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Cobalamin-dependent radical S-adenosyl-L-methionine protein functions with a partner to successively methylate tricyclic indole alkaloid for chuangxinmycin maturation and derivatization

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Science China Chemistry 2025年第1期68卷 308-316页

作者： Sili Wang Jiancheng Huang Yanan Du Wei Huang Yufeng Xue Xiaokun Xu Guannan Zhong Yuanyuan Shi Bin Hong Xiaoying Bian Wen Liu State key laboratory of Chemical Biology Shanghai Institute of Organic Chemistry University of Chinese Academy of SciencesChinese Academy of Sciences State key laboratory of Microbial Technology Shandong University national health commission (NHC) key laboratory of Biotechnology for Microbial Drugs Institute of Medicinal BiotechnologyChinese Academy of Medical Science & Peking Union Medical College

Cobalamin(Cbl)-dependent radical S-adenosyl-L-methionine(SAM) proteins constitute the largest collection of the radical SAM superfamily that has hundreds of thousands of individual members. Many of these proteins are involved in the biosynthesis of pharmaceutically important natural products to catalyze chemically demanding reactions. In the biosynthetic pathway of chuangxinmycin(CXM), a unique indole alkaloid antibiotic with potent anti-infective activity, functionalization of the characteristic thiopyrano[4,3,2-cd]indole scaffold by regio-and stereoselective C3-methylation is believed to rely on a Cbl-dependent radical process, which, however, remained to be reconstituted biochemically. We here report the dissection of this enzymatic process, which requires the incorporation of Cxm8, a Cbl-dependent radical SAM protein, with Cxm9, a DUF5825family protein that shares no homology to any proteins of known functions. Cxm8 and Cxm9 function together by forming an unexpected heterodimeric complex that selectively catalyzes C3-methylation of the tricyclic indole-S-hetero ring system in a successive manner, achieving CXM and a recently identified, C3-dimethylated congener. Detailed biochemical characterization,isotope labeling, structural simulation and bioinformatics analysis rationalized the catalysis of the Cxm8/Cxm9 complex and particularly the necessity of the DUF5825 protein for C3-methylase activity. This is the first example that a Cbl-dependent protein acts with a partner to exhibit radical SAM activity.

关键词： cbl-dependent radical SAM protein biosynthesis chuangxinmycin stereoselective methylation

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MicroRNA-132 in the Adult Dentate Gyrus is Involved in Opioid Addiction Via Modifying the Differentiation of Neural Stem Cells

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Neuroscience Bulletin 2019年第3期35卷 486-496页

作者： Meng Jia Xuewei Wang Haolin Zhang Can Ye Hui Ma Mingda Yang Yijing Li Cailian Cui Department of Neurobiology School of Basic Medical Sciences Key Laboratory for Neuroscience of the Ministry of Education and National Health and Family Planning Commission Neuroscience Research Institute Peking University

MicroRNA-132(miR-132), a small RNA that regulates gene expression, is known to promote neurogenesis in the embryonic nervous system and adult *** exposure to psychoactive substances can increase miR-132 expression in cultured neural stem cells(NSCs)and the adult brain of rodents, little is known about its role in opioid addiction. So, we set out to determine the effect of miR-132 on differentiation of the NSCs and whether this effect is involved in opioid addiction using the rat morphine self-administration(MSA) model. We found that miR-132 overexpression enhanced the differentiation of NSCs in vivo and in vitro. Similarly, speci?c overexpression of miR-132 in NSCs of the adult hippocampal dentate gyrus(DG) during the acquisition stage of MSA potentiated morphine-seeking behavior. These ?ndings indicate that miR-132 is involved in opioid addiction,probably by promoting the differentiation of NSCs in the adult DG.

