Objective To evaluate the clinical value of serum carcinoembryonic antigen(CEA),neuron specific enolization enzyme(NSE),21-1 non-small cell lung cancer associated antigen(CYFRA21-1),carbohydrate antigen 125(CA125) and ferritin(Fer) in the diagnosis of lung cancer.Methods The levels of CEA,NSE,CYFRA21-1,CA125 and Fer were measured by electrochemlium-inescence in 103 patients with lung cancer,32 patients with benign lung diseases and 40 healthy people.Results The serum levels of CEA,NSE,CYFRA21-1,CA125 and Fer in patients with lung cancer[(110.2±95.5)ng/mL,(50.6±43.4)ng/mL,(32.8±29.5)ng/L,(122.7±110.4)U/L,(854.6±497.2)ng/mL] were significantly higher than those in patients with benign lung diseases and those in healthy people(t=6.21,5.71,6.75,6.62,7.74,P<0.05;t=5.26,4.86,5.81,5.20,6.26,P<0.05).The sensitivity values of CEA,NSE,CYFRA21-1,CA125,Fer and the combined determination containing five tumor markers were 39.81%,24.27%,71.84%,68.93%,77.66% respectively.The sensitivity and specificity of the combined determination containing five tumormarkers were 96.12%,95.00%.Conclusion The joint detection in the diagnosis of lung cancer could improve the sensitivity significantly,to help for early diagnosis of lung cancer,which is value to widely applied in clinic.

Background Thyroid associated ophthalmopathy (TAO) is an autoimmune disease.Current research on the pathogenesis focuses on common autoantigen.Insulin-like growth factor-1 receptor (IGF-1R) is necessary for the function of IGF-1,also IGF-1 plays an important role in signaling pathway of thyroid stimulating hormone receptor (TSHR).Objective This study was to investigate the effects of IGF-1 on the proliferation,expression of IGF-1R and TSHR on cultured orbital fibroblasts (OFs) derived from TAO.Methods Human orbital tissue was obtained from 17 TAO patients who received orbital adipectomy and 4 normal controls who received cosmetic surgery in West China Hospital from March 2016 to June 2016.OFs were cultured by explant culture with DMEM/F12 containing 5％ fetal bovine serum and identified by immunochemistry.The OFs were treated with different concentrations of IGF-1.IGF-1 at different concentrations (0,50,100,125 μg/L) was added into the medium,respectively,and the proliferation of the cells (absorbancy) was detected by MTS.The percentages of IGF-1R and TSHR expressions in the cells were assayed by flow cytometry.Results Cultured cells appeared to be spindle-like in shape and grew well with abundant cytoplasm.The characteristics of the cells derived from TAO patients were consistant with normal ones.The cells showed the positive response for vimentin and absent respose for desmin,S-100,myoglobin and cytokeratin.The proliferative values of OFs were gradually elevated with the increase of IGF-I dose in both TAO group and normal group (Fgroup =219.639,P＜0.001;F ion =17.752,P＜0.001) with the optimal effects in 100 μg/L IGF-1.The expression levels of IGF-1R in the OFs were (0.009 1 ±0.008 7)％,(0.095 3±0.023 3) ％,(0.083 7±0.022 7) ％ and (0.070 9 ± 0.024 1) ％ in the TAO group,and those in the normal group were (0.0023± 0.0006)％,(0.0093±0.0012)％,(0.0073±0.0015)％ and (0.0083±0.0012)％ after treatment of 50,100,125 μg/L IGF-1.The expression levels of IGF-1 R were significantly higher after treatment of 50,100 and 125 μtg/L IGF-1 than those treatment of 0 μg/L IGF-1 in both TAO group and normal group,and the expression levels of IGF-1R in the OFs were significantly increased in the TAO group compared with the normal group (all at P＜0.05).No statistical difference was seen in the TSHR expression between the TAO group and normal group after treatment of 0,50,100 and 125 μg/L IGF-1 (Fgroup =0.133,P ＞ 0.05;F ion =0.004,P ＞ 0.05).Conclusions IGF-1 can promote the proliferation of OFs and up-regulate the expression of IGF-1R in OFs.However,IGF-1 dose not play a regulating effect on the expression of TSHR in OFs.

