Abstract Background Inherited antithrombin deficiency (ATD), a rare autosomal dominant disorder due to SERPINC1 gene mutations, is the most severe inherited thrombophilia. Limited literature exists that focuses on ATD and its mutations in the Chinese population. This study aimed to characterize SERPINC1 gene mutations in a Chinese cohort and to explore their relationship with thrombophilia. Methods Coagulation screening results and clinical data were meticulously collected from 23 unrelated probands with ATD and their family members. Genomic DNA was extracted and subjected to PCR amplification and direct sequencing. Putative mutations were analyzed using in silico bioinformatic tools. Mutant antithrombin (AT) proteins were expressed in HEK293 cells, and ELISA was used to detect wild-type and mutant AT. RT-qPCR was used to measure AT mRNA expression in transfected cells. Results Among the 23 probands, 15 (65.2%) exhibited concurrent reductions in both AT: A and AT: Ag (type I defects), while the remaining 8 (34.8%) had normal AT: Ag levels (type II defects). Genetic analysis revealed a spectrum of 21 distinct mutations across 87.0% (20/23) of the probands. Most were point mutations predicted to be deleterious and were primarily located in exons 5 and 3. Among the 20 mutation carriers, 15 (75%) were heterozygous and most of them experienced thrombosis with identifiable triggers. The other 5 (25%) were compound heterozygous and primarily presented with spontaneous thrombosis. Notably, the missense mutations c.1346T > A and c.442T > C were recurrent. These mutations exhibited high heterogeneity, with no ethnic-specific mutations observed. In vitro expression confirmed that synthesis and/or secretion defects in the mutant proteins are the primary mechanism underlying the antithrombin deficiency. Conclusions SERPINC1 gene analysis benefits asymptomatic family members, especially child-bearing women, by informing venous thromboembolism prevention strategies and guiding anticoagulant choice in cases involving heparin-binding site mutations. This underscores the essential role of genetic diagnosis in ATD management.

Analyze the clinical phenotype and gene mutations of a family with hereditary FXII deficiency, and preliminarily explore its phenotypic manifestations. The routine coagulation indicators and related coagulation factors were measured.Thromboelastography and thrombin generation tests simulated coagulation and anticoagulation states in vitro and in vivo. PCR direct sequencing was utilized to analyze all exons and flanking sequences of the F12 gene in the proband, confirming suspected mutations through reverse sequencing, and identifying corresponding mutation sites in family members. Using ClustalX-2.1-win to analyze the conservation of the variant, and employing online software to predict the pathogenicity of mutations. The proband exhibited significantly prolonged APTT (169.1 s) and a pronounced decrease in FXII: C to 1.0%. Thromboelastography testing indicated a diminished function of the endogenous coagulation system, while thrombin generation testing revealed a normal ability for thrombin production in the proband. Gene sequencing revealed that the proband harbored a deletion mutation c.303_304delCA in exon 5 and a substitution mutation c.800 + 1G > A in intron 8. All three bioinformatics software indicated that the mutations were pathogenic and could lead to the production of a terminator, potentially altering the structure and function of the protein. The deletion mutation c.303_304delCA and substitution mutation c.800 + 1G > A are associated with a decreased in FXII levels in this family, with the c.800 + 1G > A mutation being the first reported mutation worldwide.

