Detection of Novel BEST1 Variations in Autosomal Recessive Bestrophinopathy Using Third-generation Sequencing

Objective:Autosomal recessive bestrophinopathy(ARB),a retinal degenerative disease,is characterized by central visual loss,yellowish multifocal diffuse subretinal deposits,and a dramatic decrease in the light peak on electrooculogram.The potential pathogenic mechanism involves mutations in the BEST1 gene,which encodes Ca2+-activated Cl−channels in the retinal pigment epithelium(RPE),resulting in degeneration of RPE and photoreceptor.In this study,the complete clinical characteristics of two Chinese ARB families were summarized.Methods:Pacific Biosciences(PacBio)single-molecule real-time(SMRT)sequencing was performed on the probands to screen for disease-causing gene mutations,and Sanger sequencing was applied to validate variants in the patients and their family members.Results:Two novel mutations,c.202T>C(chr11:61722628,p.Y68H)and c.867+97G>A,in the BEST1 gene were identified in the two Chinese ARB families.The novel missense mutation BEST1 c.202T>C(p.Y68H)resulted in the substitution of tyrosine with histidine in the N-terminal region of transmembrane domain 2 of bestrophin-1.Another novel variant,BEST1 c.867+97G>A(chr11:61725867),located in intron 7,might be considered a regulatory variant that changes allele-specific binding affinity based on motifs of important transcriptional regulators.Conclusion:Our findings represent the first use of third-generation sequencing(TGS)to identify novel BEST1 mutations in patients with ARB,indicating that TGS can be a more accurate and efficient tool for identifying mutations in specific genes.The novel variants identified further broaden the mutation spectrum of BEST1 in the Chinese population.

p21^(WAF1/CIP1) Gene DNA Sequence Change and Their Relationship with the Phenotype of Human Osteosarcoma

Objective: To investigate the p21WAF1 /CIP1gene DNA sequence change and their relationship with the phenotype of human osteosarcoma. Methods: p21WAF1 /CIP1gene DNA of 36 osteosarcoma spec- imens was examined by using polymerase chain reaction-single strand conformation polymorphism (PCR- SSCP) method. The PCR products were sequenced directly. Results: In p21WAF1 /CIP1 gene exon3 of 36 cases of human osteosarcoma, the change of C→T in the p21WAF1 /CIP1gene CDNA sequence of position 609th occurred in 17 cases with the incidence being 44.4%. In 10 normal blood samples, DNA sequence analysis showed the change of C→T in the p21WAF1 /CIP1gene CDNA sequence of position 609th occurred in 8 cases with the incidence being 80%. Conclusion: The novel location of p21WAF1 /CIP1gene polymorphism of osteosarcoma, but not mutation was de?ned, and this location might provide the meaningful reference for the further research of p21WAF1/CIP1 gene.p2lWAF1/CIP1基因DNA序列分析及其与骨肉瘤表型的关系

Cloning and Sequence Analysis on IGF-1 Gene of Hubei White Swine

[Objective] The study aimed at cloning and analyzing the insulin-like growth factor-1 （IGF-1） gene from liver of Hubei white swine. [Method] The total RNA was extracted by using Trizol from the liver of Hubei white swine and used as template to amplify IGF-1 gene cDNA by RT-PCR. The cDNA product was cloned into pCRII vector, screened with blue-white colonies, digested with double enzymes and sequenced. [Result] The sequencing result indicated that the IGF-1 gene consisted of 607 nucleotides, containing 5＇-untranslated region at nucleotides 1-145, a complete ORF at nucleotides 146-538 encoding 130 amino acids, and 3＇-untranslated region at nucleotides 539-607. It shared 100% homology with the porcine IGF-1 gene reported by Muller et al. [Conclusion] The successful cloning and sequencing of the Hubei white swine IGF-1 gene confirmed that IGF-I gene was highly conserved, which provided technical basis for the use of transgenic technology for breeding of Hubei white swine.

