[ABSTRACT]OBJECTIVETo investigate the cytomegalovirus infection in neonates, characteristics of gap junction protein Connexin26 gene mutation and the hearing follow-up results, and to analyze their correlations. METHODS60 CMV-DNA positive and 40 CMV-DNA negative neonatal newborn from The Second Affiliated Hospital of Wenzhou Medical University and The first people's Hospital of Yongkang were screened, the blood biochemistry was analyzed, and the umbilical cord blood was reserved to detect the Connexin26 gene expression of mRNA with RT-PCR.PCR results was sequenced to track the newborn hearing, and analyze the correlations between neonatal cytomegalovirus types, the mutation of Connexin26 gene and hearing test results.RESULTS 26 cases from 60 CMV-DNA positive newborns were found with blood biochemical abnormalities. In all of the newborns, a total of 41 cases had 235delC mutation, 11 cases in the mutations for the development of hearing impairment. The results of correlation analysis showed that there were correlations between cytomegalovirus infection, gene mutation and hearing impairment.CONCLUSION Cytomegalovirus infection in neonates can lead to mutations in the Connexin26 gene, and may further lead to hearing impairment, and the probability of the mutation of Connexin26 gene and sensorineural hearing loss were higher in symptomatic cytomegalovirus infection neonates.

Objective To investigate the change of serum cancer antigen(CA) 125 in patients with pleural effusion.Methods One hundred and twenty-eight patients with pleural effusion were admitted to the Naval General Hospital of People's Liberation Army from January 2010 to September 2012 were selected as our subjects.The level of serum CA125 was measured.The difference of serum CA125 positive rate and level were compared according to gender,pleural effusion nature,quantity and pleural effusion chest area; And the difference of patients with malignant pleural effusion tuberculosis,inflammatory,exudative pleural effusion based on above indicators.The correlation between serum CA125 level and pleural effusion depth were analyzed.Results The positive proportions of CA125 were 83.3％ (35/42) and 76.7 ％ (66/88) of patients with malignant and benign effusion respectively,and there was no significant difference (x2 =0.74,P ＞ 0.05).The serum CA125 level of patients with malignant pleural effusion was significantly higher than benign ones ((177.8 ± 31.4) U/ml vs.(110.6 ± 13.6) U/ml,t =31.24,P ＜ 0.05).There were no significant difference in the positive proportion of serum CA125 between malignant,tuberculous,inflammatory and transudative pleural effusion(75.8％ (25/33),70.0％ (20/29),87.5％ (21/24) and 83.3％ (35/42),P ＞ 0.05).Serum CA125 levels of patients with malignant pleural effusion were significantly higher than that with inflammatory ((177.8 ± 31.4) U/ml vs.(72.5 ± 12.8) U/ml,P ＜ 0.05),but the differences were not significant among malignant,tuberculous and transudative pleural effusion group((140.6 ± 28.2) U/ml,(154.3 ± 30.5) U/ml,P ＞ 0.05).The serum CA125 levels of patients with small,moderate and large effusions were (56.4 ± 18.2) U/ml,(120.2 ± 24.5) U/ml and (185.5 ± 34.6) U/ml respectively,and the difference among these groups were significant(F =296.03,P ＜ 0.05).Serum CA125 levels was positively correlated with pleural fluid depth (r =0.56,P ＜0.01).Different gender,pleural effusion parts serum CA125 positive rate and the different levels were not statistically significant (P ＞ 0.05).Conclusion Serum CA125 increased in patients with benign and malignant pleural effusion,and serum CA125 was not severed as the diagnosis biomarker in differentiating benign and malignant pleural effusion.Serum CA125 levels is helpful in monitoring the change of pleural fluid size due to the relation with depth of pleural fluid.

Objective To investigate the application value for predicting human cytomegalovirus(HCMV) infection with viral load in urinary epithehal cell (EC).Methods Peripheral blood and urine specimens from 82 infants with HCMV latent infection and 84 infants with HCMV active infection were collected respectively.Plasma HCMV DNA load and the levels of HCMV lgM/IgG antibody were detected by real-time fluorescence quantitative polymernse chain reaction (FQ-PCR) and chemiluminescence immunsassay.HCMV pp65 antigen in peripheral blood polymorphonuclear leukocytes (PMNLs) was detected by indirect immunofluorescence assay.The urinary EC count and HCMV DNA load were detected by UF-100 automated urine sediment analyzer and FQ-PCR,respectively.HCMV DNA load in urinary EC was calculated accordingly.At the same time,the sensitivity and specificity for diagnosis of active HCMV infection were evaluated by receiver operating characteristic curve (ROC).Results The positivity of HCMV DNA in urinary EC was 94.58% (157/166),which was the highest among the urinary EC from 166 cases of HCMV infection.HCMV DNA load ranged from 5.67×102to 1.31×107 copies/103 EC There was no significantly statistical difference among urine specimens from different periods of time(P>0.05).HCMV DNA load in active infection group [5.13±0.99(copies/103EC,lg)]is significantly higher than that in latent infection group [3.92±0.82 (copies/103 EC,lg),t = 8.52,P < 0.01];According to ROC curve analysis,its sensitivity and specificity were 71.4% and 75.2% respectively when cut-off value was 4.55.The virus load was significantly decreased in urinary EC in post-treatment infants as compared with pre-trestment(t=5.44,P<0.01).Conclusion Detection of HCMV DNA load in the urinary EC is a cost-effective method and can be used to predict HCMV active infection in infants and monitor treatment of HCMV infection.