OBJECTIVE:To investigate the molecular pathogenic mechanisms of a family with hereditary factor Ⅶ (FⅦ) deficiency. METHODS:A family (3 generations, 12 members) with hereditary FⅦ deficiency, in which the proband presented with menorrhagia and was admitted to the First Affiliated Hospital of Wenzhou Medical University in April 2023, was selected as the study subject. Clinical data of the family members were collected. Peripheral venous blood samples were collected from all 12 members for routine coagulation tests and genomic DNA extraction. All exons and flanking sequences of the F7 gene were amplified by PCR and analyzed by Sanger sequencing. Thrombin generation assay was performed to evaluate the coagulation potential of the proband and her parents. Multiple online bioinformatics software tools were used to analyze the conservation and pathogenicity of candidate variants identified in the proband. The pathogenicity of variant was classified according to the Standards and Guidelines for the Interpretation of Sequence Variants released by American College of Medical Genetics and Genomics (ACMG) (hereinafter referred to as ACMG guidelines). Homology modeling of the variant FⅦ protein was performed using homology modeling (SWISS-MODEL). Amino acid sequence alignment between wild-type and variant FⅦ proteins was conducted using MEGA v7, and spatial conformational differences were analyzed using PyMOL to assess the potential impact of the F7 gene variants on the structure and function of the FⅦ protein. This study was approved by the Ethics Committee of the First Affiliated Hospital of Wenzhou Medical University (Ethics No.: KY2022-R193). RESULTS:Coagulation tests showed that the proband's prothrombin time (PT) was significantly prolonged to 33.1 s, and both factor Ⅶ activity (FⅦ:C) and antigen (FⅦ:Ag) levels were reduced to 2%. Her parents, eldest sister, second sister, younger brother, and four children all showed mildly prolonged PT, with FⅦ:C and FⅦ:Ag levels approximately 50% of normal. Genetic sequencing identified compound heterozygous variants in the F7 gene of the proband: a heterozygous missense variant c.722C>A (p.Thr241Asn) in exon 7, and a heterozygous deletion variant c.1261_1261delA (p.Ile421Ser*fs75) in exon 8. Retrieval from domestic and international databases found no previous reports of the latter variant, suggesting it is novel. Familial co-segregation analysis confirmed that these variants were inherited from her father and mother, respectively. The thrombin generation assay demonstrated that the proband had a significantly decreased peak thrombin height (peak ratio: 29.5%), significantly increased thrombin lag time ratio and time-to-peak ratio (3.03 and 2.93, respectively), but only a mildly decreased endogenous thrombin potential (ETP) ratio of 90.7%. Online bioinformatics analysis indicated that threonine-241 (p.Thr241) in the FⅦ protein was not conserved, while isoleucine-421 (p.Ile421) was highly conserved. Both the p.Thr241Asn and p.Ile421Serfs*75 variant sites in the proband's F7 gene were predicted to be pathogenic. According to the ACMG guidelines, the p.Thr241Asn (PM3+PP1+PP3+PP4+PP5) and p.Ile421Ser*fs75 (PM2+PM4 +PP1+PP3+PP4) variants were both classified as "likely pathogenic". Structural analysis of the FⅦ protein indicated that the p.Ile421Ser*fs75 frameshift variant led to the substitution of Cysteine-428 by Alanine, preventing the formation of a critical disulfide bond between amino acid residues 400 and 428 present in the wild-type FVII protein. CONCLUSION:The compound heterozygous variants p.Thr241Asn and p.Ile421Ser*fs75 in the F7 gene are likely the genetic etiology responsible for the reduced FⅦ levels in this hereditary FⅦ deficiency family.

Abstract Background Factor XII (FXII or F12) deficiency is a rare inherited disorder, typically lacking haemorrhagic symptoms. There is limited literature exists on FXII deficiency and mutations within the Chinese population. This study aimed to characterize the spectrum of F12 gene mutations in a Chinese cohort and to investigate the relationship between FXII mutations and clinical phenotypes. Methods Genetic and clinical data from 51 unrelated probands with FXII deficiency, along with their families, were meticulously collected and analysed. Results Genetic analysis revealed that 94.1% of probands carried genetic defects, with 29 mutations pinpointed in the F12 gene. Of these, 18 mutations were previously reported for the first time by our research group, including c.303_304delCA, c.1078G > A, c.1285 C > T, among others. Of the mutations, 17 are missense, constituting 58.6% of the total. Additionally, 11 are deletions or insertions, of which 8 result in frameshifts, while the remaining one is a nonsense mutation. These mutations were predominantly concentrated in two crucial regions: the catalytic domain and the kringle domain. The most frequently observed mutations were c.1681G > A, closely followed by c.1561G > A and c.1078G > A, indicating a dominance among these mutations. Additionally, a prevalent polymorphism at position 46 was observed in the majority of probands, with 47.1% having the 46T/T genotype and 13.7% having the 46 C/T genotype, which may potentially impact FXII activity. The broad spectrum of asymptomatic FXII deficiency observed within the Han population of East China. Conclusions We speculate on the potential impact of recurrent mutations on the efficacy of new drugs being developed to target FXII for thrombosis prevention and treatment. Furthermore, it is important to explore their influence on FXII-related pathways beyond the activation of the contact pathway in the coagulation cascade.

