Objectives To construct two retroviral vectors, one containing human interleukin-1 recep-tor antagonist (hIL-1Ra) gene and the other containing human interleukin-10 (hIL-10) gene and to transfect rabbit synoviocytes in vitro and detect the expression level of target genes. Methods RNA from human peripheral blood mononuclear cells were extracted and target genes were amplified by RT-PCR. The target genes were cloned into retroviral vector pLXSN, which was then transducted into GP2-293 cells to produce recombinant retrovirus. Rabbit synoviocytes were transfected and the expression of target genes was detected by RT-PCR, immunohistochemistry and western-blot. Results The retroviral vector containing hIL-1Ra gene or hIL-10 gene was constructed successfully. The hIL-1Ra gene and hIL-10 gene were transduced respectively into rabbit synoviocytes in vitro. The mRNA level of both genes reached peak in 5 days. In positive cell clones, the protein level of hIL-1Ra reached peak within 30 days and maintained at least 60 days; the protein level of hIL-10 maintained at least 40 days. Conclusion The hIL-1Ra gene and hIL-10 gene can be transduced successfully into rabbit synoviocytes by recombinant retrovirus.

Objective To explore the clinical application of FP-PCR method to detect MP-DNA.Methods Five hundred and sixty-three children suspected of MP infection were enrolled in experimental group. FP-PCR was adopted to detect MP-DNA. MP-DNA was re-detected later in 60 children. At the same time,MP-Ab (MP antibody) was detected by means of particle agglutination. MP-Ab was re-detected one or two weeks later. Also 20 healthy children were selected as the control group. Results The positive rate of MP-DNA and MP-Ab were 34. 99% and 35.52% respectively,which showed no significant difference (x2 =0. 31, P > 0. 05). The coincidence of the two methods was 97. 69%. But the positive rate of MP-DNA was significantly higher than that of MP-Ab in the early stage(30. 48% vs 10. 16%) (x2 = 74. 46, P < 0. 05).The sensitivity and specificity of FP-PCR were 96. 00% and 98.62% respectively. The result of reviewed MP-DNA was consistent with the clinical diagnosis. Conclusion FP-PCR method is very sensitive, convenient and stable. It is fit for the clinical application ,especially the diagnosis of early MP infection. It helps to identify those who had been infected with MP before.

Objective To discuss the diagnosis and early intervention treatment of fetal congenital broncho-genic cysts based on the cases reviewed. Methods The clinical features of 7 infants presenting bronchogenic cysts diagnosed antenatally from January 2013 to May 2014 in Guangdong Women and Children's Hospital and Health In-stitute were reviewed retrospectively. Pathology,the prenatal diagnosis and treatment of bronchogenic cysts experience were summarized combined with CT after birth and surgery. Results Based on the prenatal diagnosis of fetal and postnatal CT and surgical pathology,a total of 7 cases with congenital bronchial cysts were diagnosed. Of which 4 ca-ses were suggestive of congenital cystsic adenomatoid malformation by prenatal diagnosis,and the other 3 cases had fetal bronchial cysts by prenatal diagnosis,antenatal diagnosis was accurate in 42. 9%(3 / 7 cases). CT examinations were taken in 7 cases after birth,and the cyst excision was performed on them with surgery thoracic approach;the average age at surgery was(5. 3 ± 1. 7)months. Four cases had simple bronchial cystss,2 cases with congenital cystsic adenomatoid malformation,1 case with congenital pulmonary sequestration. The accuracy of CT diagnosis was 85. 7%(6 / 7 cases). All surgical treatment was effective. Conclusions Fetal bronchial cysts is always associated with the other presence of lung congenital malformations. Prenatal diagnosis of congenital bronchial cysts is difficult. Prenatal diagnosis is difficult to exclude congenital cystsic adenomatoid malformation. To avoid symptoms like oppres-sion,infection,prenatal diagnosis combined with CT examination after birth and early treatment are necessary and reliable clinically.

