1.Development of time resolved fluorescence microparticle-conjugated anti-human procalcitonin antibody for homogeneous immunoassay
jun Can WU ; yu Qing L(U) ; jie Huai HAO ; na Cheng ZHAO ; ling Yu ZHENG ; qiang Yong JIANG
Military Medical Sciences 2017;41(8):689-693
Objective To develop europium (Ⅲ) [Eu (Ⅲ)] chelated microparticles for homogeneous immunoassay.Methods Anti-human PCT antibodies were labeled with Eu (Ⅲ) chelated nanoscale microparticles as the detection antibody,and another anti-human PCT antibody was labeled with biotin as the solid-phase antibody.Magnetic microspheres labeled with streptavidin were used to separate the complexes of Eu-IgM-PCT-IgM-Biotin.Results In the homogeneous immunoassay,the standard curve fit was not linear.The quadratic curve was Y =19170.12 + 75493.74X-26.00X2(r =0.9986).According to the standard curve,the limit of detection for PCT was 0.04 ng/ml.Conclusion The homogeneous immunoassay which uses Eu (Ⅲ) chelated microparticles is highly sensitive for detection of PCT recombinant antigens and may serve as a promising method to measure serum PCT levels in the future.
2.Hapln1 promotes dedifferentiation and proliferation of iPSC-derived cardiomyocytes by promoting versican-based GDF11 trapping
Hao DING-JUN ; Qin YUE ; Zhou SHI-JIE ; Dong BU-HUAI ; Yang JUN-SONG ; Zou PENG ; Wang LI-PING ; Zhao YUAN-TING
Journal of Pharmaceutical Analysis 2024;14(3):335-347
Hyaluronan and proteoglycan link protein 1(Hapln1)supports active cardiomyogenesis in zebrafish hearts,but its regulation in mammal cardiomyocytes is unclear.This study aimed to explore the potential regulation of Hapln1 in the dedifferentiation and proliferation of cardiomyocytes and its therapeutic value in myocardial infarction with human induced pluripotent stem cell(hiPSC)-derived car-diomyocytes(CMs)and an adult mouse model of myocardial infarction.HiPSC-CMs and adult mice with myocardial infarction were used as in vitro and in vivo models,respectively.Previous single-cell RNA sequencing data were retrieved for bioinformatic exploration.The results showed that recombinant human Hapln1(rhHapln1)promotes the proliferation of hiPSC-CMs in a dose-dependent manner.As a physical binding protein of Hapln1,versican interacted with Nodal growth differentiation factor(NODAL)and growth differentiation factor 11(GDF11).GDF11,but not NODAL,was expressed by hiPSC-CMs.GDF11 expression was unaffected by rhHapln1 treatment.However,this molecule was required for rhHapln1-mediated activation of the transforming growth factor(TGF)-β/Drosophila mothers against decapentaplegic protein(SMAD)2/3 signaling in hiPSC-CMs,which stimulates cell dedifferentiation and proliferation.Recombinant mouse Hapln1(rmHapln1)could induce cardiac regeneration in the adult mouse model of myocardial infarction.In addition,rmHapln1 induced hiPSC-CM proliferation.In conclusion,Hapln1 can stimulate the dedifferentiation and proliferation of iPSC-derived cardiomyocytes by promoting versican-based GDF11 trapping and subsequent activation of the TGF-β/SMAD2/3 signaling pathway.Hapln1 might be an effective hiPSC-CM dedifferentiation and proliferation agent and a po-tential reagent for repairing damaged hearts.
3.Practical exploration on the Online Four-step Teaching of Encouraging and Sharing during COVID-19 period.
Ting-Huai WANG ; Xiao-Dan XU ; Jie-Hao LI
Acta Physiologica Sinica 2020;72(6):716-723
The "Four-step Teaching of Encouraging and Sharing" is a learner-centered teaching method that advocates teamwork and gives full play to the role of the teacher in guiding learning. It is an innovative teaching approach to realize students' self-transcendence by stimulating students' internal motivation for independent learning, applying group task-driven learning, and giving teachers' feedback to students' sharing. It consists of four steps: teachers' guiding, students' self-regulated learning, team learning and practice, experience sharing. We have applied this method to the teaching practice of physiology and experimental physiological science with a significant impact on teaching effects. This teaching method has also been implemented to other courses in other majors. To solve the problems of reduced communication and interaction, low learning enthusiasm and motivation in online teaching course during COVID-19 pandemic, we recruited 21 undergraduates from different schools and majors. Using the "Tencent Meeting" platform, the authors tried to apply the whole process of the "Four-step Teaching of Encouraging and Sharing" to the online teaching of physiology. Group tests and questionnaires were used to evaluate teaching effects. The results showed that the implementation of the "Online Four-step Teaching of Encouraging and Sharing (OFST)" was feasible and effective, and to a certain extent alleviated the problems of loneliness and low learning motivation of students during online learning caused by home quarantine, which was particularly helpful for long-distance inter-school and inter-discipline team learning.
