Objective To observe the prophylactic effects of a HSV-2gD2DNA vaccine in guinea pigs challenged with HSV-2strains.Methods Female guinea pigs were divided into3groups with10each,which was immunized intramuscularly with100?g of pc-gD plasmids(recombinant HSV-2DNA vac-cine),or with pcDNA3blank plasmids,with normal saline as control,respectively.Two booster injections were given on day7and day21.Sera were collected for virus neutralization test on day0,day28,and day56.The animals were challenged with HSV-2strain sav intravaginally,and lesions induced on the external genital skin were scored between day1and day21after challenge.Results The titer of neutralizing anti-body to HSV-2was much higher in the sera from animals immunized by pc-gD plasmids than that from ani-mals immunized by pcDNA3blank plasmids or normal saline.Furthermore,the lesion scores on external genital skin were significantly decreased in pc-gD group than those in other two groups with either primary or recurrent infections.Conclusion The constructed gD2vaccine can efficiently protect guinea pigs from genital infection and reduce recurrent infection induced by latent herpes simplex virus.

Objective Using the technology of siRNA to inhibit gene expression of T cells'nonreceptor tyrosine protein kinase Lck in asthmatic mice,and to study the effect of siRNA inhibited Lck to the function of T cells in asthmatic mice.Methods The 21 - 23 bp RNA fragments of mouse T cell Lck were made by chemosynthesis.INTERFERinTMsiRNA Transfection Reagent was used as transfection reagent to transfect the siRNA into the spleen T cells of asthmatic mice for 48 hours.Then T cells were mixed with bone marrow dendritic cells (DC) of asthmatic mice for another 48 hours.Cell culture suspension was collected and the level of IL-4,IL-13,IL-2,INF-γ were detected with respondent ELISA kits; Western Blot was used to identify if the expression of Lck was blocked.Results The expression of Lck in T cells almost could not be detected in siRNA interference group.The levels of IL-4 and IL-13 in siRNA interference group( 10.19 ± 1.66,12.34 ±0.79) were lower than no-siRNA interference(28.06 ±2.88,27.87 ± 1.61 )and control group ( 22.07 ± 2.5 1,20.47 ± 2.37 ),and the difference was statistical significant ( P ＜0.01 ).Conclusions Special siRNA could block the expression of special gene,and Lck specific siRNA could block the activation and differentiation of T cells and reduce the secretion of inflammatory cytokines in asthmatic mice.

Objective To investigate the association between serum ferritin(SF) level at early and mid trimester of pregnancy and gestational diabetes mellitus (GDM) and insulin resistance (IR).Methods A total of 192 cases of women who were accepted antenatal examination from the first trimester of pregnancy at Yueqing People Hospital from July,2012 to October,2013 was prospectively analyzed and followed up.All cases were divided into two groups:96 cases of GDM pregnant women who were diagnosed as GDM at mid trimester of pregnancy as GDM group,and 96 cases of pregnant women with normal glucose tolerance at the same time as the control,normal glucose tolerance (NGT group).The differences in clinical data were compared between two groups.The relationship between SF level and fasting plasma glucose (FPG),fasting insulin (FINS),homeostasis model assessment insulin resistance (HOMA-IR),and C-peptide was investigated with Spearman rank correlation analysis.The predictive values of SF and relative variables to GDM were calculated by receiver operating characteristic curve (ROC) or Logistic regression analysis.Results (1) There was statistically significant difference in body weight and body mass index (BMI) at prepregnancy and mid trimester of pregnancy,Hb,FPG,FINS,SF,and C-peptide at the first trimester of pregnancy,SF level and FPG and HOMA-IR and C-peptide at mid trimester of pregnancy between two groups (P ＜ 0.01 or P ＜ 0.05).(2) As showed by Spearman rank correlation analysis,there was statistical correlation between SF at mid trimester of pregnancy,and FPG,FINS,HOMA-IR and C-peptide (P ＜0.01 or P ＜ 0.05);while no statistical correlation was found between SF at the first trimester of pregnancy and the same factors above (P ＞ 0.05).(3) The area under the ROC curve of SF at mid trimester of pregnancy for GDM was 0.653.The sensitivity and specificity were 68.8％ and 59.4％ in predicting GDM at the cut-off value of 16.61 ng/ml.As showed by Logistic regression analysis,high level of SF at mid trimester of pregnancy was a independent risk factor for GDM.Odds ratio (OR) was 1.032 (95％ CI:1.008 ～ 1.058,P ＜ 0.01).Conclusions The relationship between SF at different trimester of pregnancy and GDM is variant.There is relationship between high level SF of mid trimester of pregnancy in GDM pregnant women and IR.The level of SF might predict the occurrence of GDM.

