Objective To discuss protein marks expressed differentially in placenta of Down's syndrome by means of proteomics. Methods We collected placenta of 18 patients(from March 2009 to December 2009 at Beijing Obstetrics and Gynecology Hospital), and divided them into two groups, one was 10 patients with fetal Down's syndrome, the other was normal pregnancies (normal chromosome) with other diseases. We separated proteins expressed in placentas of two groups by two-dimensional difference gel electrophoresis (2D-DIGE), and then analyzed the differential protein spots by software Decyder 6. 5, then,spots differentially expressed by 1.5 fold or more were analyzed by matrix assisted laser desorption ionizationtime of flight-mass spectrometry (MALDI-TOF-MS). In the end, the differential expressional levels of partially identified proteins were validated by western blot analysis. Results (1) Differential proteins of two groups protein spots of placentas separated by 2D-DIGE were analyzed by software Decyder 6. 5 (these colored lights scattered in the image were protein spots), a total of 56 spots out of 352 were differentially expressed (P<0. 05) in two groups. We analyzed 17 protein spots(12 protein spots were over-expressed and 5 protein spots were down-expressed) differentially expressed by 1.5 fold or more by MALDI-TOF-MS.(2) Protein matching after searching protein database, 17 protein spots turn out to be 10 proteins. Four kinds [superoxide dismutase 1 (SOD1), peroxiredoxin 6 (PRDX6), heat shock protein 27 (HSP27),endoplasmic reticulum protein 29 (ERP29)] of them were validated by western blot analysis, the group of fetal Down's syndrome were 0.74 ±0. 12,0.29 ±0. 10,0.53 ±0. 16,0.20 ±0. 09,the group of normal pregnancies were 0. 51 ±0. 08,0. 34 ± 0. 16,0. 18 ± 0. 07,0. 35 ± 0. 09, the results confirmed the observed changes in proteomics. Conclusions Compared with normal pregnancies, there were differential proteins expressed in placenta of Down's syndrome. This approach might provide new screening markers in use for prediction of Down's syndrome, however, further study should be done to make these 4 proteins (SOD1,HSP27, ERP29, PRDX6) be new screening markers.

Objective:To compare the concordance between subjective experience and clinician-rated in different treatment phrase of elderly patients with depressive disorder,and to explore the relevant factors.Methods:Sixty-nine consecutive elderly patiems with depressive disorder diagnosed with International Classification of Diseases-10 criteria were included.The Visual Scale for Depression(VSD) were used to evaluate subjective experience of depression,while the Hamilton Depression Scale (HAMD),Hamilton Anxiety Scale (HAMA),Minimum Mental State Examination (MMSE) were used by clinician to evaluate depression symptom,anxiety symptom and cognition at baseline,2-,4-and 52-week of the treatment respectively.Results:Compared with baseline,VSD scores were increased at 2 weeks and 4 weeks [(2.7 ± 1.6),(5.3 ± 2.0) vs.(7.0 ± 1.8),P ＜ 0.001],HAMD scores were decreased at 2 weeks and 4 weeks [(36 ± 11),(17 ±9) vs.(9 ±8),P ＜0.001],HAMA scores were decreased at 2 weeks and 4 weeks [(27 ± 10),(14 ±8) vs.(7 ±6),P ＜0.001].No significant differences were found between 4 weeks and 52 weeks.There was no correlation between VSD and HAMD scores at baseline and 2 weeks.There were significant correlations among scores of VSD and HAMD(r =-0.31,-0.74,Ps ＜ 0.05),HAMA (r =-0.36,-0.76,Ps ＜ 0.05) at 4 weeks and 52 weeks.Using logistic regression analysis,the concordance between VSD and HAMD related with the factors of HAMD,higher scores of anxiety/somatization factor were associated with lower concordance (OR =0.87),and higher scores of weight factor (OR =1.86),diurnal variation factor (OR =2.00),hopelessness factor (OR =1.13) were associated with higher concordance.Conclusion:Compared with acute depression phase,concordance between subjective experience and clinician-rated for depression may be higher at depression remission phase in elderly patients with depression,which suggests that characteristics of depression symptom may have a certain impact on the self-awareness for depression experience.