Cloning and Sequence Analysis of Glycoprotein D Gene of Bovine Herpesvirus-1 Strain Luojing

By means of PCR,the gene encoding gD of bovine herpesvirus-1 (BHV-1) strain Luojing was amplified,cloned and sequenced.The nucleotide sequence of this gD gene was (1 251 bp,)encoding 417 amino acids.Comparied with the published P8-2 strain,the homology of the necleotide sequence is 99.92%,and that of the deduced amino acid sequence is 100%.The results indicated that gD of BHV-1 was highly conservative.

Cloning, Sequence Analysis, and Prokaryotic Expression of the Porcine DECR1 Gene

2,4-dienoyl-CoA reductase 1 （ DECR1 ） is the key rate-limiting enzyme in the metabolism of polyunsaturated fatty acids. Although this protein has been studied in a variety of mammals, its role in por- cine is yet to be fully elucidated. However, it is a candidate determinant/indicator of meat quality, growth traits, and carcass quality. Here, we employed RT-PCR and rapid amplification of cDNA ends （RACE） analysis to amplify the full-length cDNA of DECR1 from Mashen pig liver, and cloned it into the expression vector pET-32a＋. After confirmation by sequencing and restriction analysis, the recombinant plasmid was transformed into E. coli BL21 cells. The cDNA of pig DECR1 contained 2,352 nucleotides, including a 987 bp open reading frame flanked by a 53 bp 5＇-untranslated region （UTR） and a 1,312 bp 3＇-UTR. The pig DECR1 coding sequence encoded 328 amino acid residues, which shared 99%, 88%, 87%, 87%, 87%, 87%, and 83% identity with those of Sus scrofa （predicted）, Bos taurus, Homo sapiens, Macaca mulatta, Pan troglodytes, Equus caballus, Canis, and Mus musculus, respectively. SDS-PAGE analysis revealed that the recombinant protein was expressed and that the expression level reached its highest level after 4 h induction. Western blot analysis indicated that the molecular weight of the expressed protein was the same as that predicted, ap- proximately 35 kDa. Collectively these data provide the basis for further studies into the physiological functions and molecular mechanisms of the pig DE- CR1 gene.

Isolation and sequence analysis of a Dof protein gene(PtDof1) in Populus

Both cDNA and DNA clones of PtDof1 （GenBank Accession No. FJ402844 and FJ402845） were isolated from plants grown in tissue culture ofPopulus tornentosa. The DNA sequence is 1597 bp including two exons and one intron. The cDNA is 969 bp in length with a 765 bp open reading frame which is capable of encoding 255 amino acids. The deduced amino acids sequence of the PtDofl protein shares 65%, 56% and 55% identity with Vitis vinifera （CAO48618）, Nicotiana tabacum （CAA08755） and Glycine max （ABI 16022） Dof protein by blast analysis in GenBank. Phylogenic analysis suggests PtDof1 gene could belong to the Dofgene family. PtDofl protein contains an unusual conserved single zinc finger with the pattern of C-X2-C-X21-C-X2-C, which may play a functional role in tissue-specific expression and possibly the auxin response of endogenous plant genes.

Analyses of the Sequence and Structure of the BnNIP7;1 Gene in Brassica napus

Rapeseed,the largest oil crop in China,is a boron-loving plant.Boron has an important influence on the growth and development of rapeseed.In this study,four different boron-efficient Brassica napus varieties were used as the material to analyze the structure and sequence of BnNIP7;1 gene,aiming to provide a basis for studying the role of BnNIP7;1 in the absorption and transport of boric acid in rapeseed.The results showed that BnNIP7;1 exists in multiple copies of the Brassica napus genome,consisting of five exons and four introns.There are base insertions,base deletions and more base substitutions in introns.However,there is no base insertion or base deletion in the exon,only a small number of base substitution mutations are present,bringing about three amino acid changes in the encoded protein of BnNIP7;1-ZS9b in Zhongshuang 9.The BnNIP7;1-HY7b of Huayou 7 has an ochre mutation that will lead to a protein of only 47 amino acids,thus losing its function.Bioinformatics indicates that BnNIP7;1 protein is a membrane protein with six transmembrane regions,indicating that it is involved in the absorption and transport of boric acid.

