Genetic analysis of the false positive trisomy 7 and false negative trisomy 18 by NIPT-PLUS / 中华医学遗传学杂志

OBJECTIVE@#To explore the cause of inconsistency between the results of trisomy 7 by expanded non-invasive prenatal testing (NIPT-PLUS) and trisomy 18 by prenatal diagnosis.@*METHODS@#A pregnant woman who received genetic counseling at Jiaozuo Maternal and Child Health Care Hospital on July 5, 2020 was selected as the study subject. NIPT-PLUS, systematic ultrasound and interventional prenatal testing were carried out. The middle segment and root of umbilical cord, center and edge of the maternal and fatal surface of the placenta were sampled for the validation by copy number variation sequencing (CNV-seq).@*RESULTS@#The result of NIPT-PLUS indicated that the fetus has trisomy 7. Systematic ultrasound has shown multiple malformations including atrioventricular septal defect, horseshoe kidney, and rocker-bottom feet. However, QF-PCR, chromosomal karyotyping analysis, and CNV-seq of amniotic fluid samples all showed that the fetus was trisomy 18. Validation using multiple placental samples confirmed that the middle segment of the umbilical cord contains trisomy 18, the center of the placenta contained trisomy 7, and other placental sites were mosaicism for trisomy 7 and trisomy 18. Notably, the ratio of trisomy 18 became lower further away from the umbilical cord.@*CONCLUSION@#The false positive results of trisomy 7 and false negative trisomy 18 by NIPT-PLUS was probably due to the existence of placental mosaicism. Strict prenatal diagnosis is required needed aneuploidy is detected by NIPT-PLUS to exclude the influence of placental mosaicisms.

Detection of genomic copy number variations in patients with unexplained mental retardation/developmental delay by low coverage whole-genome sequencing / 中华医学遗传学杂志

OBJECTIVE@#To detect genomic copy number variations (CNVs) among 145 children with unexplained mental retardation/developmental delay (MR/DD) by using low-depth whole-genome copy number variation sequencing (CNV-seq).@*METHODS@#Peripheral blood samples were collected from the patients and subjected to DNA extraction and CNV-seq. The results were analyzed by a combination of bioinformatic tools.@*RESULTS@#Forty-nine patients were found to carry a total of 67 CNVs with an average size of 5.27 Mb. Among these, 22 patients were assessed to carry MR/DD-related CNVs involving 21 syndromes. This gave a diagnostic rate of 15.17%(22/145) for CNVs associated with unexplained MR/DD. The corresponding regions of the 22 MR/DD-related CNVs in the human genome covered 174 MR/DD-related pathogenic genes, which have mapped to 18 sections on 10 chromosomes.@*CONCLUSION@#Genomic CNVs-related microdeletions/duplications account for a significant proportion of unexplained MR/DD, for which CNV-seq can provide an accurate diagnosis.

Clinical and pathological features of intestinal Talaromycosis marneffei infection in patients with acquired immunodeficiency syndrome / 中华传染病杂志