Objective:To investigate the signaling pathway of inhibiting macrophage phagocytosis of TIR domain-containing protein encoded by Escherichia coli (TcpC) N-terminal ubiquitin ligase active fragments of uropathogenic Escherichia coli. Methods:Bioinformatics software was used to analyze the amino acid sequences and the function of TcpC N-terminal ubiquitin ligase active fragments as well as the functional sites. PCR was performed to amplify tcpc-330, tcpc-450 and tcpc-510 genes and a prokaryotic expression system was constructed to express the target proteins. The recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 were purified by Ni-NTA affinity chromatography. LPS in the recombinant proteins was removed by Detoxi-gel chromatography. The expression of MyD88 at protein and mRNA levels in macrophages incubated with rTcpC-N110, rTcpC-N150, rTcpC-N170 or rTcpC-TIR was detected by Western blot and qRT-PCR. The activation of NF-κB signal pathway and the levels of proinflammatory factors in macrophages incubated with the above TcpC protein fragments were measured by Western blot and ELISA, respectively. Results:Cys12, Trp104 and Trp106 in the N-terminal fragment of TcpC were crucial amino acids in maintaining its ubiquitin ligase activity. The target recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 were successfully expressed and purified. After Detoxi-gel chromatography, rTcpC-N110, rTcpC-N150 and rTcpC-N170 extracts were undetectable for LPS. TcpC ubiquitin ligase fragments inhibited the expression of MyD88 at protein level, but not affect its expression at mRNA level in macrophages. LPS-induced phosphorylation of NF-κB signaling pathway-related proteins p50 and p65 was significantly inhibited in macrophages treated with TcpC ubiquitin ligase fragments. Moreover, LPS-induced production of pro-inflammatory factors was also significantly inhibited.Conclusions:The recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 could inhibit the expression of MyD88 at protein level and suppress the activation of NF-κB signaling pathway, suggesting that they were closely related to the inhibition of innate immune activity of macrophages.

Objective:To investigate the role of TcpC in uropathogenic Escherichia coli (UPEC)-induced cystitis in mice and to preliminarily analyze the pathogenic mechanism. Methods:C57BL/6 mice were injected with 10 9 CFU wild-type UPEC CFT073 (CFT073 wt) or tcpc gene-deleted mutant (CFT073 Δ tcpc) from urethra into bladder to construct the mouse model of cystitis. The mice were sacrificed 3 d after infection and the bladders were taken to observe the gross pathological changes. Histopathological changes in bladder tissues were observed after HE staining. Immunohistochemistry was used to detect TcpC in bladder tissues. Bacterial loads in urine samples of UPEC-infected mice were counted by tenfold dilution method, and the presence of tcpc gene in the genomic DNA of bacteria from the bladder and urine samples of CFT073 wt-infected mice was measured by PCR. Real-time quantitative RT-PCR (qRT-PCR) and Western blot were performed to detect the expression of TcpC at mRNA and protein levels in macrophages after CFT073 wt infection. The influence of UPEC strains on the activation of NF-κB signaling pathway in macrophages were determined by Western blot. The levels of proinflammatory factors and the bacterial and cell activity after infecting macrophages with UPEC strains were detected by ELISA, laser confocal microscope and fluorescence microscope, respectively. Results:Compared with the mice with CFT073 Δ tcpc infection, CFT073 wt-infected mice had significantly enlarged bladder and severe neutrophil infiltration and abundant TcpC in bladder tissues. The number of bacteria in the urine of CFT073 wt-infected mice was significantly greater than that of the CFT073 Δ tcpc group. PCR results showed that the bacteria in bladder or urine were CFT073 wt. The expression of TcpC at both mRNA and protein levels in macrophages increased significantly after CFT073 wt infection. Moreover, in CFT073 wt-infected macrophages, the expression of IκBα was promoted and the phosphorylation of p65 and the production of proinflammatory factors were suppressed. TcpC was instrumental in the survival and invasion of CFT073 wt in macrophages. Conclusions:TcpC expression increased significantly in mice with CFT073 wt-induced cystitis. TcpC inhibited the activation of NF-κB signaling pathway and the production of proinflammatory factors in macrophages to improve the survival rate of CFT073 wt, which was closely related to the pathogenesis and immune evasion of UPEC.

Objective:To investigate whether the hypervirulent Klebsiella pneumoniae (hvKP) induces liver abscess through activating NLRP3 inflammasome. Methods:K1-hvKP and K35-non-hvKP bacterial suspensions were intraperitoneally injected into C57BL/6 mice to establish the models of liver abscess. Human peripheral blood neutrophils were sorted by immunomagnetic beads with CD45 + and Gr-1 + , and the purity was detected by flow cytometry. The concentrations of capsular polysaccharide of K1-hvKP and K35-non-hvKP were detected by total carbohydrate assay kit. The expression of IL-18 and IL-33 by neutrophils at mRNA and protein levels was detected by real-time fluorescence quantitative PCR and ELISA, respectively. The activation of NLRP3 inflammasome in neutrophils was detected by Western blot. Neutrophil extracellular trap formation (NETosis) was observed under confocal laser scanning microscope. Results:The C57BL/6 mice with K1-hvKP infection had significantly serious liver abscess as compared with the K35-non-hvKP-infected mice. The purity of human neutrophils was more than 95%. The concentration of capsular polysaccharide in K1-hvKP was significantly higher than that in K35-non-hvKP. Compared with K35-non-hvKP, K1-hvKP significantly promoted the neutrophils to express IL-18 and IL-33 at both mRNA and protein levels, enhanced the activation of NLRP3 and induced NETosis.Conclusions:This study suggested that hvKP could promote NETosis by activating NLRP3 inflammasome to cause liver abscess.

