Alport综合征诊断中应注意的几个问题

Alport综合征(Alport syndrome, AS)为Ⅳ型胶原α链的基因突变导致的基底膜损伤,是一种表现为血尿、肾功能进行性减退的遗传性肾炎,常伴有感音神经性耳聋和眼部异常[1].国外一些资料表明AS并不罕见,估计其基因频率为1∶5000～1∶10000[2].在我国统计的2315例儿科肾活检材料中,27例(1.2%)被诊断为AS[3].AS最主要的遗传方式是X连锁显性遗传,约占85%,由COL4A5基因的突变引起;约10%～15%的AS家系呈常染色体隐性遗传,由COL4A3和/(或)COL4A4基因的突变引起;有极少部分病例呈常染色体显性遗传,可能与COL4A3和(或)COL4A4基因的突变有关[4].     

2022年美国医学遗传学与基因组学学会/分子病理学协会 Alport综合征管理和基因检测指南

基因检测致病性Ⅳ型胶原(COL4)A3-5变异是否存在常用于查找持续性血尿的原因,尤其是伴血尿家族史或肾功能损害。目前,Alport综合征专家主张对持续性血尿者进行基因检测,即使怀疑是杂合子致病性COL4A3或COL4A4变异,也建议一级家庭成员进行级联检测——这些家庭成员存在肾功能受损的风险。专家们不建议COL4A3或COL4A4杂合子作为肾脏捐赠者。疑似遗传性局灶节段性肾小球硬化(FSGS)引起的持续性蛋白尿、糖皮质激素耐药的肾病综合征、家族性IgA肾小球肾炎和未知原因的肾衰竭也应予以COL4A3-5基因变异检测。     

COL4A3基因2个新无义变异致Alport综合征的临床和遗传分析

目的探讨COL4A3基因突变所引起的常染色体隐性遗传Alport综合征的临床特点和基因变异特点。方法用基因测序和生物信息学分析方法分析1个常染色体隐性遗传Alport综合征患者的临床表型和基因型。结果男性患者,14岁发现水肿、肉眼血尿、蛋白尿、低蛋白血症伴有听力障碍,治疗、随访不规律,20岁进展至终末期肾病。应用全外显子组测序发现COL4A3基因2个新的无义变异,40号外显子上c.3508GT(p.G1170X)杂合变异,51号外显子上c.4825CT(p.R1609X)杂合变异,Sanger测序验证患者母亲携带c.3508GT(p.G1170X)杂合变异,父亲携带c.4825CT(p.R1609X);患者的哥哥未携带这2个基因变异。这2个无义变异均导致蛋白质表达提前终止,系致病变异。结论 COL4A3基因无义变异导致的常染色体隐性Alport综合征在青少年时期进展至终末期肾病;新发现2个COL4A3基因的无义变异;全外显子基因测序是确诊Alport综合征的重要手段。     

高糖环境及雷帕霉素干预对肾小球足细胞Ⅳ型胶原和MMP-9表达的影响

目的探讨高糖培养环境及雷帕霉素干预对肾小球足细胞Ⅳ型胶原和基质金属蛋白酶9（MMP-9）表达的影响。方法体外培养的小鼠肾小球足细胞分为空白对照组、高渗对照组、高糖组和高糖＋雷帕霉素干预组。Real-time PCR和Western blotting检测各组肾小球足细胞Ⅳ型胶原α3链（COL4A3）、α5链（COL4A5）mRNA及MMP-9mRNA和蛋白的表达;ELISA法检测细胞培养上清液中Ⅳ型胶原的含量。结果与对照组比较,高糖组足细胞COL4A3、COL4A5mRNA表达及培养上清液中Ⅳ型胶原含量均升高（P〈0.05）,而足细胞MMP-9mRNA和蛋白表达均降低（P〈0.05）。与高糖组比较,雷帕霉素干预组足细胞COL4A3、COL4A5mRNA表达及培养上清液中Ⅳ型胶原含量均降低（P〈0.05）,足细胞MMP-9mRNA和蛋白表达均升高（P〈0.05）。结论雷帕霉素对高糖环境中足细胞Ⅳ型胶原和MMP-9表达异常具有逆转作用。     

高糖环境及雷帕霉素干预对肾小球足细胞Ⅳ型胶原和MMP-9表达的影响

目的 探讨高糖培养环境及雷帕霉素干预对肾小球足细胞Ⅳ型胶原和基质金属蛋白酶9(MMP-9)表达的影响.方法 体外培养的小鼠肾小球足细胞分为空白对照组、高渗对照组、高糖组和高糖+雷帕霉素干预组.Real-time PCR和Western blotting检测各组肾小球足细胞Ⅳ型胶原α3链(COL4A3)、α5链(COL4A5)mRNA及MMP-9 mRNA和蛋白的表达;ELISA法检测细胞培养上清液中Ⅳ型胶原的含量.结果 与对照组比较,高糖组足细胞COL4A3、COL4A5 mRNA表达及培养上清液中Ⅳ型胶原含量均升高(P<0.05),而足细胞MMP-9 mRNA和蛋白表达均降低(P<0.05).与高糖组比较,雷帕霉素干预组足细胞COL4A3、COL4A5 mRNA表达及培养上清液中Ⅳ型胶原含量均降低(P<0.05),足细胞MMP-9 mRNA和蛋白表达均升高(P<0.05).结论 雷帕霉素对高糖环境中足细胞Ⅳ型胶原和MMP-9表达异常具有逆转作用.     