Objective:To investigate the clinical and pathological characteristics of acquired immunodeficiency syndrome (AIDS) patients with intestinal Talaromycosis marneffei (TM) infection. Methods:A total of 64 AIDS patients who underwent colonoscopy in Guangzhou Eighth People′s Hospital from January, 2010 to December, 2018 were retrospectively collected. Among them, 32 patients were co-infected with TM (AIDS with intestinal TM infection group) and 32 patients were not (AIDS without intestinal TM infection group) according to the colonic mucosa pathology. The clinical manifestations and pathological differences were compared between the two groups. Nonparametric rank sum test and Fisher exact probability method were used for statistical analysis.Results:The proportions of patients presented with fever, cough, retroperitoneal lymph nodes tume faction, nausea and vomiting, abdominal muscle tension, abdominal tenderness and rebound pain in AIDS with intestinal TM infection group were 28 (87.5%), 16 (50.0%), 13 (40.6%), 9 (28.1%), 8 (25.0%), 20 (62.5%) and 12 (37.5%), respectively, which were all significantly higher than those in AIDS without intestinal TM infection group 11 (34.4%), 6 (18.8%), 3 (9.4%), 2 (6.2%), 1 (3.1%), 8 (25.0%) and 1 (3.1%), respectively, the differences were statistically significant (Fisher exact test, all P<0.05). The median counts of peripheral blood CD4 + T lymphocyte, lymphocytes, monocytes, hemoglobin, platelet and albumin in AIDS with intestinal TM infection group were 13.5/μL, 0.30×10 9/L, 0.16×10 9/L, 88 g/L, 122×10 9/L and 23.5 g/L, respectively, which were all significantly lower than those in AIDS without intestinal TM infection group 207.0/ μL, 1.35×10 9/L, 0.35×10 9/L, 128 g/L, 201×10 9/L and 37.5 g/L, respectively, the differences were all statistically significant ( Z=-6.111, -6.191, -4.273, -5.353, -2.974 and-6.666, respectively, all P<0.05). Multivariate logistic regression analysis showed that CD4 + T lymphocytes <50/μL, hemoglobin <90 g/L and fecal occult blood positive were independent risk factors for AIDS with intestinal TM infection. The main manifestations of colonoscopy in AIDS with intestinal TM infection group were discontinuous ulcers (31.2%(10/32)), erosion (31.2%(10/32)) or co-exitance of ulcer and erosion (21.9%(7/32)), while suspected tumor-like eminence lesions were less common (15.6%(5/32)). The pathological features of colon mucosa were ulcer and/or erosion (53.1%(17/32)), chronic inflammation (46.9%(15/32)) and inflammatory granuloma (43.8%(14/32)). Oval or round spore with apparent septum could be seen by special staining. In AIDS with intestinal TM infection group, 27 patients were cured or improved, five patients died or deteriorated, while all patients in the AIDS without intestinal TM infection group improved after treatment without death. Conclusions:There are no specific gastrointestinal symptoms in AIDS patients with intestinal TM infection, while the patients present with decreased immunological cells and multiple colony pathological features. Specific fungal spores can be seen.

Chromosomal microarray analysis for the causes of miscarriage or stillbirth / 中华医学遗传学杂志

OBJECTIVE@#To assess the value of chromosomal microarray analysis (CMA) for the analysis of 824 samples from miscarriage or stillbirth.@*METHODS@#Copy number variations (CNVs) in the abortic chorionic villi or stillbirth tissues were detected by CMA.@*RESULTS@#All specimens were successfully analyzed, among which 381 (46.2%) were diagnosed with chromosomal abnormalities, which included 312 (81.9%) numerical abnormalities, 66 (17.3%) structural abnormalities and 3 (0.8%) uniparental disomies. Among numerical chromosomal abnormalities, aneuploidies was most common (92.0%), with trisomy 16 and 45,X accounting for 41 (13.1%) and 63 (20.2%) of the cases, respectively. Among the 66 structural chromosomal aberrations, there were 26 (39.4%) CNVs duplications, 20 (30.3%) CNVs deletions, and 20 (30.3%) CNVs duplication and deletions. 33 CNVs were predicted as have a high chance to lead to a disease.@*CONCLUSION@#CMA is a reliable, robust, and high-resolution method for the analysis of miscarriage or stillbirth samples. Numerical aberrations, in particular chromosomal aneuploides, are the main cause for spontaneous abortions and stillbirths.

Effect of atorvastatin on high glucose-stimulated expressions of MALAT1 and inflammatory factors in endothelial cells / 中华内分泌代谢杂志

Objective To investigate the effect of atorvastatin on the expressions of long non-coding RNA (LncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1)and inflammation factors in human umbilical vein endothelial cells (HUVECs) stimulated by high glucose. Methods The expression of MALAT1 in HUVECs incubated with high glucose(30 mmol/L) for different time periods were detected by real-time PCR. Under high glucose condition, the expressions of MALAT1, interleukin-6(IL-6), and interleukin-8 (IL-8) in HUVECs were detected after MALAT1 was silenced by siRNA or atorvastatin was added. Results (1) After HUVECs were incubated with high glucose for different time periods, the expressions of MALAT1 were increased to some extent(P<0.05), with the peak at 12h (P<0.01). The levels of IL-6 and IL-8 expression and secretion were increased after HUVECs were stimulated by high glucose for 12h (P<0.05). (2)The silence of MALAT1 markedly suppressed high glucose-stimulated expression and secretion of IL-6 and IL-8 (P<0.05). (3) Atorvastatin significantly inhibited high glucose-stimulated expressions of MALAT1, IL-6, and IL-8(all P<0.05). Conclusions High glucose induces the secretion of inflammatory factors by stimulating MALAT1 expression in endothelial cells. Atorvastatin significantly inhibits high glucose-stimulated MALAT1 expression and decreases inflammatory reaction.

