Characterized by dihydropyrimidinuria, dihydropyrimidinase (DHP) deficiency refers to a rare disorder of pyrimidine degradation, with high phenotypic heterogeneity.The disease-causing gene is DPYS, and less than 40 cases were reported worldwide.Urinary gas chromatography/mass spectrometer (GC/MS) can screen clinically suspected patients, and gene sequencing is the main means of the diagnosis of the disease.This article reviews the pathogenesis, clinical manifestation, genotype and recent research progress of the disease.

Objective To summarize the clinical features of children with viral encephalitis accompa-nied with respiratory failure,and to improve the early diagnosis and treatment. Methods The clinical data of 64 cases with viral encephalitis combined with respiratory failure in our unit from May 2005 to May 2015 were analyzed retrospectively. Results All children were characterized by sudden onset. Among them, 60 cases (93. 7%) had fever,50 cases(78. 1%) had convulsion onset,46 cases(71. 8%) had consciousness disorders, 30 cases(46. 8%) had positive pathological signs. Most of them developed respiratory failure in acute stage. Total 56 cases occurred central respiratory failure,6 cases occurred central respiratory failure with peripheral respiratory failure,2 cases occurred respiratory and circulatory failure. Total 46 cases underwent cerebrospinal fluid examination. Routine biochemical test found 32 abnormal cases,of which 10 cases had intracranial hyper-tension and 19 cases had leukocytosis,and 17 cases had increased protein content. Ten cases were positive in cerebrospinal fluid etiology examination,including herpes simplex virus positive in 8 cases,EB virus positive in 1 case,and coxsackie virus positive in 1 case. There were 6 of 64 cases with abnormal CT scans and 29 of 34 cases with abnormal MRI. The results of EEG examination were abnormal in all patients for the first time. The EEG of 48 cases showed diffuse slow waves-δ activity. EEG examination showed generalized discharges or focal discharges during treatment in 22 cases. Five cases of electrophysiological examination showed cervical spinal cord anterior horn injury. Total 24 cases were complicated with stress ulcer,4 cases with liver damage,6 cases with heart damage,4 cases with renal damage,1 case with lung damage. All cases underwent mechanical ventilation for 2-50 days. Total 33 cases(51. 5%) improved and discharged,14 cases died during hospitaliza-tion,17 cases were given up treatment. Total 25 cases had variety of neurological dysfunctions left. Total 14 cases with epilepsy were followed up for 3 months to 6 years in our department,of which 8 cases were medical-ly intractable epilepsies. Conclusion The children with viral encephalitis complicated with respiratory failure have acute onset,rapid progress,high disability and mortality. Early diagnosis and evaluation,effective mechani-cal ventilation in time,and protecting organ function,help to improve the prognosis.

[Objective]To evaluate the incidence of chromosomal abnormalities and associated abnormalities in prenatally diag?nosed clubfoot,and to determine the prognostic factors as well.[Methods]A total of 89 fetuses with clubfoot diagnosed during Janu?ary 2010 to October 2015 in prenatal ultrasound scan and confirmed postnatally or by autopsy,were selected,within which 16 (18.0%)cases were without other abnormalities and 73(82.0%)cases were with other abnormalities. The associated abnormalities were identified ,the correlation with chromosomal abnormalities were analyzed with Fisher analysis and the factors affecting the outcomes were determined with Logistic regression analysis.[Results]Among associated abnormalities ,the skeletal abnormalities besides the clubfoot were the most frequently associated anomalies (35 ,47.9%),and the central nervous abnormalities followed secondly(30,41.1%). A chromosomal abnormality,with trisomy 18 being the most frequently detected,was identified in 34%(17/49)of the clubfoot fetuses with other anomalies ,whereas none of chromosomal abnormality was identified in 11 fetuses without other anomalies,a significant different rate of aberrant chromosome noted(P < 0.001). The survival rate of clubfoot fetuses without other anomalies was higher than that of clubfoot fetuses with other anomalies(50.0%vs 1.3%,P=0.03). The conditions of with or without associated anomalies were the independent prognostic factors (P = 0.01),the clubfoot fetuses associated with other anomalies had poor outcomes[OR=11.9(95%CI:1.8,80.1)].[Conclusion]Skeletal abnormalities besides the clubfoot were the most frequently associated anomalies. The condition of with or without associated anomalies is the independent prognostic index for fetuses with clubfoot. Aneuploidy were more commonly in clubfoot fetuses with associated abnormalities than in those without other abnormalities. No indication for karyotyping suggests for the clubfoot fetuses without other abnormalities due to the low incidence of associate chromo?somal anomalies.