Objective To investigate genes involved in the mechanisms underlying the progression of severe preeclampsia.Methods We conducted a muhiregional gene expression analysis using peripheral leucocytes from patients with preeclampsia and normal controls.Total RNA was extracted from peripheral blood of six severe preeclampsia and five normotensive pregnancies.We performed genome-wide expression profiling using Affymetrix HG_U133 plus 2.0 chips to screen out differentially expressed genes of 2 fold or more and q_value ＜ 5.4%.Using Gene Ontology we identified the function of differentially expressed genes after cluster analysis.Results Among the 47 000 genes that were screened in the microarray,140 genes were found to be differentially expressed between normal and preeclamptic pregnancies. Eighty six up-regulated candidate genes were mainly involved in cysteine metabolism urea cycle and metabolism of amiogroups,proteasome,TGF-beta signaling pathway, and the ratio of calponin2 (CNN2), matrix metallopeptidase 8 (MMP8),V-set and immunoglobulin domain containing 4 (VSIG4),proteasome 26S subunit ATPase 5 (PSMC5) was evidently increased in preeclampsia patients.Among 54 down-regulatedcandidates,natural killer cell mediated cytotoxicity,antigen processing and presentation,metabolism of xenobiotics by cytochrome P450 were the main pathways.KIR3DL2,AKR1C3,CHURC1 and SLC25A13 were obviously decreased in preeclampsia patients. Conclusions The gene expression of peripheral leucocytes in preeclampsia patients is significantly different from that of uncomplicated pregnancies.CNN2,MMP8,VSIG4,PSMC5,KIR3DL2,AKR1C3,CHURC1 and SLC25A13 may be involved in the molecular mechanisms underlying the progression of severe preeclampsia.

Objective To explore the associations of the genetic polymorphisms of cytochrome P450, family 2, subfamily D, polypeptide 6 (CYP2D6) and cytochrome P450, family 2, subfamily C, polypeptide 9 (CYP2C9) with early-onset severe pre-eclampsia and the efficacy of labetalol therapy. Methods Totally 105 gravidas diagnosed with early-onset severe pre-eclampsia (experimental group) and 103 healthy gravidas (control group) were recruited from Beijing Obstetrics and Gynecology Hospital between August 2013 and July 2016. Labetalol was given to control blood pressures in gravidas with early-onset severe pre-eclampsia. If labetalol administration alone did not exceed the mean dose (100 mg, one dose per eight hours) and effectively controlled the blood pressures, it would be considered to be valid (n=75), otherwise it would be viewed as an invalid treatment. Genotype and allele frequencies of CYP2C9 gene (rs1057910 and rs4918758) and CYP2D6 gene (rs1065852, rs28371725, rs35742686 and rs3892097) in the gravidas were analyzed by TaqMan probe polymerase chain reaction. Differences in the genotype and allele frequencies were compared between the experimental and control groups, and the valid and invalid labetalol treatment groups. Chi-square test, analysis of variance and LSD test were used as statistical methods. Results The gravidas in both experimental and control groups were AA genotype in CYP2C9 gene rs1057910, TT genotype in CYP2D6 gene rs35742686 and CC genotype in CYP2D6 gene rs3892097. Frequencies of CC and CT genotypes in CYP2D6 gene rs28371725 in the experimental group were higher than those in the control group [18.1% (19/105) vs 14.6% (15/103);56.2% (59/105) vs 42.7% (44/103); χ2=6.707], and higher C allele frequency in CYP2D6 gene rs28371725 was also observed in the experimental group [46.2% (97/210) vs 35.9% (74/206), χ2=4.529] (all P<0.05). No statistical differences in maternal age, diastolic pressure, body mass index before pregnancy, serum triglyceride, creatinine and neonatal birth weight were observed among women with CC, CT or TT genotype of CYP2D6 gene rs28371725 in the experimental group (all P>0.05). Compared with the gravidas with CT or TT genotype of CYP2D6 gene rs28371725, those with CC genotype had longer gestational age [(32.5±2.1) vs (29.5±1.8) and (29.8±2.2) weeks] and higher plasma albumin [(27.2±9.3) vs (20.3±10.4) and (22.5±7.4) g/L], but lower systolic pressure and 24 hours urine protein (LSD test, all P<0.05). The G allele frequency in CYP2D6 gene rs1065852 in invalid labetalol treatment group was higher than that in valid labetalol treatment group [93.3% (56/60) vs 76.0% (114/150), χ2=8.351, P=0.004]. Conclusions The polymorphism of CYP2D6 gene rs28371725 may be associated with early-onset severe pre-eclampsia, and the allele of G in CYP2D6 gene rs1065852 may be associated with the efficacy of labetalol in treatment of early-onset severe pre-eclampsia.