蛋白C是一种由肝细胞合成和分泌的维生素K依赖性丝氨酸蛋白酶原[1], 是机体重要的生理性抗凝物质之一。蛋白C活化后能通过水解活化凝血因子Ⅴ(FⅤa)和活化凝血因子Ⅷ(FⅧa)来抑制凝血酶的产生。遗传性蛋白C缺陷症是由编码蛋白C的基因(PROC)突变引起的导致常染色体显性遗传疾病, 纯合或复合杂合突变导致的严重蛋白C缺陷症极其罕见, 发病率约为1/400万[2], 临床可表现为出生后不久发生的新生儿暴发性紫癜;单杂合型遗传性蛋白C缺陷症的发病率在普通人群中为0.14%～0.50%[3], 此类患者静脉血栓形成的风险可增加5～7倍。截至2021年4月, 人类基因组突变数据库(HGMD)共列出391种PROC基因突变, 其中错义/无义突变290种, 小部分缺失突变30种。我们近期收治2例遗传性蛋白C缺陷症患者, 并对其进行家系调查和基因突变分析。

目的 探讨一个遗传性凝血因子X(FX)缺陷症家系的临床特征与基因突变关系.方法 调查家系,采用Sanger测序法分析先证者F10基因8个外显子及其侧翼序列,并对家系成员相同突变位点进行检测.通过凝血酶生成试验评估家系成员FX的促凝活性.ClustalX-2.1-win和在线生物信息学软件(PolyPhen-2、SIFT和Mutation Taster)分别用于分析突变位点氨基酸的保守性和致病性;Swiss-Pdb Viewer软件用于蛋白质空间结构和突变氨基酸相互作用分析.结果 先证者FX活性(FX:C)和FX抗原(FX:Ag)分别降低至7％和14％(参考范围分别为:90％～114％和90％～110％);先证者平素无自发性出血现象,外伤后也无明显出血异常.基因测序发现先证者F10基因第4号外显子存在c.361T＞C(p.Cys121Arg)杂合错义突变及第8号外显子存在c.979C＞T(p.Arg327Trp)杂合错义突变;其母亲和女儿为p.Arg327Trp的杂合子,二姐和儿子为p.Cys121Arg的杂合子.先证者凝血酶生成潜力比值和峰高比值下降,而其延迟时间和达峰时间延长.保守性分析显示,p.Arg327和p.Cys121位点在同源物种间均高度保守.生物信息学软件预测两种突变均为致病突变.突变蛋白模型分析显示,p.Arg327Trp突变后,导致蛋白质空间结构和稳定性发生改变.结论 该先证者F10基因p.Arg327Trp和p.Cys121Arg复合杂合突变与其FX水平减低有关;该复合杂合突变与临床出血症状相关性不明显.

