The DNA copy-number variant (CNV) is a kind of segments of DNA ranging from 1 kb to 3 Mb that is present in a variable number of copies.CNVs widely distribute across the human genome,and dramatically increases genetic diversity.In recent years,researches have found that most CNVs are closely related to complex diseases.If a cancer gene is directly encompassed or overlapped by a CNV,it may lead to activation of oncogenes or inactivation of tumor suppressor genes,and finally results in tumorigenesis.CNVs can affect gene expression,phenotype differences and phenotypic adaptations by changing gene dosages and gene activities,and then sequentially lead to tumor or any other genetic dieases.Investigating CNVs is apparently helpful for studing chromosome recombination,genomic evolution,gene expression and the pathogenesis of multiple complex diseases especially tumor.

Objective:To investigate the impact of polysomy 17 of breast cancer on testing results of human epidermal growth factor receptor 2 (HER2) and its clinicopathologic significance. Methods:Seventy-one patients with primary invasive breast carcinoma were studied. The HER2 gene and chromosome 17 copy numbers were determined by dual-color fluorescence in situ hybridization (FISH). The testing results were expressed by absolute HER2 gene copy number or the ratio of HER2 to chromosome 17. Based on the FISH testing results and HER2 protein expression determined by immunohistochemistry the results were compared between different groups divided by related clinicopathologic parameters.Results:All patients who had doubtable FISH results, either by absolute HER2 copy number (14 of 71 patients; 19.7%) or by the ratio HER2/chromosome 17 (2 of 71 patients, 2.8%), displayed polysomy 17. Polysomy 17-positive patients had no significant difference with HER2-negative patients in tumor grade, lymph node metastasis, and estrogen receptor (ER) expression (all P>0.05); but compared with HER2-positive patients, they showed lower tumor grade (50.0% vs 81.5%, P=0.025), higher rate of negative lymph node (55.6% vs 25.9%, P=0.045), and higher rate of ER positive expression (83.3% vs 41.7%, P=0.005) and progesterone receptor(PR)positive expression (87.5% vs 44.4%, P=0.003).Conclusion:Compared with HER2 gene amplification group, polysomy 17-positive group tends to have negative HER2 gene expression. Polysomy 17 influences the testing results of HER2 and may be the main factor that caused doubtable results in FISH examination.

Objective:To investigate the efficiency of maternal serum triple screening for the genetic abnormality in second-trimester and the morbidity of adverse pregnancy outcome in false positive results of the test. Methods: A total of 4 680 pregnant women with singleton pregnancies assigned in Obs & Gyn Hospital, Fudan University, underwent triple screening test (alpha fetoprotein, AFP; human chorionic gonadotropin, HCG and unconjugated estriol, uE3) by fluorescence enzyme immunoassay between 2003 and 2005. The valid MoM (Multiples of Median) value of mid-trimester serum AFP, uE3, and hCG and risk assessments was provided by Beckman Coulter Co. When applied in the prenatal Down syndrome screening service. The study compares the incidence of chromosomal abnormalities with Down syndrome in screen positive women and compares to the MoM value established in the literature. The risks of having a fetus with congenital abnormalities or of developing obstetric complications in the screen positive women with their matched controls.Results:The MoM values for the triple tests of our study are similar to established values of literature. Only 51.01% women with pregnancies agree to receive screening. Amniocentesis utilization rate was 55.12% in the screen-positive pregnancies. The false positive rate was 6.89% and the median of maternal age of the women was 28.13 (range 19 to 49) years old. Chromosomal abnormalities were identified in 21 pregnancies, including 9 cases of trisomy 21.The detection rate was 77.77%. Pregnancies with positive screening results had a significantly higher risk of adverse outcomes than those with negative results (P< 0.05). Whereas there was no difference in the incidences of fetal congenital appearance or skeleton abnormality. Conclusion: Adjusting MoM values of local unaffected populations is limited to increasing the detection rate. Because chromosomal defects have variable exhibitions, amniocentesis utilization is still a choice for screen-positive pregnancies. Screen-positive pregnancies had increased risk of chromosomal abnormalities.