COVID-19
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Humans
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Learning
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Motivation
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Pandemics
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SARS-CoV-2
5.Proportion of Uterine Malignant Tumors in Patients with Laparoscopic Myomectomy: A National Multicenter Study in China
Yang HUA ; Li XIAO-CHUAN ; Yao CHEN ; Lang JING-HE ; Jin HANG-MEI ; Xi MING-RONG ; Wang GANG ; Wang LU-WEN ; Hao MIN ; Ding YAN ; Chen JIE ; Zhang JIAN-QING ; Han LU ; Guo CHENG-XIU ; Xue XIANG ; Li YAN ; Zheng JIAN-HUA ; Cui MAN-HUA ; Li HUAI-FANG ; Tao GUANG-SHI ; Chen LONG ; Wang SU-MIN ; Lu AN-WEI ; Huang ZE-HUA ; Liu QING ; Zhuang YA-LI ; Huang XIANG-HUA ; Zhu GEN-HAI ; Huang OU-PING ; Hu LI-NA ; Li MU-JUN ; Zhou HONG-LIN ; Song JING-HUI ; Zhu LAN
Chinese Medical Journal 2017;130(22):2661-2665
Background:The Food and Drug Administration recently announced that the use of morcellation may cause fibroids or pelvic dissemination and metastasis of uterine sarcoma;therefore,the use of morcellation is limited in the USA.A large sample study is necessary to assess the proportion of uterine malignant tumors found in patients with laparoscopic myomectomy.Methods:A national multicenter study was performed in China.From 2002 to 2014,33,723 cases were retrospectively selected.We calculated the prevalence and recorded the clinical characteristics of the patients with malignancy after morcellation application.A total of 62 cases were finally pathologically confirmed as malignant postoperatively.Additionally,the medical records of the 62 patients were analyzed in details.Results:The proportion of postoperative malignancy after morcellation application was 0.18% (62/33,723) for patients who underwent laparoscopic myomectomy.Nearly 62.9% (39/62) of patients had demonstrated blood flow signals in the uterine fibroids before surgery.And,23 (37.1%) patients showed rapid growth at the final preoperative ultrasound.With respect to the pathological types,38 (61.3%) patients had detectable endometrial stromal sarcoma,13 (21.0%) had detectable uterine leiomyosarcoma,only 3 (3.2%) had detectable carcinosarcoma,and 5 (8.1%) patients with leiomyoma had an undetermined malignant potential.Conclusions:The proportion of malignancy is low after using morcellation in patients who undergo laparoscopic myomectomy.Patients with fast-growing uterine fibroids and abnormal ultrasonic tumor blood flow should be considered for malignant potential,and morcellation should be avoided.
6.Identification of three novel SRD5A2 mutations in Chinese patients with 5α-reductase 2 deficiency.
Tong CHENG ; Hao WANG ; Bing HAN ; Hui ZHU ; Hai-Jun YAO ; Shuang-Xia ZHAO ; Wen-Jiao ZHU ; Hua-Ling ZHAI ; Fu-Guo CHEN ; Huai-Dong SONG ; Kai-Xiang CHENG ; Yang LIU ; Jie QIAO
Asian Journal of Andrology 2019;21(6):577-581
In this study, we investigated the genetics, clinical features, and therapeutic approach of 14 patients with 5α-reductase deficiency in China. Genotyping analysis was performed by direct sequencing of PCR products of the steroid 5α-reductase type 2 gene (SRD5A2). The 5α-reductase activities of three novel mutations were investigated by mutagenesis and an in vitro transfection assay. Most patients presented with a microphallus, variable degrees of hypospadias, and cryptorchidism. Eight of 14 patients (57.1%) were initially reared as females and changed their social gender from female to male after puberty. Nine mutations were identified in the 14 patients. p.G203S, p.Q6X, and p.R227Q were the most prevalent mutations. Three mutations (p.K35N, p.H162P, and p.Y136X) have not been reported previously. The nonsense mutation p.Y136X abolished enzymatic activity, whereas p.K35N and p.H162P retained partial enzymatic activity. Topical administration of dihydrotestosterone during infancy or early childhood combined with hypospadia repair surgery had good therapeutic results. In conclusion, we expand the mutation profile of SRD5A2 in the Chinese population. A rational clinical approach to this disorder requires early and accurate diagnosis, especially genetic diagnosis.
3-Oxo-5-alpha-Steroid 4-Dehydrogenase/genetics*
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Adolescent
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Adult
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Asian People/genetics*
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Child
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Child, Preschool
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China
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Disorder of Sex Development, 46,XY/genetics*
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Follicle Stimulating Hormone/blood*
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Genitalia, Male/abnormalities*
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Humans
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Hypospadias/genetics*
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Luteinizing Hormone/blood*
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Male
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Membrane Proteins/genetics*
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Mutation/genetics*
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Sequence Alignment
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Steroid Metabolism, Inborn Errors/genetics*
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Testosterone/blood*
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Young Adult