Sepsis is defined as life-threatening organ dysfunction caused by the disorder of the body′s response to infection.Sepsis is the focus of critical illness medicine and difficult problems nowadays.At pres-ent,the incidence of sepsis in children is still high around the world.If the treatment is not timely,sepsis can develop into septic shock,multiple organ dysfunction syndrome,which is a serious threat to human health. Therefore,the early identification,diagnosis and treatment of sepsis is important for reducing mortality.And the biomarkers play an important role in the early diagnosis of sepsis,pathogenetic condition and prognosis as well as efficacy evaluation.This article summarized the biomarkers of sepsis in recent years.

Three cyclotides were isolated from the whole plant of Viola yedoensis in this study. The two, vary peptide E and cycloviolacin Y5, were previously reported, and a novel cycloviolacin VY1 was characterized according to the interpretation of MS/MS fragmentation of peptides which were produced from the reduced and alkylated parent peptide with the digestion of Endo Lys-C, trypsin and chymotrypsin, separately. The stability of remarkable resistance to proteolytic degradation by trypsin and chymotrypsin, and that of thermal denaturation was confirmed again. Besides, the IC50 value of cycloviolacin VYI against influenza A H1N1 virus was (2.27 +/- 0.20) microg x mL(-1). It is the first cyclotide reported with anti-influenza A H1N1 virus activity in vitro assay.

Objective To investigate the clinical significance and changes of serum complement in lipid metabolism disorder in patients with fatty liver disease.Methods One hundred and forty patients with FLD from October 2015 to May 2017 were included in the study,in addition,120 patients with hyperlipidemia(the hyperlipidemia group)and 130 healthy subjects(the control group)in the same period were enrolled as controls.The differences in serum lipid,liver function enzymology,immunoglobulin,serum complement among the three groups were compared.Results There were significant differences in the levels of TC,TG,HDL-C,LDL-C,apoA and apoB among the three groups(TC:(5.7±1.6)mmol/L vs.(4.2±1.0)mmol/L vs.(3.5±1.1) mmol/L,F=105.01,P<0.05;TG:(2.8± 0.6)mmol/L vs.(1.5 ± 0.3)mmol/L vs.(1.1 ± 0.2)mmol/L,F=628.46,P<0.05;HDL-C:(1.2±0.3)mmol/L vs.(1.5±0.3)mmol/L vs.(1.8±0.4)mmol/L,F=107.10, P<0.05;LDL-C:(3.6±0.9)mmol/L vs.(3.0±0.8)mmol/L vs.(2.2±0.6)mmol/L,F=109.07,P<0.05;apoA:(1.0±0.2)g/L vs.(1.2±0.2)g/L vs.(1.4±0.3)g/L,F=95.20,P<0.05;apoB:(1.1±0.2)g/L vs.(0.9±0.2)g/L vs.(0.8±0.2)g/L,F=79.04,P<0.05).The levels of TC,TG,LDL-C and apoB in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of HDL-C and apoA in the FLD groups were significantly lower than those in the hyperlipidemia group and control group.The levels of TC,TG,LDL-C and apoB in the hyperlipidemia group were significantly higher than those in the control group.The levels of HDL-C and apoA in the hyperlipidemia group were significantly lower than those in the control group.There were significant differences in the levels of GGT,ALT,AST,IgG and IgM among the three groups(GGT:(77.4±15.3)U/L vs.(43.3±10.6)U/L vs.(25.5±8.2)U/L,F=668.12,P<0.05;ALT:(61.5±18.8)U/L vs.(35.7±11.2)U/L vs.(18.9±5.4)U/L,F=355.67,P<0.05;AST:(55.3±12.2)U/L vs.(32.4±12.5)U/L vs.(14.4±4.7)U/L,F=521.80,P<0.05;IgG:(15.7±3.9)g/L vs.(11.6±3.2)g/vs.,(8.5±2.6)g/L,F=162.34,P<0.05;IgM:(1.9±0.6)g/L vs.(1.2±0.4)g/L vs.(0.8±0.3)g/L,F=201.38,P<0.05).The levels of GGT,ALT,AST,IgG and IgM in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of GGT,ALT,AST,IgG and IgM in the hyperlipidemia group were significantly higher than those in the control group.There were significant differences in the levels of C3,C5,ASP and BF among the mild,moderate and severe fatty liver patients(C3:(2.1±0.4) g/L vs.(1.8±0.3)g/L vs.(1.0±0.2)g/L,F=436.37,P<0.05;C5:(92.3±10.7)mg/L vs.(71.8±8.8) mg/L vs.(58.9±6.5)mg/L,F=486.09,P<0.05; ASP:(51.4±6.8)nmol/L vs.(42.5±4.4)nmol/L vs.(32.8±5.2)nmol/L,F=369.29,P<0.05;BF:(0.48±0.13)g/L vs.(0.34±0.09)g/L vs.(0.23±0.04) g/L,F=233.39,P<0.05).The levels of C3,C5,ASP and BF in the FLD group were significantly higher than those in the hyperlipidemia group and the control group.The levels of C3,C5,ASP and BF in the hyperlipidemia group were significantly higher than those in the control group.There were significant differences in the levels of C3,C5,ASP and BF among patients with mild,moderate and severe fatty liver disease(C3:(1.8±0.3)g/L vs.(2.1±0.4)g/L vs.(2.5±0.4)g/L,F=30.85,P<0.05;C5:(80.5±9.6)mg/L vs.(92.3±10.5)mg/L vs.(100.7±8.)mg/L,F=39.39,P<0.05; ASP:(42.4±6.3)nmol/L vs.(52.8±5.7)nmol/L vs.(61.9±5.6) nmol/L,F=98.19,P<0.05;BF:(0.33±0.12)g/L vs.(0.45±0.11)g/L vs.(0.57±0.09)g/L,F=41.26,P<0.05).The levels of C3,C5,ASP and BF in the mild FLD patients were significantly lower than those in moderate and severe FLD patients.The levels of C3,C5,ASP and BF in the moderate FLD patients were significantly lower than those in severe FLD patients.Conclusion The detection of C3,C5,ASP and BF levels based on routine testes has important clinical value for the assessment of the condition,the treatment and the prognosis of FLD patients.