关键词： miR-132 Opioid addiction Neural stem cell Dentate gyrus

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H5N1 Avian Influenza Pre-pandemic Vaccine Strains in China

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Biomedical and Environmental Sciences 2014年第10期27卷 763-769页

作者： BO Hong DONG Li Bo ZHANG Ye DONG Jie ZOU Shu Mei GAO Rong Bao WANG Da Yan SHU Yue Long national Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention key laboratory for Medical Virology National Health and Family Planning Commission

Objective To prepare the 4 candidate vaccine strains of H5N1 avian influenza virus isolated in China Methods Recombinant viruses were rescued using reverse genetics. Neuraminidase （NA） and hemagglutinin （HA） segments of the A/Xinjiang/1/2006, A/Guangxi/1/2009, A/Hubei/1/2010, and A/Guangdong/1/2011 viruses were amplified by RT-PCR. Multibasic amino acid cleavage site of HA was removed and ligated into the pCIpoll vector for virus rescue. The recombinant viruses were evaluated by trypsin dependent assays. Their embnjonate survival and antigenicity were compared with those of the respective wild-type viruses. Results The 4 recombinant viruses showed similar antigenicity compared with wild-type viruses, chicken embryo survival and trypsin-dependent characteristics. Conclusion The 4 recombinant viruses rescued using reverse genetics meet the criteria for classification of low pathogenic avian influenza strains, thus supporting the use of them for the development of seeds and production of pre-pandemic vaccines.

关键词： Influenza H5N1 Pre-pandemic Vaccine strains

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A randomized, controlled, multicenter contraceptive efficacy clinical trial of the intravas device, a nonocclusive surgical male sterilization

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Asian Journal of Andrology 2014年第3期16卷 432-436页

作者： Wen-Hong Lu Xiao-Wei Liang Yi-Qun Gu Wei-Xiong Wu Li-Wei Bo Tian-Gui Zheng Zhen-Wen Chen key laboratory of Male Reproductive health National Health and Family Planning Commission National Research Institute for Family Planning and WHO Collaborating Centre for Research in Human Reproduction Beijing Institute for family planning of Guangzhou city Guangzhou Institute for family planning of Henan Province Zhengzhou Institute for family planning of Putian City Putian China.

Because of unavoidable complications of vasectomy, this study was undertaken to assess the efficacy and safety of male sterilization with a nonobstructive intravas device （IVD） implanted into the vas lumen by a mini-surgical method compared with no-scalpel vasectomy （NSV）. IVDs were categorized into two types： IVD-B has a tail used for fixing to the vas deferens （fixed wing） whereas IVD-A does not. A multicenter prospective randomized controlled clinical trial was conducted in China. The study was comprised of 1459 male volunteers seeking vasectomy who were randomly assigned to the IVD-A （n = 487）, IVD-B （n = 485） or NSV （n = 487） groups and underwent operation. Follow-up included visits at the 3rd-6TM and 12~ postoperative months, The assessments of the subjects involved regular physical examinations （including general and andrological examinations） and semen analysis. The subjects＇ partners also underwent monitoring for pregnancy by monthly interviews regarding menstruation and if necessary, urine tests, There were no significant differences in pregnancy rates （0.65% for IVD-A, 0 for IVD-B and 0.21% for NSV） among the three groups （P 〉 0.05）. The cumulative rates of complications at the 12th postoperative month were zero, 0.9% and 1.7% in the three groups, respectively. In conclusion, IVD male sterilization exhibits a low risk of long-term adverse events and was found to be effective as a male sterilization method, similar to the NSV technique. IVD male sterilization is expected to be a novel contraceptive method.

关键词： complication male sterilization nonobstructive intravas device no-scalpel vasectomy

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Sequencing the Complete Genome of COVID-19 Virus from Clinical Samples Using the Sanger Method

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China CDC weekly 2020年第25期2卷 447-452页

作者： Roujian Lu Peihua Niu Li Zhao Huijuan Wang Wenling Wang Wenjie Tan key laboratory of Biosafety National Health and Family Planning CommissionNational Institute for Viral Disease Control and PreventionChina CDCBeijingChina

What is already known on this topic?Coronavirus disease 2019(COVID-19),a disease caused by a novel human coronavirus named the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)or COVID-19 virus,was reported in December *** genomes of the COVID-19 virus from clinical samples using next generation sequencing(NGS)have been *** is added by this report?Here we provide the technical data for sequencing complete genome of COVID-19 virus from clinical samples using the Sanger *** complete COVID-19 virus genome sequences(named WH19004-S and GX0002)were obtained from clinical samples of COVID-19 patients,and two single nucleotide polymorphisms(SNPs)in ORF7a(T/C,nt 27,493)and ORF8(T/C,nt 28,253)of WH19004-S were identified by Sanger *** are the implications for public health practice?The COVID-19 virus genome sequencing by Sanger method reported here could be used to generate data of high enough quality without requirement for expensive NGS equipment,which support sequencing complete genomes from clinical samples and monitoring of viral genetic variations of COVID-19 infections.