Characterizations of Chinese isolates of Coxiella burnetii in the com1 gene sequence

Objective: To know some genetical characterizations of Coxiella burnetii Chinese isolates by comparing the coml gene sequence. Methods: com1 gene sequences of Chinese isolates were amplified, se-quenced, and analyzed by comparing our result and the previous published data. Results: Three different com1 sequences were identified in 7 Chinese isolates. Sequence comparison indicated that the isolates harboring the QpRS plasmid could be defined as a new group and, in addition, the isolates carrying the same plas-mid type showed similar com1 gene sequence. Conclusion: Study suggests that the classification of the group based on the coml gene sequence is highly associated with the plasmid type of the isolates and, however, little related to disease forms and geographical origins of the isolates.

Cloning and Sequence Analysis of Envelope Glycoprotein E1 Gene of Rubella Virus, JR23 Strain

To construct an expression vector containing the E1 glycoprotein gene of rubella virus for the study on the effect of mutation of the E1 gene glycoprotein and the analysis of phylogenetic differences of sequences, the gene encoding the E1 envelope glycoprotein was amplified from rubella virus, Jinan strain JR23, by RT-PCR and ligated into PMD-18T vector. The clones that carried the E1 gene were identified after amp r selection and analysis of restriction enzyme digestion. After sequencing this gene was analyzed by Danstar and Winstar programs, and the map of phylogenetic tree was drawn. The clone of E1 glycoprotein was thus constructed. It was found that the sequence differences between JR23 strain and the TCRB strain from Japan and those between JR23 strain and Thomas strain of England were rather small with difference values of 0.9% and 1.2% respectively. Yet those between JR23 strain and BRD2 strain from Beijing and those between JR23 strain and XG379 strain from Hong Kong were comparatively larger with difference values of 7.6% and 7.3% respectively. The sequence of JR23 strain with other strains was less than 3% except the NC strain (3.7%). It concludes that the construction of E1 glycoprotein gene offers an approach to study the relationship between structures and functions of E1 gene and its gene products. In the phylogenetic tree, it shows that there are significant differences in the sequences of rubella virus isolated in China, and this might be helpful to develop an effective subunit vaccine.

Proteolipid protein 1 gene sequencing of hereditary spastic paraplegia

PCR amplification and sequencing of whole blood DNA from an individual with hereditary spastic paraplegia, as well as family members, revealed a fragment of proteolipid protein 1 （PLP1） gene exon 1, which excluded the possibility of isomer 1 expression for this family. The fragment sequence of exon 3 and exon 5 was consistent with the proteolipid protein 1 sequence at NCBI. In the proband samples, a PLP1 point mutation in exon 4 was detected at the basic group of position 844, T→C, phenylalanine→leucine. In proband samples from a male cousin, the basic group at position 844 was C, but gene sequencing signals revealed mixed signals of T and C, indicating possible mutation at this locus. Results demonstrated that changes in PLP1 exon 4 amino acids were associated with onset of hereditary spastic paraplegia.

日本鳗鲡TBK-1基因的克隆及免疫刺激的表达模式

利用PCR技术克隆了日本鳗鲡(Anguilla japonica)TBK-1基因(AjTBK-1),其开放阅读框为2193 bp,编码731个氨基酸。序列结构分析结果显示,AjTBK-1含有4个保守结构域,分别为氨基端激酶结构域,泛素样结构域和羧基端两个卷曲-螺旋结构域。系统发育分析表明,鱼类与四足类的TBK-1各自聚为一枝。实时定量PCR(qPCR)结果显示,AjTBK-1在日本鳗鲡各组织中均有表达。Poly I:C刺激6 h后,日本鳗鲡脾脏组织中AjTBK-1的上调倍数最高,为对照组的1.63倍;迟缓爱德华氏菌(Edwardsiella tarda)感染24 h后,日本鳗鲡肝脏组织中AjTBK-1的上调倍数最高,为对照组的2.2倍:表明AjTBK-1参与了日本鳗鲡抗病毒、抗细菌免疫反应应答。

