1.Knowledge-embedded spatio-temporal analysis for euploidy embryos identification in couples with chromosomal rearrangements
Fangying CHEN ; Xiang XIE ; Du CAI ; Pengxiang YAN ; Chenhui DING ; Yangxing WEN ; Yanwen XU ; Feng GAO ; Canquan ZHOU ; Guanbin LI ; Qingyun MAI
Chinese Medical Journal 2024;137(6):694-703
Background::The goal of the assisted reproductive treatment is to transfer one euploid blastocyst and to help infertile women giving birth one healthy neonate. Some algorithms have been used to assess the ploidy status of embryos derived from couples with normal chromosome, who subjected to preimplantation genetic testing for aneuploidy (PGT-A) treatment. However, it is currently unknown whether artificial intelligence model can be used to assess the euploidy status of blastocyst derived from populations with chromosomal rearrangement.Methods::From February 2020 to May 2021, we collected the whole raw time-lapse videos at multiple focal planes from in vitro cultured embryos, the clinical information of couples, and the comprehensive chromosome screening results of those blastocysts that had received PGT treatment. Initially, we developed a novel deep learning model called the Attentive Multi-Focus Selection Network (AMSNet) to analyze time-lapse videos in real time and predict blastocyst formation. Building upon AMSNet, we integrated additional clinically predictive variables and created a second deep learning model, the Attentive Multi-Focus Video and Clinical Information Fusion Network (AMCFNet), to assess the euploidy status of embryos. The efficacy of the AMCFNet was further tested in embryos with parental chromosomal rearrangements. The receiver operating characteristic curve (ROC) was used to evaluate the superiority of the model. Results::A total of 4112 embryos with complete time-lapse videos were enrolled for the blastocyst formation prediction task, and 1422 qualified blastocysts received PGT-A ( n = 589) or PGT for chromosomal structural rearrangement (PGT-SR, n = 833) were enrolled for the euploidy assessment task in this study. The AMSNet model using seven focal raw time-lapse videos has the best real-time accuracy. The real-time accuracy for AMSNet to predict blastocyst formation reached above 70% on the day 2 of embryo culture, and then increased to 80% on the day 4 of embryo culture. Combing with 4 clinical features of couples, the AUC of AMCFNet with 7 focal points increased to 0.729 in blastocysts derived from couples with chromosomal rearrangement. Conclusion::Integrating seven focal raw time-lapse images of embryos and parental clinical information, AMCFNet model have the capability of assessing euploidy status in blastocysts derived from couples with chromosomal rearrangement.
2.Efficacy and complications of one-step and two-step percutaneous transhepatic drainage in the treatment of advanced low-level biliary obstruction
Jie GONG ; Zhenhua XU ; Zehua LEI ; Fengwei GAO ; Kangyi JIANG ; Qingyun XIE ; Xin ZHAO
Journal of Clinical Surgery 2024;32(7):688-692
Objective To investigate the efficacy and complications of one-step and two-step ultrasound-guided percutaneous transhepatic hepatic duct drainage(PTHD)in the treatment of advanced low-level biliary obstruction.Methods A total of 112 patients with advanced low-level biliary obstruction who underwent palliative surgery for reducing jaundice in the hospital from January 2019 to July 2023 were selected,and divided into the one-step group and the two-step group according to the principle of complete randomization(grouped by random number table method).The one-step group(n=56)underwent one-step PTHD,while the two-step group(n=56)underwent two-step PTHD.Surgery related indicators,serum total bilirubin(TBIL),alkaline phosphatase(ALP),alanine transaminase(ALT),the incidence of postoperative complications,and satisfaction were compared between the two groups.Results There was no difference in intraoperative blood loss[(2.96±0.50)ml vs.(3.02±0.53)ml],drainage volume in 48 hours after surgery[(555.84±90.51)ml vs.(560.52±104.23)ml]between the two groups(P>0.05).Operation time and postoperative VAS score of the one-step group were significantly shorter/lower than those of the two-step group[(32.50±4.26)min vs.(36.43±3.58)min,(3.72±1.57)points vs.(4.53±1.34)points],and the success rate of puncture was significantly higher than that in the two-step group(96.43%vs.76.69%),with statistically significant differences(P<0.05).After treatment,the levels of TBIL,ALP and ALT in the two groups were significantly reduced(P<0.05),but there was no statistically significant difference between the groups(P>0.05).There was no statistically significant difference in the incidence of complications between the two groups(3.57%vs.7.14%,Fisher's exact probability=0.679).The satisfaction rate in the one-step group was significantly higher than that in the two-step group(94.64%vs.75.00%),with a statistically significant difference(P<0.05).ConclusionThe operation time of one-step PTHD is shorter and the success rate of puncture is higher.In addition,patients undergoing one-step PTHD have milder pain,and are more satisfied.