Objective To investigate the action of chondrogenesis differentiation of bone marrow stromal cells (BMSCs) transfected with adeno-hTGF-β1. Methods In the experiment group, replication-deficient a denoviruses carrying human hTGF-β1 complementary DNA (adeno-hTGF-β1 was constructed and applied to transfect to the first generation BMSCs. As a control, each BMSCs was transduced with 200 pfu of adeno-LacZ gene. One day after transfer, BMSCs were trypsinized, counted, and 5×105 cells aliuots were spun down at 500 rpm per minute in 15 ml polypropylene conical tubes and then cultured in a defined medium in an incubator at 37℃ for 21 days. The aggregates were harvested at time points to 21 days and assessed by gross observation, histological analyses and immunohistochemical localization of type Ⅱ collagen. Results When harvested at 21 days, each pellet shrinked to spheroid tissue with apearly opalescence in gross morphology and found to be relatively firm. H.E staining showed elongate dlining cells appeared as perichon drium-like cells at the surface. Some nests of cartilage were observed at the substrate of the tissue. Mature chon drocytes were embeded in the lacuna in the experiment group. In addition, Safranin'O staining confirmed the presence of sulfated proteoglycans in the ECM of chondrogenesis region. Immunohistochemical staining revealed the presence of type Ⅱ collagen in chondrogenesis region. By contrast, HE staining showed no evidence of cartilage formation in the control group. They were fibrous tissue with no architectural feature. Safranin'O staining and Immunohistochemical staining showed no evidence of sulfated proteoglycans or typeⅡ collagen expression. Conclusion BMSCs transfected with adeno-hTGF-β1 could induce its chondro-genesis when aggregate cultured in a defined medium in vitro, laying a foundation for the application of hTGFβ1 gene-transfected BMSCs in cartilage tissue engineering.

OBJECTIVE:To systematically review therapeutic efficacy of Haikun shenxi capsule in the treatment of chronic renal failure (CRF).METHODS:Retrieved from Central,PubMed,EMBase,CJFD,CBM,VIP and Wanfang database,randomized clinical trials (RCTs) about Haikun shenxi capsule combined with routine treatment (trial group)vs.single routine treatment (control group) in the treatment of CRF were collected.Meta-analysis was performed by Rev Man 5.3 software after screening literature,extracting data and evaluating quality by using risk bias evaluation of Cochrane collaboration network.RESULTS:A total of 10 RCTs were included eventually,involving 704 patients.The results of analysis showed that compared to general therapy,Haikun shenxi capsule could improve total effective rate [RR =4.42,95 ％ CI (2.70,7.22),P＜ 0.001],and could reduce SCr[MD =-140.37,95 ％ CI (-191.72,-89.03),P＜ 0.001],BUN[MD =-5.49,95 ％ CI (-8.36,-2.63),P＜ 0.001] and 24 h-Upro [MD =-0.43,95 ％ CI (-0.62,-0.23),P＜ 0.001],with statistical significance.CONCLUSIONS:The clinical efficacy of Haikun shenxi capsule in the treatment of CRF is good and significantly improve related indexes of renal function.