镉胁迫下中辽1号杨转录组分析

【目的】分析镉胁迫下中辽1号杨的转录水平,筛选差异表达基因,挖掘与镉胁迫相关的功能基因,为深入探索中辽1号杨对镉胁迫响应的分子机制提供理论依据。【方法】以中辽1号杨为试验材料,采用盆栽试验的方法将其扦插于土壤Cd含量为20 mg/kg(M20)的花盆中,以不加Cd为对照(CK),80 d后采集不同处理中辽1号杨叶片进行转录组测序,使用DESeq2软件筛选CK与M20处理中辽1号杨的差异表达基因,将得到的差异基因在基因本体数据库(gene ontology,GO)、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)、蛋白相邻类聚簇数据库(clusters of orthologous groups of proteins,COG)中进行注释,分析差异表达基因在不同数据库中的注释信息。随机挑选6个差异表达基因(超氧化物歧化酶(SOD)、丝氨酸/苏氨酸蛋白激酶(STK)、脱落酸(ABA)、T复合蛋白1(TCP1)、MYB基因(MYB)、丝裂原活蛋白激酶12(MAPK12)),利用Primer 5软件设计特异性引物进行实时荧光定量PCR(RT-qPCR)试验,检测这6个基因表达水平的变化,验证转录组结果的准确性。【结果】在CK和M20处理的中辽1号杨中,共发现3812个差异表达基因,其中上调表达的有2209个,下调表达的有1603个。GO分析发现,差异表达基因注释到3大类49个功能组中,其中与代谢过程有关基因最多。转录组KEGG代谢通路分析发现,差异表达基因主要在信号转导途径、代谢途径和生物合成途径富集。COG分析发现,1455个差异基因被注释到22种分类中,其中一般功能预测基因注释最多,其次是信号转导机制基因。结合各数据库分析结果发现,Cd胁迫下中辽1号杨转录组的注释结果中代谢过程和信号转导过程相关基因较多。镉胁迫下差异表达基因最多的家族是ABC、MYB、WRKY、bHLH和NAC基因家族,挖掘出与镉胁迫相关基因WRKY家族基因47(WRKY47)、硝酸盐转运蛋白基因(NRT)、ABC家族转运蛋白基因2(ABC2)、苹果酸脱氢酶(MDH)、谷胱甘肽S-转移酶基因(GSTs)、NAC家族基因2(NAC2)的表达量显著上调,MYB家族基因44(MYB44)、重金属相关异戊二烯化植物蛋白基因39(HIPP39)的表达量显著下调。RT-qPCR结果显示,随机挑选6个差异表达基因表达量的变化趋势与转录组测序结果一致。【结论】镉胁迫后中辽1号杨转录组的差异表达基因主要富集在代谢过程和信号转导过程,挖掘出与镉胁迫相关基因,分别是WRKY47、NRT、ABC2、MDH、GSTs、NAC2、MYB44、HIPP39,为深入探索杨树对镉胁迫响应的分子机制提供了理论依据。

Mental retardation,seizures and language delay caused by new SETD1B mutations:Three case reports

BACKGROUND The SETD1B gene is instrumental in human intelligence and nerve development.Mutations in the SETD1B gene have been linked in recent studies to neurodevelopmental disorders,seizures,and language delay.CASE SUMMARY This study aimed to analyze the clinical manifestations and treatment of three patients suffering from mental retardation,epilepsy,and language delay resulting from a new mutation in the SETD1B gene.Three individuals with these symptoms were selected,and their clinical symptoms,gene test results,and treatment were analyzed.This article discusses the impact of the SETD1B gene mutation on patients and outlines the treatment approach.Among the three patients(two females and one male,aged 8,4,and 1,respectively),all exhibited psychomotor retardation,attention deficit,and hyperactivity disorder,and two had epilepsy.Antiepileptic treatment with sodium tripolyvalproate halted the seizures in the affected child,although mental development remained somewhat delayed.Whole exome sequencing revealed new mutations in the SETD1B gene for all patients,specifically with c.5473C>T(p.Arg1825trp),c.4120C>T(p.Gln1374*,593),c.14_15insC(p.His5Hisfs*33).CONCLUSION Possessing the SETD1B gene mutation may cause mental retardation accompanied by seizures and language delay.Although the exact mechanism is not fully understood,interventions such as drug therapy,rehabilitation training,and family support can assist patients in managing their symptoms and enhancing their quality of life.Furthermore,genetic testing supplies healthcare providers with more precise diagnostic and therapeutic guidance,informs families about genetic disease risks,and contributes to understanding disease pathogenesis and drug research and development.