Objective To study the clinicopathologic characteristics of triple-negative breast cancer (TNBC) and its value in the prediction of prognosis. Method In this study,500 cases of female breast cancers were examined immunohistochcmically for the TNBC. The clinicopathologic characteristics of the 243 TNBC cases were inspected. Results TNBC accounted for 17.6% (88/500) of the 500 breast cancers. The histological types of the TNBC included mainly infihrative ductal carcinoma, metaplastic carcinoma and medullar carcinoma. Among those, histological grade Ⅲ accounted for 72.7% (64/88) of all the TNBC and was more common than that in hormone receptor positive breast cancers (HR+ group ) and Her-2 overexpression breast cancers (Her-2 group)(P=0.000). The positive rates of CK5/6 and EGFR in the TNBC were 30.7% (27/88) and 34.1% (30/88), respectively. The positive rates of ERCC1 and KIT in the TNBC were 28.4% (25/88) and 34.1% (30/88), respectively, Both of which were higher than those in the HR + group and Her-2 group, respectively (P=0.032 and P=0.026). 3-year survival rate of the TNBC was 71.5% and it was lower than that of HR group (P=0.021) and not significantly different from that of Her-2 group (P=0.474). Conclusions TNBC is the breast cancer with high aggressive pathologic futures and poor prognosis. EGFR and ERCC1 expression were positive in a portion of TNBC cases.

Objective To compare the metabolic characteristics, dosages and blood concentrations of tacrolimus (Tac) in patients subject to cadaveric liver transplantation (CLT) vs living-donor partial liver transplantation (LDLT) in order to investigate the usage of Tac in patients undergoing LDLT. Methods The clinical data of 85 patients undergoing liver transplantation from April 2007 to September 2009 were analyzed retrospectively. Thirty-four underwent LDLT (group A)and the remaining 51 underwent CLT (group B). Results The time to reach therapeutic window was shorter in group A (3. 4 ± 1.0 days) than in group B (4. 5 ± 2. 0 days, P = 0. 002). The Tac dosage in group A was significantly less than in group B during the first 28 days post-transplantation. However,the Tac dosage approached gradually and tended to be consistent after 28 days. On the postoperative day7, 14, 21 and 28 days, the Tac dosage in group A was 72.74 ％, 82.26 ％, 83.92 ％ and 88. 87 ％ of that in group B respectively. Correlation analysis revealed that graft-recipient body weight ratio (GR/WR) was significantly correlated with the Tac dosage on the day 7 (mg·day-1 · kg-1) (r =0. 728, P＜0. 01) and Tac concentration/dosage ratio (ng/ml)/(mg/kg) (r = - 0. 644, P＜0. 01 ).Conclusion The early Tac dosages in patients subject to LDLT were correlated significantly with the volume of graft. The early Tac dosages in patients undergoing LDLT were about 70 ％ of those in patients undergoing cadaveric liver transplantation. Moreover, with the regeneration of the liver, they tended to be consistent after 28 days.

Objective To investigate the value of DWI using 3.0 T MRI to predict response to radiotherapy(RT) and concurrent chemoradiotherapy(CCRT) in patients with advanced non-small cell lung cancer (NSCLC).Methods From January 2014 to May 2015, 40 patients with stageⅢ(Ⅲa orⅢb) NSCLC underwent DWI using 3.0 T MRI before RT/CCRT were enrolled. The imaging quality of diffusion-weighted images were evaluated on 3-level grades as good, moderate and non-diagnostic.The patients with good or moderate image quality were underwent DWI at 2 weeks after starting therapy(total dose of 20 Gy), and at the end of therapy (total dose of 60 Gy). Apparent diffusion coefficient(ADC) of lung cancer with good and moderate image quality were calculated by Funtool. The following quantitative parameters were recorded and calculated: the mean pretreatment ADC value(ADCpre), the mean mid-treatment ADC value (ADCmid), the mean post-treatment ADC value(ADCpost), the rate of changes inmean ADC value at 2 weeks post therapy (ΔADCmid) and the rate of changes inmean ADC value at the end of therapy(ΔADCpost). The patients were classified into response group and non-response group according to the tumor response, which was assessed with revised response evaluation criteria in solid tumors (RECIST1.1) after CCRT. The Mann-Whitney U test was used to compare parameters between the two groups.The relationship between these obtained parameters and tumor response was evaluated by Spearman correlation analysis. The value of parameters on predicting tumor response was calculated by receiver operating characteristic curve.Results 96.4%(80/83) DW images were graded as good or moderate image quality. The responders had lower median ADCpre[1.32 (0.77—1.96) × 10- 3 mm2/s] than non-responders[1.60(1.12—2.33) × 10- 3 mm2/s], which had statistically significant difference (Z=-2.934,P=0.003).Tumor regression rate after treatment had negative correlation with ADCpre(r=-0.386, P=0.018).The responders had increased ADC [ΔADCmid: 38.9%(12.8%—139.0%),ΔADCpost: 48.3% (25.6%—148.1%)] than non-responders [ΔADCmid: -2.5% (-15%—29.4%), ΔADCpost:14.2% (- 28.1% —71.3% )], which had statistically significant difference (Z=- 2.847, - 2.221, respectively;P<0.05). Tumor regression rate after treatment had positive correlation with ΔADCmid(r=0.637, P=0.001) and ΔADCpost(r=0.631, P=0.005).From ROC analysis,when setting threshold on pretreatment ADCpre=1.38 × 10-3 mm2/s, ΔADCmid=21.6%, ΔADCpost=38.8%, the area under curve was 0.782, 0.838 and 0.813.Conclusion The mean ADC value before RT/CCRT and its changes during treatment is likely to be a valuabletool for predicting the response after RT/CCRT in advanced NSCLC, which may be helpful to clinical decision on individualized therapy.

