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BACKGROUND@#Sweat secreted by eccrine sweat glands is transported to the skin surface through the lumen. The eccrine sweat gland develops from the initial solid bud to the final gland structure with a lumen, but how the lumen is formed and the mechanism of lumen formation have not yet been fully elucidated. This study aimed to investigate the mechanism of lumen formation of eccrine gland organoids (EGOs).@*METHODS@#Human eccrine sweat glands were isolated from the skin for tissue culture, and the primary cultured cells were collected and cultured in Matrigel for 14 days in vitro. EGOs at different development days were collected for hematoxylin and eosin (H&E) staining to observe morphological changes and for immunofluorescence staining of proliferation marker Ki67, cellular motility marker filamentous actin (F-actin), and autophagy marker LC3B. Western blotting was used to detect the expression of Ki67, F-actin, and LC3B. Moreover, apoptosis was detected using a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) apoptosis assay kit, and the expression of poly (ADP-ribose) polymerase and Caspase-3 was detected by Western blot. In addition, 3-methyladenine (3MA) was used as an autophagy inhibitor to detect whether the formation of sweat glands can be effectively inhibited.@*RESULTS@#The results showed that a single gland cell proliferated rapidly and formed EGOs on day 4. The earliest lumen formation was observed on day 6. From day 8 to day 14, the rate of lumen formation in EGOs increased significantly. The immunofluorescence and Western blot analyses showed that the expression of Ki67 gradually decreased with the increase in days, while the F-actin expression level did not change. Notably, the expression of autophagy marker LC3B was detected in the interior cells of EGOs as the apoptosis signal of EGOs was negative. Compared with the control group, the autophagy inhibitor 3MA can effectively limit the formation rate of the lumen and reduce the inner diameter of EGOs.@*CONCLUSION@#Using our model of eccrine gland 3D-reconstruction in Matrigel, we determined that autophagy rather than apoptosis plays a role in the lumen formation of EGOs.
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Humans , Apoptosis , Autophagy , Eccrine Glands , Epithelial Cells , OrganoidsABSTRACT
Objective:To explore the clinical application value and accuracy of cell-free fetal DNA (cff-DNA) technique in prenatal screening.Methods:The results of quantitative fluorescent PCR (QF-PCR) and karyotype of amniotic fluid cells were analyzed retrospectively in 2 398 monocyesis pregnant women who had been amniocentesis at the First Affiliated Hospital of Zhengzhou University from May 2013 to December 2019, and the results of 359 cases who had been examined by single-nucleotide polymorphism array (SNP array).Results:Cff-DNA test of 2, 398 cases indicated 987 cases of trisomy 21, 351 cases of trisomy 18, 135 cases of trisomy 13, 566 cases of sex chromosome abnormality, and 359 cases of other chromosome abnormality. Chromosome karyotype analysis detected 826 cases of trisomy 21, 213 cases of trisomy 18, 17 cases of trisomy 13, 221 cases of sex chromosome abnormality, and 26 cases of other chromosome abnormality. The detection rate were 83.69% (826/987), 60.68% (213/351), 12.59% (17/135), 39.04% (221/566) and 7.24% (26/359), respectively. QF-PCR detected 1 046 cases of trisomy and 188 cases of sex chromosomes abnormality, and the detection rate was 99.05% (1 046/1 056) and 85.07% (188/221), respectively. Compared with the abnormal number detected by chromosome karyotype analysis, 10 cases of trisomeric chimerism and 24 cases of sex chromosome were missed by QF-PCR. Among the 359 other chromosomal abnormalities detected by SNP array, 64 cases were consistent with the results of cff-DNA, and the detection rate was 17.83% (64/359), which was 10.59% higher than the karyotype result.Conclusions:Karyotype analysis is the gold standard for diagnosing chromosomal abnormalities. QF-PCR could diagnose common chromosome aneuploidy rapidly and accurately, and it could be used as an auxiliary detection technique for karyotype analysis. The incidence of sex chromosome chimerism is high, so missed diagnosis should be warned. SNP array could be given priority to verify chromosome microdeletion or microduplication detected by cff-DNA.