Objective To assess the longitudinal mitral annular plane systolic excursion ( M APSE) of different directions in normal fetuses during mid‐late pregnancy based on two‐dimensional speckle tracking imaging ( ST I) . Methods Seventy‐six normal fetuses during middle and late pregnancy were selected at 26－32 weeks of gestation . T he peak M APSE was measured by free angle M‐mode echocardiography ( FAM ) perpendicular to the lateral annulus in the mitral annular plane . The time‐displacement curves of interventricular septal mitral annulus in three different directions including points A ,B and C through transverse level of apex were recorded by STI . T he peak M APSE of interventricular septal mitral annulus ( SEPT‐M APSE‐A ,SEPT‐M APSE‐B ,SEPT‐M APSE‐C) in three different directions including points A ,B and C and the time to peak ( T T P :SEPT‐T T P‐A ,SEPT‐T T P‐B ,SEPT‐T T P‐C) were recorded respectively . T he time‐displacement curves of lateral mitral annulus in three different directions including points A ,B and C through transverse level of apex were recorded by STI . T he peak M APSE of lateral mitral annulus ( LAT‐M APSE‐A ,LAT‐MAPSE‐B ,LAT‐MAPSE‐C) in three different directions including points A ,B and C ,the time to peak( LA T‐T T P‐A ,LA T‐T T P‐B ,LA T‐T T P‐C) were recorded respectively . Finally ,the data were analyzed statistically . Results T he peak M APSE of the lateral mitral annulus in 3 different directions including points A ,B and C[ LA T‐M APSE‐A ( 3 .62 ± 1 .01) mm ,LA T‐M APSE‐B ( 3 .95 ± 1 .04) mm ,LAT‐M APSE‐C ( 4 .45 ± 1 .05) mm ] were greater than those of the interventricular septum mitral annulus[ SEPT‐MAPSE‐A (3 .41 ± 0 .63)mm ,SEPT‐MAPSE‐B (3 .07 ± 0 .50) mm ,SEPT‐MAPSE‐C (2 .82 ± 0 .51) mm] . LAT‐M APSE‐C and SEPT‐M APSE‐A were the largest longitudinal excursions of mitral annulus . T he differences were statistically significant in points B and C ( P ＜0 .05) . T here was no significant difference in point A ( P ＞0 .05) . LA T‐M APSE‐C was less than FAM‐M APSE [ ( 6 .06 ± 1 .35 ) mm ] . T here was a significant difference between them ( P ＜0 .05 ) . Strong correlation was found between them ( r ＝0 .896 , P＜0 .05) . T here were no significant differences in the time to peak of interventricular septal mitral annulus [ SEPT‐T T P‐A ( 0 .210 ± 0 .008 ) s ,SEPT‐T T P‐B ( 0 .213 ± 0 .008 ) s ,SEPT‐T T P‐C ( 0 .210 ± 0 .005 ) s] in directions including points A ,B ,C ( P＞ 0 .05 ) . T here were no significant differences in time to peak of lateral mitral annulus [ LAT‐T T P‐A ( 0 .210 ± 0 .008 ) s , LAT‐T T P‐B ( 0 .213 ± 0 .006 ) s , LAT‐T T P‐C ( 0 .210 ± 0 .007) s] in directions inclucling points A ,B ,C ( P ＞0 .05) . Conclusions Longitudinal systolic motion of fetal left ventricular wall during mid‐late pregnancy has good synchronization . Longitudinal motion of fetal mitral annulus is a comprehensive movement of multiple directions and different degrees of displacement ,with the movement perpendicular to the annulus as the maximum displacement direction . T he displacement parameters of mitral annulus measured by ST I can reflect the left ventricular longitudinal systolic function and have clinical application value in evaluating the left ventricular longitudinal systolic function of fetuses .

