Aim To investigate the synergistic effects and possible molecular mechanism of hepatitis B virus (HBV) and alcohol on liver injury in HBV transgenic mice(HBV-Tg mice).Methods 20 HBV-Tg mice and 20 wild-type mice were randomly divided into 4 groups:alcohol-fed Tg mice and alcohol-fed Wt mice, and they were given intragastric administration with alcohol. Control Tg mice and control Wt mice received intragastric administration with saline.All groups were rasied for 10 weeks.The levels of ALT and AST in serum, the degree of inflammation, the degree of fibrosis, the mRNA expression of TGF-β_1, Smad3, Smad7, CTGF and the protein expression of TGF-β_1, CTGF, α-SMA in liver tissue were detected.Results The serumlevel of ALT and AST, the mRNA expression of TGF-β_1, Smad3, Smad7, CTGF and the protein expression of TGF-β_1, CTGF, α-SMA in liver all increased markedly in alcohol-fed Tg mice. Alcohol consumption induced hepatocyte steatosis and hepatic inflammation in alcohol-fed Tg mice, but the change of liver fibrosis was not remarkable.Conclusion HBV and alcohol have synergistic effects on early liver injury, possibly by enhancing the expression of TGF-β_1, Smad3, CTGF, α-SMA and inducing unbalanced expression of Smads.

Objective:To observe the role of lamividine and silymarin preventing and curing liver fibrosisrelevant factors induced by alcohol drinking in hepatitis B virus (HBV) transgenic mice (Tg mice).Methods:Forty HBV-Tg BALB/C mice with 1.3 copy were randomly divided into 4 groups:a control group,a model group,a lamivudine group and a silymarin group.Tg mice in control group were treated with normal saline via intragastric administration;Tg-mice in the model group were treated with 50％ alcohol (5 mL/kg) once a day via intragastric administration;while Tg-mice in lamivudine group and silymarin group were treated with alcohol (5 mL/kg) plus laminvudine (100 mg/kg) and silymarin (200 mg/kg) once a day via intragastric administration respectively.All groups were raised for 10 weeks.The levels of HBV-DNA copy number,ALT,AST in serum,the degree of inflammation,the degree of fibrosis,the mRNA expression levels of TGF-β 1,Smad3,Smad7 and connective tissue growth factor (CTGF),and the protein expression levels of TGF-β1,CTGF and α-SMA in liver tissue were detected.All the images were scanned with electronic computer and the data were analyzed with SPSS13.0 software.Results:Compared with the control group,liver injury were significantly aggravated,while HBVDNA copies,mRNA levels ofTGF-β1,Smad3,Smad7 and CTGF as well as the protein levels of TGF-β1,CTGF and α-SMA were significantly increased (P＜0.05).Compared with the model group,liver injury were significantly attenuated in silymarine group and lamivudine group,while mRNA levels of TGF-β 1,Smad3 and CTGF as well as the protein levels of TGF-β1,CTGF and α-SMA were significantly decreased;mRNA level of Smad7 was further increased (P＜0.05);the levels of ALT and AST in serum were decreased in the silymarine group (P＜0.05).Conclusion:Lamivudine and silymarin relieve the histological damage in the liver of alcohol-fed Tg mice.The mechanisms for the beneficial effects of lamivudine or silymarin might be related to inhibiting the expression of TGF-β 1,Smad3 and CTGF,modulating the expression of Smads and suppressing the activation of HSC.