Objective:To analyze the phenotype and genotype of two propositi with inherited hypodysfibrinogenaemia caused by compound heterozygous mutations, and investigate the molecular mechanism.Metheds:Two propositi and their family members(7 person in 3 generations and 10 person in 3 generations,respectively) were investigated. The activity of plasma fibrinogen (Fg:C) and thrombin time (TT) were analyzed by coagulation method, the antigen of plasma fibrinogen (Fg:Ag) was detected by immunoturbidimetry. All of the exons and flanking sequences of FGA,FGB,FGG of two propositi were amplified by PCR, followed by direct sequencing. The ClustalX-2, 1-win software was used to analyze the conservatism of mutated gene locus. PROVEAN and Mutation Taster were applied to analyze the pathogenicity of mutated amino acid. The changes of the protein spatial structure and intermolecular interaction were analyzed by Pymol.Results:Fg:C and Fg:Ag of proposita A and B were both significantly decreased (0.74 and 0.78 g/L, 0.96 and 0.94 g/L, respectively). Gene analysis revealed that proposita A and B both carried a heterozygous mutation c.2185G>A(p.AαGlu710Lys) in exon 6 of FGA. Furthermore, proposita A also carried a heterozygous mutation c.701G>T(p.γTrp208Leu) in exon 7 of FGG, and proposita B carried a heterozygous mutation c.1015A>C(p.γSer313Arg) in exon 8 of FGG. Phylogenetic analysis suggested that p.AαGlu710,p.γTrp208 and p.γSer313 were highly conserved among homologous species. All variants were predicted to be deleterious by two online bioinformatic softwares. The protein model analysis indicated that protein spatial structure and intermolecular hydrogen bonds were changed by these variants, which destroyed the stability of Fg.Conclusion:The compound heterozygous mutations of p.AαGlu710Lys and p.γTrp208Leu,p.AαGlu710Lys and p.γSer313Arg might account for the hypodysfibrinogenemia in two propositi.

目的 分析1个遗传性凝血因子Ⅻ缺乏症家系表型及基因突变情况,探讨其分子致病机制.方法 对先证者及其家系进行活化部分凝血活酶时间(APTT)、凝血酶原时间(PT)、纤维蛋白原(Fib)、凝血因子Ⅻ活性(FⅫ:C)和凝血因子Ⅻ抗原(FⅫ:Ag)测定.用DNA直接测序法进行FⅫ基因突变检测.采用ClustalX-2.1-win软件分析突变氨基酸的保守性,用4个生物信息学评分软件分析突变对蛋白质功能的影响.结果 先证者APTT结果明显延长,达179.2 s,先证者父亲、长子及女儿APTT略延长.先证者FⅫ:C及FⅫ:Ag明显减低,均为1％;其父亲、长子、女儿及次子FⅫ:C及FⅫ:Ag减低,约为50％.基因测序发现先证者FⅫ基因存在复合杂合突变,包括第1外显子启动子区为46T/T型,第9外显子存在c.797G>A(Cys247Tyr)及第14外显子存在c.1681G>A(Gly542Ser).先证者父亲及长子表现为c.797G>A杂合突变,女儿及次子表现为c.1681G>A杂合突变.ClustalX-2.1-win软件保守性分析表明Cys247和Gly542在同源物种间高度保守.生物信息学评分软件分析结果表明突变可能影响蛋白质的功能.结论 先证者FⅫ基因46T/T,c.797G>A和c.1681G>A复合杂合变异导致其凝血因子Ⅻ水平极度减低.c.797G>A为首次报道,丰富了人类基因突变数据库(HGMD).

To analyse F11 gene mutations in a Chinese pedigree with hereditary factor XI (FXI) deficiency and investigate the molecular mechanism. The plasma FXI activity and FXI antigen of the proband and the family members were detected by clotting assay and ELISA, respectively. The F11 gene was amplified by PCR and sequenced directly. Online bioinformatics software were needed to analyse the mutations. The proband showed a prolonged activated partial thromboplastin time (93.3 s), whose FXI activity and FXI antigen were low to 2, 4.5%, respectively. A novel mutation c.233T>C (p.Leu60Pro) in exon 4 and a previously described mutation c.1253G>T (Gly400Val) were found in the proband. Protein Leu60 is conserved highly among homologous species. Bioinformatics software indicated that Leu60Pro mutation might affect the protein function. Other coagulation abnormalities were not found. We preliminarily considered the mutations Leu60Pro and Gly400Val were responsible for the decrease FXI level in the family. Leu60Pro mutation in the F11 gene has not been described elsewhere.