Objective To calculate Z-score for mitral and tricuspid color blood flow widths in normal fetuses and fetuses with dilated coronary sinuses ( CS ) using fetal echocardiography ,and explore the application value of Z-score of the color flow widths of atrioventricular valves in normal fetuses and fetuses with dilated CS . Methods Two hundred and thirty-eight normal fetuses (control group) with a gestational age of 16 to 38 weeks were studied by color Doppler echocardiography . Gestational age ( GA ) ,biparietal diameter (BPD) ,femoral length (FL) ,aortic inner diameter (AOd) ,pulmonary artery diameter (PAd) ,and heart area ( HA) were measured as independent variables ,and mitral and tricuspid valve color flow widths were measured as the dependent variables . Z-score models were established by regression analysis . Thirty fetuses with dilated CS (dilated CS group) from 22 to 33 weeks'gestation were involved . The Z-score of the CS fetus was calculated based on the established Z-score models and were compared with those of the normal fetuses . Results The independent sample t-test showed that there were no significant differences in the Z-scores of the blood flow width of the fetal mitral and tricuspid valves between dilated CS group and control group ( P >0 .05) . Conclusions The simple dilated CS does not affect the mitral valve diastolic blood flow ,so there is no significant effect on the filling of left ventricular blood flow .

Objective: To explore the relationship between HCM pathogenic gene mutations and clinical phenotypes by analyzing the prenatal diagnosis and genetic characteristics of a pregnant woman from a family with hypertrophic cardiomyopathy (HCM). Methods: The clinical data of the proband and her family members was collected. The DNA was extracted from the peripheral blood, amniotic fluid cells and cultured amniotic fluid cells of proband. Next generation sequencing (NGS) was utilized for screening pathogenetic loci of the proband. The suspected mutation sequences of HCM pathogenic candidate genes MYH7 and MYBPC3 were directly sequenced after PCR. Pathogenicity prediction of amniotic fluid cells was performed by using genetic data and bioinformatics software, such as Mutation taster, PolyPhen-2 and ANTHEPROT. Results: The sequencing results showed that heterozygous mutations of MYH7 c.1988G＞A (p.Arg663His) and MYBPC3 c.151G＞A (p.Ala51Thr) were found in the proband. The phenotype of her father was normal, and no abnormal mutations were detectable. Her mother also showed normal phenotype but carried MYBPC3 c.151G＞A heterozygous mutation. Only MYH7 c.1988G＞A heterozygous mutation was found in the fetus and no abnormal variation of MYBPC3 was showed. The prediction of mutation effect and analysis of protein structure and function revealed that the two missense mutations could affect the hydrophobicity and antigenicity of the protein. The genetic data demonstrated MYH7 c.1988G＞A was defined as a pathogenic mutation. Conclusion MYH7 c.1988G＞A should be a newly generated pathogenic mutation in the proband, or caused by reproductive chimerism of her parents. MYBPC3 c.151G＞A mutation may promote the occurrence of HCM. Although the fetus only carries MYH7 c.1988G＞A, her phenotype may still display as HCM.

Objective: To explore the genetic basis for a pedigree affected with Marfan syndrome (MFS). Methods: Clinical data of the patients was collected.With genomic DNA extracted from peripheral blood samples, potential mutation was detected by targeted exome sequencing.Candidate variants were validated by Sanger sequencing and bioinformatic analysis. Results: Targeted exome sequencing and Sanger sequencing revealed a missense c. 649T>C(p.Trp217Arg) variant in the exon 7 of FBN1 gene, which was unreported previously.Bioinformatics analysis suggested that the variant can cause amino acid replacement and affect the structure and function of fibrillin-1. Conclusion A novel missense variant of the FBN1 gene was identified, which probably underlies the autosomal dominant MFS in this pedigree.

OBJECTIVE: To assess the clinical application of single nucleotide polymorphism microarray (SNP array) in patients with intellectual disability/developmental delay(ID/DD). METHODS: SNP array was performed to detect genome-wide DNA copy number variants (CNVs) for 145 patients with ID/DD in Women's Hospital, Zhejiang University School of Medicine from January 2013 to June 2018. The CNVs were analyzed by CHAS software and related databases. RESULTS: Among 145 patients, pathogenic chromosomal abnormalities were detected in 32 cases, including 26 cases of pathogenic CNVs and 6 cases of likely pathogenic CNVs. Meanwhile, 18 cases of uncertain clinical significance and 14 cases of likely benign were identified, no significant abnormalities were found in 81 cases (including benign). CONCLUSIONS SNP array is effective for detecting chromosomal abnormalities in patients with ID/DD with high efficiency and resolution.
Chromosome Aberrations ; DNA Copy Number Variations ; Genome-Wide Association Study ; Humans ; Intellectual Disability ; diagnosis ; genetics ; Oligonucleotide Array Sequence Analysis ; standards ; Polymorphism, Single Nucleotide