On four closely related hypotrichous ciliates(Protozoa,Ciliophora,Spirotrichea):molecular characters,interspecific relationships and phylogeny defined with multigene sequence information

In order to clarify the phylogeny and relationships of the most confused hypotrichous ciliates,Holosticha-complex,four closely related holostichids（five populations）,Holosticha bradburyae,H.diademata,Anteholosticha sp.,and A.manca,were compared and analyzed using ITS2 secondary structures,ITS1-5.8S-ITS2 region and SSrRNA gene sequences.The ITS1-5.8S-ITS2 region sequences of these four species were first sequenced,and they shared sequence identities ranging from 68.0% to 90.1%,while two populations of Anteholosticha sp.differed in three nucleotides（sequence identity 99.8%）.There were several minor differences among ITS2 secondary structures of these species,while two populations of Anteholosticha sp.had the identical secondary structure.Phylogenetic trees inferred from the ITS1-5.8S-ITS2 region sequences of stichotrichs using multiple algorithms（Neighbor-Joining,Maximum Parsimony and Bayesian） revealed similar topologies.The results show that：（1） Holosticha bradburyae and H.diademata firmly clustered together with strong bootstrap supports,forming a sister clade with Anteholosticha sp.,（2） Anteholosticha appeared to be a paraphyletic assemblage,in which the morphotype A.manca was more closely related to Diaxonella trimarginata than to its congener Anteholosticha sp.Phylogenetic analyses based on the SSrRNA gene and the combined sequences of SSrRNA gene and ITS1-5.8S-ITS2 region revealed the similar relationships between Holosticha and Anteholosticha,nevertheless their positions within the subclass Stichotrichia differed from each other inferred from different genes.

水牛SFRP 1基因序列分析、真核表达载体构建及组织表达分析

【目的】获取水牛分泌型卷曲相关蛋白1(SFRP1)基因CDS区序列,并预测其编码蛋白的结构功能,构建SFRP 1基因真核表达载体,检测SFRP 1基因在水牛不同组织中的表达情况,为探索SFRP 1基因在水牛生长发育中的作用奠定基础。【方法】以水牛卵巢组织cDNA为模板,通过RT-PCR对SFRP 1基因CDS区序列进行扩增并测序,利用生物信息学在线分析软件对水牛SFRP 1基因与不同物种进行比对和系统进化树构建,并预测SFRP1蛋白理化性质、信号肽、跨膜结构等。将获得的目的基因连接至pCMV-HAhyPBase-mcheery载体并转染至水牛颗粒细胞,检测转染后荧光和基因表达情况。通过实时荧光定量PCR检测水牛不同组织中SFRP 1基因表达情况。【结果】水牛SFRP 1基因CDS区长927 bp,共编码308个氨基酸。多重序列比对结果显示,水牛SFRP 1基因氨基酸序列与黄牛、牦牛、山羊、虎鲸、黑猩猩、北极狐、猫、人、小鼠的相似性分别为100%、100%、99.65%、98.58%、98.23%、98.23%、98.58%、98.23%、96.45%,存在CRD_FZ、NTR_like和DUF3367结构域。水牛SFRP 1基因核苷酸序列与黄牛、绵羊、山羊、牦牛、野牛、马鹿、野骆驼、猪、人的相似性分别为97.3%、95.9%、95.8%、94.5%、94.2%、93.7%、80.4%、78.5%和77.3%。系统进化树结果显示,水牛与黄牛、牦牛、野牛聚为一支。生物信息学分析结果显示,SFRP1蛋白呈碱性,为不稳定蛋白;第1—15位氨基酸处存在信号肽,为分泌型蛋白,存在跨膜结构,主要定位于细胞外;存在21个磷酸化位点和8个O-糖基化修饰位点。二级结构主要由α-螺旋、延伸链和无规则卷曲构成;水牛、黄牛、人的SFRP1蛋白三级结构高度相似。成功构建pCMV-mcheery-SFRP1真核表达载体并转染水牛颗粒细胞,转染72 h后pCMV-mcheery-SFRP1重组质粒组细胞内SFRP 1基因表达量极显著高于对照组(P<0.01)。实时荧光定量PCR结果显示,SFRP 1基因在水牛不同组织中均有表达,且在脾脏中表达量最高,极显著高于其他组织(P<0.01)。【结论】SFRP 1基因在不同物种以及遗传进化过程中具有高保守性,在水牛不同组织中广泛表达。研究结果为今后探究SFRP 1基因在水牛生长发育中的功能及分子机制奠定基础。

