1.Identification of novel pathogenic variants in genes related to pancreatic β cell function: A multi-center study in Chinese with young-onset diabetes.
Fan YU ; Yinfang TU ; Yanfang ZHANG ; Tianwei GU ; Haoyong YU ; Xiangyu MENG ; Si CHEN ; Fengjing LIU ; Ke HUANG ; Tianhao BA ; Siqian GONG ; Danfeng PENG ; Dandan YAN ; Xiangnan FANG ; Tongyu WANG ; Yang HUA ; Xianghui CHEN ; Hongli CHEN ; Jie XU ; Rong ZHANG ; Linong JI ; Yan BI ; Xueyao HAN ; Hong ZHANG ; Cheng HU
Chinese Medical Journal 2025;138(9):1129-1131
2.The perioperative application of erector spinae plane block
Daoyi LIN ; Peng YE ; Cansheng GONG ; Danfeng WANG ; Ting ZHENG ; Xiaochun ZHENG
Journal of Clinical Surgery 2024;32(6):657-660
The erector spinae plane block(ESPB)technique proposed by Forero et al.has drawn increasing attention due to its reliable effects and fewer complications in simple and safe procedures.It has been successfully applied in anesthesia management during the perioperative period.This article mainly introduces the anatomical basis,possible mechanism of action,common operating methods,and clinical applications during the perioperative period of ESPB.Starting from the advantages,disadvantages,and complications of ESPB,it compares ESPB with other common thoracolumbar analgesic methods,providing better evidence support for the perioperative application and future development of ESPB.
3.A study on KIF1A gene missense variant analysis and its protein expression and structure profiles of an autism spectrum disorder family trio.
Yan HUANG ; Jian JIAO ; Manxue ZHANG ; Mingjing SITU ; Danfeng YUAN ; Peng LYU ; Sixun LI ; Zhuo WANG ; Yanping YANG ; Yi HUANG
Chinese Journal of Medical Genetics 2021;38(7):620-625
OBJECTIVE:
To analyze the pathogenic variants of the KIF1A gene and its corresponding protein structure in an autism spectrum disorder (ASD) family trio carrying harmful missense variants in the KIF1A gene.
METHODS:
The peripheral blood DNA of the patient and his parents was extracted and sequenced using whole exome sequencing (WES) technology and verified by Sanger sequencing. Bioinformatics software SIFT, PolyPhen-2, Mutation Taster, and CADD software were used to analyze the harmfulness and conservation of variants. The Human Brain Transcriptome (HBT) database was used to analyze the expression of the KIF1A gene in the brain. PredictProtein and SWISS-MODEL were further used to predict the secondary structure and tertiary structure of KIF1A wild-type protein and variant protein. PyMOL V2.4 was utilized to investigate the change of hydrogen bond connection after protein variant.
RESULTS:
The WES sequencing revealed a missense variant c.664A>C (p.Asn222His) in the child's KIF1A gene, and this variant was a de novo variant. The harmfulness prediction results suggest that this variant is harmful. By analyzing expression level of KIF1A gene in the brain. It is found that KIF1A gene widely expressed in various brain regions during embryonic development. By analyzing the variant protein structure, the missense variant of KIF1A will cause many changes in the secondary structure of protein, such as alpha-helix, beta-strand, and protein binding domain. The connection of hydrogen bond and spatial structure will also change, thereby changing the original biological function.
CONCLUSION
The KIF1A gene may be a risk gene for ASD.