OBJECTIVE:To explore the molecular pathogenesis for a pedigree affected with hereditary coagulation factor XII (FXII) deficiency.METHODS:Potential variant of the F12 gene was analyzed by PCR and Sanger sequencing. Expression plasmids were constructed by site-directed mutagenesis based on the wild-type and transiently transfected into 293T cells. FXII:C and FXII:Ag of the expression products were determined in the supernatant and cell lysate. Western blotting was used to verify the identify of the protein.RESULTS:Gene sequencing revealed that the proband has carried 46TT genetype and heterozygous p.Glu502Lys variants in exon 13, and a heterozygous p.Gly542Ser variant in exon 14 of the F12 gene. Transfection experiment suggested that the FXII:C and FXII:Ag of p.Glu502Lys variant in the supernatant were 28% and 24%, compared with the wild-type (100%) and FXII:Ag of cell lysates was 39% compared to the wild-type (100%). The FXII:C and FXII:Ag of p. Gly542Ser variant in the supernatant were 32% and 17% and the FXII:Ag of cell lysates was 59%.CONCLUSION:The 46TT genetype, p.Glu502Lys and p.Gly542Ser variants of the F12 gene probably underlie the low FXII level in the proband. As shown by in vitro experiment, the p.Glu502Lys and p.Gly542Ser variants can both inhibit the synthesis and secrection of the FXII protein.

Coagulation factor XII deficiency is a rare autosomal recessive disorder, which could be found in a consanguineous family. We studied a Chinese family in which the activated partial thromboplastin time (APTT) of the proband had clearly prolonged up to 101.7 s, associated with low FXII activity of 3% and FXII antigen < 1%. To analyze the gene mutation in this FXII-deficient patient, we performed FXII mutation screening, and analyzed the DNA sequence of the F12 gene. A ClustalX-2.1-win and four online bioinformatics software services were used to study the conservatism and effects of the mutation. A transient in vitro expression study was performed to elucidate the possible pathological mechanism. Sequence analysis revealed a homozygous c.1681 G > A point mutation in exon 14, causing a novel Gly542Ser mutation in the catalytic domain. The results of the conservatism and bioinformatics analyses both indicated that the mutation likely affects the function of the protein. Additional expression studies in COS-7 cells showed that the antigen level of mutant FXII (FXII-Gly542Ser) was lower than wild type in culture medium, whereas the corresponding level of FXII antigen in cell lysates was equivalent. These results suggest that the Gly542Ser mutation causes FXII deficiency through intracellular degradation.

Objective To investigate the expression of related molecules of lymphotoxin β receptor(LTβR) signal pathway in join fluid of patients with rheumatoid arthritis(RA),to undretand the relevance of LTβR signal pathways with rheumatoid arthritis.Methods The real-time fluorescent quantitative PCR(Q-RT-PCR) technology was used to detect LTβR mRNA,LTα mRNA,LTβ mRNA,p65 mRNA and RelB mRNA expression in articular cavity effusion specimens with rheumatoid arthritis (RA,22 cases) patients and osteoarthritis(OA,25 cases) patients.The expression differences of LTβR signal pathway related molecules with RA and OA patients were analyzed.Results The LTβ mRNA and RelB mRNA expression in RA group patient were higher than those in OA group.The difference was statistically significant(Z values were 24.399,-2.167,respectively,P ＜ 0.05).The expressions of LTα mRNA,LTβ mRNA and p65 mRNA in RA group were higher than those in OA group,but the difference was not statistically significant(Z values were-1.595,-1.461,-1.493,respectively,P ＞ 0.05).Condusion LTβR mRNA and RelB mRNA were highly expressed in the articular cavity effusion of patients with rheumatoid arthritis,and they have certain value in diagnosis.It showed that it may play a role in the occurrence and development of rheumatoid arthritis and the LTβR signaling pathway may be involved in the activation of the NF-κB pathway after ligand binding.