3.Regulatory role of m6A methylation modification in osteoarthritis
Junhao LIU ; Wei WANG ; Song CHEN ; Qingyun XIE
Chongqing Medicine 2024;53(21):3329-3334,3339
N6-methyladenosine(m6A)is produced by methylation of the 6th nitrogen atom of ade-nine.The m6A methylation modification is the most common internal modification of RNA,and the modifica-tion process mainly relies on m6A-related enzymes,including methyltransferases,demethylases and binding proteins.The core structure of the methyltransferase complex(MTC)is methyltransferase-like 3(METTL3).The pathogenesis of osteoarthritis(OA)is intricate and complex,and various factors interact with each other.This paper summarizes the mechanism of action of m6A and related enzymes,and elaborates the pathogenesis of os-teoarthritis from the aspects of chondrocytes,proteases,cytokines and signaling pathways,emphatically intro-duces the regulatory role of METTL3 on OA in m6A methylated modification.The aim is to provide new per-spectives on the pathogenesis of osteoarthritis in order to provide new ideas and approaches for the treatment of osteoarthritis.
4.miR-18a ameliorates inflammation and tissue injury in a mouse model of allergic rhinitis via blocking TLR4/NF-κB pathway.
Jun YANG ; Qingyun LI ; Lu WANG ; Hui XIE
Chinese Journal of Cellular and Molecular Immunology 2023;39(8):680-685
Objective To investigate the role of microRNA-18a (miR-18a) in the pathogenesis of allergic rhinitis in mice. Methods Twenty-two BALB/c mice were randomly divided into a blank group, a model group and a miR-18a group. Mice in the model group and the miR-18a group were injected intraperitoneally with obumin (OVA) suspension to prepare allergic rhinitis models, and mice in the miR-18a group were simultaneously given lentiviral vector plasmid for overexpression of miR-18a. Allergy symptoms were evaluated by the behavioral score and HE staining. The plasma levels of interleukin-1β (IL-1β), IL-6 and tumor necrosis factor α (TNF-α) were measured by ELISA. The distribution of CD45+ cells in nasal mucosa was measured by immunofluorescence histochemistry, and CD45+ cells in nasal lavage fluid were measured by flow cytometry. The mRNA expression levels of IL-1β, IL-6 and TNF-α in nasal mucosa tissues were measured by fluorescence quantitative PCR, and the protein expressions of Toll like receptor 4 (TLR4), nuclear factor κB p65 (NF-κB p65), inhibitor of NF-κB α (IκBα) and phosphorylated IκBα (p-IκBα) in nasal mucosa were measured by Western blot analysis. Results Compared with the blank group, the plasma levels of IL-1β, IL-6, and TNF-α in the model group increased significantly. The number of CD45+ cells in both nasal mucosa tissue and nasal irrigation fluid increased, and the mRNA levels of IL-1β, IL-6 and TNF-α and the protein expression levels of TLR4, NF-κB p65 and p-IκBα in nasal mucosa increased. Compared with the model group, the plasma levels of IL-1β, IL-6 and TNF-α in the miR-18a group decreased significantly. The number of CD45+ cells in both nasal mucosa tissue and nasal lavage fluid decreased, and the mRNA levels of IL-1β, IL-6 and TNF-α and the exprotein expression levels of TLR4, NF-κB p65 and p-IκBα in nasal mucosa decreased. Conclusion miR-18a can inhibit the occurrence and development of allergic rhinitis, and its molecular mechanism is related to the inhibition of TLR4/NF-κB pathway activation.
