ObjectiveTo explore the relationship between male adolescents' childhood abuse and externalizing behaviors,and the moderating effect of monoamine oxidase A (MAOA) gene promoter region variable number tandem repeat (VNTR) polymorphism.MethodsThrough random cluster sampling,352 Han male middle school students from Changsha were tested by Achenbach child behavior checklist-youth self reports (CBCL-YSR)and childhood trauma questionnaire-28 item short form (CTQ-SF),their MAOA genotypes were also identified.A hierarchical multiple regression analysis was used to test the moderating effect.Results ( 1 ) Compared participates with high activity MAOA gene and that with low activity MAOA gene,there were no significant differences on age ( ( 15.82 ± 1.52) vs ( 15.94 ± 1.62 ) ),externalizing behaviors ( ( 15.13 ± 10.14 ) vs ( 14.33 ± 9.70 ) ),total abuse ( (52.59 ± 10.46) vs (51.39 ± 7.56 ) ),emotional abuse( ( 7.63 ± 3.31 ) vs ( 7.11 ± 2.68 ) ),physical abuse ( ( 6.40 ± 2.82) vs (6.12 ± 2.05 ) ),sexual abuse ( ( 6.42 ± 3.24 ) vs ( 5.94 ± 1.72 ) ),emotional neglect ((13.44±5.12) vs (13.16 ±4.83) ),physical neglect( (10.27 ±2.64) vs (10.44±2.53))(t=-1.789～0.678,P ＞ 0.05 ).(2)Except emotional neglect and physical neglect,emotional abuse,physical abuse and sexual abuse could predict externalizing behaviors( Sβ =0.141 ～0.347,P ＜ 0.01 ).(3) MAOA gene was not directly related to externalizing behaviors( Sβ =- 0.023,P ＞ 0.05 ).There was a significant interaction between MAOA gene and emotional abuse( Sβ =-0.148,P ＜ 0.01 ).The interaction between MAOA gene and physical abuse or sexual abuse showed no statistical significance( Sβ =- 0.067,- 0.005,P ＞ 0.05 ).ConclusionMAOA gene polymorphism can moderate the relationship between male adolescents'childhood emotional abuse and externalizing behaviors.

Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.

Human herpesvirus 7 (HHV-7) infection is dependent on the functions of structural glycoproteins at multiple stages of the viral life cycle. These proteins mediate the initial attachment and fusion events that occur between the viral envelope and a host cell membrane, as well as cell to cell spread of the virus. To characterize the HHV-7 glycoproteins that can mediate cell fusion, a cell-based fusion assay was used. 293T cells expressing the HHV-7 glycoproteins of interest along with a luciferase reporter gene under the control of the T7 promoter were cocultivated with SupT1 cells transfected with T7 RNA polymerase. HHV-7 glycoproteins gB, gH, gL and gO can mediate the fusion of 293T cells with SupT1 cells, and the fusion can be inhibited by anti-CD4 mAbs. Thus, the coexpression of HHV-7 gB, gO, gH and gL is sufficient and necessary for HHV-7 induced membrane fusion, and one of these glycoproteins or protein complex formed by these glycoproteins might be the ligand(s) of CD4 molecule.

Comprehensive biochemistry experiment,which is interlocked and has difficulties in a certain degree,requires considerable knowledge,multiple techniques and long time.In order to ensure the smooth progress of the experiment,biochemistry and molecular biology department of Hebei medical university has taken following measures in teaching preparation and teaching implementation:building a specialized laboratory;performing collective lesson preparation and pre-experiments;technical teaching;teaching with multimedia equipments;students submitting experimental preparatory reports before class and then completing the experiments in groups.These measures achieved the intended purpose of setting up a comprehensive experiment.