Objective To investigate the nutritional status of vitamin D in preterm infants after birth and further explore its possible influencing factors, so as to guide clinical vitamin D therapy and to screen the preterm infants who are at high risk of vitamin D deficiency.Methods Retrospective analysis was conducted in the neonatal department of our hospital from April 21st, 2014 to February 5th, 2016.The serum 25(OH)D level in preterm infants were measured 2 weeks after birth.Data including gender, season of birth, time to initiation of breastfeeding were collected.According to the 25(OH)D levels[25(OH)D≤37.5 nmol/L, 37.5 nmol/L≤50.0 nmol/L, and 25(OH)D>50.0 nmol/L], all the preterm infants were divided into three groups: vitamin D deficiency, insufficiency, and sufficiency groups.The influencing factors of vitamin D in preterm infants were screened by using statistical method.Results The mean 25(OH)D level of 172 preterm infants was (43.1±16.7)nmol/L.In vitamin D deficient, insufficient, and sufficient groups, there were 68 (40％), 50 (29％) and 54(31％) cases of preterm babies, respectively.The mean values of 25(OH)D in these three groups were (27.8±16.7)nmol/L, (42.4±3.4)nmol/L, and (63.0±11.7)nmol/L, respectively.Only the season of birth had significant difference among three groups (P=0.013): 44.2％ of the preterm infants born in winter had vitamin D deficiency, which was higher than those in spring (41.7％), summer(33.3％), and autumn (38.1％);44.2％ of the preterm infants born in winter had vitamin D insufficiency, which was much higher than those in spring (30.6％), summer (25.1％), and autumn (19.0％);furthermore, only 11.6％ of the preterm infants born in the winter had vitamin D sufficiency, which was much lower than those in spring (27.8％), summer (41.2％), and autumn (42.9％) (OR=4.655, 95％ CI=1.716-12.627, P=0.003).Conclusion Vitamin D deficiency in preterm infants 2 weeks after birth is prevalent, and winter birth is a risk factor of vitamin D deficiency and insufficiency in preterm infants.

目的 分析2006-2015年温州市人民医院住院新生儿病因及死亡变化趋势,为新生儿疾病防控提供指导.方法 回顾性分析2006-2015年温州市人民医院住院及死亡新生儿病例资料,分为2006-2010年、2011-2015年2个时间段,统计新生儿时期常见病类型,分析死亡趋势变化及死因构成变化.结果 ①2006-2010年新生儿病因以新生儿高胆红素血症(黄疸)(31.78％)、新生儿窒息(23.38％)、新生儿肺透明膜病(14.68％)为主要病因;2011-2015年病因以黄疸(38.91％)、新生儿肺炎(16.31％)、新生儿窒息(13.02％)为主要病因;黄疸仍为主要病因,新生儿窒息、新生儿肺透明膜病均有减少,前后5年新生儿窒息及新生儿肺透明膜病在疾病构成所占比例对比差异有统计学意义(P＜0.05).②10年间该院新生儿死亡率为1.20％.死亡新生儿以男性多见,出生≤7 d死亡率高于出生＞7 d,流动人口死亡率高于常住人口,早产儿死亡率高于足月新生儿(P＜0.05).③2010年新生儿死亡率最高,其次为2006年,2007年位列第3;2006年后新生儿死亡率缓慢下降,2009年有所上升,2010年后逐渐下降,随后2年下降趋势明显.④2006-2010年新生儿肺透明膜病为新生儿主要死因,其次为早产儿、新生儿肺炎、新生儿窒息;2011-2015年早产儿病死率有所下降,但仍为主要病因,其次为新生儿肺透明膜病、新生儿肺炎、新生儿窒息.2011-2015年新生儿肺炎、早产儿死亡率对比差异有统计学意义(P＜0.05).结论 2006-2015年,该院新生儿住院病因以黄疸、新生儿肺炎、新生儿肺透明膜病为主,10年间住院新生儿死亡率逐渐下降,后5年下降明显,早产儿、男性、流动人口死亡率较高,早产、新生儿肺透明膜病、新生儿肺炎、新生儿窒息为其主要死因.为降低新生儿死亡率,必须强化产前筛查,加强孕期宣教,引进新生儿复苏技术.

