Along with the development of screening, diagnostic and therapeutic technologies, the spectrum of fetal abnormalities has been constantly expanded. This brings increasing challenges to the clinical diagnosis and treatment, including but not limited to optimizing multidisciplinary cooperation, options of prenatal genetic testing methods, the uncertainty in the transition period of new technology implementation, and the comprehensiveness of genetic counseling before and after testing. We discuss the above issues aiming to meet the dilemma and achieve the leap of fetal medicine to the advanced level through multidisciplinary collaboration resulting in the improvement of diagnosis and treatment efficiency.

Objective To explore the metabolic phenotype of the adult offspring rats with maternal severe hyperglycemia during pregnancy and overfeeding in early life. Methods The adult Wistar rats were administered intraperitoneally with 20% Streptozotocin (STZ, 50 mg/kg) on day 5 of pregnancy to induce severely gestational diabetes mellitus (SDM) model (blood glucose ≥20 mmol/L). Early postnatal overfeeding models were established through reduction of the number of newborn rats fed by one mother rat. Offsprings were divided into 3 groups according to maternal blood glucose level during the pregnancy and feeding patterns during the lactation period (1) control pups (CP) :maternal euglycemia was achieved during pregnancy and eight pups fed by one mother rat; (2)SDM-normal feeding group: SDM mothers and 8 pups fed by one mother rat;(3)SDM-overfeeding group:SDM mother and 4 pups fed by one mother rat. All pups were fed by standard laboratory chow adlibitum after weaning at 3 weeks of age. The body weight of all pups were measured weekly after weaning. At the age of 26 weeks, the systolic blood pressure (SBP), diastolic blood pressure (DBP) ,heart rate (HR) and metabolic markers, including fasting plasma glucose (FPG), fasting insulin (FINS), total triglyceride (TG), total cholesterol (TC), low density lipoprotein-cholestrol (LDL-C)and high density lipoprotein-cholestrol (HDL-C) were measured in all 3 groups. ANOVA and LSD test were applied in statistical analysis. Results The average plasma glucose level was significantly higher in the SDM mothers than in the controls [(28.34±5.14) mmol/L va (6.25±1.41) mmol/L,P＜0.05]. After weaning at 3 weeks, the weight of SDM-overfeeding group and SDM-normal feeding group was lighter than that of CP group [(43.63±4.83) g, (31.45±10.21) g vs (55.75±8.41) g,P＜0. 05], meanwhile, that of the SDM-overfeeding group were heavier than that of the SDM-normal feeding group (P＜0. 05). Pups in SDM-overfeeding group exhibited catch-up growth during the lactation period, and those in SDM-normal group showed catch-up growth at both lactation period and childhood. At 26 weeks of age, the systolic blood pressure and TG level of pups in SDM-normal feeding and overfeeding group were (153.31 ± 13.91) mm Hg and (147.21 ± 12.29) mm Hg, and (0. 73±0. 22) mmol/L and (0. 71±0.49) mmol/L, respectively, which were higher than those of the CP group [( 132.21 ± 11.26) mm Hg, and (0. 37 ± 0. 08) mmol/L, P＜0.05], but no significant difference was found between the two SDM groups (P＞0.05). There was no difference in FPG levels among the three groups, but the FINS level and HOMA-IR in the SDM-overfeeding group were higher than in the SDM normal feeding and CP group [( 12. 552 ± 3.260) mU/L vs (9.067 ± 1.782) mU/L and (8.590± 0.806) mU/L, 2.400± 0.624 vs 1.797 ± 0.508 and 1.729 ± 0.246; P＜0.05].Conclusions Fetal exposure to maternal severe hyperglycemia during pregnancy may lead to low birth weight infant who will exhibit postnatal catch-up growth. This may lead to the increased risk of metabolic syndrome in the offspings when they grow up, and this process would be accelerated by early overfeeding.

Objective To evaluate the expression and distribution of Toll-like receptor 4 (TLR4) in term placentas of women with gestational diabetes mellitus (GDM). Methods Placental samples were collected from 36 normal full-term singleton pregnant women (control group) and 33 full-term singleton pregnant women with GDM who had elective cesarean section in Peking University First Hospital between November 2014 and June 2015. Immunohistochemical assay was used to detect the expression and distribution of TLR4 in placental tissues. T test was used to compare the expression of TLR4 in various cell types of placenta by semi-quantitative analysis. Results (1) TLR4 was expressed in the cell membrane, cytoplasm or nuclei of trophoblast, decidual cells, vascular endothelial cells and amniotic epithelial cells of term placentas. (2) Compared with the control group, the expression of TLR4 was significantly enhanced in trophoblast and decidual cells of GDM women (0.38±0.01 vs 0.31±0.01, 0.39±0.01 vs 0.34±0.01, t=5.218 and 4.525, all P<0.01). Moreover, the change of TLR4 average optical density was most significant in trophoblast. (3) There was no difference in the expression of TLR4 in vascular endothelial cells and amniotic epithelial cells in term placentas of GDM women compared with the control group (all P>0.05). Conclusions The expression of TLR4 is different in various cell types of GDM term placentas.