Objective To investigate the efficacy of the combination of bone marrow stromal cells (BMSCs) and bcl-2 gene in the treatment cerebral ischemia taxi its effect on the expression of basic fibroblast growth factor (bFGF) in rats.Methods Forty Wistar rats were used to establish middle cerebral artery occlusion model. They were randomly divided into 4 groups: saline control, bcl-2, BMSCs and BMSCs + bcl-2 groups (n = 10 in each group). Every group was redivided into 3- and 14-day after reperfusion subgroups (n = 5 in each subgroup). Neurological scores of the experimental rats were assessed. BMSCs were labeled with bromodeoxyuridine (BrdU). The distribution and numbers of BMSCs, the expressions of Bcl-2 and bFGF were detected by immunohistochemistry, and apoptotic cells were detected with TUNEL staining in rat brain. Results The neurological score at day 3 after reperfusion in the BMSCs + bcl-2 group was significantly lower than that in the saline control group (P < 0. 05), and at day 14, it was significantly lower than that in the other 3 groups (all P <0. 05). A large number of BrdU-positive BMSCs were observed in the infarcted hemisphere in the BMSCs + bcl-2 and BMSCs groups. The numbers of BrdU-positive BMSCs at day 3 and 14 after reperfusion in the BMSCs + bcl-2 group were significantly higher than those in the BMSCs group (all P <0. 05). The expressions of Bcl-2 in the infarcted hemisphere at day 3 and 14 after reperfusion in BMSCs +hel-2 group wre significantly higher than those in the other 3 groups (all P <0. 05). The expressions of Bcl-2 at all time points were increased more significantly than those in the other 3 groups (all P <0. 05). The numbers of apoptosis in brain at all time points in the BMSCs + bcl-2 group were decreased more significantly than those in the other 3 groups (all P <0. 05). Conclusions Both BMSCs and bcl-2 genes have the therapeutic effect on cerebral ischemia. The efficacy of combination of both ,of them is significantly superior to monotherapy. They may significantly improve the neurological function and increase the expression of bFGF in rats. Its mechanism may be that bcl-2 genes have inhibited BMSCs apoptosis at the same time of anti-apoptosis in brain.

To screen interaction proteins of CVB3 VP3 from cDNA library of human heart ,yeast two hybridization was conducted in this study .The bait plasmid pGBKT7-VP3 was constructed ,VP3 fusion protein and its self-activation in AH109 yeast cells was then detected .The positive clones were confirmed by PCR amplification of cDNA inserts ,Alu I digesting ,DNA sequencing ,and Blasting were used to sort positive colonies to eliminate duplicates .Positive clones were confirmed by one-to-one yeast two hybridization ,and them were sequenced and analyzed for homology .Theα-galactosidase assay was performed to detect the interaction strength .Totally ,10 positive proteins interacting with VP3 of CVB3 were obtained by homology analy-sis,namely,EIF4A2,HADHB,GAPDH,ASPG,ACTA1,TNNI3,CKM,LMOD3,ERGIC1,and ALDH2.The strength of interactions between VP3 and 10 candidate proteins were proved byα-galactosidase assay .This study will contribute to explore the CVB3 VP3 function on molecular level and provides some new clues to explain the pathogenic mechanism of myo-carditis and cardiomyopathy .