Animals
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Mice
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Disease Models, Animal
;
Inflammation
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Interleukin-6/genetics*
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MicroRNAs/genetics*
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NF-kappa B/metabolism*
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NF-KappaB Inhibitor alpha
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Rhinitis, Allergic
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RNA, Messenger
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Toll-Like Receptor 4/metabolism*
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Tumor Necrosis Factor-alpha/genetics*
5.Non-selective versus highly selective partial splenic embolization in treating patients with hypersplenism secondary to liver cirrhosis: a comparative study
Qingyun XIE ; Fengwei GAO ; Zehua LEI ; Kangyi JIANG ; Jie GONG ; Xin ZHAO
Chinese Journal of Hepatobiliary Surgery 2021;27(12):917-922
Objective:To compare the clinical outcomes of treatment using non-selective versus highly selective partial splenic embolization in patients with hypersplenism secondary to liver cirrhosis.Methods:The clinical data of patients with hypersplenism secondary to hepatitis B cirrhosis who underwent splenic embolization at the Department of Hepatobiliary and Pancreatic Surgery of Leshan People's Hospital from July 2017 to July 2020 were analyzed retrospectively. Of 65 patients with hypersplenism, there were 42 males and 23 females, with age of (58.5±9.8) years. Twenty-six patients underwent splenic artery non-selective partial splenic embolization (the non-selective group) and 39 patients underwent partial splenic embolization using highly selective intubation (the highly selective group). The postoperative peripheral hematological indexes, liver function, operation-related complications and portal vein color Doppler ultrasonography were compared between the two groups.Results:The white blood cell count and platelet count of patients in the 2 groups were significantly higher than those before operation. The white blood cell count at 4, 12 and 24 weeks after operation and the platelet count at 12 and 24 weeks after operation in the highly selective group were significantly higher than those in the non-selective group ( P<0.05). Compared with the non-selective group, the total bilirubin, ICG-R15, portal vein diameter and portal vein blood flow in the highly-selective group significantly lower ( P<0.05). The incidences of 0/Ⅰ/Ⅱ/Ⅲ pain in the non-selected group was significantly higher when compared with that in the highly selected group (5/10/11/1 vs. 12/22/7/0), ( P<0.05). There were no significant differences in the incidences of postoperative complication between the two groups ( P>0.05). Conclusion:Compared with non-selective partial splenic embolization, highly selective partial splenic embolization gave more stable and lasting treatment outcomes in patients with hypersplenism caused by liver cirrhosis with better recovery of blood-related indicators, better improvement in postoperative liver function and relief of portal hypertension symptoms.
6.Establishment of kinetic detection method for PKA content
Jiafei CHEN ; Xiuliang CHEN ; Qingyun LIAN ; Lu YANG ; Ying XIE ; Xiaoming LIANG
Chinese Journal of Blood Transfusion 2021;34(3):227-231
【Objective】 To develop and verify a kinetic method for the determination of prekallikrein activator (PKA) content. 【Methods】 The optimal reaction conditions were determined by comparing the factors of pH and ionic strength of different sample dilution buffers, incubation time of each procedure, and incubation temperature. The accuracy, specificity, precision, linearity, stability and durability of the method were validated. 【Results】 The sample was diluted with 0.05 mol/L Tris-HCl buffer (pH8.5, containing 0.15 mol/L NaCl) and incubated by prekallikrein (PK) at 37℃ for 20 min. After that, the substrate S-2302 was added. Within 10 min before the measurement, the absorbance change rate reached △A405/min. The validation results indicated that the linear range of the method was (0.5~4.0)IU/mL, while the recovery of calibration standard was 96.9%~103.7% with the R2 value more than 0.99. The specificity test showed that human serum albumin, excipients of intravenous human immunoglobulin (pH4), low pH and protein content had no significant effect on the detection of PKA, The recovery rates of standard sample solution in the specificity experiment were 98.0% (0.9% sodium chloride solution), 95.3% (0.46% sodium caprylate solution), 96.7% (10% maltose solution, pH4.0), 94.0%(20%BSA), and 94.0%(5%BSA, pH4.0), respectively. The accuracy and precision of the method can meet requirements in the range between 0.5 and 4.0 IU/mL. The inter-batch recovery rate of quality control samples were between 96.4%~109.5% with the coefficients of variation(CV) between 0.2%~6.9%, while the intra-batch recovery rate were between 101.5%~102.9% with the CV between 2.6%~5.9%. The linearity, accuracy and precision of the assay can meet the requirements when PK and S-2302 were placed at room temperature for less than 6 hours, with the recovery rate of quality control samples between 94.9%~109.9%. The end-point method and kinetic method were used to determine the PKA in 20 batches of human serum albumin, and the consistency showed that there was no significant difference between the two methods(P>0.05). 【Conclusion】 A kinetic method for determination of PKA content with good linearity, specificity, accuracy, precision, stability and durability has been established. Compared with the method in ChP, the new method is more convenient, accurate and rapid to determine the content of PKA in human albumin and human immunoglobulin (pH4) for intravenous injection.
7.A study on the timing and modality of surgery for pancreatic sinistral portal hypertension
Zehua* LEI ; Fengwei GAO ; Xin ZHAO ; Tao WANG ; Kangyi JIANG ; Qingyun XIE ; Jianping WU ; Jinqiang FU ; Bo DU ; Zhixu WANG ; Yu LIU ; Yuantao GAN
Chinese Journal of General Surgery 2018;33(7):556-558
Objective To investigate the opportunity and skill of surgery for pancreatic sinistral portal hypertension.Methods Clinical data were retrospectively analyzed on 15 cases of pancreatic sinistral portal hypertension admired from Dec 2015 to Dec 2017.Results All fiften cases underwent surgical treatment,among them three cases were initially treated conservatively in the early stage and treated surgically for gastrointestinal bleeding,12 cases with definite pancreatic disease and pancreatic sinistral portal hypertension treated in the first stage.Three patients underwent second surgery for recurrent gastrointestinal bleeding.The patients were followed up for 6 to 18 months with symptoms significantly impioved without deaths.Conclusions Splenectomy combined with esophagogastric devascularization is the basic surgical treatment for pancreatic sinistral portal hypertension.