Inhibitory effect of Saxagliptin on high glucose-induced overexpression of LncRNA-MALAT1 in endothelial cells / 医学研究生学报

Objective Saxagliptin regulates the level of blood glucose by selectively inhibiting high-performance dipeptidyl peptidase 4, but its action mechanism is not yet clear .This study was to investigate the effect of the novel hypoglycemic agent Saxaglip -tin on the expression of long non-coding RNA (LncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) and its target gene products transforming growth factor-β1 ( TGF-β1 ) in human umbilical vein endothelial cells ( HUVECs) stimulated by high glucose. Methods HUVECs were cultured in with D-glucose (D-GS) at the concentrations of 5.5, 10, 20, and 30 mmol/L and Saxagliptin at 0, 0.01, 0.1, 1, and 10μmol/L.The best concentrations of D-GS and Saxagliptin were determined as 30 mmol/L and 1 μmol/L, respectively.The HUVECs were divided into four groups:control (5.5 mmol/L D-GS), Saxagliptin (5.5 mmol/L D-GS+1 μmol/L Saxagliptin ) , high glucose ( 30 mmol/L D-GS ) , and high glucose +Saxagliptin (30 mmol/L D-GS +1μmol/L Saxaglip-tin), all cultured for 24 hours.Then the expressions of MALAT1 and TGF-β1 mRNA in the cells were detected by qRT-PCR, that of the TGF-β1 protein determined by Western blot , and the level of TGF-β1 in the supernatant measured by ELISA . Results The expressions of LncRNA-MALAT1 and TGF-β1 were significantly increased in the high glucose group as compared with the control ( 8.65 ±0.70 vs1.00 ±0.00 and 1.36 ±0.07 vs 1.00 ±0.00, P<0.01) but markedly inhibited in the high glucose +Saxagliptin group in compari-son with the high glucose group (2.17 ±0.24 vs 8.65 ±0.70 and 1.15 ±0.02 vs 1.36 ±0.07, P<0.05). Conclusion High glu-cose can induce the overexpression of LncRNA-MALAT1 and its target gene products TGF-β1 in HUVECs and cause damage to the cells, while Saxagliptin can significantly suppress this effect .

Effects of Foot Three-needle Therapy on Colonic Cystathionine Gamma-lyase and Myeloperoxidase in Postoperative Rats / 广州中医药大学学报

Objective To observe the effect of acupuncture on cystathionine gamma-lyase(CSE)and myeloperoxidase (MPO) in the colonic tissues of rats and to explore its mechanism for recovering the function of interstitial cells of Cajal(ICCs)in rats after colonic anastomosis . Methods Thirty SD rats were randomized into normal control group, model group(receiving colonic anastomosis) and acupuncture group. The acupuncture group received foot three-needle therapy on bilateral Zusanli(ST36), Sanyinjiao(SP6) and Taichong(LR3), once a day for 3 days after colonic anastomosis. And then the propulsive rate of the small intestine was measured. The count of ICCs with positively expressive c-kit in rat colonic tissues was measured by immunohistochemistry, the activity of CSE was observed by enzyme-linked immunosorbent assay (ELISA) and the activity of MPO was tested by biochemical method. Results Compared with the normal control group, the small intestinal propulsive rate in the model group was decreased, the number of ICCs with positively expressive c-kit was reduced, while the activities of CSE and MPO were increased(P<0.05). The acupuncture group had higher intestinal propulsive rate, more ICCs with positively expressive c-kit, and lower CSE and MPO activities than the model group(P<0.05).Conclusion Acupuncture can promote the recovering of postoperative gastrointestinal function, and its mechanism may be related to the regulation of CSE and MPO activities in the colonic tissues and to the restoration of ICCs function in the focus with positive c-kit.