Objective To understand the antimicrobial resistance of Acinetobacter baumannii isoaltes in the First Affiliated Hospital of Wenzhou Medical University,to monitor the nosocomial infection of Acinetobacter baumannii by random amplified polymorphic DNA( RAPD) technology and to study the molecular epidemiological characteristics of Acinetobacter baumannii. Methods The disk diffusion method was applied to test the susceptibility of 100 Acinetobacter baumannii strains. Random amplified polymorphic DNA was used to determine the homology of these isolates. Phylogenetic tree was drew by NTSYSpc software to study the distribution of Acinetobacter baumannii in each department. Results In addition to cefoperazone/ sulbactam and levofloxacin,these Acinetobacter baumannii isoaltes showed drug resistance above 60% to other common antibotica. The 100 Acinetobacter baumannii strains can be divided into six genotypes,A,B,C,D,E and F in the light of the number and size of DNA fragment,in which B and C had the closest phylogenetic relationship. The most popular genotypes in the hospital were A and B, 28 isolates belonged to A and 24 isolates belonged to B. Conclusion The antimicrobial resistance of Acinetobacter baumannii is very serious in the hospital,with A and B as the popular genotypes. Some departments are found small-scale epidemic of the 6 genotypes of Acinetobacter baumannii.

目的：了解临床分离肠球菌的分布特点及耐药特性，为临床治疗提供参考。方法统计2304株肠球菌的耐药情况，采用 K - B 琼脂扩散法进行抗菌药物敏感性试验。结果2304株肠球菌中以尿液标本所占比例最高（37.3％），菌种分布以粪肠球菌为主（占33.6％）。不同菌种在不同来源标本中的分布比例存在差异。肠球菌对大部分临床常用抗菌药物表现为高度耐药，并且屎肠球菌、鹑鸡肠球菌、铅黄肠球菌的耐药谱与粪肠球菌的耐药谱存在明显差异：粪肠球菌对喹诺酮类抗菌药物和β-内酰胺类抗菌药物的耐药率明显低于其它三种肠球菌，而对奎奴普丁/达福普汀和四环素的耐药率明显高于其它三种肠球菌（P＜0.05）。并且分离到耐万古霉素的肠球菌（VRE）96株（占4.2％）。结论临床上肠球菌感染仍以粪肠球菌为主，屎肠球菌的检出率已接近粪肠球菌。肠球菌可引起临床各类感染，呈多重耐药趋势。

目的 研究万古霉素耐药肠球菌(VRE)的耐药表型和基因特性.方法 采用浓度梯度法(E-test)测定VRE对万古霉素(VA)、替考拉宁(TP)的耐药表型;采用聚合酶链反应检测vanA、vanB、vanC1、vanC2、vanC3基因,并对van基因产物进行测序.结果 万古霉素耐药基因检测结果:VRE经PCR扩增,均获得vanA基因阳性片段.未检测到vanB、vanCl及vanC2/3基因.结论 万古霉素耐药肠球菌的耐药表型与耐药基因型一致,菌株之间有较高的同源性.住院患者肠道中VRE携带率高,是医院感染的危险因素.

目的 分析65例腹膜透析患者腹膜透析液常规及细菌培养结果.方法 取腹膜透析液两管,一管行常规分析,包括有核细胞计数和沉渣细胞分类,另一管做细菌培养.结果 65例患者中37例表现为腹膜透析相关感染性腹膜炎,占56.9％;45例患者送检细菌培养,阳性率为62.2％,主要为凝固酶阴性葡萄球菌,占20.0％,其次为链球菌和革兰阴性杆菌,分别占17.8％和15.6％;35例腹膜炎患者有8例表现为顽固性腹膜炎,2例出现腹膜透析液性质改变,其转归情况明显差于其他患者(P＜0.05).结论 腹膜透析相关感染性腹膜炎是腹膜透析最常见的严重并发症,以细菌性感染多见,因此对腹膜透析液进行细胞学常规和病原学相结合的分析,能及时了解透析情况和变化,更好地为临床服务.

