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Drosophila models used to simulate human ATP1A1 gene mutations that cause Charcot-Marie-Tooth type 2 disease and refractory seizures

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Neural Regeneration Research 2025年第1期20卷 265-276页

作者： Yao Yuan Lingqi Yu Xudong Zhuang Dongjing Wen Jin He Jingmei Hong Jiayu Xie Shengan Ling Xiaoyue Du Wenfeng Chen Xinrui Wang Institute of Life Sciences College of Biological Science and EngineeringFuzhou UniversityFuzhouFujian ProvinceChina NHC Key Laboratory of Technical Evaluation of Fertility Regulation for Non-human Primate(Fujian Maternity and Child Health Hospital) FuzhouFujian ProvinceChina Medical Research Center Fujian Maternity and Child Health HospitalCollege of Clinical Medicine for Obstetrics&Gynecology and PediatricsFujian Medical UniversityFuzhouFujian ProvinceChina department of Neurology and Institute of Neurology of The First Affiliated Hospital Institute of Neuroscienceand Fujian Key Laboratory of Molecular NeurologyFujian Medical UniversityFuzhouFujian ProvinceChina

Certain amino acids changes in the human Na^(+)/K^(+)-ATPase pump,ATPase Na^(+)/K^(+)transporting subunit alpha 1(ATP1A1),cause Charcot-Marie-Tooth disease type 2(CMT2)disease and refractory *** develop in vivo models to study the role of Na^(+)/K^(+)-ATPase in these diseases,we modified the Drosophila gene homolog,Atpα,to mimic the human ATP1A1 gene mutations that cause *** located within the helical linker region of human ATP1A1(I592T,A597T,P600T,and D601F)were simultaneously introduced into endogenous Drosophila Atpαby CRISPR/Cas9-mediated genome editing,generating the Atpα^(TTTF)*** addition,the same strategy was used to generate the corresponding single point mutations in flies(Atpα^(I571T),Atpα^(A576T),Atpα^(P579T),and Atpα^(D580F)).Moreover,a deletion mutation(Atpα^(mut))that causes premature termination of translation was generated as a positive *** these alleles,we found two that could be maintained as homozygotes(Atpα^(I571T)and Atpα^(P579T)).Three alleles(Atpα^(A576T),Atpα^(P579)and Atpα^(D580F))can form heterozygotes with the Atpαmut *** found that the Atpαallele carrying these CMT2-associated mutations showed differential phenotypes in *** heterozygous for Atpα^(TTTF)mutations have motor performance defects,a reduced lifespan,seizures,and an abnormal neuronal *** Drosophila models will provide a new platform for studying the function and regulation of the sodium-potassium pump.

关键词： ATP1A1 Atpα bang-sensitive paralysis Charcot-Marie-Tooth disease type 2 CRISPR/Cas9 homology-directed repair Na^(+)/K^(+)-ATPase point mutation seizures sodium pump

Inhibition of Expression of Hypoxia-inducible Factor-1α mRNA by Nitric Oxide in Hypoxic Pulmonary Hypertension Rats

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2004年第1期24卷 5-8页

作者：敖启林黄磊朱朋成熊密王迪浔 department of Pathology and Pathophysiology department of obstetrics and gynecology

In order to study the effect of nitric oxide (NO) on the expression of hypoxia inducible factor 1 alpha (HIF 1α) mRNA in hypoxic pulmonary hypertension (HPH) rats, 30 healthy male Wistar rats were randomly divided into normoxic control group, chronic hypoxic group and hypoxia plus L argine (L Arg) group. The animal model of HPH was developed. The mean pulmonary arterial pressure (mPAP) was measured by inserting a microcatheter into the pulmonary artery. The HIF 1α mRNA expression levels were detected by in situ hybridization (ISH) and semiquantitative RT PCR. It was found that after 14 days hypoxia, the mPAP in normoxic control group (17.6±2 7 mmHg,1 mmHg=0 133 kPa) was significantly lower than that in chronic hypoxic group(35.8±6.1 mmHg, t =0.2918, P <0.05) and mPAP in chronic hypoxic group was higher than that in hypoxia plus L argine group(24.4±3.8 mmHg, t =0.2563, P <0.05). ISH showed that the expression of HIF 1α mRNA in the intraacinar pulmonary arteriolae (IAPA) in normoxic control group (0.1076±0.0205) was markedly weaker than that in chronic hypoxic group (0.3317±0.0683, t =3.125, P <0.05) and that in chronic hypoxic group was stronger than that in hypoxia plus L argine group (0.1928±0.0381, t =2.844, P <0.05). RT PCR showed that the content of HIF 1α mRNA in chronic hypoxic group (2.5395±0.6449) was 2.16 times and 1.75 times higher than that in normoxic control group (1.1781±0.3628) and hypoxia plus L argine group (1.4511±0.3981), respectively. It is concluded that NO can reduce the mPAP by the inhibition of the expression of HIF 1α mRNA, which may be one of the mechanisms through which NO affects the pathogenesis of HPH.