Background Our previous study demonstrated that hyperglycemia aggravate the choroidal neovascularization (CNV) by promoting the chemotaxis process of bone marrow-derived cells (BMCs).Bioluminescence imaging (BLI) can dynamically monitor CNV in vivo.However,how diabetes mellitus (DM)participate in CNV is still in research.Objective This study was to dynamically observe the influence of BMCs to CNV under hyperglycaemia by using BLI combined with histopathology.Methods BMCs from luciferase-green fluorescent protein (Fluc-GFP) double transgenic mice were injected to adult wild type C57BL/6J mice (nine mice per group) via caudal vein to create the chimera models with a chimerism degree higher than 85％,and the chimeric mice were randomized into the control group and DM group based on randomized number table.Streptozotocin [60 mg/(kg · d)] was intraperitoneally injected daily for 5 days to establish the DM models in the chimeric mice of the DM group.CNV was induced in the chimeric mice of both control group and DM group with 532 nm laser photocoagulation.BLI signal of BMCsFluc+GFP+ was in vivo examined by IVIS Kinetics system 1,3,5,7,14,21 and 28 days after CNV modeling.At the seventh day after laser,part of mice were sacrificed,and choroidal and retinal sections were prepared for histopathological examination.The length and thickness of CNV were compared between the control group and DM group.The use and care of experimental followed Statement of ARVO.Results The chimerism degree of the chimeric mice was (88.85 ± 2.46) ％ 28 days after BMCs transplantation,and the blood glucose concentration in the DM group was (17.88±0.86)mmol/L.Histopathological examination revealed that CNV broke through the Bruch membrane toward subretinas.The length of the CNV was (338.67±33.17) μm in the DM group and (180.33±24.68)μm in the control group,showing a significant difference between the two groups (t =8.943,P＜0.05).However,no significant difference was seen in the CNV thickness between the two groups (t =1.790,P＞0.05).Light signals appeared 1 day and reach strongest 7 days after CNV modeling in both groups.The Light signals were stronger in the DM group than those in the control group on 5,7,14 and 21 days after CNV modeling (t =3.411,5.594,5.067,2.663,all at P＜0.05).Conclusions Hyperglycemia can promote more BMCs to participate in the pathogenesis and aggravation of CNV.The behavior of BMCs in CNV can be evaluated using BLI in vivo.

OBJECTIVE To determine the feasibility of clindamycin-loaded calcium phosphate cement(CLCPC) as a local antibiotic delivery system.METHODS The initial setting time(tI) and the final setting time(tF) were measured for 0%,2% and 5% CLCPC according to ASTM C266-89 method.Clindamycin concentrations eluting from the samples of 2% and 5% CLCPC in PBS were analyzed by HPLC at different times.The bacteriostasis tests were done by plate diffusion method for 2% and 5% CLCPC and 2% Palacos R-40 bone cement(PMMP) samples,and the diameters of the bacteriostasis ring and bacteriostasis duration were observed.The setting product and crystal size of 0%,2% and 5% CLCPC were analyzed and observed by X-ray diffraction(XRD) and scanning electron microscopy(SEM).RESULTS The setting time could be shortened by adding clindamycin(tI,tF) of 2% and 5% CLCPC.Clindamycin was with burst-release from CLCPC within the intial 6-hour period and the release rate slowed down on 4th day.Clindamycin could still release until to the 42th day.The ring of 2% Palacos R-40 bone cement(PMMP) bacteriostasis was smaller than that of 2% CLCPC,and the ring of 2% CLCPC bacteriostasis was smaller than that of 5%CLCPC.The bacteriostasis still existed to the 42th day of test for 2% and 5%CLCPC and 2% Clindamycin-loaded Palacos R-40 bone cement(CLPMMP).From the 30th day,many bacterial colonies were seen in the culture media laying 2%CLPMMP sample.On the contrary,bacterial colony was not found in the media putting 2% and 5%CLCPC.XRD and SEM showed that clindamycin did′t have an influence on setting product,crystal size and structure of CPC.CONCLUSIONS Clindamycin-loaded calcium phosphate cement can be used as a local antibiotic delivery system.

Objective To evaluate the value of superb micro-vascular imaging(SMI) in the detection of TI-RADS 4 thyroid nodules . Methods A retrospective analysis of 61 patients( 68 thyroid nodules) with grade 4 TI-RADS diagnosed by routine ultrasound and undergone SMI were performed ,and the differences between CDFI and SMI were compared . The TI-RADS level were corrected by CDFI and SMI technology , combined with the pathological results to compare the diagnostic efficacy before and after the correction . At the same time ,the related factors had been collected to evaluate the risk factors and to compare the value of SMI in benign and malignant nodules . Results SMI was more likely to detect the type Ⅲ blood flow ( P =0 .001) of malignant nodules . The area under the ROC curve before TI-RADS correction ,after CDFI correction and SMI correction were 0 .66 ,0 .69 and 0 .78 ,respectively . Compared with before TI-RADS correction and after CDFI correction ,the area under the curve of SMI correction was significant different ( P=0 .002 ,0 .009) . There was no significant difference in the area under the curve between after CDFI correction and before correction ( P = 0 .4196 ) . The sensitivity of SMI was higher ,but there was no significant difference ( P >0 .05) . After single factor analysis and multivariate analysis ,central blood flow and perforating blood flow found by SMI were not independent risk factors for thyroid cancer . Conclusions SMI is more effective than CDFI in detecting TI-RADS 4 thyroid nodules ,and it is expected to be an auxiliary tool for the diagnosis of benign and malignant thyroid nodules .