Autism Spectrum Disorder/genetics*
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Child
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Female
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Humans
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Kinesin/genetics*
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Mutation
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Mutation, Missense
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Pregnancy
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Protein Domains
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Whole Exome Sequencing
4.Preparation of chitosan-gelatin porous microspheres by high voltage electrostatic method combined with freeze-drying and ionic cross-linking method
Fang HUANG ; Wenbin RUI ; Danfeng XU ; Yu ZHU ; Baiyong SHEN ; Chenghong PENG
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(7):885-890
Objective·To prepare chitosan-gelatin porous microspheres by high voltage electrostatic method combined with freeze-drying and ionic cross-linking method and investigate the factors that influence the formation of porous medium.Methods·Porous chitosan microspheres and chitosangelatin porous microspheres were prepared using high voltage electrostatic method combined with freeze-drying and ionic cross-linking method,with sodium tripolyphosphate (STPP) as crosslinking agent.Factors that affect the porous structure and pore size of porous microspheres were compared,such as different chitosan-gelatin ratio,freezing temperature,curing time with saturated STPP in 85% ethanol solution.The morphology,surface and internal structure,particle size of the porous chitosan microspheres and chitosan-gelatin porous microspheres were observe by using light microscope,scanning electron microscope and hematoxylin-eosin staining.Results·Microspheres prepared by freeze-drying with an electrostatic and ionic cross-linking method have open,interconnected and highly macroporous,with good spherical surface.Saturated STPP ethanol solution (85% ethanol) was chosen as the crosslinking agent to prevent destruction of the porous structure.The order of freeze and crosslinking,cross-linking time and the second freezing temperature,can influence the pore size of porous microspheres.Gelatin and chitosan can form polyelectrolyte complexes,and can also be used as porogen in porous structure.Conclusion·The preparation of porous chitosan-gelatin microspheres via this method has a large pore size (diameter 100-200 μm),suitable for cell growth and the migration.
5.Antimicrobial Resistance and Molecular Characteristics of Nasal Staphylococcus aureus Isolates From Newly Admitted Inpatients.
Xu CHEN ; Kangde SUN ; Danfeng DONG ; Qingqiong LUO ; Yibing PENG ; Fuxiang CHEN
Annals of Laboratory Medicine 2016;36(3):250-254
Staphylococcus aureus, or methicillin-resistant S. aureus (MRSA), is a significant pathogen in both nosocomial and community infections. Community-associated MRSA (CA-MRSA) strains tend to be multi-drug resistant and to invade hospital settings. This study aimed to assess the antimicrobial resistance and molecular characteristicsof nasal S. aureus among newlyadmitted inpatients.In the present study, 66 S. aureus isolates, including 10 healthcare-associated MRSA (HA-MRSA), 8 CA-MRSA, and 48 methicillin-sensitive S. aureus (MSSA) strains, were found in the nasal cavities of 62 patients by screening 292 newlyadmitted patients. Antimicrobial resistance and molecular characteristics of these isolates, including spa-type, sequence type (ST) and SCCmec type, were investigated. All isolates were sensitive to linezolid, teicoplanin, and quinupristin/dalfopristin, but high levels of resistance to penicillin and erythromycin were detected. According to D-test and erm gene detection results, the cMLSB and iMLSB phenotypes were detected in 24 and 16 isolates, respectively. All 10 HA-MRSA strains displayed the cMLSB phenotypemediated by ermA or ermA/ermC, while the cMLSB CA-MRSA and MSSA strains carried the ermB gene. Molecular characterization revealedall 10 HA-MRSA strains were derived from the ST239-SCCmec III clone, and four out of eight CA-MRSA strains were t437-ST59-SCCmec V. The results suggest that patients play an indispensable role in transmitting epidemic CA-MRSA and HA-MRSA strains.