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Objective To investigate the value of prenatal diagnosis in identifying the etiology and predicting the prognosis of fetal pleural effusion (FPE).Methods Forty-two cases of FPE were recruited in this study from January 2012 to September 2016.Ultrasound scan and genetic tests were performed on all fetuses.Seven fetuses with severe FPE were given pleurocentesis.Pregnancy outcomes of all the fetuses were followed up.Results FPE was commonly accompanied with other abnormalities,such as ascites,hydrops,hydramnion,hygroma colli,abnormal posturing,joint contractures,arrhythmia and micromandible.Chromosomal abnormality was detected in 11 fetuses (26.2％),of which ten were further confirmed by karyotype analysis,including six with 45,X,three trisomy 21 and one trisomy 18,and one was detected with a 9.83 Mb uniparental disomy (UPD) located at 12q24.21q24.31 by gene chip.One fetus was diagnosed with--SEA/--SEA thalassemia.All of the 12 families decided to terminate the pregnancies after genetic counseling.Among the other 30 fetuses,seven with severe FPE and normal karyotype underwent pleurocentesis.Five of the seven cases were with favorable outcomes,one with progressive hydrops was aborted and one neonate with severe hydrops died after birth.Spontaneous regression of FPE with good outcome was found in two cases.Parents of the other 21 fetuses chose to terminate the pregnancies.Conclusions Prenatal diagnosis is important to identify the etiology and predict the outcome of FPE.Chromosomal abnormality is a relatively common cause of FPE,and 45,X and trisomy 21 are the most common abnormalities.Intrauterine intervention is beneficial for FPE without chromosomal or other definite genetic abnormalities.Genetic test may be of great value for pregnant counseling.
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Objective To investigate the levels of serum AFP and CEA in healthy physical examination people and their sexual difference.Methods (1 )Healthy physical examination people were divided into the group A(510 males)and group B(477 fe-males);(2)these two groups were subdivided into the group A1 (<40 years old),A2 (40 - <50 years old),A3 (50 - <60 years old),A4(60-<70 years old)and A5 (70 -87 years old),and group B1 (<40 years old),B2(40 -<50 years old),B3 (50 -<60 years old),B4(60-<70 years old)and B5(70-87 years old).The serum AFP and CEA in 987 healthy physical examination peo-ple were measured by the electrochemiluminesence immunoassay,and then the detection results were compared among various groups.Results (1)The detection results of serum AFP and CEA had statistically significant differences between the group A and B (P <0.01);(2)there was no statistical differences among the group A and among the group B.Conclusion The serum AFP and CEA levels in healthy physical examination people in the local area have difference between males and females,which in males are significantly higher than those in females,indicating that the reference range of AFP and CEA should be repectively established ac-cording to different sexes;while the serum AFP and CEA levels have no differences among various age groups in males and fe-males.
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Objective To investigate the effects of functional electrical stimulation (FES) on motor function and the expression of bromodeoxyuridine (Brdu) + and glial fibrillary acid protein (GFAP) + in the subventricular zone (SVZ) of rats with acute cerebral infarction,and to explore it's mechanism. Methods A rat model of cerebral infarction was established using Longa's technique for middle cerebral artery occlusion (MCAO) with an intraluminal filament.The rats were randomly divided into a FES group,a placebo stimulation group and a control group.In each group,rats were randomly allocated into 1 d,3 d,7 d and 14 d subgroups (6 rats/subgroup).Superficial electrodes were pasted on the paralyzed forelimbs of rats in the FES group for connecting with the FES instrument,and FES treatment was carried out with a current of 4-5 mA for 15 min on the third day after the MCAO operation to produce extension of the wrist and the digits of the paralyzed forelimb.The rats in the placebo stimulation group were pasted with electrodes,but no FES was administered and they received no other treatment.Neurological deficits were evaluated using the modified neurological severity score (mNSS) before treatment and on the 1 st,3rd,7th,and 14th day after treatment. BrdU and GFAP positive cells in the SVZ were detected by immunofluorescence techniques.Results After 7 or 14 days the motor function of rats in the FES group had improved significantly compared with the placebo stimulation and control groups.Compared with the other two groups,the expression levels of BrdU+ and GFAP+ cells in the ischemic SVZ in the FES group were significantly higher at the 3rd,7th and 14th day.Conclusion FES can improve motor function after acute cerebral infarction and also promote the proliferation and differentiation of neural stem cells in the SVZ.
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Background and purpose:MMMT is known as a rare malignancy in gynecological tumor. Because of its difficulty in diagnosis and treatment, the prognosis is extremely poor. This study was to discuss the clinicopathologic feature and PRA,PRB expressions of uterine MMMT. Methods:We analysed clinicopathologic data and the expressions of PRA,PRB by immunohistochemical staining on 17 cases of uterine MMMT, and 11 patients were followed up. Results:①The patients usually presented with abnormal vaginal bleeding with no specifi city clinically. ②The morphological changes were various and complicated, including epithelial and mesenchymal components and a variety of inter-permeated and transitional tissue elements.③The positive rate of PRA in stageⅠ and stageⅡ were 66.7% and 40%, and PRB in stage Ⅰ and stageⅡwere 55.6% and 20%, respectively. ④The mean survival time in stageⅠ,Ⅱ and Ⅲ were 43.8 months (32-59), 34.25 months (19~41) and 5 months, respectively. Conclusion:The diagnosis of uterine MMMT was mainly based on tissue morphology; the development of uterine MMMT might be related with the loss of PRA and PRB ; the clinical stage and the expression of PRA and PRB might be the prognostic factors for uterine MMMT.