OBJECTIVETo apply single nucleotide polymorphism (SNP) microarray for delineation of small supernumerary marker chromosome (sSMC) in two newborns.

METHODChromosome karyotyping was performed on newborns who were born in Jan. 2013 and Jan. 2014 in Haidian Maternal and Child Health Hospital because of the abnormalities found in pregnancy checkups. SNP microarray analysis was carried out on 2 newborns with de novo sSMCs (one was mos 47,XY, + mar[45]/46,XY[5] and the other was mos 47, XY, + mar [30]/46, XY [20]), which could not be determined by conventional banding techniques. Genomic DNA was extracted from cord blood samples, amplified, tagged and hybridized following the manufacturer' s protocol. Data were collected and analyzed.

RESULTThere was a 78. 6 Mb duplication in chromosome 8 for Newborn A, which was associated with 8p22 duplication syndrome; and a 32. 7 Mb duplication in chromosome 13 for Newborn B, which was not yet reported definitely as pathogenic. The newborn A was identified with agenesis of the corpus callosum, obvious right eyelid drooping, the onset of low muscle tone and mental developmental lag behind their peers, while the newborn B had normal findings on physical and mental evaluation.

CONCLUSIONSNP-array can identify sSMCs of newborns at the DNA level, and can be used as an important supplement to the conventional karyotype analysis, but the pathogenicity of positive outputs need further verification.

Objective To investigate the value of copy number variation analysis based on next-generation sequencing (NGS-CNVA) in the genetic analysis of missed abortion chorionic villi. Methods From August 2012 to May 2014, chorionic villi from 74 cases of missed abortion at 6-13 gestational weeks in Haidian Maternal and Child Health Hospital were collected and analyzed by karyotype analysis and NGS-CNVA. The results of the two methods were compared. Results (1) Karyotype analysis was carried out for the villi from the 74 missed abortion patients. Thirty cases were euploid, 26 cases were aneuploid, while 18 cases had structural abnormalities. The resolution of the karyotyping was 320 bands and the average report time was 22 days. (2) All of the 74 samples obtained NGS-CNVA results and the report time was 7-10 days. (3) The NGS-CNVA results of 56 cases were consistent with karyotype. Among them, 28 cases (28/56, 50%) had no copy number variants (CNV), and 19 cases (19/56, 34%)had CNV between 1 Mb and 10 Mb. 9 cases (9/56,16%) had CNV≥10 Mb found by NGS-CNVA, but not found by karyotyping. (4) According to the results of NGS-CNVA, karyotype were reviewed. The reviewed results found 7 cases with CNV<10 Mb and 3 cases with CNV≥10 Mb in 30 cases which got normal karyotype results at the first analysis. (5) Among the 18 cases of structural abnormalities, 6 cases were Robertsonian translocation. Sequencing technology could confirm the specific area of chromosome deletion/duplication in 8 cases, but could not locate them. Conclusions NGS-CNVA has lower failure rate, higher resolution, lower specimen requirement and shorter report time than karyotype analysis when used for the genetic analysis of missed chorionic villi . NGS-CNVA could be a useful genetic analysis method for the missed abortion villi.

Objective To explore the relationship between bone mineral density(BMD) and body tat disrribution in the aged in Ningbo area of China. Methods The BMD of lumbar vertebra and total body bones as well as body composition were measured by dual-energy X-ray absorptiometry (DXA) in 61 aged males, and also in 63 aged females as control group. Results In the aged males, a negative correlation was found between BMD and age, while BMI, muscle, trunk fat were positively correlated with BMD. As for the aged female, age, BMI, muscle and fat were all positively correlated with BMD. Conclusions The correlation between body fat and BMD is closer in females than in males in the aged. There is a positive correlation between trunk fat and lumbar spine BMD, and it indicates that central obesity may have a protective effect on lumbar spine BMD.