AIMTo prepare the PEGylated liposomes modified with cell penetrating peptides, which protect siRNA from nuclease degradation and deliver efficiently siRNA into cells to facilitate silencing of target gene.

METHODSThe purity of R8-PEG-PE and pNP-PEG-PE was detected by HPLC; the quantity of R8, PEG-DPPE modified R8, and R8 attached to the out membrane surface of the liposomal siRNA by transfer from R8-PEG-DPPE micelles to the liposomes was tested by fluorescence; Size and size distribution of siRNA loaded liposomes with and without attached R8 were determined by Zetasizer 5000; A comparison of mediated siRNA transfection efficiency between R8-liposomes and lipofectamine 2000 was examined by individual inside cell fluorescence intensity; The growth inhibition of small cell lung carcinoma NCI-H446 cells treated with R8-liposomal hdm2-siRNA or lipofectamine 2000-hdm2-siRNA complex was tested by MTT assay.

RESULTSThe retention times of PEG-DPPE and R8-PEG-DPPE were 9.0 min and 7.8 min, respectively. Fluorescence scanning indicated that lipids composed of liposomes and siRNAs didn't interfere to the determination of R8 when it was attached to the liposomal siRNA. The cells treated with R8-liposomal hdm2-siRNA significantly enhanced the cellular uptake of hdm2-siRNA and facilitated the functions of hdm2-siRNA through silencing of target gene which, in turn, inhibited tumor cell growth, compared with lipofectamine 2000.

CONCLUSIONThe R8 attached liposomes are shown to be powerful carriers for delivery siRNAs into cell to silence targeted gene.

Carcinoma, Small Cell ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gene Silencing ; Gene Targeting ; Humans ; Liposomes ; Lung Neoplasms ; metabolism ; pathology ; Oligopeptides ; administration & dosage ; pharmacology ; Phosphatidylethanolamines ; Polyethylene Glycols ; Proto-Oncogene Proteins c-mdm2 ; genetics ; metabolism ; RNA, Small Interfering ; administration & dosage ; metabolism ; pharmacology ; Transduction, Genetic

ObjectiveTo surnmarize the experience of tacrolimus or cyclosporine A-based immunosuppression after pediatric living-donor liver transplamation.Methods The clinical data of 30 children undergoing living-donor liver trarsplantation from October 2006 to January 2010 were analyzed retrospectively.In 30 patients,7 were given Tac-based immunosuppression (group A),10 given CsA-based immunosuppression (group B),and 13 switched from CsA to Tac for complications or adverse effects of drugs.Dosages and blood concentrations of immunosuppressants were recorded.Changes of liver and kidney functions were monitored.Incidence of rejection,infection and adverse effects of drugs were observed.ResultsIn the premise of the stable concentration and liver and kidney functions,the weight of children was increased by about 50％ and the per- kilogram dosage of CNIs was decreased significantly 1year postoperatively.There was no case of rejection in group A and 4 cases of rejection in group B(40％,4/10),and the original symptoms were gradually alleviated after the increased dosage in immunosuppressants.During the first 3 months,there was 1case of abdominal infection in group A (1/7) and 3 cases of lung infection in group B (3/10),and the original symptoms were gradually alleviated after anti-infective therapy.There was 1CMV lgM-positive case in group A (1/7) and 2 CMV IgM-positive cases in group B (2/10),and the original symptoms were gradually alleviated after using ganciclovir.The original symptoms of the 13 children switched from CsA to Tac were gradually alleviated.ConclusionThe two CNIs can be safely used in children undergoing pediatric livlng-donor liver transplantation.Both of them show the same effect in promoting the restoration of liver and kidney functions,but tacrolimus has more satisfactory effect in inhibiting the rejection and it has leas adverse effects.