Objective: To evaluate the impacts of the prognostic factors of T2N0M0 upper tract urothelial carcinoma (UTUC) for Chinese patients.Methods: A retrospective study was conducted including 235 patients who were diagnosed with T2N0M0 UTUC in our hospital and received radical nephroureterectomy (RNU) or partial ureterectomy during January 2000 and December 2013.The 3 and 5-year can-cer-specific survival rates and bladder recurrence-free survival rates of all the patients were valued using Kaplan-Meier method, and the survival curves with statistical significance between the two were compared using the Log-rank test.Variables with significant differences in the univariate analysis were subjected to the multivariate analysis by Cox regression model.Results: A total of 235 patients were included in this study, including 95 (40.4%) male patients and 140 (59.6%) female patients.The mean age was 66.73±10.49 years.The median follow-up time was 53 (rang: 3-142) months, and during the follow-up, 74 (31.5%) patients died of UTUC after a median of 35 months,and 96 (40.9%) patients developed intravesical recurrence after a median of 19.5 months.The 3 and 5-year cancer-specific survival rates of all the patients were 89.1% and 85.9%, respectively;the bladder recurrence-free survival rates were 85.5% and 80.2%, respectively.The independent prognostic factors of cancer-specific mortality were tumor age elder than 55 years (HR=3.138, 95%CI: 1.348-7.306, P=0.008) and diameter larger than 5 cm (HR=3.320, 95%CI: 1.882-5.857, P<0.001).The independent prognostic factors of bladder recurrence-free survival were ureter tumor (HR=1.757, 95%CI: 1.159-2.664, P=0.008) and lower tumor grade (HR=1.760, 95% CI: 1.151-2.692, P=0.009).Conclusion: T2N0M0 UTUC has a better cancer-specific survival.The intravesical recurrence was equivalent to non-muscle invasive UTUC but earlier.The tumor diameter larger than 5 cm and the patient age elder than 55 years were independently associated with cancer-specific mortality;the primary tumor located in ureter and lower tumor grade were more likely to develop intravesical recurrence.

Objective To investigate the relationship of the expression of vascular endothelial growth factor (VEGF)and the mi-crovessel density (MVD)with multi-slice spiral CT perfusion imaging.Methods 80 patients with solitary pulmonary nodules under-went perfusion scan by 16-slice spiral CT.Among them,45 diagnosed as lung cancer by pathology were enrolled in the study.After surgery,the slice of the pecimen was selected similar to the corresponding slice of CT images,and the immunohistochemical staining was performed to determine the expression of VEGF and the MVD.Spearman correlation analysis was used to determine the rela-tionship of expression of VEGF and the MVD with CT perfusion parameters.Results There was more expression of VEGF and the MVD in NSCLC.There were positive correlations between VEGF,MVD and BF (both P<0.05).The peak enhancement image (PEI)and TTP have significant correlations with MVD (P<0.05).Conclusion Some parameters of lung CT perfusion imaging are correlated with MVD and VEGF.

Objective:To compare the dose distribution among CyberKnife, Tomotherapy, Edge, Triology and γ-knife in stereotactic body radiation therapy (SBRT) for pancreatic cancer.Methods:Clinical data of 10 panreatic cancer patients receiving CyberKinife treatment were retrospectively analyzed. The treatment plans were designed by five apparatuses from five centers according to the uniform requirement. All plans were transferred to MIM system for the extraction of parameters, which mainly included D min, D mean and D max of PTV, conformity index (CI), new conformity index (nCI), homogeneity index (HI), gradient index (GI), coverage, D max and dose-volume of the stomach and bowel. Results:The best CI and nCI were obtained in Triology ( P<0.001), and the worst HI was found in γ-knife ( P<0.001). The best GI was found in CyberKnife, followed by γ-knife and Tomotherapy, and Edge showed the worst GI ( P<0.001). The highest D min of PTV was found in both Edge and Triology, while lower D min of PTV was found in CyberKnife and Tomotherapy ( P<0.001). Additionally, γ-knife provided the highest D mean and D max of PTV ( P<0.001). Regarding the organs at risk, the lowest D max and D 5cm 3 of the bowel ( P<0.001), D max of the stomach ( P=0.003), D max( P=0.001), D 5cm 3 ( P<0.001) and D 10cm 3 ( P=0.005) of the duodenum, D max( P<0.001) and D 0.35cm 3 ( P<0.001) of the spinal cord were found in CyberKnife. The highest D max of the bowel was found in γ-knife. Furthermore, the highest D 5cm 3 of the duodenum was demonstrated in Edge ( P<0.001) and Tomotherapy provided the highest D max( P<0.001) and D 0.35cm 3 of the spinal cord ( P<0.001). Conclusions:All five radiotherapy apparatuses can meet the requirement of SBRT for pancreatic cancer. More rapid dose fall-off could be obtained via CyberKnife and γ-knife. Triology and Edge provide better target conformity. CyberKnife can better protect the gastrointestinal tract.