Chemical constituents from stems and leaves of Buddleja lindleyana Fort / 国际药学研究杂志

Objective To investigate the anti-H5N1 activities and chemical constituents from stems and leaves of Buddleja lindleyana Fort.. Methods Constituents were separated through AB-8 macroporous resin, chromatography of silica gel, Sephadex LH-20 and recrystallization. Structures of the compounds were identified by analysis of physicochemical properties and spectral data. Results Ten compounds were isolated and identified as linarin(1), rutin(2), luteolin(3), quercetin(4), apigenin(5), hesperetin (6), salidroside (7), oleanolic acid (8),β-sitosterol (9), and daucosterol (10), respectively. Conclusion Compounds 2-7 were obtained from this plant for the first time.

Histopathological characteristics of colonoscopic biopsy specimens of AIDS patients / 中华临床感染病杂志

Objective To investigate the histopathological characteristics of colonoscopic biopsy specimens from AIDS patients .Methods A total of 310 clinically confirmed AIDS patients with abdominal pain, diarrhea or tenesmus were enrolled from Guangzhou NO .8 People’s Hospital during 2010 and 2014. All patients underwent colonoscopic examination , and the biopsy specimens were collected .Conventional HE staining, special stainings including Gomori’s methenamine silver ( GMS), Periodic Acid-Schiff stain (PAS), acid-fast staining, and immunohistochemical staining of cytomegalovirus (CMV) were performed. Results The biopsy specimens showed mucosa chronic inflammation (93.9%), epithelial degeneration and necrosis;the local erosion and ulcer formation were observed in severe cases .Among 310 patients, the infective pathogens were identified in 139 ( 44.8%) cases, including 47 ( 15.2%) cases with CMV infections, 36(11.6%) cases with mycobacterium infections , 21(6.8%) cases with penicillium marneffei infections, 10(3.2%) cases with Cryptococcus infections, 3(1.0%) cases with candida infections, 2(0.6%) cases with cryptozoite infections and the rest 20(6.5%) cases were with uncertain pathogens . Conclusion Chronic inflammatory lesions are common in patients with AIDS , and colonoscopic mucosal biopsy can help to identify the pathogens of intestinal opportunistic infection .

A new coumarin and eight known polyphenolics from Radix Campylotropidis Hirtellae / 军事医学

Objective To investigate the chemical constituents of the Radix Campylotropidis Hirtellae ( RCH ) . Methods 60%ethanol extract of the RCH was separated by column chromatography on silica gel, ODS and Sephadex LH-20 to obtain aimed compounds.Structures of the compounds were investigated by modern spectroscopy.Results Nine compounds were separated from Campylotropis hirtella ( Franch.) Schindl, and the structure of the new compound 1 was determined by spectral analysis and named hedysarimcoumestanⅠ( 1 ) , while the others were identified as the known isotrifoliol(2), hedysarimcoumestan B(3), umbelliferone(4), orobol(5), quercetin(6),(+) catechin(7), kaempferol (8) and dihydrokaempferol(9) .Conclusion Compound 1 is a new compound and 3-5 are separated from C.hirtella for the first time.The screening results of inhibition of H5N1 virus activity show that compound 5 has some inhibitory activity at a level of 50.0 μg /ml.

Screening for methylation-silenced genes in acute myeloid leukemia HL-60 cell line by a quantitative proteomic approach / 中南大学学报(医学版)

Objective To screen for new methylation association genes in HL-60 to reveal the pathogenesis of leukemia, and provide important theoretical and scientific basis for the prevention and cure of leukemia. Methods Two-dimensional fluorescence difference gel electrophoresis (F-2D-DIGE) was performed to separate the total proteins from acute myelogenous leukemia (AML) cell line HL-60 cells with or without 5-aza-2-deoxycytidine (5-aza-2-dC) treatment. Imaging software Decyder 6.5 and PDQuest were used to detect the differential expression protein spots, and matrix-assisted laser desorption/ionizaion time-of-flight mas spectrometer (MALDI-TOF MS) was adopted to identify the differential expression proteins. Results F-2D-DIGE maps of 5-aza-2-dC-untreated HL-60 and-treated HL-60 cells were established. A total of 53 differential protein spots were detected, and 35 differential proteins were successfully identified. Of the identified proteins, 32 proteins were up-regulated, and 3 proteins were down-regulated in HL-60 cells after 5-aza-2-dC treatment.Conclusion Thirty-five differential proteins may be associated with methylation in HL-60 cell line, which provides the important clues for epigenetic study of leukemia.