Objective To evaluate the efficacy of Epipolis laser in situ keratomileusis(Epi-LASIK)and laser in situ keratomileu- sis(LASIK)on high myopia patients.Design Prospective,case-controlled study.Participants 62 patients(123 eyes)with high myopia. Methods 62 patients(123 eyes)underwent Epi-LASIK or LASIK surgery for high myopia:28 patients(56 eyes)underwent Epi-LASIK and 34 patients(67 eyes)underwent LASIK.The differences in postoperative uncorrected visual acuity(UCVA),best corrected visual acuity(BCVA),refraction,and root mean square(RMS)of high-range wavefront aberration were compared one-week,one-month, three-months and six-months postoperatively.Main Outcome Measures UCVA,BCVA,refraction,and RMS of high-range wavefront aberration.Results There was no serious complication during and after the operation.The recovery of postoperative UCVA after Epi-LASIK was slower than that of LASIK.One week postoperatively,the proportion of UCVA≥0.8 of Epi-LASIK(46.4%)was less than that of LASIK(77.6%)(P=0.0003).No significant differences were found in those of Epi-LASIK(85.7%,94.6%,91.1%)and those of LASIK(92.5%,95.5%,94.0%)one-month,three-months and six-months postoperatively(P=0.590,0.822,0.530).BCVA was same after Epi-LASIK and LASIK.The proportion of mean spherical equivalents within?0.50D for Epi-LASIK(42.9%,51.8%,60.7%,64.3%)had no difference with those for LASIK(53.7%,59.7%,71.6%,73.1%)one-week,one-month,three-months and six-months postoperatively (P=0.230,0.378,0.200,0.290).The postoperative RMS increased after both surgeries,especially after LASIK.At postoperative one-month,three-months and six-months RMS of Epi-LASIK(1.51?0.77)?m,(1.32?0.76)?m,(1.18?0.71)?m were much higher than the (0.87?0.27)?m preoperative ones(P=0.016,0.019,0.026).At postoperative one-month,three-months and six-months RMS of LASIK (2.41?0.81)?m,(2.17?0.63)?m,(1.89?0.87)?m were also much higher than the preoperative(0.91?0.22)?m(P=0.011,0.008,0.006). There were significant differences between the RMS of Epi-LASIK and LASIK one-month,three-months and six-months postoperatively (P=0.039,0.035,0.033).The I grade haze was found in two eyes after Epi-LASIK.Conclusion Epi-LASIK has better visual quality re- sult than LASIK on correcting high myopia.(Ophthalmol CHN,2007,16:336-339)

Objective To develop a quantitative scoring system for the House -Tree -Person (H -T -P)test.Methods By literature review and expert consultation,preliminary evaluation items were defined.After pilot test,the scoring system for assessing depression were defined.A total of 21 6 cases of patients with depressive mood disorder from the Tong De hosptial of the Zhejiang Province were tested.The reliability and validity of a scoring system of house -tree -person test for depression were analyzed.Results The scoring system of depression evaluation including 23 items and 5 factors,could explain 77.50% of the cumulative total variance.The split -half reliability of the system was 0.87 and 0.88,and the internal consistency reliability of the system was 0.81 .The Kappa coefficient of each item was 0.71 -0.89 (P <0.01 ). The standard validity of the scale was 0.65,and the criterion validity of each factor was 0.34 -0.60 (P <0.01 ).The critical point of positive demarcation was 4,and the critical point of positive demarcation of each factor was 1 .The Kappa value of the consistency test was 0.85 with the PHI Depression Inventory.Conclusion This study demonstrates H -T -P scoring system has acceptable validity and internal consistency.