Objective To carry out a molecular screening of Chinese common deafness gene mutations in Chinese pregnant women group,so as to expatiate on the content,provide molecular epidemiological data,reduce the birth rate and provide a theoretical basis to the deaf children. Methods The molecular detection was done to the pregnant women underwent normal antenatal care in our hospital,using gene chips to screen the four com?mon deaf genes(GJB2,GJB3,SLC26A4 and mitochondrial 12S rRNA)in China;then,the newborn infants carrying mutations were treated with the hearing screening,using the methods of Otoacoustic Emissions(OAE)and Brainstem Auditory Evoked Potentials(BAEP),and the husbands of mutation carrying pregnant women were adopted molecular testing of the deaf susceptibility genes in order to investigate the correlation of the rate of pregnant women carrying the mutant genes and newborn infants deafness. Results Totally 2 067 cases of pregnant women were accepted to do the molecular screening,there were 110 cases of deafness mutations detected(5.320%),in which GJB2 gene(67 cases),GJB3 gene(6 cases), SLC26A4gene(33 cases),mitochondrial 12SrRNAgene(4 cases)mutation detection rates were 3.240%,0.290%,1.600%and 0.190%,respec?tively;especially:GJB2gene 235 del C,GJB2gene 299 del AT double mutant 1 case;GJB2gene 299 del AT,GJB3gene 538 C>T double mutant 1 case;GJB2 gene 235 del C,SLC26A4 gene IVS7?2 A>G double mutant 1 case. About 108 cases children newborn accepted to do the hearing screening,in which 3 cases had problems with the left ear,3 cases with the right ear,and 4 cases with the double ears. Conclusion The use of ge?netic deafness gene chip to do the molecular diagnostics in pregnant women can be convenient,fast and efficient for prenatal diagnosis of deafness, which provides a theoretical basis and good method for reducing the birth rate of deaf children and should be popularized more widely.

With the development of high-throughput sequencing technology, non-invasive prenatal testing (NIPT) has been rapidly used for fetal chromosomal aneuploidy screening and gradually become a first-line test. With the increase of sequencing depth and improvement of bioinformatic analysis ability, NIPT technology may also be used to detect chromosomal copy number variations and certain single gene diseases in fetuses. However, it has also encountered some problems such as insufficient understanding by different groups, false-positive and false-negative results, and selection of appropriate subjects and gestational age. This article has given a summary for such problems.

Objective:To evaluate the value of modified magnetic bead screening for enrichment of cell-free fetal DNA (cffDNA) in non-invasive prenatal testing (NIPT).Methods:This study retrospectively recruited 31 cases with low concentration of cffDNA (<6.00%), Z value in the gray zone (3.00-4.00) at the first detection, or false-positive (confirmed by invasive prenatal diagnosis) or false-negative (confirmed by postnatal chromosome test) results among 11 000 pregnant women who underwent routine NIPT in Beijing Haidian District Maternal and Child Health Care Hospital from October 2017 to December 2019. Plasma samples collected for the first-time routine NIPT were used to enrich cffDNA using modified magnetic beads for NIPT (modified NIPT). Wilcoxon rank sum test was used to compare the modified NIPT with the routine NIPT in detecting the cffDNA concentrations of male fetuses.Results:Among the 31 pregnant women, there were 13 cases with low cffDNA concentration in routine NIPT, 11 having false-positive results in the routine NIPT (three for trisomy 13, four for trisomy 18 and four for trisomy 21, all were confirmed by invasive prenatal diagnosis), six with gray-zone Z values in the first-time NIPT (retesting indicating low risk) and one having false negative result for trisomy 21 (confirmed by postnatal chromosome test). Cell-free DNA (cfDNA) fragments less than 150 bp were effectively enriched using the modified magnetic bead screening and the concentration of cffDNA was increased from 4.43% (2.45%-17.61%) in routine NIPT to 13.46% (7.75%-36.64%) in the modified NIPT ( Z=-14.22, P<0.01). Results of the modified NIPT indicated that 13 cases with low cffDNA concentration of routine NIPT were successfully detected as low risk, as well as the risks in the six cases with gray-zone Z value and six of the 11 false-positive cases in the routine NIPT were low, which were consistent with the retest results of the routine NIPT, while high risk was found in one false-negative case. Conclusions:The modified NIPT could reduce the false positive rate by lowering the failure rate caused by low concentration of cffDNA and is able to identify false-negative cases. Compared with the routine NIPT, it shows a higher success rate and a lower false positive rate.

Objective:To design a replaceable corneal scaler and explore its effect on improving the positive detection rate of corneal bacteria and the cure rate of patients with infectious keratitis.Methods:Totally 100 patients with infectious keratitis treated in the Eye&Ent Hospital of Fudan University in Shanghai from June 2020 to October 2021 were conveniently selected as the application object, 50 patients from June 2020 to January 2021 were selected as the control group, and 50 patients from February 2021 to October 2021 were selected as the experimental group. The control group used the traditional corneal scalpel for corneal scraping, and the experimental group used the replaceable corneal scalpel for corneal scraping. The positive detection rate of bacteria, the cure rate of patients as of the date of data collection, the comfort status of patients and the operation difficulty score of medical staff were compared between the two groups.Results:The 28 cases (56%) were positive in the experimental group, and 30 cases (60%) were cured as of the date of data collection; 16 cases (32%) were positive in the control group and 18 cases (36%) were cured as of the date of data collection. The difference between the two groups was statistically significant ( χ2=-2.47, -2.20, all P<0.05). The score of comfort in the experimental group was (83.92 ± 6.40) and that in the control group was (54.04 ± 10.69). The difference between the two groups was statistically significant ( t=17.14, P<0.01). The operation difficulty score of medical staff in the experimental group was (1.46 ± 0.54) and that of medical staff in the control group was (3.24 ± 0.72). The difference between the two groups was statistically significant ( t=-13.50, P<0.01). Conclusions:The use of replaceable corneal scalpel can not only improve the positive detection rate of patients, improve the treatment effect of patients, improve the comfort and satisfaction of patients, but also improve the work efficiency of medical staff and reduce the operational risk of medical staff, so as to improve the real professional value of medical staff.