Antigen Gene Cloning and Expression of HIV-1 Toward an AIDS Vaccine Design Ⅰ.Amplification and Sequencing of HIV-1 Antigen Genes

Objective:To amplify antigen genes from patients with human immunodeficiency virus type 1 (HIV-1) in Guangdong Province for candidate AIDS vaccine design. Methods:Viral nucleic acid was isolated from 10 HIV-1 infected individuals' peripheral blood collected during 1995-2000 in Guangdong Province. The viral gag p24 gene and env gp120 gene were amplified by nested-PCR and sequenced. The homologies among HIV-1 isolates were compared with HIV-BLAST. Results: Among 10 HIV-1 isolates, nine are homologous to viruses of subtype B, and one is homologous to viruses of subtype E. Conclusion: Subtype B viruses of HIV-1 are predominantly present in Guangdong Province.

长链非编码RNA CASC15对肝细胞癌增殖、侵袭和迁移的影响及作用机制

目的探究长链非编码RNA(lncRNA)癌症易感性基因15(Cancer susceptibility candidate 15,CASC15)在肝细胞癌(Hepatocellular carcinoma,HCC)中的分子调控机制。方法通过生物信息学方法预测目的基因表达,并分析目的基因表达与患者生存时间的关系;收集临床HCC患者肝癌组织和癌旁组织;CCK-8、Transwell和流式细胞术实验检测HCC细胞SMMC7721和Huh-7的增殖、侵袭、迁移以及凋亡;双荧光素酶实验检测miR-144-3p与CASC15和富含亮氨酸的重复序列蛋白1(Leucine rich repeat containing protein 1,LRRC1)的靶向关系;qRT-PCR和Western blot检测mRNA和蛋白表达情况;免疫荧光实验用于蛋白定位研究;回复实验验证CASC15/miR-144-3p/LRRC1对HCC进展的影响。体内实验验证CASC15对HCC进展的影响。结果TCGA数据库与qRT-PCR检测显示HCC组织和细胞中CASC15高表达、miR-144-3p低表达、LRRC1高表达(P<0.05)。增殖、侵袭、迁移的细胞功能实验结果表明在肿瘤发展中CASC15和LRRC1起到促进作用,miR-144-3p则是抑制作用,与凋亡实验结果一致(P<0.05)。细胞功能实验表明CASC15抑制miR-144-3p功能,miR-144-3p抑制LRRC1,CASC15与miR-144-3p结合,并导致LRRC1上调。回复实验结果表明CASC15通过抑制miR-144-3p促进LRRC1的表达从而促进HCC细胞增殖、侵袭和迁移并抑制细胞凋亡。结论CASC15可能通过调节miR-144-3p/LRRC1轴,从而促进HCC进展。