关键词： nitric oxide hypoxia-inducible factor-1 alpha hypoxic pulmonary hypertension

Abnormal regulation of chemokine TECK and its receptor CCR9 in the endometriotic milieu is involved in pathogenesis of endometriosis by way of enhancing invasiveness of endometrial stromal cells

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Cellular & Molecular immunology 2010年第1期7卷 51-60页

作者： Yun Wang Jing Yu Xuezhen Luo Xiaoqiu Wang Mingqing Li Ling Wang Dajin Li Laboratory for Reproductive immunology Hospital and Institute of Obstetrics and GynecologyShanghai Medical College of Fudan UniversityShanghai 200011China department of obstetrics and gynecology Affiliated Hospital of Hainan Medical CollegeHaikou 570102China

The chemokine thymus-expressed chemokine(TECK),which regulates T-cell development and tissue-specific homing,has been identified as a potential contributor to the pathogenesis and progression of ***(2,3,7,8-tetrachlorodibenzo-p-dioxin,TCDD),an air pollutant,and estrogen also appear to be involved in *** endometrial stromal cells(ESCs)and the combination of 17b-estradiol and TCDD increase the secretion of TECK in the endometriosis-associated cells and promote the invasiveness of ESCs by increasing expression of matrix metalloproteinase(MMP)-2 and ***-TECK neutralizing antibodies can effectively inhibit the invasiveness of ESCs and the expression of MMP-2 and MMP-9 in the ***,the expression of chemokine C receptor 9(CCR9)and its ligand TECK increases significantly in the endometriotic milieu of patients with ***,the over-expressed TECK interacts with CCR9 on the ESCs in the endometriotic milieu,which may contribute to the onset and progression of endometriosis.

关键词： TECK CCR9 DSC invasiveness endometriosis

Inhibitory effect of lanthanum chloride on migration and invasion of cervical cancer cells

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Journal of Rare Earths 2013年第1期31卷 94-100页

作者：陈宏伟刘丝荪缪丽芳俞玲芳汪泱郭菲 Burns Institute The First Affiliated Hospital of Nanchang University Postgraduate department of Nanchang University department of gynecology and obstetrics The First Affiliated Hospital of Nanchang University department of gynecology and obstetrics Jiujiang Maternal and Children Health Hospital

Tumor metastasis remains the main reason for treatment Jhilure and death of patients with cervical cancer. The prcsent study was designed to explore the effects of lanthanum chloride （LaCl3 on the invasion and migration of cervical cancel- cells and the underlying mechanisms. The migration and invasion of tumor cells was evaluated by a modified Transwell/Boyden chamber assays. It is well known that MMPs （Matrix metalloproteinases） and NF-κB （Nuclear factor-κB） pathway play important roles in migration and invasion of tumor cells, and also the expression of MMPs were regulated by NF-κB signaling. The expression on MMP-1 and MMP-9 was detected by reverse transcription polymerase chain reaction （RT-PCR）; Western blot and the NF-KB-DNA-binding ac tivity assay were used to analyze the NF-KB activity. The results indicated that LaCl3 was capable of inhibiting the cell invasion and migration of human cervical cancer Hela cells by decreasing the expression of MMP-1 and MMP-9 via blocking NF-κB pathway

关键词： lanthanum chloride (LaCl3) cervical cancer invasion migration: NF-~B rare earths

Decline of semen quality during IVF is not associated with subjective male stress

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Asian Journal of Andrology 2014年第4期16卷 597-601页

作者： Kazem Nouri Brigitte Litschauer Michael Sator Denise Tiringer Johannes Ott Katherina Walch Lukas A Hefler Clemens B Tempfer department of obstetrics and Gynecolog department of Clinical Pharmacology Medical University of Vienna Vienna department of obstetrics and gynecology Krankenhaus Barmherzige Schwestern Linz Austria department of obstetrics and gynecology Ruhr University Bochum Bochum Germany.

