Objective To investigate the effects of physical and chemical factors in the environment for dried blood sample (DBS) preparation of neonatal screening assay.Methods A total of 60 normal and 120 positive DBS were prepared under control and 10 different conditions.Another 30 normal and 80 positive DBS were prepared under control and 7 different concentration gradients of formaldehyde.The levels of phenylalanine (Phe),glucose-6-phosphate dehydrogenease (G6PD),thyroid stimulating hormone (TSH) and 17α-hydoxyprogesterone (17α-OHP) were tested by time-resolved fluorescence immunoassay or fluorescence assay.Statistical analysis was performed using SPSS 22.0 software.Results Compared with the control group,the results of Phe were not significantly different (P ＞ 0.05) when the samples were dried under the formaldehyde sensitive threshold (4.62 to 6.95 ppm for 18 hours).G6PD levels were significantly lowered when the samples were dried under all the conditions except for fast cold drying (2 to 8 ℃ overnight and formaldehyde condition,0.30 to 0.38 ppm for 4 hours or 0.21 to 0.24 ppm for 18 hours).TSH and 17α-OHP levels were lowered obviously when the samples were dried under the conditions of humidity,UV and formaldehyde condition (TSH:0.32 to 0.52 ppm for 4 hours,0.38 to 0.45 ppm for 18 hours,17α-OHP:4.37 to 4.62 ppm for 4 hours,0.38 to 0.45 ppm for 18 hours).The results of Phe,G6PD,TSH and 17α-OHP were not statistically different with the control group when the samples were dried under the fast cold drying and 2 to 8 ℃ overnight.Conclusion The physical and chemical factors in the environment of DBS preparation should be related to the accuracy of neonatal disease screening closely.The necessary control factors including formaldehyde,ethanol,glacial acetic acid,ultraviolet irradiation,heat,humidity and decoration pollution may exhibit significant effects on the preparation of DBS.Fast cold drying and overnight at 2 to 8 ℃ could be available for DBS preparation.

Objective To evaluate the protective effects of Sipunculus nudus preparation(SNP) on mice irradiated by 60Co.Methods The mice were divided into control group,model group and 0.5,1.0,and 2.0g?kg-1?d-1 SNP groups.SNP was administrated by intra-gastric infusion with the volume of 0.4mL per 20g.In the fifteenth day,the model group and the treated groups were irradiated by ?-ray of 60Co(0.83Gy/h) with the dose of 5 Gy.Peripheral blood WBC,RBC,PLT,the spleen(SI) and thymus index(TI),the BMNC,SOD and MDA in serum,testicle index,germ cells were detected.Results Comparing with the model group,TI,SI and peripheral blood WBC,RBC and BMNCs were higher.The activities of SOD in serum were increased,while MDA decreased.Testicle index and germ cells were increased,and germ abnomality rate was decreased in SNP groups.Conclusion SNP has protective effects on mice irradiatied by 60Co.

Objective:To introduce the method and clinical effect of inferior pedicle flap used in reduction mammoplasty.Methods:From January 2010 to December 2019, 19 patients with moderate to severe macromastia who underwent reduction mammoplasty with inferior pedicle flap were enrolled in this study. The method of inferior pedicle flap was based on Robbins vertical inferior pedicle flap. New position of nipple areola and inferior pedicle flap were designed in terms of the breast anatomy.Results:A total of 19 patients (38 breasts) were included in this study. The average tissue reduction of one side breast was 570 g (385 g to 1 525 g). After operation, all patients were satisfied with natural appearance and nipple erection effect. There were no operation-related complications including nipple-areola necrosis. After 6 months to 8 years following-up, No inverted T hypertrophic scar occurred. Hyperpigmentation occurred on two patients with dark-skinned. The position of bilateral nipples and areola was higher in 1 patient after operation. The other 18 patients were satisfied with the position and shape of nipples and areola.Conclusions:It is a good choice for patients with moderate to severe macromastia to receive reduction mammoplasty with inferior pedicle flap. Through the method, large amount of breast tissue reduction and stable blood supply of nipple and areola complex could be obtained.

