DNA polymerase γis the only known DNA polymerase in human mitochondria,and is essential for mitochondrial DNA replication and repair.DNA polymerase γ is encoded by POLG gene.POLG-related disorders resulted from mutations of POLG gene comprise a continuum of overlapping phenotypes including Alpers Huttenlocher syndrome and other five subtypes, with high prevalence rate at patients with intractable seizure.Genetic testing for POLG mutations in patients with intractable epilepsies is very important for clinical diagnostics, genetic counseling, and treatment decisions because of the increased risk for VPA-induced liver failure in patients with POLG mutations.

Objective To investigate the in vitro effects of human bone marrow derived mesenchymal stem cells (hBMSC) on Th17 cells of the human peripheral blood.Methods The density gradient centrifugation combined with lymphocyte separation medium was used to isolate hBMSC,which were then cultured.Cytokine IL-17 in the peripheral blood from a healthy person was measured by enzyme-linked immunosorbent assay (ELISA).Proportion of Th17 cells was evaluated by flow cytometry.Results The expression level of IL-17 in spent culture supernatant of the healthy person PBMC and AML hBMSC was (292.32±37.25) pg/ml,and was significantly higher than that of the healthy person PBMC and healthy hBMSC [(169.64±17.47) pg/ml,P ＜ 0.01].There was no significant difference between the expression level of IL-17 in spent culture supernatant of the healthy person PBMC and ALL hBMSC [(159.89±23.71) pg/ml] and the level of the healthy person PBMC and hBMSC.The percentages of Th17 cells of co-culture systems from hBMSC,ALL-hBMSC,and AML-hBMSC and PBMC were (10.13±2.19) ％,(13.77±4.04) ％ and (21.53±5.05) ％,respectively,with the result between AML patients and healthy people being statistically different (P ＜ 0.01).ALL patients and healthy people showed no difference (P ＞ 0.05).Conclusion AML-hBMSC promotes the CD~ T cells to generate Th17 cells,which suggests that the MSC from AML marrow may play a role in the regulation of immune suppression.

To investigate the prevalence of mental health problems in male violent offenders of Hunan and Sichuan Provinces; and to compare the types and severity of problems between the violent and nonviolent offenders.Methods:Ninety-one violent juvenile offenders and 64 nonviolent juvenile offenders in the juvenile detention centers of Hunan,and 81 violent juvenile offenders in the juvenile detention centers of Sichuan; 39 high school students from a middle school of Hunan Province and 49 from a middle school of Sichuan Province were investigated using the Investigation Screening Inventory for Child Mental Disorder and the Kiddy Schedule for Affective Diseases and Schizophrenia (K-SADS-PL),and classified according to the Diagnostic and Statistical Manual of Mental Disorders.Results:In the Hunan violent group,86.6％ met the criterion of conduct disorder (CD),17.6％ of substance abuse,9.9％ of substance dependence,while in the Hunan nonviolent group,75.0％ met the criterion of CD,and 11.7％ of substance abuse; these were all significantly higher than those of the control group (P＜0.05).In the Sichuan violent group,17.3％ met the criterion of attention deficit/hyperactivity disorder (ADHD),18.5％ of oppositional defiant disorder (ODD),69.1％of CD,and 22.2％ of substance abuse; these were all significantly higher than those of the Sichuan control group.Moreover,63.7％ of Hunan-violent,55.6％ of Sichuan-violent,and 45.0％ of Hunannonviolent offenders had a previous offence records.Conclusion:Mental and behavioral disorders among delinquent youth is becoming a serious problem,and there is an urgent need to develop and implement effective assessment and treatment approaches for juvenile offenders with the aim of reducing offence and recidivism in this population.

Objective To determine the margins of planning target volume (MPTV) in primary cervical cancer patients with tomotherapy and evaluate the importance of automatic registration(AR) plus manual registration.Methods The setup errors of 29 primary cervical cancer patients receiving external radiation from June 2012 to Dec 2014 were measured by megavoltage computed tomography (MVCT),which were performed at least two times weekly before treatment and were registered with the planning CT.The setup errors between automatic registration and total shift (TS) including both AR and manual registration were compared MPTV was calculated.Results Setup errors were collecte from 443 sets of MVCT in 29 patients.AR and total shift (TS) values in the x,y,z directions and rotation angle were (-0.9±2.3),(0.0±3.1),(1.0±2.6) mm,0.2° ±0.8° and (-0.8±1.8),(-0.4±3.4),(l.4 ± 2.5) mm,0.1° ± 0.5°,respectively.There were statistically significant differences between the two groups in all directions except for the x axis (t =5.1,-5.2,3.2,P ＜ 0.05).MPTV were 4.6,5.7,3.3 mm in the x,y,z directions,respectively.Conclusions Manual registration is necessary after automatic registration in cervical cancer patients with tomotherapy.For patients with cervical cancer treated by tomotherapy,planning target volume MPTV parameters are suggested to be 5,6,4 mm in the x,y,z directions.