Objective To prepare 18F-5-fluoro-N-(2-(diethylamino) ethyl) picolinamide (18F-5-FPN) and evaluate its binding affinity with melanin.Methods 5-bromo-N-(2-(diethylamino) ethyl)picolinamide (precursor) was synthesized and the structure was characterized by ultraviolet (UV),nuclear magnetic resonance (NMR) and mass spectrometry.18F-5-FPN was prepared with FX-XN module through nucleophilic substitution reaction.The product was purified and identified by HPLC.The binding specificity of 18F-5-FPN with melanin was demonstrated by in vitro study of cellular uptake,and in vivo static PET imaging of pigmented B16F10 and amelanotic A375m allografts.Biodistribution study was performed to evaluate the pharmacokinetics of 18F-5-FPN in vivo.Two-sample t test was used for data analysis.Results The structure of precursor was characterized by UV,1H NMR,13C NMR and mass spectrometry.18 F-5-FPN was successfully prepared with radiochemical yield of 5％-8％,radiochemical purity ＞95％ and the specific activity of 100-120 GBq/μmol.The cell uptake study showed that the uptakes of 18F-5-FPN in B16F10 cells at 30,60 and 120 min ((9.80±0.46) ％,(10.34±0.32) ％,(7.27±0.26)％) were significantly higher than those in A375m cells ((1.36±0.14)％,(1.75±0.12)％,(1.54±0.09)％;t =30.3,46.8,38.4,all P＜0.05),and the optimal uptakes were observed at 60 min for both cells.In static PET imaging,the tumors in B16F10-bearing mice were clearly visible with the uptake value of (18.20±3.21) ％ID/g and the T/B ratio of 19.17±10.03 at 1 h postinjection,while no tumor uptake was seen in A375m-bearing mice.The main clearance pathway of 18F-5-FPN was the renal system,which cleared the unbound tracer rapidly.Conclusions 18F-5-FPN can specifically target the melanin in vitro and in vivo with favorable pharmacokinetics and good T/B ratio.18F-5-FPN may be an ideal molecular probe for diagnosis of malignant melanoma.

Objective To prepare a kind of 99Tcm-labeled estrogen receptor (ER) SPECT imaging agent,and evaluate its binding characteristics with ER by in vitro and in vivo studies.Methods EDL was prepared from estrone and then reacted with GAP to synthesize GAP-EDL.Then,GAP-EDL was labeled with 99Tcm to obtain 99Tcm-GAP-EDL.Cell uptake and blocking assays were performed in vitro on MCF-7and MDA-MB-231 cells.The biodistribution study of 99Tcm-GAP-EDL was performed on normal BALB/c mice at 30,60,120,180 and 240 min post injection.Nude mice bearing either MCF-7 or MDA-MB-231derived tumors were injected with 99Tcm-GAP-EDL,and subjected to γimaging at 1,2,4 h post injection.The mice injected with excess unlabeled GAP-EDL or estradiol were used as the blocking control.Two-sample t test was used for data analysis.Results The radiolabeling yield of 99Tcm-GAP-EDL was (98.8 ±0.5) ％ and the radiochemical purity after 24 h was over 90％.The cell uptake rate of MCF-7 cells at 240min was (4.84± 0.21) ％,which was significantly higher than that of MDA-MB-231 cells ((2.11±0.21) ％;t =15.96,P＜0.05).99 Tcm-GAP-EDL uptake rate of MCF-7 cells in blocking groups ((2.31 ± 0.28) ％ and (2.05±0.35) ％) decreased significantly compared to that of non-blocking group(t=11.52,11.16,both P＜0.05).Biodistribution studies showed that 99Tcm-GAP-EDL was mainly metabolized by the liver and kidneys,and exhibited quick blood clearance.Gamma imaging showed high uptake in MCF-7 tumors 1 h post injection and the uptake reached the highest at 4 h,while there was little 99Tcm-GAP-EDL uptake in MDA-MB231 tumors and blocked MCF-7 tumors.Conclusions 99Tcm-GAP-EDL may be prepared under mild conditions with high labeling purity and stability.The in vitro and in vivo characteristics of 99Tcm-GAP-EDL suggest that it may be a promising probe for ER positive tumor imaging.

Objective To evaluate the influences of FBP and OSEM (with different iteration numbers or subset numbers) on cardiac function parameters with MPI,and to identify a suitable clinical method.Methods The MPI of 66 patients (33 males,33 females,age range: 18-77 years) from January 2012 to December 2014 were retrospectively analyzed.The imaging data was reconstructed with FBP and 5 different OSEM (4-16 subsets and 2-4 iterations) in order to evaluate if there was any difference of cardiac function parameters calculated by QGS between the different image processing methods.In addition,the results were compared with those of cardiac ultrasonography.One-way analysis of variance was used for data analysis.Results The statistical differences of LVEF,EDV,and ESV(F values: 10.73,4.89 and 5.97,all P<0.05) were found among 6 reconstructed methods.The values of LVEF were (66.14±11.12)％,(75.05±12.10)％,(70.09±11.27)％,(66.88±10.38)％,(64.97±10.25)％,and (62.58±9.84)％.Those of EDV were (77.32±27.58),(67.97±27.56),(75.10±27.89),(81.03±28.11),(84.94±29.07),and (89.98±29.71) ml,and the ESV values were (28.71±10.04),(19.71±16.51),(25.13±17.66),(29.01±18.47),(32.10±19.63),and (35.83±20.41) ml respectively.The cardiac function parameters measured by OSEM (with 2 iterations,12 subsets) were much similar to that measured by cardiac ultrasonography.Conclusion Compared with other 5 processing methods of MPI,the OSEM (with 2 iterations,12 subsets) method may be more suitable for practical clinical application.