The aim of the present study was to assess if semen quality declines during in vitro fertilization （IVF） and whether or not this phenomenon is triggered by chronic male stress. In order to test this hypothesis, we first investigated a retrospective cohort of 155 male IVF patients （testing cohort）. Subsequently, we started a prospective cohort study in men undergoing their first IVF and assessed semen quality and subjective male chronic stress using a validated tool, i.e. the Fertility Problem Inventory （FPI） questionnaire. The association between stress and sperm quality decline measured 4-6weeks before the start of IVF （T1） and at the day of oocyte retrieval （T2） was the primary outcome. Live birth rate, first trimester abortion and rate of poor responders were secondary outcomes. In the testing cohort, mean progressive motility, but not mean sperm density significantly declined. There were 78/154 （51%） men who showed a decline in semen density and 50/154 （32%） men who showed a decline in progressive motility. In the validation cohort, progressive motility declined, whereas, sperm density increased from T1 to T2. Of 78 men, 27 men had increased stress （FPI-score 〉 146）. Sperm density and progressive motility were not significantly different in men with and without stress. However, in the presence of male stress, couples had a higher rate of poor responders, miscarriages and a lower rate of live births. Subjective stress is not associated with a decline in semen quality observed during IVF but may be associated with adverse ore^nancv outcome.

关键词： in vitro fertilization questionnaire semen quality stress

Ovarian cancer presenting with hypercalcemia:two cases with similar manifestations but different mechanisms

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Cancer Biology & Medicine 2018年第2期15卷 182-187页

作者： Xuegong Ma Yingmei Wang Xuhong Zhang Mengting Dong Wen Yang Fengxia Xue department of gynecology and obstetrics Tianjin Medical University General Hospital

Hypercalcemia presenting in ovarian cancer is uncommon in the ***,two cases of ovarian epithelial carcinoma that presented with severe hypercalcemia were reported,with a review of the *** laboratory findings and stepwise clinical investigations of these two cases differed,indicating distinct underlying causes of *** case one,the serum levels and immunostaining for parathyroid hormone-related protein(PTHr P)verified humoral hypercalcemia of malignancy(HHM).In case two,the high level of parathyroid hormone(PTH)and the scintigraphy scan showing parathyroid gland adenoma confirmed primary hyperparathyroidism-induced *** patients received optimal cytoreductive operation and adjuvant chemotherapy but showed different outcomes *** article focused on differential diagnosis of ovarian cancerassociated hypercalcemia,by stepwise imaging and laboratory investigation,and the appropriate therapy should be considered based on the different etiologies.

关键词： Ovarian cancer hypercalcemia PTHrP hyperparathyroidism

Transmission of the Ychromosome microdeletion to a baby boy conceived after in tracytoplasmic sperm injection

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Chinese Medical Journal 2001年第1期114卷 97-99页

作者： Tse JYM Yeung WSB Lau EYL Ng EHY Tam PC So WWK Tang MHY Lau ET Ho PC department of obstetrics and gynecology The University of HongKongHongKongChina

Subfertility can be caused by acquired or genetic factors. Y chromosome microdeletion is one of the genetic factors associating with male infertility.1 Azoospermia factors (AZFa, AZFb and AZFc) have been mapped to different subregions in Yq11.2 So far, two gene families, RNA-binding motif (RBM) and deleted in azoospermia (DAZ) from interval 6, were proposed as candidate spermatogenesis genes for AZF.3,4 Recent studies demonstrated that microdeletions were detected at a frequency of 5% to 18% in the AZF region of oligospermic and azoospermic men.5-7 With the development of assisted reproductive technologies, particularly intracytoplasmic sperm injection (ICSI), these men can now father a child and the genetic abnormalities in defective spermatozoa could be transmitted to future offspring. To examine the possible transmission of the Y-chromosome microdeletion to the offspring via ICSI treatment, we performed both cytogenetic and molecular analyses of the Y chromosome on both an infertile patient with Y chromosome microdeletion and his offspring.