Objective To investigate the prevalence and plasmid size of qnrD determinant in Morganella morganii (M.morganii) isolates.Methods A total of 100 non-duplicated M.morganii clinical isolates were collected from inpatients.Standard ager dilution method was used to determine the minimum inhibitory concentrations (MICs) of fluoroquinolones against M.morganii isolates.PCR were performed to detect plasmid-mediated quinolone resistance determinants (PMQRs) in M.morganii isolates and the prevalence of extended-spectrum β-lactamase (ESBL) genes and AmpC β-lactamase genes in PMQRs-positive M.morganii strains.The homology analysis among qnrD-positive M.morganii strains were conducted by using pulsed-field gel electrophoresis (PFGE).The location of qnrD gene and the size of plasmid carrying it were determined by southern hybridization.The transferability of qnrD gene was determined by conjugation experiment.Results Thirty out of 100 M.morganii isolates (30％) were found carrying PMQRs including 17 qnrD-positive strains,14 aac (6')-Ib-cr-positive strains and 5 qepA-positive strains.PCR and sequencing confirmed that thirty PMQRs-positive isolates carried blaDHA-1.Among them,six isolates were positive for ESBLs genes (four for blaCTX-M-14,one for blaCTX-M-3 and one for blaCTX-M-24) and four isolates were positive for blaTEM-1.Almost all PMQRs-positive M.morganii isolates showed reduced susceptibility to fluoroquinolones.Moreover,seventeen qnrD-positive M.morganii isolates harbored blaDHA-1 including five (29.4％) harboring aac(6')-Ib-cr gene,four (23.5％) harboring blaCTX-M-14,two (11.8％) harboring blaTEM-1 and one harboring aac(6')-Ib-cr gene,blaCTX-M-14 and blaDHA-1.PFGE analysis showed that the 17 qnrD-positive M.morganii isolates were divergent from each other and not clone-related.Southern hybridization analysis showed that qnrD genes of all M.morganiiis isolates were mainly located in a 2.7 kb plasmid,but only a few of them were located in a size of 5.1 kb plasmid.M.morganiiis isolates failed to transfer qnrD gene to E.coli EC600 through conjugation.Conclusion PMQRs were widely distributed in M.morganiiis isolates.qnrD gene was the predominant determinants with a high prevalence rate of 17.0％,followed by aac(6')-Ib-cr gene.qnrD gene was located on a non-conjugative plasmid of approximately 2.7 kb or 5.1 kb.One qnrD-positive M.morganii isolate carrying aac(6')-Ib-cr gene,blaCTX-M-14 and blaDHA-1 was detected.

Objective To investigate the clinical infection and drug resistance in Serratia marcescens,and analysis its resistance characteristics to carbapenems antibiotic.Methods Retrospective survey the distribution of 109 Serratia marcescens clinical isolated from 2004 to 2010,VITEK-60 automatic microorganism analyzer was used to detect the susceptibility of antibiotics,and minimum inhibitory concentration(MIC) of imipenem,meropenem and ertapenem were detected by agar dilution method.Results The number of Serratia marcescens strains isolated had a upward trend year after year.And the specimen which was from the sputum occupy the most,accounting for 73.4%,followed by urine 12.8% and blood 6.4%;The sensitivity rates of ciprofloxacin,sulfamethoxazole,amikacin,levofloxacin and ceftriaxone were all above 80%,the resistance rates were 8.3%(9/109),5.5%(6/109) and 22.0%(24/109) to imipenem,meropenem,ertapenem respectively.Conclusion Clinical isolates of Serratia marcescens has a fine overall sensitivity to antimicrobial drugs,but there is a high proportion of carbapenem antibiotics resistant,and there is a discrepancy in different carbapenem antibiotics.