应用16S rRNA基因测序比较分析不同品系1型糖尿病小鼠肠道菌群的异同

目的比较C57BL/6(B6)和FVB两个品系小鼠建立的1型糖尿病(T1DM)模型肠道菌群组成和多样性的异同,为探索两模型在T1DM相关肠道菌群研究中的进一步应用提供背景数据。方法采用8周龄SPF级雄性B6和FVB小鼠各20只,两组小鼠每天腹腔注射40 mg/kg的链脲佐菌素(STZ)连续5 d,小鼠空腹血糖连续2周≥11.1 mmol/L为T1DM建模成功,建模6周后每组分别收集10份洁净粪便,进行16S rRNA基因V3-V4区测序,分析粪便中肠道菌群的Alpha多样性、Beta多样性、优势菌科及肠道菌群相关的功能通路。结果B6与FVB组T1DM小鼠肠道菌群的Alpha多样性和丰富度没有显著差异(P>0.05),两组小鼠的菌群Beta多样性存在统计学上的差异(P<0.05)。B6组T1DM小鼠肠道的优势菌科为Muribaculaceae、Lactobacillaceae、Lachnospiraceae、Prevotellaceae、Desulfovibrionaceae、Akkermansiaceae;FVB组T1DM小鼠肠道的优势菌科为Lactobacillaceae、Muribaculaceae、Lachnospiraceae、Prevotellaceae、Clostridiales_unclassified、Saccharimonadaceae、Marinifilaceae;两组中共有的优势菌科在比例上差异很大。B6与FVB组的厚壁菌门与拟杆菌门的比值都显著增加。两组T1DM小鼠肠道菌群的功能通路集中在以氨基酸、糖代谢及核苷酸的代谢途径中。结论两组T1DM小鼠肠道菌群Alpha多样性无差异,但肠道菌群的组成及比例差异较大,优势菌群在不同模型中的组成比例发生了改变。

Updated Genetic Testing of Primary Hyperoxaluria Type 1 in a Chinese Population:Results from a Single Center Study and a Systematic Review

Primary hyperoxaluria type 1(PH1)is a rare but devastating autosomal recessive inherited disease caused by mutations in gene AGXT.Pathogenic mutations of AGXT were mostly reported in Caucasian but infrequently in Asian,especially in Chinese.To update the genotypes of PH1 in the Chinese population,we collected and identified 7 Chinese probands with PH1 from 2013 to 2017 in our center,five of whom had delayed diagnosis and failed in kidney transplantation.Samples of peripheral blood DNA from the 7 patients and their family members were collected and sequencing analysis was performed to test the mutations of gene AGXT.Western blotting and enzyme activity analysis were conducted to evaluate the function of the mutations.Furthermore,a systematic review from 1998 to 2017 was performed to observe the genetic characteristics between Chinese and Caucasian. The results showed that a total of 12 mutations were identified in the 7 pedigrees.To the best of ourknowledge,2 novel variants of A GXT,p.Gly41 Trp and p.Leu33Met,were first reported.Bioinformatics and functional analysis showed that only 7 mutations led to a reduced expression of alanine-glyoxylate amino transferase (AGT)at a protein level.The systematic review revealed significant population heterogeneity in PH1.In conclusion,new genetic subtypes and genetic characteristics of PH1 are updated in the Chinese population. Furthermore,a genotype-phenotype correlation is found in PH1.

A POTENTIAL TUMOR SUPPRESSOR GENE: Doc-1R

Objective: To detect the expression and the genomic sequence of Doc-1R gene in mice. Methods: The gene specific primers were designed and synthesized according to the cDNA sequence of Doc-1R gene. The sequence of Doc-1R gene was cloned by nested PCR. The expression of Doc-1R gene was examined by RT-PCR in thirteen kinds of tissues of mice. Results: The mouse Doc-1R gene has been obtained by two times genomic walking, which spans 2787 bp and contains four exons and three introns. All of the splice donor/acceptor site sequences are in accordance with the consensus 揋T-AG?rule. There was expression of Doc-1R gene in the thirteen tissues. Conclusion: The mouse Doc-1R gene was cloned successfully. The expression pattern suggests that Doc-1R gene is a housekeeping gene, which is important to keep the function of tissues and organs.