Anti-Bacterial Agents/*pharmacology
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Bacterial Proteins/genetics
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Drug Resistance, Multiple, Bacterial/genetics
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Humans
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Inpatients
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Methicillin-Resistant Staphylococcus aureus/*drug effects/genetics/isolation & purification
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Methyltransferases/genetics
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Microbial Sensitivity Tests
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Nasal Cavity/*microbiology
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Staphylococcal Infections/diagnosis/microbiology
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Staphylococcus aureus/*drug effects/genetics/isolation & purification
6.Clinical study of acute spinal spontaneous hematoma
Deqing PENG ; Guangyu YING ; Liangliang ZHU ; Danfeng YU ; Yongjian ZHU
Chinese Journal of Emergency Medicine 2016;25(4):486-490
Objective To evaluate the etiology, pathogenesis, clinical manifestation, imaging features, treatments and factors related to prognosis of acute spinal spontaneous hematoma.Methods The clinical data of 38 patients with acute spinal hematoma treated in our hospital from 2011 till now were analyzed retrospectively.Duration of follow-up was 6 months.The factors influencing the prognosis were analyzed.Results Acute epidural hematomas (n=29) were much more common than subdural (n=5), subarachnoid (n=1) and intramedullary (n=3).Most hematomas were located in the cervical and thoracic vertebra regions.The etiology of acute spinal spontaneous hematoma was unknown in most patients.Twenty-nine patients were dealt with surgical intervention and 9 patients were treated conservatively.After 6-month follow up, recovery rate measured by JOA score in patients of spinal injury ASIA level A and B was (51.26 ±38.97), and level C, D and E was (80.33 ±25.83), P<0.05.Recovery rate in patients with hematoma discovered in less then 24 hours treated with surgical decompression was (64.79 ±36.10), and that in those with hematoma present over 24 hours was (34.54 ±30.17), P<0.05.Conclusions Acute spinal hematoma always caused by unknown etiology, and usually manifests itself in a sudden onset of pain and neurological deficits.The early diagnosis mainly depends on MRI.Patients presenting with severe neurologic dysfunction or showing signs of progressive deficit should have immediate surgical intervention. The status of neurological deficits before surgery and the length of interval between onset and surgical intervention are associated with recovery.
7.Clinical and microbiological characterization of Clostridium difficile infection in a tertiary care hospital in Shanghai, China.
Danfeng DONG ; Yibing PENG ; Lihua ZHANG ; Cen JIANG ; Xuefeng WANG ; Enqiang MAO
Chinese Medical Journal 2014;127(9):1601-1607
BACKGROUNDOver the last decade, Clostridium difficile infection (CDI) has emerged as a significant nosocomial infection, yet little has been reported from China. This study aimed to characterize the clinical and microbiological features of CDI from a hospital in Shanghai.
METHODSPatients with CDI seen between December 2010 and March 2013 were included in this study, of which clinical data were retrospectively collected. The microbiological features of corresponding isolates were analyzed including genotype by multi-locus sequence typing (MLST), antimicrobial susceptibility, toxin production, sporulation capacity, biofilm formation, and motility.
RESULTSNinety-four cases of CDI were included during this study period, 12 of whom were severe cases. By reviewing the clinical data, all patients were treated empirically with proton pump inhibitor or antibiotics or both, and they were distributed widely across various wards, most frequently to the digestive ward (28/94, 29.79%). Comparing the severe with mild cases, no significant differences were found in the basic epidemiological data or the microbiological features. Among the 94 isolates, 31 were toxin A-negative toxin B-positive all genotyped as ST37. They generated fewer toxins and spores, as well as similar amounts of biofilm and motility percentages, but exhibited highest drug resistance to cephalosporins, quinolones, macrolide-lincosamide and streptogramin (MLSB), and tetracycline.
CONCLUSIONSNo specific clinical genotype or microbiological features were found in severe cases; antimicrobial resistance could be the primary reason for epidemic strains leading to the dissemination and persistence of CDI.