关键词： Sanger clinical acute

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Disruption of dopamine D1 receptor phosphorylation at serine 421 attenuates cocaine-induced behaviors in mice

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Neuroscience Bulletin 2014年第6期30卷 1025-1035页

作者： Ying Zhang Ning Wang Ping Su Jie Lu Yun Wang Neuroscience Research Institute and Department of Neurobiology The Key Laboratory for Neuroscience of the Ministry of Education/National Health and Family Planning CommissionPeking University Health Science Center PKU-IDG/Mc Govern Institute for Brain Research Peking University

Dopamine D1 receptors（D1Rs） play a key role in cocaine addiction, and multiple protein kinases such as GRKs, PKA, and PKC are involved in their phosphorylation. Recently, we reported that protein kinase D1 phosphorylates the D1 R at S421 and promotes its membrane localization. Moreover, this phosphorylation of S421 is required for cocaineinduced behaviors in rats. In the present study, we generated transgenic mice over-expressing S421A-D1 R in the forebrain. These transgenic mice showed reduced phospho-D1R（S421） and its membrane localization, and reduced downstream ERK1/2 activation in the striatum. Importantly, acute and chronic cocaine-induced locomotor hyperactivity and conditioned place preference were significantly attenuated in these mice. These findings provide in vivo evidence for the critical role of S421 phosphorylation of the D1 R in its membrane localization and in cocaine-induced behaviors. Thus, S421 on the D1 R represents a potential pharmacotherapeutic target for cocaine addiction and other drug-abuse disorders.

关键词： protein kinase D1 dopamine D1 receptor phosphorylation cocaine addiction conditioned place preference locomotor activity

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Three Novel Real-Time RT-PCR Assays for Detection of COVID-19 Virus

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China CDC weekly 2020年第25期2卷 453-457页

作者： Peihua Niu Roujian Lu Li Zhao Huijuan Wang Baoying Huang Fei Ye Wenling Wang Wenjie Tan key laboratory of Biosafety National Health and Family Planning CommissionNational Institute for Viral Disease Control and PreventionChina CDCBeijingChina

What is already known on this topic?A novel human coronavirus,known as SARS-CoV-2 or 2019-nCoV,is the causative agent of the coronavirus disease 2019(COVID-19).We have released the primers and probes of real-time reverse transcription polymerase chain reaction(rRT-PCR)assays for the laboratory detection of COVID-19 *** is added by this report?Here we provide detailed technical data and evaluate the performance of three novel rRT-PCR assays targeting the ORF1ab,N,and E genes for detection of COVID-19 *** application of rRT-PCR assays among four types of specimens(alveolar lavage,sputum,throat swabs,and stool)from patients with COVID-19 indicated that the mean viral loads detected in sputum were higher than other *** are the implications for public health practice?These rRT-PCR assays reported here could be used for laboratory diagnosis of COVID-19 infection with high sensitivity,specificity,and *** rather than throat swabs and stool should be a priority for specimen collection for laboratory detection of COVID-19.

关键词： diagnosis specificity stool

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Biosynthesis of antibiotic chuangxinmycin from Actinoplanes tsinanensis

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Acta Pharmaceutica Sinica B 2018年第2期8卷 283-294页

作者： Yuanyuan Shi Zhibo Jiang Xingxing Li Lijie Zuo Xuan Lei Liyan Yu Linzhuan Wu Jiandong Jiang Bin Hong key laboratory of Biotechnology of Antibiotics the National Health and Family Planning Commission (NHFPC) Institute of Medicinal Biotechnology Chinese Academy of Medical Sciences & Peking Union Medical College