目的：分析Claudin-2蛋白在食管鳞癌（esophageal squamous cell carcinomas，ESCC）组织中的表达及其与患者临床病理特征、5年生存率的关系，探索其对ESCC细胞KYSE450的增殖、迁移和侵袭的影响。方法：选取河南省肿瘤医院2010至2013年间初治的ESCC患者手术切除肿瘤组织52例及其中20例对应的癌旁组织标本，采用免疫组化和WB法检测Claudin-2的表达并分析其与患者临床病理特征和5年生存率的关系。WB法检测ESCC细胞（KYSE450、KYSE150、KYSE510、KYSE140）和人永生化食管上皮细胞SHEE中Claudin-2的表达，构建Claudin-2 shRNA慢病毒载体并转染KYSE450细胞构建敲低Claudin-2表达的细胞系，进一步通过克隆形成实验、细胞划痕实验及Transwell实验检测敲低Claudin-2对KYSE450细胞增殖、迁移和侵袭的影响。结果：ESCC组织中Claudin-2阳性率显著高于癌旁组织（P<0.05），ESCC组织中Claudin-2的表达与淋巴结转移有关（P<0.05）。Claudin-2表达阳性患者5年生存率显著低于阴性者（P<0.05）。成功构建敲低Claudin-2表达的KYSE450细胞系。sh-Claudin-2组细胞的克隆形成数、伤口愈合率和侵袭细胞数均显著低于sh-NT组和对照组（P<0.05）。结论：ESCC组织中Claudin-2的表达高于癌旁组织，且与患者5年生存率呈负相关，Claudin-2能够增强KYSE450细胞的增殖、迁移和侵袭能力。

