Application of flat-sided culture tubes during prenatal diagnosis / 中华医学遗传学杂志

OBJECTIVE@#To assess the value of using flat-sided culture tubes for preparing chromosomes through chorionic villi (CV) and amniotic fluid (AF) cell cultures during prenatal diagnosis.@*METHODS@#From February to March 2020, 157 CV samples and 147 AF samples subjected to prenatal diagnosis at the Maternal and Child Health Care Hospital of Guangxi Zhuang Autonomous Region were selected as the study subjects. For each sample, one flat-sided tube and one flask culture were set up by following the standard protocols. The methods were evaluated by comparing the cell growth, experimental process, quality of chromosome preparation and costs.@*RESULTS@#The success rates for the culturing of CV and AF samples by the flat-sided culture tube method were 97.45% (153/157) and 97.96% (144/147), respectively. By contrast, the success rates for the conventional flask method were 98.72% (155/157) for CV and 98.64% (145/147) for AF samples. No significant difference was found between the two methods (P > 0.05). The average harvest time required by the flat-sided culture tube method was 8.45 days for CV and 9.43 days for AF cultures, whilst the average harvest time for conventional flask method was 9.05 days and 9.54 days, respectively. The flat-sided culture tube method for CV had required significantly shorter average harvest time than the conventional method (P < 0.001). No statistical significant difference was found in the average harvest time for AF by the two methods (P > 0.05). The conventional culturing method had required three containers with two sample transfers. By contrast, the flat-sided culture tube method was carried out in one tube without any sample transfer. The average total amount of medium used was 3.91 mL for each flat-sided culture tube and 6.26 mL for each conventional flask.@*CONCLUSION@#The flat-sided culture tube method can provide a simple, cost-effective and error-reducing procedure for the CV and AF samples culture during prenatal diagnosis.

Effects of different thyroid stimulating hormone cut-off values on the screening of congenital hypothyroidism in newborns in Guangxi / 中华地方病学杂志

Objective To analyze the effects of different thyroid stimulating hormone (TSH) cut-off values on the screening of congenital hypothyroidism (CH) in newborns in Guangxi.Methods The TSH results of 83 608 newborns tested by Genetic Screening Processor (GSP) from the Genetic Metabolism Center of the Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region from May 2017 to April 2018 were collected.Using the percentile method and the receiver operating characteristic (ROC) curve method,the TSH cut-off values were calculated and compared with the assumed cut-off values 9.00 or 10.00 mU/L,to analyze the effects of four different TSH cut-off values on CH screening.Results Using GSP,the TSH results of 83 608 newborns showed a positive skewed distribution,TSH cut-off value of the percentile method (P99) was 7.96 mU/L,836 cases were suspicious,43 cases were diagnosed with CH (6 cases were missed diagnosis),and 65 cases were high TSH (21 cases were missed diagnosis);TSH cut-off value of the ROC curve method was 6.45 mU/L,1 480 case were suspicious,49 cases were diagnosed with CH,and 86 cases were high TSH,both were no missed diagnosis;when TSH cut-off values were 9.00 or 10.00 mU/L,the suspicious were 478 and 305 cases,respectively,and the confirmed CH were 37 and 35 cases (missed diagnosis were 12 and 14 cases,respectively),high TSH were 46 and 33 cases (missed diagnosis were 40 and 53 cases,respectively).The CH incidence of the ROC curve method was compared with the percentile method and using the cut-off values 9.00 and 10.00 mU/L,the differences were statistically significant (P ＜ 0.05).Conclusions The GSP and ROC curve method were used to successfully establish the TSH cut-off value on the screening of CH in newborns in Guangxi.The cut-off value can not only ensure the accuracy of screening,but also avoid missed diagnosis and reduce birth defects.

Gene screening of neonatal non-syndromic hereditary hearing loss in Guangxi / 重庆医学

Objective To use the matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) technique for detecting the mutation gene of neonatal non-syndromic hereditary hearing impairment gene in Guangxi and to investigate its effectiveness and feasibility in clinical application.Methods A total of 7 100 newborns were performed the hearing preliminary screening and secondary screening by adopting AABR.The genomic DNA was extracted by the heel blood spot.Twenty mutation characteristics of 4 deaf predisposing genes were detected by MALDI-TOF-MS.Results The pass rate of hearing screening in 7 100 newborns was 97.11％ (6 895/7 100),the positive rate of neonatal gene mutation was 3.54％ (251/7 100),in which the GJB2 gene mutation was in 131 cases,the carrying rate was 1.84％,235delC heterozygous mutation was in 108 cases.SLC26A4 gene mutation was in 93 cases,which dominated by 1229C＞T heterozygous mutation and IVS7-2A＞G heterozygous mutation,mtDNA12SRNA gene mutation was in 16 cases and GJB3 gene mutation was in 11 cases.Conclusion Adopting the MALDI-TOF-MS screening technique can increase the detection rate of hot point mutation in common deaf related genes and discover neonatal genetic NSHI from molecular level and provides the corresponding geneticconsulting guidance for early finding and predicting deaf occurrence,and formulating the interventional measures.

Analysis of mass spectrometric detection in 21 neonatal with neonatal intrahepatic cholestasis disease / 实用医学杂志

Objective To analyze blood Met、 Phe 、Tyr、 Arg、 Cit、 Orn、 Ser、 Thr、 C0、 C2、 C3、 C14、C14 ∶ 1 , C16 , C16 ∶ 1 , C18 , C18 ∶ 1 and urine 4-OH-PHPLA , 4-OH-PHPPA level of NICCD patient and discuss the application value of diagnosis NICCD. Methods From May 2011 to May 2015, 21 NICCD patient were diagnose in Guangxi Newborn Screening Center. Meanwhile, 100 normal children were selected as the control group. Blood Met, Phe, Tyr and other factors and urine 4-OH-PHPLA, 4-OH-PHPPA level were analyzed by SPSS. Results In the experimental group, blood Met, Phe, Tyr and many other indexes and urine 4-OH-PHPLA, 4-OH-PHPPA level were higher and blood Orn/Cit were lower than the control group(P 0.05). Conclusion NICCD patient has abnormal biochemical index. Blood test by TMS and urine test by GC-MS are very important in NICCD diagnosis.