Human acute premyeloid leukemia cell cDNA expression library was constructed to screen acute premyeloid leukemia tumor antigen. Total RNA and purified mRNA were extracted from human premyeloid cell line NB4. First and second strands of cDNA were synthesized by reverse transcription. After blunting, the cDNA fragments were ligated with EcoR I adapters. Then the cDNAs were digested with Xho I, and less than 400 bp cDNA fragment was removed by Sephacryl-S400 spin column, the remaining were ligated with lambdaZAP vector. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli XL1-Blue-MRF' for titration. The recombinants were examined by color selection. In order to evaluate the size of cDNA inserts and the diversity of library, the pBK-CMV phagemid was excised from the ZAP express vector by using ExAssist helper phage with XLOLR strain, and then the pBK-CMV phagemid was digested by Xho I and EcoR I. The results showed that the NB4 cell line cDNA library consisting of 1.65 x 10(6) recombinant bacteriophages was constructed with the recombinant ratio of 99.6%. The average length of the recombinant exogenous inserts was about 1.7 kb. It was concluded that the constructed cDNA library are deserved to screen target clones.
Bacteriophages/genetics ; DNA, Complementary/*biosynthesis ; *DNA, Neoplasm/biosynthesis ; DNA, Recombinant/biosynthesis ; *Gene Library ; Genetic Vectors ; Leukemia, Promyelocytic, Acute/*genetics ; Leukemia, Promyelocytic, Acute/metabolism ; Leukemia, Promyelocytic, Acute/pathology ; RNA-Directed DNA Polymerase/metabolism ; Transcription, Genetic/genetics

Objective:To investigate the effect of deviation from the isocenter point on the quality of CT images at the same radiation dose.Methods:A 160-layer CT scanner was used to scan the phantom at isocenter and deviations of 3, 6, 9, 12 and 15 cm. CT was performed with the following parameters: 120 kVp; 400 mAs; slice thickness, 1 mm; and slice increment, 1 mm. Images were reconstructed using the filtered back projection algorithm. Noise power spectrum (NPS), task transfer function (TTF) and detectability index (d′) were measured. NPS peak was used to quantify the noise magnitude and TTF 50% was used to quantify the spatial resolution. NPS, TTF and d′ were compared using one-way ANOVA. Results:The NPS average spatial frequency, spatial resolution and d′ values gradually decreased as the offset distance increased and the amount of noise increased. NPS peak at isocenter and deviations of 3 cm, 6 cm, 9 cm, 12 cm and 15 cm were (94.31±1.48), (104.25±1.46), (131.44±1.96), (171.86±1.91), (224.05±1.37), (286.51±2.09)HU 2·mm 2, respectively ( F=37 241.91, P<0.001). And d′ values of 2 mm low-contrast lesions were 3.51±0.06, 3.31±0.04, 3.01±0.04, 2.59±0.06, 2.21±0.03, 1.88±0.03, respectively. The reduction in spatial resolution was more pronounced for high contrast, and the d′ values decreased to a similar extent for various types of lesions. The noise was increased by about 82%, the high contrast spatial resolution was decreased by about 12%, and the d′ value was decreased by about 26% at 9 cm from the isocenter point (all P<0.05). The noise was increased by about 204%, the high contrast spatial resolution was decreased by about 27%, and the d′ value was decreased by about 45% at 15 cm from the isocenter (all P<0.05). Conclusion:The CT image quality was decreased with the increase of the offset distance from the CT isocenter point. The image quality was severely compromised at offset distances greater than 9 cm.