赖氨大黄酸对快速老化小鼠SAMP 10肾组织COL1A1、COL3A1和COL4A1表达的影响

目的 研究赖氨大黄酸(rhein lysinate,RHL)对快速老化小鼠(senescence accelerated mouse prone 10,SAMP 10)肾组织Ⅰ型胶原α1链(COL1 A1)、Ⅲ型胶原α1链(COL3A1)和Ⅳ型胶原α1链(COL4A1)蛋白表达的影响.方法 选取7月龄SAMP 10小鼠18只,并根据随机数字表分为空白对照组、低剂量RHL组(25 mg/kg)和高剂量RHL组(50 mg/kg),每组6只;另选抗快速老化小鼠(senescence accelerated mouse resistance 1,SAMR 1)6只作为青年对照,给药时间6个月.采用HE染色观察肾脏组织病理改变,Masson染色观察肾脏组织纤维化改变,Westem blot方法检测肾组织COL1A1、COL3A1、COL4A1和NF-κB蛋白的表达.结果 与SAMP 10空白对照组相比,25和50 mg/kg RHL组治疗后SAMP 10小鼠精神状态、活动度、毛色等方面较好;肾脏指数升高.HE染色显示SAMP 10空白对照组小鼠肾组织有节段性肾小球萎缩、硬化和明显的间质纤维化,SAMR 1组和RHL组小鼠则无肾小球萎缩、硬化和明显的间质纤维化.RHL能够抑制SAMP 10小鼠衰老过程中出现的肾组织COL1A1、COL3A1和NF-κB蛋白过度表达(P＜0.05).结论 RHL可通过抑制COL1A1、COL3A1和NF-κB蛋白过度表达,减轻衰老过程中SAMP 10小鼠肾组织纤维化程度,发挥肾脏保护作用.     

老年男性骨密度与Ⅰ型胶原α1链及α2链基因多态性的研究

[目的]探讨Ⅰ型胶原α1链(Collagen typeⅠ alpha 1,COL1A1)及α2链(COL1A2)基因型与老年男性骨密度(BMD)相互关系.[方法]选取年龄≥65岁男性个体193例,采用双能X线吸收法(DEXA)测定其全身、腰椎2～4(L2～4)、股骨颈(Neck)、Ward'三角和大转子区的BMD值,并分别采用AS-PCR及PCR-RFLP方法检测外周血白细胞基因组COL1A1及COL1A2基因型.[结果]193例受试对象中,COL1A1基因型均为SS型;COL1A2基因型分别为EE 81例、Ee83例、ee29例.[结论]受试人群Ⅰ型胶原α1链Sp1结合位点不存在G→T突变,骨密度与COL1A1未见基因多态性无相关性;对于COL1A2基因而言,携带EE基因型的老年男性个体具有较低的骨密度值.     

风湿性心脏瓣膜病合并心房颤动中胶原蛋白表达与分布

目的 探讨胶原纤维在风湿性心脏瓣膜病心房颤动(房颤)患者中的异常表达以及与房颤发生的相关性.方法 33例风湿性心脏病接受换瓣患者分为窦性心律组与房颤组.于手术中获取右心耳组织.采用转录组学进行差异表达基因的功能分析;RT-PCR检测Ⅰ~Ⅳ型胶原纤维RNA在右心房组织中的表达;免疫组化观察心肌组织各型胶原纤维的表达与分布.结果 与窦性心律组相比,房颤组左心房内径明显扩大.转录组学分析显示COL1A2、COL2A1、COL3A1以及COL4A3与COL5A2、COL6A3、COL11A1等胶原纤维存在相互关联之外,与多种细胞因子也存在相互作用;RT-PCR结果示COL1A2、COL2A1、COL3A1和COL4A3均在房颤患者右心房组织中上调表达,其中COL3A1、COL4A3上调表达显著(P<0.05).免疫组化结果显示Ⅰ型胶原纤维分布在血管内皮细胞及血管基底膜;Ⅱ型胶原纤维分布在心肌细胞肌节;Ⅲ型胶原纤维分布在间皮细胞及心肌细胞浆;Ⅳ型胶原纤维分布在心肌细胞外膜.结论 心房组织中COL3A1与COL4A3表达异常,与多种细胞因子相互作用引发房颤的发生和发展.     