Objective To investigate the effect of NAIF1 in gastric cancer cell lines MKN45. Methods We constructed pLVX-Tight-Flag-NAIF1-puro plasmid with Tet-on system. DOX was added to induce NAIF1 expression in MKN45 cells. The cells were collected at 0, 6, 12 and 24 hours after DOX addition for gene expression microarray detection and biological analysis of differentially expressed genes. qRT-PCR and Western blot were used to verify the changes in mRNA and protein levels of the selected target differential genes. Results The biological analysis of gene microarray hybridization results showed that IFIT1, IFIT2 and IFIT3 expression significantly increased at 24h, qRT-PCR also showed this change, and Western blot further verified the change in protein level. However, IFIT5 showed no significant change in mRNA and gene expression. Conclusion Over-expression of NAIF1 in gastric cancer cells can promote the expression of some immune system-related IFIT family proteins.

Objective:To analyze the construction and operation of the 19th Hangzhou Asian Games designated hospitals, and to discuss the medical emergency security work of large-scale sports events, so as to provide references for the planning of designated hospitals in future large-scale sports events.Methods:Retrospective analysis was made on the establishment principles, requirements, selection of medical support personnel, and training exercises of the designated hospitals, focusing on the key links such as organizational system, staffing, designated areas, and drug management.Results:Total of 40 designated hospitals have successfully completed the task of medical security by rebuilding the medical security area of the Asian Games, elevating the process, equipping facilities, and strengthening staff training. During the Asian Games, 349 people were transferred to designated hospitals by ambulance, 54 people were hospitalized, 19 people underwent surgery, and 1022 people went to designated hospitals by themselves.Conclusion:The construction of the designated hospitals during the 19th Hangzhou Asian Games was of high quality, efficient and smooth operation. It is suggested that efforts should be made in the reconstruction of the medical security area for the Asian Games to be "relatively independent". The treatment process of self-visiting patients should be fully considered and the flat urgent emergency response mechanism needs to be established.

Objective:To assess the efficiency of modified enrichment method for cell-free fetal DNA (cffDNA) through purified superparamagnetic beads during non-invasive prenatal testing (NIPT).Methods:A total of 26 252 pregnant women undergoing NIPT at the Maternal and Child Health Care Hospital of Haidian District from December 2017 to September 2022 were recruited and randomly assigned into the conventional group ( n = 10 573) and the modified enrichment group ( n = 15 679), who were then subjected to the screening and enrichment of the cffDNA using a conventional and modified technique, respectively. High-risk pregnant women detected by NIPT were subjected to invasive prenatal diagnosis. All women were followed up for their pregnancy outcomes, and the detection efficacy of the two methods was compared in terms of fragment size, concentration of cffDNA, duplicate detection rate, and indices of clinical laboratory tests. Results:The fragment size of the main peak of the cell-free DNA library of the modified enrichment group was significantly lower than that of the conventional group [267 (264, 269) bp vs. 294 (292, 296) bp, P<0.01], while the concentration of cffDNA was significantly higher [21.86% (17.61%, 26.36%) vs. 9.08% (6.87%, 11.87%), P<0.01]. In addition, the duplicate detection rate (0.740% vs. 2.02%, χ2=83.90, P<0.01) and detection failure rate (0.006% vs. 0.057%, P<0.05) in the modified enrichment group were significantly lower than those of the conventional group. The combined positive predictive value (PPV) in both high-risk (64.3% vs. 76.1%) and low-risk (35.3% vs. 45.5%) pregnant women from the modified enrichment group was slightly lower than those from the conventional group, though no significant difference was detected. There was one false negative case for trisomy 21 among the high-risk pregnant women from the conventional group, and no false negative case was found in the modified enrichment group. Conclusion:The modified technique to screen and enrich the cffDNA has significantly enhanced the relative concentration of cffDNA and reduced the failure and duplication detection rate of NIPT, which has significantly reduced the incidence of false negative cases due to the low concentration of cffDNA, and greatly increased the overall detection efficacy of NIPT.