Chuangxinmycin is an antibiotic isolated from Actinoplanes tsinanensis CPCC 200056 in the1970 s with a novel indole-dihydrothiopyran heterocyclic skeleton. Chuangxinmycin showed in vitro antibacterial activity and in vivo efficacy in mouse infection models as well as preliminary clinical *** the biosynthetic pathway of chuangxinmycin has been obscure since its discovery. Herein, we report the identification of a stretch of DNA from the genome of A. tsinanensis CPCC 200056 that encodes genes for biosynthesis of chuangxinmycin by bioinformatics analysis. The designated cxn cluster was then confirmed to be responsible for chuangxinmycin biosynthesis by direct cloning and heterologous expressing in Streptomyces coelicolor M1146. The cytochrome P450 CxnD was verified to be involved in the dihydrothiopyran ring closure reaction by the identification of seco-chuangxinmycin in S. coelicolor M1146 harboring the cxn gene cluster with an inactivated cxn D. Based on these results, a plausible biosynthetic pathway for chuangxinmycin biosynthesis was proposed, by hijacking the primary sulfur transfer system for sulfur incorporation. The identification of the biosynthetic gene cluster of chuangxinmycin paves the way for elucidating the detail biochemical machinery for chuangxinmycin biosynthesis, and provides the basis for the generation of novel chuangxinmycin derivatives by means of combinatorial biosynthesis and synthetic biology.

关键词： Chuangxinmycin Actinoplanes tsinanensis Biosynthesis gene cluster Heterologous expression Cytochrome P450 Seco-chuangxinmycin C–S bond formation Sulfur incorporation

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Influencing factors of inpatient expenditure pattern for cancer in China, 2015

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Chinese Journal of Cancer Research 2017年第1期29卷 11-17页

作者： Lan Lan Yue Cai Tao Zhang Ruixian Wu Ming Xue Qun Meng West China School of Public health Sichuan University Center for health Statistics and Information National Health and Family Planning Commission

Objective： We analyzed the proportion of cancer-caused hospitalization expenses in total hospitalization expenses from national authoritative data and explored influencing factors of the proportion so as to provide effective data information for more rational utilization of health ***： Two-level lineal regression model was used to explore influencing factors of ratios of the cancer inpatient expenditure over the total inpatient expenditure of hospitals in China in 2015. A total of 40.76 million inpatient medical records were used to generate the outcome variables, while the explanatory variables were from hospital information database and China health and family planning Statistical Yearbook and ***： Inpatient expenditure pattern for cancer（IEPC） varied largely across provinces, ranging from 3.03% to19.61%. The major sources of variability were from the differences of hospital level and number of beds. There was homogeneity within a province, while heterogeneity between the provinces. Rising one level of the hospital led to the increase of 0.475 natural logarithm units of IEPC averagely. The number of beds increasing 1,000 each made the natural logarithm of IEPC increase one unit ***： Our study showed that a considerable proportion of IEPC variation could be explained by the differences of hospital level and number of beds. It implied that it is possible to estimate disease-specific ratio of inpatient expense taking into account key influencing factors in China. Furthermore, this study is an input to economic and financial analyses and provides evidence for future study on the national economic burden of cancer.

关键词： Cancer expenditure pattern multilevel China

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Protein kinase D: a new player among the signaling proteins that regulate functions in the nervous system

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Neuroscience Bulletin 2014年第3期30卷 497-504页

作者： Gang Li Yun Wang Neuroscience Research Institute and Department of Neurobiology School of Basic Medical Sciences Key Laboratory for Neuroscience Ministry of Education/National Health and Family Planning Commission Peking University PKU-IDG/McGovern Institute for Brain Research Peking University

Protein kinase D （PKD） is an evolutionarily-conserved family of protein kinases. It has structural, regulatory, and enzymatic properties quite different from the PKC family. Many stimuli induce PKD signaling, including G-protein-coupled receptor agonists and growth factors. PKD1 is the most studied member of the family. It functions during cell proliferation, differentiation, secretion, cardiac hypertrophy, immune regulation, angiogenesis, and cancer. Previously, we found that PKD1 is also critically involved in pain modulation. Since then, a series of studies performed in our lab and by other groups have shown that PKDs also participate in other processes in the nervous system including neuronal polarity establishment, neuroprotection, and learning. Here, we discuss the connections between PKD structure, enzyme function, and localization, and summarize the recent findings on the roles of PKD-mediated signaling in the nervous system.

关键词： PKD neuronal polarity pain modulation neuroprotection learning

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