Methylation and expression of glioma pathogenesis-related protein 1 gene in acute myeloid leukemia / 中南大学学报(医学版)

OBJECTIVE@#To detect the methylation and expression of glioma pathogenesis-related protein 1(GLIPR1) gene in the acute myeloid leukemia (AML) cell lines and bone marrow cells from AML patients, and to determine the relationship between promoter methylation and expression of GLIPR1.@*METHODS@#Five leukemia cell lines, 54 bone marrows from the newly diagnosed AML patients, 48 bone marrows from the acute lymphoblastic leukemia (ALL )patients, 40 bone marrows from the chronic myeloid leukemia (CML) patients,35 bone marrows from control patients, and 8 bone marrows from the complete remission AML patients were collected. RT-PCR and methylation-PCR (MSP) were used to detect the mRNA expression and promoter methylation of GLIPR1, respectively, and the relationship between them was analyzed.@*RESULTS@#The level of GLIPR1 mRNA in the AML cell lines was lower than that in the chronic myeloid leukemia (CML) and ALL cell lines, whereas the methylation level of GLIPR1 in the former was higher than that in the latter. The level of GLIPR1 mRNA in the AML cell lines was significantly increased, but had no obvious changes in the CML and ALL cell lines after 5-aza-2dC treatment. The mRNA level of GLIPR1 in the AML bone marrows (0.38+/-0.20)was obviously lower than that in the ALL bone marrows (0.76+/-0.18), CML bone marrows (0.80+/-0.14), and control bone marrows(0.85+/-0.12). The level of GLIPR1 mRNA in the bone marrows with complete remission AML was obviously higher than that in the AML bone marrows before the treatment (0.78+/-0.13 vs. 0.36+/-0.20); but there was no obvious difference between the ALL bone marrows and the control bone marrows, and the CML bone marrows and the control bone marrows (both P>0.05). The positive rate of GLIPR1 gene methylation in the AML bone marrows (81.5%) was obviously higher than that in the ALL bone marrows(37.5%), CML bone marrows (27.5%) and the control bone marrows(14.3%). The positive rate of GLIPR1 gene in the bone marrows with complete remission AML was obviously lower than that in the bone marrows before the treatment (12.5% vs. 75.0%), but there was no obvious difference between the ALL bone marrows and between the control bone marrows,and the CML bone marrows and the control bone marrows (both P>0.05). There was a negative correlation between the mRNA level and methylation status of GLIPR1 in the AML bone marrows.@*CONCLUSION@#GLIPR1 expression is downregulated or even lost by promoter methylation in AML, and the expression and methylation level of GLIPR1 gene may have some significance in evaluating the curative effect of AML patients.

A Frequency Weighting Contour for Chinese Young Males / 航天医学与医学工程

Objective To obtain the frequency weighting contour for seated Chinese young males in z axis.Method 10 male volunteers served as the subjects.They were exposed to standard vibration stimulus (a sinusoidal vibration of 1.0 m/s2 at 8 Hz) and 14 kinds of sinusoidal experimental vibration stimuli at frequencies from 4 Hz to 80 Hz.The magnitude of the experimental vibration stimulus was changed by the subject through a signal condition and switching device made by ourselves to obtain an equal vibration sensation to the standard vibration.The data were fitted to the same form of the frequency weighting Wk of ISO 2631-1 with the least square method and optimization of parameters.Result Compared with ISO 2631-1,the frequencies weightings of Chinese young males at 16～80 Hz are significantly higher than those of ISO 2631-1(P<0.005),but there are no significant difference between two frequency weighting contours at other frequencies. Conclusion Chinese young males are more sensitive to vibration at 16～80 Hz in Z axis.