Objective To investigate the relationship between Klotho and autophagy in sepsisinduced acute kidney injury mice model.Methods The male healthy Balb/c mice were used to establish the model of sepsis-induced acute kidney injury by using cecal ligation and puncture (CLP).Mice were sacrificed at 3 h,6 h,12 h,1 d,2 d,3 d,and 5 d after CLP (n =12 for each interval) and on 1 d 6 mice in sham group as well as 6 mice in normal group were sacrificed at the same time.Scr and BUN in the blood serum were detected.The HE and PAS staining were employed for observation on the histopathological changes in kidney tissues under light microscope.The autophagosomes were observed under transmission electron microscope (TEM).The renal protein of Klotho,LC3 and P62 were detected by using Western blot and Immunohistochemistry.Statistical analyses were performed using Student's t-test by SPSS 23.0.software.Results Scr and BUN increased significantly after CLP,especially on 1 d,respectively (165.64 ± 20.56) μmol/L and (45.51 ± 4.05) mmol/L.HE and PAS staining showed renal tissue was damaged obviously 1 d after CLP,as indicated by desquamation of the brush border of proximal tubular epithelial cells,appearance of bare basement membrane,and interstitial inflammatory cell infiltration.Under TEM,autophagosomes and phagocytosis were observed.Compared with sham group,the expression of Klotho protein decreased gradually from 3 h to 1 d and dropped to the trough at 1 d (t =51.851,P ＜0.01),then resumed gradually from 2 d to 5 d.On the contrary,the activation of autophagy increased as indicated by the expression of LC3-Ⅱ/L3-Ⅰ and p62.Autophagy was induced gradually from 3 h to 1 d and reached peak at 1 d,then declined gradually from 2 d to 5 d (P ＜ 0.01).The protein of Klotho and LC3-Ⅱ mainly distributed in renal tubular cytoplasm,and Klotho was reduced significantly (t =-8.371,P ＜ 0.01) and LC3-Ⅱ appeared in high density remarkably (t =4.995,P =0.001) on 1 d after CLP.Conclusions Klotho protein reduction and autophagy protein increase were observed in sepsis-induced acute kidney injury,and the expressions of Klotho and autophagy acted out in certain extent of time dependence.

Objective:To explore the relationship between serum matrix metalloproteinase-8 (MMP-8), myeloperoxidase (MPO), human cytomegalovirus (HCMV) infection and the type of carotid atherosclerosis in patients with ischemic cerebrovascular disease (ICVD).Methods:From February 2017 to May 2019, 90 patients with ischemic cerebrovascular disease diagnosed by Peking University Medical Rehabilitation Hospital were selected as study group and 90 healthy adult persons who underwent health checkup as control group, and the study group was divided into non-stable plaque type group (30 cases) and stable plaque type group (30 cases) according to the ultrasonic imaging data and referring to the type of carotid plaque of the patients. The patients whose plaque property were between the two groups were were enrolled as the middle type group (30 cases). The relationship between human cytomegalovirus PP65 antigen (HCMV-PP65) in the serum, MMP-8, MPO and peripheral blood white blood cells and carotid atherosclerotic plaque in patients with ischemic cerebrovascular disease was analyzed.Results:The serum MMP-8, MPO level and HCMV-PP65 positive rate in the 3 study groups were significantly higher than that of the control group ( P<0.05), and the serum MMP-8 in different plaque types groups was significantly higher in the study group and the control group than in the control group. The higher the level of MPO and the positive rate of HCMV-PP65, the higher the serum MMP-8 and MPO level in the middle plaque type group, and the positive rate of HCMV-PP65 in the middle plaque type group was higher than that of the stable plaque group and the unstable plaque group ( P<0.05). Conclusions:The serum MMP-8, the MPO level and the HCMV infection are related to the unstable type of the carotid atherosclerosis plaque in the patients with the ischemic cerebrovascular disease, and the recognition of the carotid atherosclerosis is improved; it is an important link to control the severity of patients with ischemic cerebrovascular disease and to take timely and effective prevention and treatment measures.

