Application value of modified magnetic bead screening for enrichment of cell-free fetal DNA from maternal peripheral blood in non-invasive prenatal testing / 中华围产医学杂志

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.

Clinical application of upper arm central venous port in cancer patients / 中国实用护理杂志

Objective:To investigate the clinical effect of upper arm central venous port in cancer patients.Methods:A total of 500 patients with tumor were selected as the study subjects from March 29, 2018 to January 19 2020. Complications such as catheter-related thrombosis and catheter-related bloodstream infection during the indwelling period were recorded.Results:One patient with severe superior vena cava syndrome failed to be intubated. The other 499 patients were successfully intubated with a success rate of 99.8% (499/500). The rate of complications was 2.6% (13/499), and the rate of catheter-related bloodstream infection was 0.029‰ (4/139 614) in 4 cases, the incidence of catheter-related thrombus was 0.057‰ (8/139 614) in 8 cases. There were 1 case of body turnover, 1 case of median nerve injury, 1 case of local tissue necrosis and 1 case of lymphatic leakage. Extubation was planned in 15 patients, and unplanned extubation in 6 patients due to complications.Conclusions:The upper arm port has less complications and is suitable for patients with tumor chemotherapy and long-term need of infusion. It is worthy of clinical promotion.

Screening for hotspot mutations associated with genetic hearing impairment in pregnant women and subsequent prenatal diagnosis in high risk pregnancies / 中华耳鼻咽喉头颈外科杂志

Objective@#To screen for hotspot gene mutations associated with genetic deafness in Chinese pregnant women, and to perform risk assessment and prenatal diagnosis in high-risk families.@*Methods@#Between November 2012 and October 2017, 26 117 pregnant women were screened by molecular hybridization microarray for 9 hot-spot mutations in 4 hereditary deafness related genes (GJB2 c. 35 del G, c. 176_191 del 16 bp, c. 235 del G, c. 299_300 del AT, GJB3 c. 538 C>T, SLC26A4 c. 2168 A>G, IVS 7-2 A>G, mitochondrial DNA 12S rRNA m. 1494 C>T, m. 1555 A>G). Genotype analysis was carried out in husbands of women carrying mutations, and prenatal diagnosis was carried out in the fetuses with high risk of deafness.@*Results@#Among all women tested, 1 208(4.63%) were carriers of genetic deafness mutations, 7 with hearing impairment were affected by homozygous or compound heterozygous mutations, 51 were mitochondrial gene mutation carriers, 103 were carriers of GJB3 c. 538 C>T heterozygous mutation, 1 026 were carriers of GJB2 or SLC26A4 heterozygous mutations, and 21 carried heterozygous mutations in two genes simultaneously. In 394 families, the husbands accepted gene sequence testing, and 27 in which were determined as carriers of mutations in identical genes as their wives. Among which, 18 families received prenatal diagnosis, and 5 fetuses were diagnosed as hereditary deafness. In 9 families who did not receive prenatal diagnosis, 1 neonate was diagnosed as compound heterozygote after delivery.@*Conclusion@#In order to prevent birth defects with congenital hearing problems, it is effective to provide screening for hotspot mutations in pregnant women and to perform prenatal diagnosis on high risk pregnancies.

Application of copy number variation analysis based on next-generation sequencing in the genetic analysis of missed abortion chorionic villi / 中华妇产科杂志

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.

Detection of small supernumerary marker chromosome by single nucleotide polymorphism microarray chips / 中华儿科杂志

<p><b>OBJECTIVE</b>To apply single nucleotide polymorphism (SNP) microarray for delineation of small supernumerary marker chromosome (sSMC) in two newborns.</p><p><b>METHOD</b>Chromosome 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.</p><p><b>RESULT</b>There 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.</p><p><b>CONCLUSION</b>SNP-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.</p>

Analysis of the Rate of Common Genetic Mutations of Deaf in Pregnant Women / 中国医科大学学报

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.

Molecular adjuvant C3d can improve humoral immunity of hCG? DNA immunization / 中国免疫学杂志

Objective:In the past study, a plasmid of pcDNA3 hCG? C3d3 had been constructed. It was suggested that the fusing protein be expressed by the pcDNA3 hCG? C3d3 plasmid both in the transient expression system in COS 7 cells, and the stable expression system in CHO cells. In the present research, it would be testified that the C3d molecular adjuvant could improve the immunogenicity of the hCG? DNA immunization or not. Methods:BALB/C mice of 6 weeks old were immunized intramuscularly two times at interval of 3 weeks by the plasmid pcDNA3, pcDNA3 hCG?, pcDNA3 hCG? C3d3 at dosage of 5, 10, 20 pmol, respectively. The anti hCG? antibody titers were determined by indirect ELISA in 6 weeks of last immunization. Results:The result showed that the C3d molecular adjuvant could increase significantly the titer of anti hCG? antibody in a dose dependent manner after DNA immunization.Conclusion:The C3d molecular adjuvant does improve the humoral immunity of the hCG? DNA immunization, which would be helpful for technical progress and clinical trial of the contraceptive vaccine. [

