-free survival rates for the patients who underwent lobectomy were 88% and 85%，respectively, compared with 97% and 96% for the patients who underwent lobectomy(P＜0.05). Cox Regression analysis showed that tumor size and pathological subtype were the independent prognostic factors on recurrence(RR=18.219，95％CI：2.484～133.652，P=0．004；RR=2.107，95％CI：1.403～3.163，P<0.001) and death(RR=12.765，95％CI：1.332～122.37，P=0．027；RR=2.223，95％CI：1.376～3.499，P=0．001). Three-year overall survival rates and progression-free survival rates were 98% and 97%, 88% and 88%, 78% and 78%, and 75% and 58%(P<0.05) for lepidic, acinar, papillary, micropapillary and solid predominant pulmonary adenocarcinoma tumors, respectively. Conclusion：Tumor size and pathological subtype have significant influence on the prognosis of patients in stage IA pulmonary adenocarcinoma. For the patients in stage IA pulmonary adenocarcinoma, the prognosis of thoracoscopy segmentectomy is not second to that of lobectomy on the premise of strictly mastering the surgical indication.

Abstract:Objective: To evaluate the clinicopathological characteristics of gastric cancer patients with perioperative serum carbohydrate antigen 19-9 (CA19-9)-positive and alpha-fetoprotein (AFP)-positive, and combined effect of perioperative serum CA19-9 and AFP on clinicopathological characteristics of gastric cancer patients. Methods: The clinicopathological characteristics of 900 gastric cancer patients with serum CA19-9 and AFP detection perioperatively from the First Affiliated Hospital of NJMU were analyzed retrospectively. Results: Patients with elevated serum CA19-9 had the characteristics of more advanced T stage (T4 stage, 94.2% vs. 65.5%), later clinical stage (Ⅲ＋Ⅳstage ,89.2% vs. 61.5%), more common lymph node metastasis (81.5% vs. 59.1%) and more remarkable vascular as well as neural invasion compared with those with normal level of serum CA19-9 (32.5% vs. 23.3%; 42.5% vs. 27.1%); Compared with AFP-negative patients, the AFP-positive patients suffered later clinical stage (Ⅲ＋Ⅳstage ,88.9% vs. 60.6%), more common lymph node metastasis (81.5% vs. 59.1%) and more marked neural invasion (44.4% vs. 27.0%) (P＜0.05). Patients with one or two positive tumor markers suffered more advanced T stage, later clinical stage, more common lymph node metastasis and more remarkable vascular as well as neural invasion compared with those with double-negative tumor markers (P＜0.05). Two-category logistic regression analysis showed that CA19-9(+) was the independent risk factor for GC T stage; compared with CA19-9(-)-AFP(-), CA19-9(+)-AFP(+) was the independent risk factor for GC vascular invasion, CA19-9(+)-AFP(-) was the independent risk factor for GC T stage (P＜0.05). Conclusion: Perioperative serum levels of CA19-9 and AFP are closely correlated with clinicopathological features for patients with gastric cancer, and the two tumor markers are highly recommended detecting perioperatively for offering information of clinicopathological characteristics.

Abstract:Objective: To explore the expression pattern of ADAM12 gene in patients with colorectal cancer and analyze its correlation with the clinicopathological parameters and the tumor markers. Methods: ADAM12 expression in mRNA level was detected using quantitative real-time PCR in the colorectal cancer tissues and corresponding adjacent normal tissues from 50 patients. The correlations of ADAM12 expression profile with the clinicopathological parameters and prognosis were evaluated. The expression pattern of ADAM12 gene in protein level was detected using Western blot in the colorectal cancer tissues and corresponding adjacent normal tissues from 5 patients. Results: ADAM12 expression level was significantly higher in the colorectal cancer tissues than in the corresponding adjacent normal tissues,in both protein and mRNA level. Of the 50 patients, 38 (76%) showed up-regulated ADAM12 expression, which correlated significantly with lymphatic metastasis (P=0.005), TNM stage(P=0.002) and survival time(P<0.05) .During the 3-year follow up, 57.89%（22/38）patients in up-regulated died while in normal group,only 17.6%（2/12）died for colorectal cancer(P =0.013). Conclusion: Up-regulation of ADAM12 in colorectal cancer is correlated with lymphatic metastasis and progression and prognosis, suggesting the value of ADAM12 as a potential marker for colorectal cancer.

Abstract:Objective: This article aimed to study the value of tumor abnormal protein (TAP) and serum tumor markers (including CEA, CA125 and CA 15-3) in the evolution of efficacy of neoadjuvant chemotherapy (NCT) in locally advanced breast cancer (LABC). Methods: In this retrospective study, 68 LABC patients received NCT were enrolled. They were classified into pathologic complete response (pCR), partial response (PR), stable disease (SD) and progression of disease (PD) according to the therapy efficacy (some patients received targeted therapy at the same time). We set group pCR and PR as reaction group, SD and PD as no reaction group. All patients received tests for TAP, CEA, CA125 and CA15-3 before and after NCT. We compared results of these tests and their changes to finds out whether there exists statistical difference between reaction group and no reaction group. Results: In accordance with the comparison among test figures before and after NCT, there were statistical significance in TAP after NCT between two groups (P<0.05). And comparison among the numerical changes showed that there were statistical significance in TAP and CA15-3 before and after NCT. Conclusion: TAP and CA15-3 may play a role in the evolution of NCT efficacy in LABC patients.

Abstract:Objective: To explore the radiological anatomy of classification of internal iliac arteries (IIAs) as well as variant origins of obturator artery (OA), prostatic artery (PA) and uterine artery (UA) in adults based on CT angiography (CTA). Methods: A total of 163 patients undergoing pelvic CTA for vessel evaluation before kidney transplantation were retrospectively enrolled. Multiplanar reformation (MPR), maximum-intensity-projection (MIP), and volume-rendered (VR) based on axial images were performed on an off-line workstation. The origins of OA, PA and UA were studied. Results: Type A，B and C accounted for 70.2%, 28.3%, and 1.5% respectively. In addition, the most frequent OA origin was inferior gluteal and internal pudendal trunk (GPT), accounting for 27.6%. The most frequent PA and UA origins were IPA , accounting for 38.5% and 44.8% respectively. Conclusion: Pelvic CTA examination can clearly display the classification of IIA, and variant origins of OA, PA and UA.

Abstract:Objective: On the basis of previous systematic investigation on a Chinese pedigree with dominantly inherited auditory neuropathy, we aims to further search the deafness-causing genes (mutations) involved in this family. Methods: Whole-exome sequencing was conducted in three affected members and one spouse to determine the candidate genes related to the pedigree, and cosegregation analysis was performed on other members of the family. Finally, fifty normal-hearing individuals acting as controls were recruited for candidate variant detection. Results: A total of 41 variations were identified by whole-exome sequencing analysis. These variations were verified in the kernal pedigree members and 2 normal-hearing individuals by PCR-Sanger sequencing. The results showed that there was only ALOX15B gene co-segregated with the phenotype of the family. This gene mutation, however, was also detected in 2 of 50 normal-hearing individuals, which excluded the causative effect of ALOX15B mutation in this family. Conclusion:Whole-exome sequencing combined with Sanger sequencing does not reveal novel gene mutation in this family, which excludes the causative effects of Indels or mutations in coding sequence.