关键词： sperm

Realgar-Induced Apoptosis of Cervical Cancer Cell Line Siha via Cytochrome C Release and Caspase-3 and Caspase-9 Activation

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Chinese Journal of Integrative Medicine 2012年第5期18卷 359-365页

作者：程艳香刘嵘王琴李秉枢许学先胡敏陈璐付琼濮德敏洪莉 department of gynecology Renmin Hospital of Wuhan University department of obstetrics and gynecology No.161 Air Force Hospital department of obstetrics and gynecology Tongji HospitalMedical College of Wuhan University

Objective： To explore the molecular mechanism of realgar-induced apoptosis of cervical cancer cells. Methods： The cervical cancer cell line Siha was used to determine the cell viability and apoptosis after treatment with realgar using MTT assay and flow cytometry. The activities of caspase-3, -8, and -9 were detected by fluorescence resonance energy transfer technology and colorimetric assay, while the levels of Bcl-2, cytochrome c, and Bax were detected by Western blot method. Results： Induction of apoptosis by realgar was detected in Siha cell line in a dose-dependent manner. The apoptosis was accompanied by a significant increase in cytochrome c release and activation of caspase-3 and caspase-9 but not caspase-8. Further, the realgar-induced apoptosis was inhibited by a broad-spectrum caspase inhibitor, a caspase-3 inhibitor, and a caspase-9 inhibitor but not by a caspase-8 inhibitor. Bcl-2 and Bax protein expressions were not changed by realgar. Conclusion： The induction of apoptosis by realgar is mediated through a cytochrome c-dependent pathway, which sequentially activates caspase-9 and caspase-3.

关键词： realgar cervical cancer apoptosis cytochrome c caspases

Prognostic effect analysis of molecular subtype on young breast cancer patients

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Chinese Journal of Cancer Research 2015年第4期27卷 428-436页

作者： Hong-Liang Chen Ang Ding Fu-Wen Wang department of Breast Surgery Obstetrics and Gynecology Hospital of Fudan University

Objective： To make a prognostic effect analysis of molecular subtype on young breast cancer ***： Totally 187 cases of young breast cancer patients less than 40 years old treated in obstetrics and gynecology Hospital of Fudan University between June 2005 and June 2011 were included in our study. We described their clinical-pathological characteristics, disease-free survival（DFS） rate, and overall survival（OS） rate after a median follow-up period of 61 months. The factors associated with prognosis were also evaluated by univariate and multivariate ***： All patients were premenopausal, with an average age of 35.36±3.88 years old. The mean tumor size was 2.43±1.53 cm. Eighty-one cases had lymph node metastasis（43.3%）, 126 cases had lymphovascular invasion（67.4%）, and 125 cases had histological grade III（66.8%） disease. Twenty-seven cases（14.4%） were Luminal A subtype, 99 cases（52.9%） were Luminal B subtype, 29 cases（15.5%） were human epidermal growth factor receptor 2（HER-2） overexpression subtype, while 32 cases（17.1%） were triple negative breast cancer（TNBC） subtype according to 2013 St Gallen expert consensus. One hundred and thirty-five cases underwent mastectomy whereas 52 cases had breast-conserving surgery. One hundred and seventy-eight cases underwent adjuvant or neoadjuvant chemotherapy. Recurrence or metastasis occurred in 29 cases, 13 of which died. The 5-year DFS and OS rates were 84% and 92%. Multivariate analysis showed that nodal status（P=0.041） and molecular subtype（P=0.037） were both independent prognostic factors of DFS, while nodal status（P=0.037） and TNBC subtype（P=0.048） were both independent prognostic factors of OS. Conclusions： Molecular subtype is an independent prognostic factor of young breast cancer patients. TNBC has a high risk of relapse and death.

关键词： Molecular subtype young breast cancer prognosis