Anti-Bacterial Agents ; pharmacology ; Biofilms ; drug effects ; Cephalosporins ; pharmacology ; China ; Clostridium difficile ; drug effects ; genetics ; isolation & purification ; Genotype ; Multilocus Sequence Typing ; methods ; Quinolones ; pharmacology ; Tertiary Healthcare ; statistics & numerical data ; Tetracycline ; pharmacology
8.Mutation of P927S in PRD1 gene mediates azole resistance in Candida glabrata
Cen JIANG ; Danfeng DONG ; Lihua ZHANG ; Xuefeng WANG ; Yibing PENG
Chinese Journal of Infectious Diseases 2014;32(6):325-329
Objective To investigate the role of PDR1 gene in azole-resistant Candida glabrata (C.glabrata).Methods Thirty-eight clinical isolates of C.glabrata were collected from five different hospitals.The minimal inhibitory concentrations (MIC) of azole antifungals including fluconazole,itraconazole and voriconazole against C.glabrata were determined by broth microdilution.Sequencing and amplification of PDR1 gene was achieved by real-time quantitative polymerase chain reaction (PCR).The mutation was cloned into an expression plasmid and then transferred into C.glabrata.The efflux of rhodamine 6G and drug sensitivity test were performed,and expressions of CDR1 and CDR2 were examined to verify function of mutation.Results Among these 38 isolates of C.glabrata,17 were resistant to at least one of azole antifungals.Moreover,mutations of PDR1 gene existed in every resistant isolates.Results of phenotyping test showed that in the isolate that expressed PDR1P927S,the expression of CDR1 and CDR2 were increased by 20.53 and 4.03 fold,respectively.And the fluorescence intensity of rhodamine 6G was decreased to 0.62 in efflux experiment.Conclusion P927S mutation of PDR1 gene could induce azole resistance of C.glabrata by increasing the expressions of CDR1 and CDR2,which results in drug resistance due to enhanced effect of efflux pump.
9.Mechanisms of fluconazole resistance in clinical and experimental induced isolates of Candida glabrata
Cen JIANG ; Danfeng DONG ; Beiqin YU ; Xuefeng WANG ; Yibing PENG
Chinese Journal of Microbiology and Immunology 2012;32(6):537-541
Objective To investigate the mechanisms of fluconazole resistance in clinical and experimental induced isolates of C.glabrata.Methods Efflux of rhodamine 6G was performed to evaluate the effects of efflux pumps.The expression levels of transporter genes CDR1,CDR2,SNQ2 and ERG11 were examined by real-time RT-PCR.Meanwhile,sequence of PDR1 was determined by PCR based DNA sequencing.Results Efflux pumps of all fluconazole-resistant isolates had stronger effects than that of susceptible isolates,consistently with significant upregulation of CDR1,but no obvious difference was found in CDR2 or SNQ2.Also,no notable change in the expression level of ERG11 between susceptible and resistant isolates.PDR1 mutations existed in both clinical and experimental induced isolates of C.glabrata,among which P927S,L543P and S947L haven't been reported previously.Conclusion Mutations of PDR1 were induced by fluconazole both in vivo andin vitro,which will result in overexpression of CDR1 and strengthen the effect of efflux pump.
10.Genotyping of candida glabrata clinical isolates by repetitive sequence-based PCR and multilocus sequence typing
Danfeng DONG ; Cen JIANG ; Beiqin YU ; Qishi FAN ; Yibing PENG
Chinese Journal of Laboratory Medicine 2011;34(9):810-813
Objective To assess the application value of REP-PCR in genotyping of candida glabrata strains in clinical pratice. MethodsFrom 2009 to 2010, thirty-eight candida glabrata strains were isolated from Shanghai Ruijin Hospitals, Shanghai Renji Hospital, Shanghai Huashan Hospital, Anhui Medical University Hospital, Shenzhen People's Hospital. Six loci in housekeeping genes (FKS, LEU2,NMT1, TRP1, UGP1 and URA3 ) were amplified and sequenced. The sequences were compared with the MIST database and allele profile and sequence type (ST) were obtained. With primers Ca21, Ca22 and Com21 used to amplify the adjacent variable gene regions,the amplicons were analyzed through electrophoresis to generate different REP-PCR types. Finally, the results of these two genotyping methods were compared. ResultsFor REP-PCR, Ca22-Com21 has the best genotyping effect. REP-PCR and MLST have the same genotyping results. Five REP-PCR types were found in 38 candida glabrsta isolates. Type A,B, C, D and E strains from REP-PCR were genotyped as ST 7, 3, 19, 45 and new type respectively byMIST. REP-PCR saves time compared with MIST. Conclusions REP-PCR offers a simple and rapid method for molecular typing, with similar discriminatory power with MIST. Therefore, REP-PCR can be the preferred choice in laboratory, especially for a large number of isolates.

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