Objective To construct a fusion protein that used for treatment of resistance and palindromia in leukemia and studied its biological activity. Methods IL-3 and LP gene fragments were amplified by PCR. After enzymatic digestion and T4 ligation, the fusion gene was cloned into expression vector pAYZ. The product was purified by exchange chromatography and anti-Etag affinity chromatography. IL3-G4SLP fusion protein was analyzed by SDS-PAGE and Western blot. Protein biological activity was detected by FACS. Results The fusion protein was expressed as soluble protein by E.Coli 16C9. The protein expression level was about 1 mg/L, its purity was over 95 %, and the expression level was about 1 mg/L. The fusion protein can combined specificely with CD123 on leukemia stem cells. Conclusion Fusion protein IL-3-G4S-LP can target on leukemia stem cells and maybe as a potential drug used for treatment of resistance and palindromia in leukemia.

Objective To compare the sensitivity and diagnostic features of mammogram (MG) and magnetic resonance imaging (MRI) on diagnosis of breast ductal carcinoma in situ (DCIS) with or without microinvasion (DCIS-MI).Methods From Jan 2012 to Nov 2013,results of MG and MRI from 72 cases of DCIS or DCIS-MI were retrospectively analyzed.Results The sensitivity of MG was 52.8％ (38/72).The sensitivity of MRI was 87.5％ (63/72),among those 76.2％ (48/63)lesions presented as non-mass-like enhancement.Sensitivity of MRI was significantly higher in DCIS-MI than DCIS (84.6％ vs 100％,P =0.027).Logistic regression analysis showed calcifications was an independent factor influencing the sensitivity of MG (OR =23.785,P ＜ 0.001).Conclusions The sensitivity of MRI is higher than MG for the diagnosis of DCIS and DCIS-MI.

[Objective] To investigate the effect of P-selectin on the intestinal tumorigenesis in the 15,24-week-old ApcMin/+ mice.[Methods] Male ApcMin/+ mice were mated with female P-selectin knockout (P-sel-/-) mice to generate AMin/+Ps-/-mice.The diameters and numbers of the intestinal tumors in the intestines of ApcMin/+ and AM/+ps-/-mice were measured using an inverted microscope.The tumor volumes were measured using the following equation:volume =0.52 × (length × width2).[Results] There were no significant difference between the volume and number of intestinal tumor in 15-week-old ApcMin/+ mice and 15-week-old A+ P-mice.The volume and number of intestinal tumor were significantly increased in 24-week-old A+P-mice compared to 24-weekold ApcMin/+ mice.P-selectin deletion significantly prolonged the survival time of ApcMin/+ mice.[Conclusions] P-selectin deletion promotes the intestinal tumorigenesis in the 24-week-old ApcMin/+ mice.

Background and purpose:Bispecific antibody is a special pattern of genetic engineering antibody.It possesses 2 antigen binding sites, one directed at the effecter cells and the other at the target cells which are always the tumor cells.This enables the effecter cells to recruit to the targets efficiently and exert cytotoxic effect.The experiment was aimed to construct the expression vector of anti-CD3?anti-CD19 bispecific antibody, purification and analyzing the activity of the diabody.Methods:Applyed PCR and overlap PCR to construct the expression vector of anti-CD3?anti-CD19 bispecific antibodies, transformed it into E.coli 16C9 for expression.Purified the diabody by His-tag affinity chromatography and confirmed by 12% SDS-PAGE and Western blot.Antigen binding activity of the diabody was analyzed by FACS.Results:The DNA sequence of the expression vector containing anti-CD3?anti-CD19 fragment was comfirmed.The yield of purified diabody was up to 5 mg/L.The molecular weight of diabody was correctly indicated by SDS-PAGE and Western blot.The diabody could specifically bind to CD19+ Raji cells and CD3+ Jurkat cells while competitively inhibit binding of HIT3a and HIT19a to the cells.Conclusion:Successfully constructed the expression vector of anti-CD3?anti-CD19 bispecific antibodies, the diabody acquired by the method could be produced with high level expression and also had high specific affinity with CD19+ Raji cells and CD3+ Jurkat cells.

Objective To evaluate the difference of Doumas′method and 15 commercial serum total protein ( TP ) methods based on EP9-A3.Methods Serum panels were quantified for TP with Doumas′method and measured in parallel with 15 commercial methods.The linear regression analyses were performed, followed by calculating relative deviation and 95%CI between commercial method and Doumas′method at three different medical decision levels (45 g/L, 60 g/L, 80 g/L).We also calculated relative deviation, 95% limit of agreement ( LoA ) and 95% CI based on classical and improved Bland-Altman method at three different medical decision levels.If both the relative deviation and 95%CI were within 5%, we conside red the commercial serum total protein method was comparable to Doumas′method.Results (1) All assays presented high correlation ( r>0.975, P<0.001) with the Doumas′method.All assays showed that the relative deviations and 95%CIs were within the biological total error goal (5%) at medical decision levels based on regression analysis.(2) Based on classical and improved Bland-Altman method, fourteen of 15 commercial methods showed that the relative deviations and 95%CIs were within +/-5%. Conclusions All commercial assays are comparable to Doumas′method at medical deviation levels.There is no difference between regression analysis and Bland-Altman method for comparison study.

The study,based on Huizhou Chronicle,on official medical services in Huizhou and local medical officers in Xin'an in the Ming and Qing Dynasties finds out that the major affairs of Xin'an local medical officials were to rescue and cure poor and sick military and civilians,and dealt with plague in early Ming Dynasty.But official medical services and local medical officers have been gradually abolished since the Jiajing period of the Ming Dynasty,and stepped down gradually from the historical stage after the Qing Dynasty except of Wuyuan County.And then folk multiple forces have risen and become undertakers of social medical insurance in Huizhou.