Objective To evaluate the imaging quality of the non-contrast enhanced MR angiography of sampling perfection with application optimized contrasts using different flip angle evolutions (SPACE)in showing portal system and compared it with that of the contrast enhanced MR angiography of volumetric interpolated breath-hold examination (VIBE),and study its diagnostic ability in the detection of portosystemic and portohepatic collaterals.Methods Thirty consecutively cirrhotic patients with suspected of portosystemic and portohepatic collaterals were enrolled,and underwent SPACE followed by VIBE at 1.5 T MR scanner.The diagnostic accuracy of SPACE for the portal vein disease was evaluated by two doctors and compared it with that of VIBE.The contrast-to-noise ratio(CNR) and signal-to-noise ratio(SNR) of two MRA techniques were compared by using the Wilcoxon signed rank test.The quality assessment including scores of the portal vein segments and overall image quality were used the paired t test.Results Twenty-one patients were diagnosed as portal hypertension,including five types of portosystemic collaterals: esophageal varices (n =5),gastric fundic varices (n =11),splenic varices (n =5),paraumbilical varices (n =5) and cavemous transformation (n =2),and one patient was diagnosed as portal vein tumor thrombus.The diagnostic efficiency of SPACE was equivalent to that of VIBE.In SPACE,the SNR were 291 ± 57,301 ± 74,344 ±76 and the CNR were 231 ±59,242 ±73,286 ±76 at main portal vein,the left branch of portal vein and the right branch of portal vein,respectively.However in VIBE,the SNR were 185 ± 56,176 ± 52,182 ±52 and the CNR were 57 ±23,50 ±21,57± 19 at,respectively.Both SNR and CNR of portal vein segments in the former were better than those in the latter (t values were 7.691,7.418,7.946,15.746,13.508 and 13.880,respectively,P ＜ 0.05).There were no significant difference for the scores of displaying main,left branch and right branch portal vein and overall image quality in VIBE and SPACE (Z values were -1.496,-1.895,-1.496,-2.138,-2.324 and-1.328,respectively,P ＞ 0.05).The scores of displaying the distal branches of left and right portal vein were 2.08 ± 0.78,2.08 ± 0.78 in SPACE,and 1.75 ± 0.53,1.71 ± 0.55 in VIBE,respectively.It was better (Z =-2.138,-2.324,P ＜ 0.05) in SPACE than that in VIBE.Conclusion The SPACE has better visualization of portal vein distal branches than VIBE,and it can be applied for the diagnosis of the portal vein disease.

Objective To explore the inhibitory effect of Oldenlandia diffusa extract on colorectal cancer angiogenesis in BALB/c mice. Methods Thirty-two BALB/c mice with subcutaneous CT26 colon cancer animal model were randomly equally divided into four groups,including the control group (groupⅠ,saline 0.1 mL/(10. d), O. diffusa ethanol extract of 90 mg/(kg.d) (groupⅡ), O. diffusa ethanol extract of 180 mg/(kg.d) (groupⅢ) and O. diffusa ethanol extract of 360 mg/(kg.d) (group Ⅳ) . Each group of mice were treated with intragastric administration of law administration 12 days after vaccination, then stopped and continue fed to 32 days, and the mice were killed. Micro-vascular dense ( MVD) was observed and countered under the microscopy by immunohistory chemistry. Results The murine colon tumor volumes of GroupⅡ,ⅢandⅣwere significantly less than that of groupⅠ,with significant difference ( <0.05) . The tumor microvessel density values of four groups was (7.83±2.87), (5.32±1.27), (1.77±0.70) and (1.87±0.68),respectively. The number of tumor blood vessels in GroupⅡ,Ⅲ and Ⅳ were significantly less than that of Ⅰ group, with significant difference ( <0.05) . Conclusion Within a certain dose range, the ethanol extract of O. diffusa can significantly inhibit the mouse colon cancer and the mechanism may be realated to inhibiting tumor angiogenesis.

OBJECTIVETo identify the Cynomorii Herba and its analogues species using DNA barcoding technique.

METHODTotal genomic DNA extracted from all materials using the DNA extraction kit. The internal transcribed spacer 2 (ITS2) regions were amplified using polymerase chain reaction (PCR), and purified PCR products were sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner 3.7.1. The Kimura 2-Parameter (K2P) distances and GC content were computed using MEGA 5. 0. Species identification analyses were conducted through the species identification system for traditional Chinese medicine and neighbor-joining (NJ) trees.

RESULTThe ITS2 sequence lengths of Cynomorii Herba were 229 bp. The average intra-specific genetic distances of Cynomorii Herba were 0.003. The average inter-specific genetic distances between Cynomorii Herba and its adulterants species were 0.760. The results showed that the minimum inter-specific divergence is larger than the maximum intra-specific divergence. The species identification system for traditional Chinese medicine and NJ trees results indicated that Cynomorii Herba and its adulterants species can be easily identification.

CONCLUSIONThe ITS2 region is an efficient barcode for identification of Cynomorii Herba, which provide a new technique to ensure clinical safety in utilization of traditional Chinese medicine.