Objective To assess the variation in patient setup corrections for three different anatomic treat-ment sites using daily pretreatment megavoltage CT(MVCT)in helical tomotherapy,and to analyze alternative refer-ence margins for specific tumor site.Methods Sixty patients treated for three anatomical sites on helical tomotherapy were analyzed.Daily MVCT was carried out for registration and setup corrections before each treatment fraction.Setup errors and rotational setup corrections from 587 head and neck,500 thoracic,371 abdomen and gynecology MCVT imaging were input to statistical analysis.Incidences of three dimensional vector error lengths were calculated for each anatomic site.Overall distribution histograms of the three -dimensionally error was presented using GraphPad Prism 5.The distributions of systematic and random setup errors were quantitative analyzed and the additional margins required were also taken into account.Results 1 458 MVCT scans were implemented for sixty patients.Head and neck had lower frequencies of translational setup errors than others.Frequency of at least 3mm three -dimensional setup errors for head and neck,thoracic,abdomen and gynecology was 55.3%,70.8%,79.8%,respectively.This fre-quency decreased to 17.5%,40.6%,47.2% if 3D vector distance ≥5mm was scored.Overall systematic errors ranged from -1.4mm to 2.7mm,abdomen and gynecology had the largest setup errors in the vertical direction which was statistically significant(χ2 =19.3,P <0.05).The suggested margins should be increased by 4 -7 mm in three -dimensional direction for head and neck,9 -14mm for thoracic,10 -17mm for abdomen and gynecology.Conclusion Differences in setup corrections are perceived between head and neck,thoracic,abdomen and gynecology.The accura-cy of patient positioning can be improved if pre -treatment daily MVCT scans are put into use.Results from setup cor-rection can provide evidence for tumor treatment margin and improve the accuracy of regular radiotherapy.

ObjectiveUsing the technology of siRNA to inhibit the gene expression of no-receptor　tyrosine protein kinase Lck in T cells of asthmatic mice,and to study the therapeutic effect of Lck specific　siRNA in asthmatic mice.MethodsReceptor tyrosine protein kinase Lck specific siRNA fragments were　taken from chemosynthesis.In vivo-jetPEITM was used to transfect the siRNA into mice body through tail　vein injection.The mice were killed 48 hours later,and the levels of IL-4,IL-17 in bronchoalveolar lavage　fluid (BALF) were detected with respondent ELISA kits.The change of inflammatory histopathology in lung　was observed with H.E.staining.The expression of Lck in lung was detected with immunohistochemistry　(IHC),and the level of Lck in lung tissue homogenate was detected with Western Blot.Results Compared with asthmatic group[ (234.68 ± 11.15 ) pg/ml,( 96.76 ± 8.28 ) pg/ml],the levels of IL-4,IL-17　[ (234.68 ± 11.15)pg/ml,(96.76 ±8.28) pg/ml] in the BALF of siRNA interference group decreased,　and the inflammation in the lung relieved.IHC indicated that the expression of Lck in lung decreased and　the level of Lck in lung tissue homogenate decreased ( P ＜ 0.05 ).Conclusions Lck specific siRNA　could reduce the level of IL-4,IL-17 in the lung tissues of asthmatic mice,and relieve the inflammatory reaction in lung.

Objective To explore the inter-fraction setup errors and affecting factors from data of daily fan-beam megavoltage computed tomography(MVCT). Methods A total of 37consecutive NPC patients treated with tomotherapy were hospitalized during the period of February 2015 to September 2015. For each patient,one MVCT scan was obtained after conventional positioning ,online correction and tomotherapy delivery daily ,and the scans were put into the planning computed tomography to determine inter-fraction setup errors. The MPTV was calculated with the equation:MPTV=2.5∑+0.7σ(∑:systematic error;σ:random error). Results The average absolute errors of the inter-fraction were(2.102 ± 0.0406)mm,(1.490 ± 0.0348)mm,(1.306 ± 0.335)mm and(1.392 ± 0.0384)° at three dimensions. The total MPTV accounting for inter-error was 3.4675 mm,2.9795 mm,and 2.8885 mm. Gradual increases in both inter-fraction three-dimensional displacement were observed with time and treatment(P < 0.05). Univariate analysis revealed that weight loss and retraction of neck lymph nodes were affecting factors of set-up errors. Conclusions 3 mm margins uniformly expended from clinical target volume to planning target volume may not be suitable. The personalized margin should be adopted for the design of IMRT planning. Displacement increases as a treatment course is prolonged.