Objective To investigate the effect of multidisciplinary diagnosis and treatment including ex-utero intrapartum treatment (EXIT) procedure to improve the prenatal survival rate of fetus with neck mass.Methods Multidisciplinary diagnosis and treatment model were carried out in four pregnancy women with fetal neck mass from September 2007 to February 2010.The model included prenatal assessment and monitoring,EXIT procedure during cesarean section,neonatal reassessment and surgical treatment by the cooperation of obstetricians,neonatologists,children surgeons,sonographers and anesthetists.Results All patients underwent cesarean section after 37gestational weeks.Mean delivery time was 37+4 weeks (37-38+3 weeks); mean birth weight was 2972 g (2600-3250 g); mean operation time was 4 min (2-7 min).The gestational age of primary diagnosis of fetal neck mass was 24-34 gestational weeks.After delivery,the size of neck mass was from 3.0 cm × 2.0 cm × 1.0 cm to 6.2 cm× 5.8 cm × 6.8 cm.The tracheal compression and displacement were found by color doppler ultrasound scan and magnetic resonance imaging in all cases.Two of them were completed with polyhydramnios and the others with normal volume of amniotic fluid.EXIT procedure was successfully carried out during cesarean section.Neonatal reassessment showed the trachea of three infants were obviously compressed and lapsed by enhanced CT; the infants relied on mechanical ventilation after birth and underwent operation on day 6 to 8.Tracheal impression was not presented in one infant and trachea cannula was removed on the second day,operation was not performed.All of those infants had good outcomes.Conclusions The multidisciplinary diagnosis and treatment model,including EXIT procedure,is a safe,efficient and feasible strategy,which is necessary for fetus with neck mass.

Objective To explore the high risk factors of stillbirth. Methods 176 cases of stillbirth were collected in the Obstetrics and Gynecology Hospital of Fudan University from January 1st, 2010 to December 31st, 2016. All cases were analyzed retrospectively, including general profile, high risk factors of stillbirth in different years and pregnancy periods. Results (1) The incidence of stillbirth was 0.178%(176/98 785). Stillbirth occured mostly at 28-28+6gestational weeks (10.8%,19/176), and the second peak was 29-29+6weeks(10.2%,18/176),while the third common period was 37-37+6weeks(9.1%,16/176).After 39 weeks,it maintained at a low level.(2)The top 5 high risk factors of stillbirth were infection (18.2%,32/176), unexplained (13.6%,24/176), hypertention disorders in pregnancy (13.1%, 23/176), umbilical cord torsion(12.5%,22/176)and fetal malformations(10.2%,18/176).(3)From 2010 to 2012,the top 3 high risk factors were unexplained, the umbilical cord torsion and infection, while hypertention in pregnancy,infection and fetal malformation became the top 3 high risk factors after 2013.(4)Early stillbirth (20-27+6weeks)accounted for 21.6%(38/176);and unexplained(47.4%,18/38),fetal edema(13.2%,5/38), infection(13.2%,5/38),umbilical cord torsion(5.3%,2/38)were the top 4 high risk factors.Late stillbirth(≥28 weeks)accounted for 78.4%(138/176),with infection(19.6%,27/138),hypertention in pregnancy(15.9%, 22/138), umbilical cord torsion (14.5%,20/138) and fetal malformation(12.3%,17/138)being the top 4 high risk factors. Conclusions More attention should be paid to maternal complications, especially infection and hypertension in pregnancy. Antenatal fetal monitoring, timely termination of pregnancy, standard management of stillbirth and looking for the causes may help reduce the incidence of stillbirth.