Effects of human urotensin II on ischemia/reperfusion injury in isolated perfused rat hearts / 中国病理生理杂志

AIM: To investigate the effects of human urotensin II (hUII) on ischemia/reperfusion (I/R) injury in isolated rat hearts. METHODS: In the ischemia/reperfusion (I/R) model of isolated perfused rat hearts,the effects of hUII pretreatment on cardiac function was monitored with cardiac function software of MFL Lab200. ATP,total calcium,and malondialdehyde (MDA) content in myocardium were detected. The coronary perfusion flow (CPF) and lactate dehydrogenase (LDH) activity in coronary effluent were measured during reperfusion. RESULTS: In the hUII pretreated group,the release of LDH from myocardium was lower [(78.3?18.1)U/L] than I/R group [(109.3?23.9) U/L,P

Effects of adrenomedullin 2 on proliferation of microvascular endothelial cells from the rat cerebral cortex / 中国病理生理杂志

0.05).10% FBS induced -TdR incorporation increased by 87.5%(vs control,P

Effects of human urotension Ⅱ on in vivo mesenteric microcirculation in rats / 中国病理生理杂志

AIM: To investigate the effects of human urotensin Ⅱ (hUII) on in vivo mesenteric microcirculation in rats. METHODS: For recording of microcirculation images in the mesentery, the intestinal loop was mounted on the stage of an intravital microscope equipped with a TV camera. Video images of microcirculation were stored by a video cassette recorder. Temporal changes in internal diameter and microcirculatory velocity of microvesseles were measured by computer using the ImagePro software. The blood flow in intestinal wall was measured with PIMII laser Doppler perfusion Imager (Lisca Sweden). RESULTS: The internal diameters of arterioles and venules in control group were (21.4?2.3) ?m and (38.1?3.6) ?m,respectively. In UII group, the arterioles and venules contracted immediately after treated with UII and up to the peak at 1 min [(14.1?1.4) ?m and (22.2?5.2) ?m vs control,P0.05). The blood flow in intestinal wall increased 1 min after treated with UII and up to high peak at 5 min(6.4?1.1 perfusion unit vs control 4.2?0.9,P

Role of hypoxia- inducible factor-1 in cardioprotection of hypoxic preconditioning / 中国病理生理杂志

AIM: To investigate the effects of hypoxia-inducible factor-1 (HIF-1) on cardioprotection of hypoxic preconditioning (HPC) and its mechanisms. METHODS: In the model of hypoxia/reoxygenation (H/R) of cardiomyocytes from neonatal Spargue-Dawley rats, delayed cardioprotective effects of HPC on lethal H/R were observed. Whole cell extracts were prepared for determining activities of extracellular signal-regulated protein kinases (ERKs) and HIF-1? phosphorylation. RESULTS: HPC significantly promoted survival and membrane integrity of cardiomyocytes subjected to sustained H/R. SDS-PAGE mobility shift experiments showed increased phophorylation level of HIF-1? in hypoxic preconditioned cardiomyocytes. ERK1/2 were activated at 10 min after HPC and returned to the control level at 60 min. CONCLUSION: HPC protects neonatal cardiomyocytes from H/B injury. HIF-1? phosphorylation induced by ERKs might contribute to the cardioprotection of HPC.

Protective effect of ischemic preconditioning on rat brain with ischemia/reperfusion injury / 中国病理生理杂志

AIM:To study whether ischemic preconditioning(IPC) has a protective effect against ischemia/reperfusion(I/R) injury in brain, and the possible relationship between IPC and the regulating function of microcirculation. METHODS: The I/R models were established both in I/R and IPC groups of Sprague-Dawley rats. Additional procedure was performed of short term cerebral ischemic preconditioning in IPC group 24 hours before I/R. Skull windows were performed through which microcirculation features were measured before ischemia, during ischemia, and reperfusion. Finally, brains were cut into slices and stained with red tetrazoline(TTC). RESULTS: Most TTC stained brains in I/R group presented irregular palely red areas which were few in IPC group. Compared with I/R group, IPC group presented relatively increase in accumulated length of capillaries, mean cerebral microcirculatory perfusion, and microcirculatory velocity in ischemic and reperfusion phase. There was no-reflow phenomenon in I/R group in reperfusion phase, which was substituted by the course of increasing reperfusion in IPC group. CONCLUSIONS:IPC could relieve the reduction of tissue perfusion during ischemia and the no-reflow phenomenon during reperfusion by improving the regulating function of microcirculation, which relatively promote the opening of capillaries and accelerating of microvascular flow, therefore protect brain from I/R injury.