Objective To investigate the abnormality of intrahepatic biliary epithelial cells autophagy in primary biliary cirrhosis (PBC) mice,and explore the mechanism of UC-Mesenchymal stem cell (MSCs) in treating PBC.Methods After establishing the PBC model,we divided them into the PBC model group;the UC-MSCs treatment group and the Stattic group [Signal transducer and activator of transcription 3 (STAT3) inhibitor group].Six mice were used as the control group.Liver pathology and the serum pyruvate dehydrogenase complex E2 subunit (PDC-E2) antibody titers were detected.Autophagosome of the intrahepatic biliary epithelial cell was observed by electronic microscope.Protein levels of STAT3/pSTAT3,Beclin-1 were detected by western blot.We cultured human intrahepatic biliary epithelial cells in vitro,and down regulated STAT3.After stimulated by GCDC,we co-cultured them with UC-MSCs,and collected the cells in order to detect LC3 Ⅱ.The measurement data were compared with t test or single factor analysis of variance.Results Compared with the control group,periportal inflammatory cell infiltration and granuloma formation were observed in the PBC group.MSCs treatment decreased the infiltration of inflammatory cells.The level of antiPDC-E2 of the PBC group (107±18) ng/ml was higher than that of the control group (42±6) ng/ml (q=6.326,P＜0.01),MSCs treatment down regulated anti-PDC-E2 level (43±4) ng/ml (q=5.801,P＜0.01).More autophagosomes in the PBC group (5.00±1.29) than the control group (1.75±0.25) were observed (q=4.061,P＞0.05).Western blot showed that the level of Beclin-1 was higher in PBC group (1.80±0.36) than the control group (0.40±0.20) (q =6.757,P＜0.01),MSCs reduced the expression of Beclin-1 (0.86±0.06)(q=4.536,P＜0.05) as well as Stattic (0.72±0.03) (q=5.226,P＜0.05).PBC group had a higher expression level of STAT3 (1.80±0.42) (q=5.730,P＜0.05) and pSTAT3 (2.04±0.29)(q=6.492,P＜0.01) than the control group (0.50±0.05)(0.91±0.14).MSCs treatment decreased the expression of STAT3 (0.51±0.13)(q =5.703,P＜0.01) and pSTAT3 (0.76±0.07) (q =7.388,P＜0.01) in intrahepatic biliary epithelial cells.After down regulated STAT3 of HiBECs,MSCs reduced the expression of LC3 Ⅱ of HiBECs.Conclusion The intrahepatic biliary epithelial cells autophage of PBC mice is abnormal,MSCs can alleviate PBC by down regulating the autophage of intrahepatic biliary epithelial cells via STAT3.

AIM: To clone EBV-LMP2A gene, construct and identify the recombinant retroviral vector and stable cell strains expressing EBV LMP2A. METHODS: The full-length EBV LMP2A gene was generated by RT-PCR amplification from B95.8 cells which contain complement nucleotide sequence of EBV LMP2A gene. The gene was ligated to T-vector and sequenced to construct retroviral vector consisting with LMP2A. To produce retroviral virus, packing cells, 293T cells were co-transfected with recombinant retroviral expression vector pGEZ-LMP2A and two auxiliary viral vectors pHIT456 and pHIT60 by lipofectAMINE2000. Viral titration was performed according to the instructions of the manufacturer. To establish L929 cell line stable expressing LMP2A, L929 cells were infected with recombinant retrovirus three times and selected by Zeocine. The Zeocine-resistant clones (L929/LMP2A) were screened for LMP2A expression by RT-PCR and Western blot. RESULTS: The recombinant retrovirus vector carrying LMP2A gene was constructed successfully. Transfection yield a titer of 5×10~8 infectious particles/L. The infected L929 cells were selected by Zeocine. Results of RT-PCR and Western blot indicated that L929 transgenetic cells could stably express EBV-LMP2A. CONCLUSION: The L929 cell line stably expressing LMP2A provides suitability for extraction of the LMP2A protein and preparations of the vaccine for the therapy of EBV-associated diseases.

Objective To evaluate the clinical outcome and relative factors of intervention treatment for atherosclerotic renal artery stenosis in elderly patients.Methods The clinical data of 79 patients diagnosed as atherosclerotic renal artery stenosis by angiography and treated by revascularization were analyzed.Results There were 55(69.6%)successes and 24(30.4%)failures in decreasing blood pressure and 28(35.4%)successes and 51(64.6%)failures in improving renal function after intervention treatment.Predictors of favorable outcome of intervention treatment in decreasing blood pressure were related to lower urine protein,higher glomerular filtration rate,higher systolic and diastolic blood pressure before treatment,lower resistance index(RI)of renal artery,and no complication of cerebral vascular diseases.Predictors of favorable outcome of intervention treatment in improving renal function were related with percentage of angiographic stenosis,category of antihypertension and lower urine protein.The logistic regression analysis showed that the percentage of angiographic stenosis was the most important predictor of intervention treatment for blood pressure control,age and level of serum creatinine before intervention treatment were the most important predictors of intervention treatment for improving renal faction.Conclusion Percentage of stenosis(≥85%),age(133 ?mol/L)can be used as the predictors of therapeutic success for renovascular stenosis in older patients.