Objective To investigate the clinical performance of ultrasound screening for fetal structural anomalies at 11-13+6 weeks of gestation and to evaluate the relation of structural anomalies with karyotypes and copy number variations. Methods A retrospective analysis was conducted on fetuses with structural anomalies detected by ultrasound examination at 11-13+6 gestational weeks in First Affiliated Hospital of Sun Yat-Sen University from January 2013 to December 2017. Karyotype and chromosomal microarray analysis(CMA) were offered to these fetuses and ultrasound scans were repeated at 16-18 gestational weeks. All fetuses were followed up to termination or birth. Fisher's exact test was used for statistical analysis. Results A total of 362 fetuses with structural anomalies were studied including 101 (27.9%) fatal malformations, 253 (69.9%) major malformations and eight (0.2%) minor malformations. Cardiac malformation (32.6%, 118/362), central nervous system anomalies (24.9%, 90/362) and anterior abdominal wall defects (20.9%, 76/362) were the three most common abnormalities. Invasive prenatal test was performed in 107 cases including 25 fatal, 79 major and three minor malformations. Thirty (28%) out of the 107 cases had abnormal karyotypes, which were chromosomal aneuploidies (n=28) and chromosomal fragment abnormalities (n=2). Among the 99 cases received CMA, 25 had abnormal karyotypes, and copy number variations were identified in eight [three (4.05%) were pathogenic variations] out of the rest 74 with normal karyotypes. The incidence of chromosomal abnormalities in fetuses with major malformations was higher than that of fetuses with fatal malformation [32.9% (26/79) vs 12.0% (3/25), P=0.045]. Altogether, 117 cases repeated second-trimester ultrasound among which 16 (13.7%) were normal; 19 (16.2%) had cardiac defect which was discordant with the first-trimester evaluation and five (4.2%) were found to have additional malformations. Diagnosis of the other 77 cases were consistent with the first-trimester ultrasound findings. After the second-trimester ultrasound scanning, 49 pregnancies were terminated; 39 twin pregnancies and four triplet pregnancies underwent selective fetal reduction; 25 continued to delivery with good neonatal outcomes. Out of the 23 699 cases without abnormal ultrasound findings at 11-13+6 gestational weeks, 20 182 (85.2%) were successfully followed up, among which structural abnormalities were found in 178 during the second trimester and in 31 after birth. Conclusions A detailed ultrasound examination at 11-13+6 weeks of gestation is important to identify fetal structural defects. However, it could not replace the second-trimester ultrasound. There is a high risk of chromosomal abnormalities in fetuses with early-detected structural defects. CMA is able to identify pathogenic copy number variations with a relatively low detection rate.

OBJECTIVE: To explore the genetic basis for a child with dihydropyrimidase (DHP) deficiency. METHODS: High-throughput sequencing was carried out for the child. Suspected variants were verified by using Sanger sequencing. RESULTS: The proband was found to carry compound heterozygous variants of the DPYS gene, namely c.1468C>T (a missense variant) and c.1339-1363del (a frameshifting variant). CONCLUSION The compound heterozygous variants of the DPYS gene probably underlie the DHP in this child. Above result has enabled genetic counseling and prenatal diagnosis for his parents.

Objective: To analyze the clinical characteristics and gene mutations of early-onset epileptic encephalopathy(EOEE) caused by ion channel gene mutation, to identify the etiology, to guide the treatment and to provide the basis for genetic counseling. Methods: The clinical data from 17 children with EOEE caused by ion channel gene mutation and the peripheral blood of the children and their parents were collected from June 2014 to May 2018 at the Department of Neurology, Tianjin Children′s Hospital.Epilepsy gene sequencing was performed by using disease gene targeting second generation sequencing technology.The mutation of pathogenic ion channel gene was found.The confirmed mutations were verified by Sanger sequencing and the source of the mutation was identified. Results: Among 17 case with EOEE, 3 cases had genetic mutation, and 14 cases had denovo mutations.Dravet syndrome was found in 8 cases (47.1%), there were SCN1A gene missense mutation in 5 cases, SCN1A gene nonsense mutation in 3 cases, KCNQ2 gene missense mutation in 1 case (5.9%) and non-specific epileptic encephalopathy in 8 cases (47.1%). SCN2A gene missense mutation, SCN4A gene missense mutation, SCN8A gene missense mutation, KCNQ2 gene missense mutation and KCNH gene missense mutation were found in suspected pathogenic mutations.There were 1 missense mutation out of 5 genes, 1 missense mutation of CACNA1A gene, 1 missense mutation of GRIN2A gene and 1 missense mutation of GRIN3A gene.Seventeen patients were treated with 2 or more antiepileptic drugs, 4 with ketogenic diet and 1 with vitamin B₆ supplementation.During 11 to 96 months of follow-up, seizures were completely controlled in 3 cases (17.6%), decreased in 7 cases (41.2%) by more than 50%, and decreased in 7 cases (41.2%) by less than 50%. Conclusions The clinical phenotypes for children with unexplained EOEE are varied, and gene mutations of ion cha-nnel are most common.Some gene sites are denovo mutations which have not been reported such as missense mutation for 3 case SCN1A gene, 1 case SCN2A gene, 1 case CACNA1A gene, 1 case KCNH5 gene, and nonsense mutation for 2 case SCN1A gene, which have enriched the mutation spectrum of EOEE.