8.Accurate authentication of and its closely related species by comparative analysis of complete plastomes.
Shuying ZHU ; Zhitao NIU ; Qingyun XUE ; Hui WANG ; Xuezhu XIE ; Xiaoyu DING
Acta Pharmaceutica Sinica B 2018;8(6):969-980
Owing to its great medicinal and ornamental values, is frequently adulterated with other species on the market. Unfortunately, the utilization of the common DNA markers ITS, ITS2, and + is unable to distinguish from 5 closely related species of it (, , , and ). Here, we compared 63 plastomes comprising 40 newly sequenced plastomes of the 6 species and 23 previously published plastomes. The plastomes of and its closely related species were shown to have conserved genome structure and gene content. Comparative analyses revealed that small single copy region contained higher variation than large single copy and inverted repeat regions, which was mainly attributed to the loss/retention of genes. Furthermore, the intraspecific sequence variability among different species was shown to be diversified, which necessitates a cautious evaluation of genetic markers specific for different species. By evaluating the maximum likelihood trees inferred from different datasets, we found that the complete plastome sequence dataset had the highest discriminatory power for and its closely related species, indicating that complete plastome sequences can be used to accurately authenticate species.
9.Association of variants in signal transducer and activator of transcription 4 in Chinese Han population with rheumatoid arthritis
Meng WEI ; Qingyun XIE ; Tao WANG ; Yue CHENG ; Fan ZHANG ; Dongyang GUO
Chinese Journal of Rheumatology 2016;(2):83-87
Objective To investigate the associations of variants in signal transducer and activator of transcription 4 (STAT4) in Chinese Han population with rheumatoid arthritis (RA). Methods The study was performed in 228 RA cases and 228 controls. Haplotypes from the HapMap database Chinese population were used to select tag-single-nucleotide polymorphisms (SNPs) (r2=0.9) in STAT4 gene. Twenty-three SNPs located in STAT4 gene were examined by multiplex polymerase chain reaction (PCR) and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). The Hardy-Weinberg equilibrium (HWE) was tested by a chi-square and Fisherˊs analysis. Differences in genotypes of the STAT4 polymorphism variants were evaluated using a Chi-square test. All statistical analyses were done with Haploview 4.1 software. Results Significant difference was detected in three SNPs (rs11685878, rs129888 and rs16833437) in allele frequencies analysis (χ2=6.014 8, 4.024 8, 5.539 1, P<0.05). Association was detected for three SNPs (rs11685878, rs16833437 and rs12988825) in genotype analysis ( χ2=6.814 9, 6.098 7, 6.691 7, P<0.05). Conclusion STAT4 may associate with susceptibility in Chinese Han population.
10.Exercise intensity and the expression of toll-like receptor 4 and nuclear transcription factor-κB in the diabetic liver
Yi WANG ; Rongqiu MAO ; Qingyun CHEN ; Ying CHEN ; Hong LIU ; Xie CHEN
Chinese Journal of Physical Medicine and Rehabilitation 2016;38(8):566-571
Objective To investigate the changes in toll-like receptor 4 (TLR4) and nuclear transcription factor-κB (NF-κB) expression in the livers of diabetic rats after exercise at different intensities.Methods Fifty Sprague-Dawley rats were randomly divided into a control group (NC),a diabetic group (DM),a low intensity exercise group (LIE),a medium intensity exercise group (MIE) and a high intensity exercise group (HIE),each of 10.The control group was given a normal diet.Type 2 diabetes was induced in the others through feeding a high-fat and high-sugar diet,as well as injecting a small dose of streptozotocin intraperitoneally.The three exercising groups swam to 40%,70% and 100% of their exhaustion times once a day,6 d/w for 6 weeks.Due to infection,drowning and other reasons,only 8 rats in each of the DM,LIE and MIE groups,as well as 7 in the HIE group survived.After the intervention,body weight,liver weight and fasting blood glucose (FBG) were tested.The expression of TLR4 and NF-κB mRNA in the livers was tested using RT-PCR technology,and the expression of TLR4 and NF-κB protein in the liver was tested using immunohistochemistry.Results Compared with the NC group,the average body weight and liver weight of the other four groups decreased significantly,while the FBG and the expression of TLR4 and NF-κB mRNA and protein increased significantly.There were significant differences among the exercise groups with the lowest values in the MIE group.Conclusion Six weeks of aerobic exercise can reduce FBG and the level of expression of TLR4 and NF-κB mRNA and protein in the liver,at least in rats.The effect of exercise at medium intensity is best.

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