Objective To observe the dynamic changes of plasma mitochondrial DNA (mtDNA) in children with sepsis in order to explore the clinical value of it in evaluation of these cases. Methods A total of 37 sepsis children admitted from June 2016 to January 2017 in the intensive care unit of Hunan Children's Hospital were enrolled for this prospectively study. And another 27 healthy children with similar age and gender were randomly selected as the control group. The venous blood samples were taken on the 1st, 3rd and 7th day after admission. Fluorescence quantitative PCR was used to detect the plasma mtDNA level. Meanwhile, the laboratory examinations such as detections of CRP and PCT were carried out. The diagnosis of sepsis and septic shock was based on Sepsis Criteria 3. Patients with genetic metabolic disease, liver and kidney disease, end-stage of tumors were excluded. Data of two groups were analyzed with Mann-Whitney U test. The sensitivity and specifi city were assessed by receiver operating curve (ROC). Results The plasma mtDNA level in the sepsis group 3 384.4(1 368.5, 6 857.5) pg/mL was higher than that in the healthy control group 1 904.8(1 267.9, 2 395.5) pg/mL with statistical signifi cance (P<0.05). The levels of plasma mtDNA in the sepsis multi-organ dysfunction group were higher than those in the single organ disorder group at day 1,3,7, with statistically signifi cant in three intervals (P <0.05). The level of plasma mtDNA in sepsis group were signifi cantly higher than those in non-shock group at day 1,3,7, with statistical difference (P <0.05). The plasma mtDNA levels in the non-survival group were higher than those in survival group 13 515.1(4 832.7,152 348.5)vs.2 780.0(1 226.8,5 261.2)on day 1;5 842.4(3 402.8,101 937.5)vs.1 450.5(710.6,2 481.6)on day 3 with statistical difference(P<0.05).there was no significant difference in mtDNA on day 7 was 1 045.1 vs.741.8(334.0,1 254.6)between survival group and non-survival group (P >0.05). In respect of diagnostic effi cacy of plasma mtDNA, PCT and CRP in predicting sepsis MODS, the largest area under the ROC curve of plasma mtDNA was 0.848 occurring on the 1st day, when the critical value was 2 176.2 pg/mL, the sensitivity and specifi city was 89% and 72%, respectively, and the difference was statistically signifi cant(P<0.01). When the critical value of CRP was 71.3 mg/L, the sensitivity and specifi city was 58% and 67% respectively. When the critical value of PCT was 7.24 ng/L, the sensitivity and specifi city were 84% and 61%, respectively. The plasma mtDNA peaked on day 1 followed by a downward changes. Conclusions The elevated level of plasma mtDNA in sepsis children was associated with organ dysfunction, indicating that it can be used as one of biomarkers for the diagnosis of sepsis MODS in children. The level of plasma mtDNA in children with sepsis was significantly high on the first and third day after admission, which was positively correlated with the severity of sepsis and it has certain value in assessment of the disease.

Objective:To investigate the effect of long-term application of prostaglandin analog drops on bulbar conjunctival thickness in rabbits.Methods:Twenty-four healthy New Zealand white rabbits were randomly divided into latanoprost group, carteolol group and blank control group using the random number table method, with 8 rabbits in each group.The left eyes of rabbits were taken as experimental eyes.The rabbits in the latanoprost group and carteolol group were given latanoprost eye drops or carteolol eye drops once a day for 2 months according to grouping.The bulbar conjunctival thickness of left eyes of the latanoprost group and carteolol group were measured by optical coherence tomography (OCT) at baseline and two months after administration, respectively.The conjunctival tissue of the three groups were extracted to investigate the protein and mRNA expression level of matrix metalloproteinases-1 (MMP-1) and MMP-3 by Western blot and real-time fluorescence quantitative polymerase chain reaction (PCR). The study protocol was approved by an Ethics Committee of Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine (No.2017-0014). The use and care of the experimental animals complied with the ARVO Statement.Results:In the latanoprost group, the conjunctival thickness was significantly reduced from baseline (178.88±5.23)μm to (124.19±11.29)μm at 2 months after administration ( P<0.01). In the carteolol group, there existed no significant difference in the conjunctival thickness between baseline (184.94±11.85)μm and (183.31±8.71)μm at 2 months after administration ( P>0.05). The conjunctival thickness at 2 months after administration of the latanoprost group was significantly thinner than that of the carteolol group ( P<0.01). The protein and mRNA expression levels of MMP-1 and MMP-3 in conjunctival tissue of the latanoprost group were significantly higher than those of the blank control group and carteolol group (all at P<0.01). Conclusions:The long-term topical use of prostaglandin analog drops can significantly reduce the bulbar conjunctival thickness in rabbits.The mechanism may be related to the elevated expression levels of MMP-1 and MMP-3 in the bulbar conjunctival tissue.