Karyotype analysis has been considered as the key tool for prenatal diagnosis .Although it is cost-effective, it has great challenge to meet the growing demand of efficiency and quality in clinical settings.To improve the effeiciency and detection quality , cytogenomic microarray analysis ( CMA ) is developed, with high detection rate.However, traditional karyotype analysis at different resolution should also be used as the reference for CMA .

Objective To study the clinical significance of chromosome karyotyping in pregnant women with a history of abnormal pregnancy. Methods The fetal chromosome karyotypes of 1 193 pregnant women with a history of abnormal pregnancy in Peking University First Hospital from January 4, 2005 to December 31, 2013 were analyzed. According to the etiology of their previous abnormal pregnancy, these women were divided into four groups: 273 women had children with inherited metabolic disorders or single-gene genetic diseases (group A), 81 women had children or fetuses with chromosome abnormalities (group B), eight cases had an abnormal chromosomal karyotype in either husband or wife (group C), and 833 women had abnormal pregnancy of unknown causes(group D). Results Forty-eight [4.0%(48/1 193)] and fetuses were found to have abnormal chromosomal karyotypes, including 26 cases of chromosome polymorphism variations and 22 cases of numerical and structural abnormalities (four cases of trisomy 21, four cases of numerical sex chromosome abnormalities, three cases of trisomy 18, three cases of extra small chromosome mosaicism, three cases of reciprocal translocation, one case of Robertsonian translocation, one case of chromosome six inversion between the arms, one case of chromosome three inversion between the arms, one case of mosaicism of trisomy 14 and one case of structural abnormality of chromosome 14). In group A, four cases (1.5%) of chromosomal abnormalities of clinical significance and four cases of chromosome polymorphism variations were detected. Meanwhile, 61 fetuses with inherited metabolic disorders or single-gene genetic diseases and two cases of gene mutation carriers were detected in group A, but among whom, there were no abnormal chromosome karyotype cases. In group B, two cases (2.5%) of chromosomal abnormalities were found. In group C, two cases (2/8) of reciprocal translocation were found, whose karyotypes were the same as the parents. In group D, three cases of trisomy 21, three cases of trisomy 18, two cases of extra small chromosome mosaicism and two cases of numerical sex chromosome abnormalities were found. All the mothers in this group were of advanced age. Four cases of structural abnormalities and 22 cases of chromosome polymorphism variations were also found in this group, chromosomal analysis was subsequently performed in those couples, and found that the abnormal chromosomal karyotypes in the fetuses were the same as those in the parents. Conclusions Appropriate prenatal cell genetic diagnostic methods should be chosen according to the causes of abnormal pregnancy history.

Objective To investigate the role of cerebrospinal fluie( CSF)cytology on eiagnosis of meningeal cancer. Methods Retrospectively analyzee the clinical eata of 23 cases with meningeal cancer. Results (1)Of 23 patients,CSF eetection of 17 cases were showee with cancer cells at the first lumbar puncture,3 cases at the 2ne lumbar puncture,2 cases at the 3re eetection,ane 1 case at 4th eetection.(2)Of 23 cases with cerebrospinal fluie cytology positive patients,17 cases were carriee cranial MRI scan. The MRI showee that 9 were normal ane 8 cases were brain parenchyma ane meningeal image enhance. Ten cases were performee the enhancee MRI scan,5 cases were with extensive meningeal thickening,3 cases were showee meningeal extensive enhancement of brain surfer ane intracranial cerebral sulci,1 case was the circular shaeow strengthen at eifferent size at next to the eouble lateral parietal,occipital ane left cerebella hemisphere,ans 1 case was with tough brain surface ane abnormally long T1,long T2 signal at lateral ventricles,thire ventricle, fourth ventricle epeneymal.(3)The relationship between the primary tumor ane cancer eetection in cerebrospinal fluie:19 patients were foune with primary tumor lesions,inclueing 5 cases leukemia(3 cases with acute lymphoblastic leukemia,2 cases with acute non-lymphocytic leukemia),4 cases with lung cancer,3 cases with breast cancer,2 cases with brain lymphoma,1 case with melanoma,1 case with Hoegkin′s lymphoma,l case with nasopharyngeal carcinoma, 1 case with vaginal squamous cell carcinoma, 1 case with gastric carcinoma. Conclusion Multiple cerebrospinal fluie cytology eetection may improve meningeal cancer eetection rate. CSF cytology eetection can improve eiagnosis rate of meningeal cancer. No relationship between the eetection of cancer cells in cerebrospinal fluie ane brain MRI meningeal lesions,ane the further research neee to be eone with expaneing the sample size.