OBJECTIVE: To explore the genetic basis for a child with facial dysmorphism and multiple malformations. METHODS: The child, born at 34⁺⁶ weeks' gestation due to premature rupture of amniotic membrane, dichorionic diamniotic twinning and gestational diabetes, was subjected to chromosomal karyotyping analysis and copy number variations sequencing (CNV-seq). RESULTS: The child was found to have facial dysmorphism, hypospadia, cryptorchidism and hypotonia. He was found to have a 46,XY,del(3)(p26) karyotype in addition with a 9.80 Mb deletion (chr3: 60 000-9 860 000) encompassing 33 protein coding genes. CONCLUSION The 3p26.3p25.3 deletion probably underlay the multiple malformations in this child. Continuous follow-up is required to improve his quality of life.
Humans ; Male ; Chromosome Deletion ; DNA Copy Number Variations ; Quality of Life ; Abnormalities, Multiple/genetics* ; Phenotype

OBJECTIVE: To explore the molecular pathogenesis of a Chinese pedigree affected with inherited protein C (PC) deficiency. METHODS: The protein C activity (PC:A) and protein C antigen (PC:Ag) of the proband and his family members were determined by a chromogenic substrate method and enzyme-linked immunosorbent assay, respectively. The proband was subjected to whole exome sequencing. Candidate variants were verified by Sanger sequencing of other members of the pedigree. RESULTS: The PC:A and PC:Ag of proband were reduced to 15% and 11%, respectively. The above parameters of his parents and elder sister were also decreased to approximately 50% of reference values. Next generation sequencing has revealed that the proband has harbored a heterozygous c.572_574delAGA (p.Glu191_Lys192delinsGlu) variant in exon 7 and a missense c.752C>T (p.Ala251Val) variant in exon 8 of the PROC gene. His father was heterozygous for the c.572_574delAGA variant, while his mother and elder sister were heterozygous for the c.752C>T variant. According to the American College of Medical Genetics and Genomics Standards and Guidelines, the c.572_574delAGA (p.Glu191_Lys192 delinsGlu) variant was predicted to be likely pathogenic (PS1+PM4+PP3). c.752 C>T (p.Ala251Val) variant was also likely pathogenic (PS1+PM1+PP3). CONCLUSION The deletional variant of c.572_574delAGA (p.Glu191_Lys192delinsGlu) in exon 7 and missense variant c.752C>T (p.Ala251Val) in exon 8 of the PROC gene probably underlay the inherited protein C (PC) deficiency in this pedigree. Above finding has enriched the spectrum of PROC gene variants and provided a basis for genetic counseling for this pedigree.
Humans ; China ; Mutation ; Pedigree ; Protein C/genetics* ; Protein C Deficiency/genetics* ; Male ; Female

OBJECTIVE: To explore the genetic basis for a Chinese pedigree with 6q26q27 microduplication and 15q26.3 microdeletion. METHODS: A fetus with a 6q26q27 microduplication and a 15q26.3 microdeletion diagnosed at the First Affiliated Hospital of Wenzhou Medical University in January 2021 and members of its pedigree were selected as the study subject. Clinical data of the fetus was collected. The fetus and its parents were analyzed by G-banding karyotyping and chromosomal microarray analysis (CMA), and its maternal grandparents were also subjected to G-banding karyotype analysis. RESULTS: Prenatal ultrasound had indicated intrauterine growth retardation of the fetus, though no karyotypic abnormality was found with the amniotic fluid sample and blood samples from its pedigree members. CMA revealed that the fetus has carried a 6.6 Mb microduplication in 6q26q27 and a 1.9 Mb microdeletion in 15q26.3, and his mother also carried a 6.49 duplication and a 1.867 deletion in the same region. No anomaly was found with its father. CONCLUSION The 6q26q27 microduplication and 15q26.3 microdeletion probably underlay the intrauterine growth retardation in this fetus.
Female ; Humans ; Pregnancy ; East Asian People ; Fetal Growth Retardation/genetics* ; Karyotype ; Pedigree ; Prenatal Diagnosis ; Sequence Deletion ; Chromosome Duplication

Objective:To explore the genetic basis for a proband with Shprintzen-Goldberg syndrome (SGS).Methods:Whole exome sequencing was carried out to detect potential variants associated with the relevant phenotypes. Candidate variants were verified by Sanger sequencing of the patient and her family.Results:DNA sequencing revealed that that the proband has carried a de novo heterozygous missense c. 94C>G (p.Leu32Val) variant in exon 1 of the SKI gene (NM_003036), which has been reported previously. The same variant was not detected in either parent. Based on the guidelines of the American College of Medical Genetics and Genomics, the variant was predicted to be pathogenic (PS1+ PS2+ PM1+ PM2+ PP2+ PP3). Conclusion:The SKI c. 94C>G (p. Leu32Val) variant probably underlay the autosomal dominant SGS in this patient.