Objective: To investigate the prevalence of galactosemia(GAL), and the characteristics of genotype and phenotype of newborns who were confirmed with GAL in newborn screening in Zhejiang province. Method: The number of all live births, newborn screened infants and all clinical data of confirmed newborns with GAL from October 2013 to March 2015 were retrospectively analyzed by reviewing the data of Zhejiang Province screening center database. And the characteristics of genes and the clinical data of GAL cases who were confirmed by correlative gene test and enzyme activity measurement were analyzed. Result: The prevalence of GAL in Zhejiang province was 1/189 857. Among them, there was 1 case confirmed with GAL typeⅠ (prevalence, 1/759 428), with mutations of c. 904+ 1G>T and c. 687G>A, the enzyme activity of galactose-1-phosphate uridyltransferase (GALT) was 56.4% of controls. And there was 1 case of GAL typeⅡ(prevalence, 1/759 428), with mutations of c. 85G>T and c. 502G>A. There were 2 cases confirmed with GAL type Ⅲ(prevalence, 1/379 714), with mutations of c. 505C>T, c. 452G>A, c. 280G>A and c. 925G>A, the enzyme activity of UDP-galactose-4′-epimerase (GALE) were 42% and 38% of controls, respectively. All cases had different abnormal biochemical marks of liver function, and 1 case had combined hyperlactacidemia or hyperammonemia or increase of multiple kinds of amino acids, respectively. The newborn of GAL type Ⅱ had phacoscotasmus before treatment. All the cases were fed with lactose free milk powder, and all the abnormal parameters were improved during following up. Conclusion The disease of GAL is rare in Zhejiang province, and its genotype distribution is scattered with comparatively mind clinical manifestations, and the cases with early treatment with lactose free milk powder have good prognosis. All cases needed to be treated and followed up for a life-long time. It is recommended that the high risk cases with GAL should be screened as soon as possible.

OBJECTIVE: To explore the genetic basis for a child with mental retardation. METHODS: The child was subjected to next generation sequencing (NGS). Candidate variant was analyzed with bioinformatic software. RESULTS: NGS revealed that the child has carried a de novo heterozygous c.4035G>C (p.Gln1345His) variant of the ARID1B gene. The variant was unreported previously and may cause instability of the protein structure. CONCLUSION The de novo missense variant of ARID1B gene may underlie the mental retardation in the child. Above result has enabled genetic counseling and prenatal diagnosis for her family.

Objective: To explore procedures and clinical effect of the technique, combined reposition of rectus abdominis with abdminioplasty, for treating postpartum women with diastasis rectus abdominis and lax abdominal wall. Methods: Postpartum women with diastasis rectus abdominis and lax abdominal wall who underwent reposition of rectus abdominis and abdminioplasty were enrolled in this study. This combined technique was based on abdominoplasty, including reposition of rectus abdominis, umbilical transposition and tightening abdominal wall. Results: A total of 12 patients were included in this study. Umbilical transposition was performed in 9 patients. Skin was resected by 8 to 14 centimetre in width.There were no infection, skin necrosis, and delayed wound healing.Natural abdominal contour has been found in all patients and the abdominal perimeter as well as the waistline have shrunk.Further more, this combined technique has been shown to improve quality of life by reducing pain, restoring function, and increasing satisfaction with appearance.After 3 months to 2 years following up, all patients were satisfied with operative outcomes. Conclusions This combined technique has been shown to be a good choice for postpartum women with diastasis rectus abdominis and lax abdominal wall.

Objective To investigate the prevalence of galactosemia ( GAL) , and the characteristics of genotype and phenotype of newborns who were confirmed with GAL in newborn screening in Zhejiang province .Method The number of all live births , newborn screened infants and all clinical data of confirmed newborns with GAL from October 2013 to March 2015 were retrospectively analyzed by reviewing the data of Zhejiang Province screening center database .And the characteristics of genes and the clinical data of GAL cases who were confirmed by correlative gene test and enzyme activity measurement were analyzed.Result The prevalence of GAL in Zhejiang province was 1/189 857.Among them, there was 1 case confirmed with GAL typeⅠ(prevalence,1/759 428), with mutations of c.904+1G>T and c.687G>A,the enzyme activity of galactose-1-phosphate uridyltransferase ( GALT) was 56.4% of controls.And there was 1 case of GAL typeⅡ( prevalence,1/759 428),with mutations of c.85G>T and c.502G>A. There were 2 cases confirmed with GAL type Ⅲ(prevalence, 1/379 714),with mutations of c.505C>T, c. 452G>A, c.280G>A and c.925G>A,the enzyme activity of UDP-galactose-4′-epimerase ( GALE) were 42% and 38% of controls, respectively .All cases had different abnormal biochemical marks of liver function, and 1 case had combined hyperlactacidemia or hyperammonemia or increase of multiple kinds of amino acids, respectively.The newborn of GAL type Ⅱhad phacoscotasmus before treatment .All the cases were fed with lactose free milk powder , and all the abnormal parameters were improved during following up . Conclusion The disease of GAL is rare in Zhejiang province , and its genotype distribution is scattered with comparatively mind clinical manifestations , and the cases with early treatment with lactose free milk powder have good prognosis .All cases needed to be treated and followed up for a life-long time.It is recommended that the high risk cases with GAL should be screened as soon as possible .