Objective To analyze the feasibility of rapid prenatal diagnosis in the advanced maternal age women with or without positive serologic screening results.Methods We conducted a retrospective study of the women who underwent a mid-trimester amniocentesis in Peking University First Hospital from January 1,2001 to December 31,2012.Maternal age,indication for invasive prenatal diagnosis,karyotyping and pregnancy outcome were documented.Using a young population with high risk in serologic screening (S) as the standard,chromosome abnormalities in the advanced maternal age (A) group and the advanced maternal age with high risk in serologic screening (A+S) group were compared with the S group.Chromosome abnormalities were divided into detectable (D) and undetectable (U) during rapid prenatal diagnosis.Results Of 9 606 cases,222 (2.3％,222/9 606) cases with chromosome abnormalities were detected,23.0％ (51/222) of which were undetectable by rapid prenatal diagnosis.The detection rate of detectable chromosome abnormalities was 1.8％ (57/3 177) in group A,1.4％(13/925) in group A+S,and 1.8％(57/3 250) in group S (x2=0.662,P＞0.05).The rate of undetectable chromosome abnormalities was 0.5％ (15/3 177) in group A,0.3％ (3/925) in group A+S,and 0.5％ (16/3 250) in group S (x2=0.452,P＞0.05).The most common indications for undetectable chromosome abnormalities in the young population were abnormal history of pregnancy,abnormal family history and chromosome abnormality history (16.4％,9/55),and abnormal ultrasound in the advanced maternal age population (4.4％,3/68).Conclusions The performance of rapid prenatal diagnosis in the advanced maternal age population with or without high risk in screening without abnormal findings in ultrasound,was similar to the young population with high risk in screening.Fluorescent in situ hybridization may be integrated into the strategy of prenatal diagnosis for this group of women.

Objective:To determine synchronously protocatechuic aldehyde and danshensu in Compound Danshen Lozenge and Dropping Pills.Methods:HPLC method was used.Results:The average recoveries(n=5) were 100.47% for Danshensu of Compound Danshensu Lozenge(RSD=1.08%) and 99.27% for protocatechuic aldehyde(RSD=1.22%), respectively. And the average recoveries(n=3) were 100.40% for Danshensu of Compound Danshen Dropping Pills(RSD=0.65%) and 98.47% for protocatechuic aldehyde(RSD=1.76%).Conclusion:This method is simple, quick with a good separation.

Aim To compare the cytotoxicity of geni-poside (GS)and its metabolite genipin (GP)on hu-man hepatocelluar HepG2 cells and explore the sub-stance and mechanism of hepatotoxicity induced by Fructus Gardeniae.Methods The cytotoxic effect of GS and GP was analyzed by MTT method;the antioxi-dant enzyme activities of manganese superoxide dis-mutase (Mn-SOD),catalase (CAT)and levels of glu-tathione (GSH)were detected by respective kits;the change of intracellular reactive oxygen species (ROS ) was measured by 2′,7′-dichlorofluorescin diacetate (DCFH-DA)staining method;multiparameter cytotox-icity analysis (cell loss,nuclear size and morphologi-cal changes,DNA content,cell membrane permeabili-ty,mitochondrial membrane potential changes,cyto-chrome C release ) were measured simultaneously by high content screening (HCS)assays.Results No cytotoxicity was found in GS (20~1 000 μmol·L-1 ) groups (P>0.05 ),but GP was found to exert obvi-ous cytotoxic effect,and 50μmol·L-1 GP could obvi-ously inhibit HepG2 cell proliferation (P<0.05 ),and the IC50 value was (450.00 ±26.15)μmol·L-1;GS showed no obvious effects on Mn-SOD, CAT, GSH ,ROS (P >0. 0 5 ),GP could significantly decrease the activity of Mn-SOD,CAT and the level of GSH,and obviously increase the content of ROS (P<0.05 or P <0.01 );treatment with 50,500,1 000μmol·L-1 GP resulted in loss of mitochondrial mem-brane potential,cytochrome C release and increased cell permeability (P<0.05 or P<0.01),500,1 000μmol· L-1 GP could also show abvious nuclear con-densation,increased total nuclear intensity and cell loss (P<0.0 1 ).Opposed with GP,GS had no signif-icant effect on the cytotoxic parameters (P>0.05 ). Conclusion GP is the direct substance of hepatotox-icity induced by Fructus Gardeniae,and the mecha-nism might be associated with oxidative stress,mito-chondria injury and apoptosis.

Objective To analyze the clinical application of fluorescence in situ hybridization ( FISH) and karyotype analysis in prenatal diagnosis of chromosomal abnormalities .Methods The prenatal diagnosis of chromosome aneuploidies by FISH analysis of chromosome-specific probes (chromosome 13,18, 21,X,Y) in interphase amniocytes of 1 809 pregnant women of 17-38 weeks of gestational age was used , comparisons with the karyotyping results was done simultaneously.All the 1 809 cases came from Peking University First Hospital from July 1,2012 to December 31,2013, and the relevant clinical data and birth follow-up information were collected.Results All the 1 809 cases had been successfully examined by FISH , including 1 767 normal cases and 42 cases of numerical abnormality (39 cases of aneuploid, 1 case of triploid and 2 cases of mosaicism),which were consistent with the karyotyping analysis .What′s more,34 cases of chromosomal structural abnormalities , 5 cases of chimera and 12 cases of normal variant were failed to detected by FISH.Conclusion With the advantages of high-speed,simplicity,high accuracy,etc,FISH can be an effective tool in clinical applications , and had great significances in cytogenetic prenatal diagnosis combined with karyotyping analysis.