Objective To evaluate the inter-fraction setup error during the treatment with megavoltage computed tomography (MVCT) and provide theoretical basis for clinical target volume-planning target volume (CTV-PTV) margins for nasopharyngeal carcinoma (NPC) patients treated with tomotherapy.Methods Thirty-seven consecutive NPC patients treated with tomotherapy were prospectively enrolled for the study between February 2015 and September 2015.For each patient,one MVCT scan was obtained after conventional positioning,online correction and tomotherapy delivery daily,and the scan was registered to the planning CT to determine inter-fraction setup error.The expanding margin for PTV (MPTV) was calculated with the recipe:MPTV =2.5∑ + 0.76 (∑:systematic error;6:random error).Results The average absolute errors of the inter-fraction were (2.102 ± 0.040 6) mm,(1.490 ± 0.034 8) mm,(1.306 ± 0.335) mm and (1.392 ± 0.038 4) ° in the three dimensions.Gradual increases in both inter-fraction three-dimensional displacement were observed with time and treatment (P ＜ 0.05).The total MPTV ac counting for inter-error were 3.467 5 mm,2.979 5 mm and 2.888 5 mm.Conclusions Tomotherapy irradiation technology personalized MPTV should be adopted for the design of tomotherapy plan.Displacement increased as a function of time.

Objective: To observe the integration frequency of aadA2 resistance cassette at attI site of the integron under different concentration of streptomycin. Methods: Class 1 integron with known gene sequence was cloned into plasmid pACYC184 to produce recombinant plasmid pACIDA, meanwhile the integrase gene was cloned into plasmid pET28a to construct recombinant plasmid pETINT. These two recombinant plasmids were consecutively transformed into E. coli BL(DE3). These transformed bacteria was cultured in the LB medium at 37 ℃ overnight with addition of different concentration of streptomycin. The copy number of total integrons and the copy number of integrated aadA2 at attI site of integrons were determined by using real-time PCR. and the integration frequency is the result of the former divided by the latter. Results: The resulting frequencies were (1.97±0.24)×10^-3, (3.23±1.77)×10^-3, (3.27±0.67)×10^-3, 0.45±0.13 and 1.32±0.11, with respective streptomycin concentrations of 0, 20, 30, 40 and 50 μg/ml. The background frequency of integration without integrase overexpression was less than (1.75±0.33)×10^-7. Conclusion These findings indicate that antibiotic concentration significantly increase recombination frequency of aadA2 resistance cassette at attI site of the integron, catalyzed by integron integrase.(Chin J Lab Med, 2018, 41: 531-535)

Objective To compare the dosimetric differences in heart and lung among three radiotherapy techniques in the treatment of thoracic esophageal cancer. Methods A total of 15 thoracic esophageal cancer patients treated in Zhejiang Cancer Hospital during the period of January 2015 to February 2016 were enrolled in this retrospective dosimetric study. Three radiotherapy treatment plans, including intensity-modulated radiotherapy ( IMRT) , volumetric intensity modulated arc therapy ( VMAT) with Raystation 4. 0v TPS, and tomotherapy (TOMO) plans with TomoTTM v2. 0. 5 TPS were generated for each patient with a prescribed dose of 60 Gy at 2 Gy per fraction. Dosimetric differences in planning target volume ( PTV) , heart, cardiac subunits and lung were compared. Results The mean volumes of PTV, heart and lung were (399 ±355), (671 ±274) and (3907 ±1717)cm3, respectively. Compared with VMAT and IMRT, TOMO reduced the maximum dose of PTV, heart, left atrium and lung ( H=10. 889, 7. 433, 12. 080, 11. 401, P <0. 05). No difference was observed in conformity or homogeneity among these three plans. Conclusions TOMO reduced the maximum dose to PTV, heart, left atrium and lung compared with VMAT and IMRT, However, no difference in conformity and homogeneity was observed. The impact of dosimetric advantage of TOMO needs further verification due to the interaction between heart and lung for thoracic esophageal cancer patients.

Objective: This study aimed to systematically analyze the effect and stability of miniscrew-assisted rapid palatal expansion (MARPE) to provide a reference for the clinical treatment of patients with maxillary transverse deficiency (MTD). Methods: We searched PubMed, Science Direct, Web of Science, Embase, Cochrane Library, CNKI, and Wanfang Database for relevant studies published before February 18, 2021 and selected them according to the eligibility criteria. The Cochrane Handbook for Systematic Reviews (version 5.1.0) criteria were used for the quality assessment of randomized controlled trials, while the scoring protocol of the methodological index for non-randomized studies was used for non-randomized controlled trials. Statistical analysis was performed using the RevMan5.3 software. Results: All the included studies showed a relatively high success rate of expansion. The changes in both the intermolar and alveolar widths after MARPE were statistically significant. MARPE exhibited greater skeletal expansion effects than did conventional RPE. The midpalatal suture was opened in parallel after MARPE. A small amount of relapse was observed 1 year after expansion. MARPE caused tooth inclination and a decrease in alveolar height, but it was less significant than in conventional RPE. Conclusions MARPE may be an effective treatment modality for patients with MTD. It causes great transverse skeletal expansion in late adolescence. In comparison to conventional RPE, MARPE has lower detrimental periodontal effects and has certain clinical advantages.