Objective:To develop a list of basic and expanded medical laboratory tests in community health service centers in Shanghai.Methods:The status quo of human and equipment resource allocation, the test items and quality control currently performed, the perspectives of various stakeholders, the capacity building of community clinical laboratory in community health service centers in Shanghai were investigated by quantitative survey and qualitative interview; and the rating scores of each test item were assessed by expert consultation using Delphi method. The expert focus discussion was conducted, and each test item was rated and classified. Finally a list of the basic tests and expanded tests in clinical laboratories of community health service center was developed.Results:A total of 247 questionnaires were distributed and 192 (77.7%) were answered. A list of 94 laboratory test items was screened out based on the questionnaire survey of the laboratories of the community health centers. Thirty one experts in the relevant areas were invited to rate the test items, the average authority coefficient of experts was 0.90, with which the weighted average of the expert ratings was made. There were 45 (47.9%) items scored 7 or higher, 38 (40.4%) scored between 5 and 7, and 11 (11.7%) scored less than 5. Based on the results of the expert focus discussion, 48 items were recommended as the basic tests and 46 items as the extended tests.Conclusion:In this study a list of tests recommended to clinical laboratories in Shanghai community health service centers has been developed, which contains 48 basic tests and 46 extended tests.

Objective:To explore the value of detecting plasma donor-derived free DNA (dd-cfDNA) fraction in distinguishing antibody mediated-rejection (ABMR) and T cell-mediated rejection (TCMR) of renal allografts.Methods:Patients with acute rejection confirmed by allograft biopsy in the First Affiliated Hospital of Medical College of Zhejiang University from December 1, 2017 to July 18, 2019 were retrospectively included. Based on pathological classification of Banff renal allograft rejection in 2017, the patients were divided into ABMR group and TCMR group, and the latter was subdivided into TCMR Ⅰ subgroup and TCMR Ⅱ subgroup. The second generation sequencing and target region capture were used to detect candidates' peripheral blood dd-cfDNA. The demographic and clinicopathological data of the two groups were compared. The receiver operating characteristic curve (ROC) was used to evaluate the differential value of plasma dd-cfDNA and serum creatinine levels in two kinds of acute renal allograft rejection.Results:A total of 60 patients with acute rejection of renal transplantation were enrolled in this study, including 42 patients in TCMR group and 18 patients in ABMR group. The plasma dd-cfDNA percentage (%) in the ABMR group was significantly higher than that in the TCMR group [2.33(1.19, 4.30)% vs 0.98(0.50, 1.82)%, P=0.001]. The absolute value of dd-cfDNA in ABMR group was obviously higher than that in TCMR group [0.94(0.60, 2.27) ng/ml vs 0.43(0.20, 0.96) ng/ml, P=0.003]. ROC analysis to discriminate TCMR from ABMR showed that, the area under the curve ( AUC) of dd-cfDNA% was 0.76(95% CI 0.64-0.88), when the threshold was 1.11%, the sensitivity and specificity were 88.89% and 59.52%, respectively; the AUC of absolute value of dd-cfDNA was 0.74(95% CI 0.61-0.86), when the threshold was 0.53 ng/ml, the sensitivity was 88.89% and the specificity was 54.76%. TCMR subgroups were further analyzed, there was no significant difference between TCMR subgroups on the absolute value and percentage of dd-cfDNA (both P>0.05); dd-cfDNA% in ABMR group was apparently higher than that in TCMRⅠ subgroups ( P=0.008) and TCMRⅡsubgroup ( P=0.030). The absolute value of dd-cfDNA in ABMR group was significantly higher than that in TCMRⅠsubgroups ( P=0.003). Conclusion:Plasma dd-cfDNA level may help to distinguish between ABMR and TCMR rejection.