Objective:To investigate the application feasibility of Region 4 Stork (R4S) system, an international collaborative newborn screening data platform, combined with cut-off value analysis in the neonatal screening for very long chain acyl-CoA dehydrogenase deficiency (VLCADD) by tandem mass spectrometry (MS/MS).Methods:The retrospective study was performed in 2, 040 072 neonates screened by MS/MS in Neonatal Screening Center of Zhejiang Province, China from October 2013 to July 2018. Nine hundred and ten cases were determined and identified as suspected positive VLCADD neonates by traditional cut-off method of tandem mass spectrometry. The original data of these 910 screened neonates were further analyzed by R4S system. Based on clinical diagnosis and ACADL gene test results, the screening efficiency between two methods was statistically compared.Results:The data of 910 suspected VLCADD-positive cases interpreted by cut-off method were further analyzed by R4S system, and the positive interpretation was reduced to 238 cases (including 9 confirmed positive cases). A total of 16 different mutations were found in ACADL gene sequencing among the confirmed children. The screening false positive rate (FPR) declined from 0.44‰ (901/2 040 072) to 0.11‰ (229/2 040 072), the rate of positive predictive value (PPV) increased from 0.99% (9/910) to 3.78% (9/238), and the specificity increased from 99.96% (2 039 162/2 040 063) to 99.99% (2 039 834/2 040 063). There was a statistically significant difference between cut-off method alone and cut-off method combined R4S system analysis (χ2=393.5, P<0.05). Conclusions:The R4S system combined with cut-off method applied in VLCADD neonatal screening by MS/MS can effectively improve screening performance, reduce false positive rate, and has certain value in clinical application.

Objective To investigate the feasibility of Region 4 Stork(R4S)project used for newborn screening by tandem mass spectrometry in China.Methods This retrospective study was performed among 362 822 neonates screened by tandem mass spectrometry from May 2015 to April 2016 in Zhejiang newborn screening center.Infants were grouped by screening result category: 83 true positive cases,360 554 true negative cases and 2 185 false positive cases.Raw data was uploaded into R4S website to perform postanalytical interpretive tools, then results were analyzed with interpretation rules.The comparisons of normal population percentiles were done at five selected percentiles between Zhejiang newborn screening center and R4S project with min-max normalization.Results Compared with cutoff system by using R4S project with interpretation rules,the positive predictive value increased from 3.7%to 8.3%,the specificity increased from 99.40%to 99.75%, and the false positive rate declined from 0.6% to 0.2%. The two cases of true positive hyperprolinemia were reported negative, and one case of β-ketothiolase deficiency was misdiagnosis.Totally 311 638 cases in true negative group were resolved by postanalytical interpretive tools,and the remaining 48 916 cases were excluded with interpretation rules.False positive cases were reduced to 897 cases.Results of percentiles comparison showed that levels of some markers were significantly different between zhejiang newborn screening center and R 4S project.Conclusions R4S project effectively improved the newborn screening performance, whereas leaded to a small number of misdiagnosis and missed diagnosis.Besides,many true negative cases should be excluded with interpretation rules.Optimization should be achieved based on local normal population.(Chin J Lab Med,2018,41:300-304)

Objective:To investigate the expression of long non-coding RNA (lncRNA) COX10-AS1 in renal cell carcinoma tissues and cell lines and its effect on proliferation and migration of renal cancer cells.Methods:Fluorescence real-time quantitative PCR (qRT-PCR) was used to detect the expression of COX10-AS1 in surgical specimens that have been diagnosed as renal cancer tissues and adjacent tissues by pathology, renal cancer cell lines (786-O, CaKi-1, A498, ACHN) and normal renal tubular epithelium cell line (HK-2). The ACHN cells with the lowest expression were divided into a control group (transfected with a negative control plasmid carrying nonsense sequences) and an experimental group (transfected with a plasmid carrying COX10-AS1 sequences). The expression level of COX10-AS1 was detected by qRT-PCR in two groups of cells. The proliferation and migration ability of ACHN cells were detected by MTS assay and cell scratch assay. The expression of MFN2 mRNA was detected by qRT-PCR. The expressions of MFN2 and Ras-NF-κB signaling pathway proteins were detected by Western blotting. The measurement data were expressed as mean±standard deviation ( Mean± SD), the comparison between the two groups used the t-test, and the comparison among multiple groups adopts the one-way analysis of variance. Results:The expression of COX10-AS1 in renal cell carcinoma was significantly lower than that in adjacent tissues ( P<0.01), The expression of COX10-AS1 in renal cell carcinoma cells was significantly lower than that in renal tubular epithelial cells ( P<0.05), the expression of COX10-AS1 was the lowest in ACHN cells( P<0.01), the above differences were statistically significant compared with the control group, the expression of COX10-AS1 in ACHN cells of experimental group was significantly increased ( P<0.01), the above differences were statistically significant compared with the control cells, the proliferation of ACHN cells in the experimental group was significantly decreased ( P<0.05), and the cell migration ability was significantly decreased ( P<0.01). Compared with the control cells, the expression of MFN2 mRNA in ACHN cells of experimental group was significantly increased ( P<0.01). The expression levels of MFN2 were significantly up-regulated ( P<0.01), and Ras-NF-κB signaling pathway proteins were significantly down-regulated ( P<0.05), the above differences were statistically significant. Conclusions:The expression of COX10-AS1 is decreased in renal cell carcinoma tissues and cell lines. COX10-AS1 may inhibit the proliferation and migration of ACHN cells by promoting the expression of MFN2 gene.