检测皮肤成纤维细胞cDNA确定Alport综合征COL4A5基因突变

目的:检测、分析Alport患者Ⅳ型胶原α5链mRNA及其序列,试图建立一种简单、快捷的检测COL4A5基因突变的方法,同时明确基因突变对COL4A5基因转录的影响.方法:取21例X连锁显性遗传型Alport综合征患者和2例对照的皮肤标本培养后提取成纤维细胞总RNA,应用逆转录-聚合酶链反应(RT-PCR) 和直接测序的方法分析Ⅳ型胶原α5链mRNA.对于cDNA序列异常者,进而应用PCR和直接测序的方法分析COL4A5基因.结果:成功进行了原代、传代皮肤成纤维细胞的培养并提取了总RNA;直接测序揭示对照组Ⅳ型胶原α5链mRNA序列及已检测的8例患者Ⅳ型胶原α5链3′端mRNA 序列与所发表的序列完全一致;通过RT-PCR和直接测序发现了4 例新突变,并与从基因组DNA样本中检测到的突变相一致.结论:通过分析皮肤成纤维细胞中Ⅳ型胶原α5链mRNA序列,可用以检测Alport综合征患者COL4A5 基因突变,且本文报道的4例突变均为新突变;COL4A5基因突变后存在异常的转录产物.     

血清COL4A3蛋白与非小细胞肺癌(NSCLC)患者临床病理特征及预后的相关性

 目的 探索血清COL4A3蛋白与非小细胞肺癌（non-small cell lung cancer,NSCLC）患者临床病理特征及预后的相关性。方法 ELISA试剂盒检测90例NSCLC患者和58名健康对照血清中COL4A3蛋白含量。ROC曲线分析血清COL4A3水平以预测NSCLC患者肿瘤转移的敏感性与特异性。Kaplan-Meier生存分析研究血清COL4A3蛋白表达与患者生存期的相关性。对NSCLC患者预后的相关性因素进行单因素和多因素分析。采用MTT实验和Transwell实验分别检测血清COL4A3蛋白对NSCLC细胞增殖和迁移能力的影响。结果 NSCLC患者血清COL4A3水平明显高于健康对照（P<0.001）,且高表达COL4A3的患者总生存期（overall survival,OS）低于低表达患者（P=0.021）。在单因素分析中,NSCLC患者预后与肿瘤大小（P=0.015）、病理组织分级（P=0.042）、淋巴结转移（P=0.015）、临床分期（P<0.01）以及COL4A3表达（P<0.01）密切相关;在多因素分析中,NSCLC患者的预后与患者淋巴结转移（P=0.032）、临床分期（P<0.01）以及血清COL4A3水平（P=0.012）密切相关。COL4A3蛋白可显著增强体外NSCLC细胞增殖和迁移能力,并呈浓度依赖性。结论 血清COL4A3蛋白水平与NSCLC进展密切相关,可能是一种NSCLC患者预后不良的独立标志物。     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

Dr.Zhang Ren''''s Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

Luo Lingjie''''s Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

猪肺磷脂注射液联合经鼻持续气道正压通气对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性的影响

目的 探讨猪肺磷脂注射液联合经鼻持续气道正压通气(NCPAP)对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性(CK-MB)的影响.方法 选取呼吸衰竭早产儿80例,分为观察组和对照组各40例.对照组采用NCPAP给氧治疗,观察组给予NCPAP给氧联合猪肺磷脂气管内给药.观察两组患儿治疗前及治疗12h、24 h后PaO2、PaCO2、血氧饱和度(SaO2)、pH的变化情况,检测治疗前及治疗5d后血清CK-MB水平;评估两组患儿的临床治疗效果.结果 两组患儿PaO2、PaCO2、SaO2、pH比较,差异均有统计学意义(P＜0.05),其中观察组治疗后的PaO2、SaO2、pH均高于对照组,PaCO2则低于对照组.两组的PaO2、SaO2、pH均随观察时间延长而升高(P＜0.05),PaCO2均随观察时间的延长而降低(P＜0.05).观察组治疗有效率为87.5％,显著高于对照组的70.0％ (P ＜0.05).治疗5d后两组患儿血清CK-MB水平均较前降低(P＜0.05),且观察组明显低于对照组(P＜0.05).结论 猪肺磷脂注射液气管内给药联合NCPAP可以显著降低呼吸衰竭早产儿CK-MB的含量,提高治疗有效率,起到很好的呼吸循环支持作用.     

Survey and practice of reporting quality of randomized controlled clinical trials on traditional Chinese medicine

Evidence obtained from randomized controlled trials （RCTs） has been generally accepted as the gold standard in the evaluation of clinical effectiveness. Readers need to understand the trial design, implementation, results, analysis and interpretation, so as to fully Jnderstand the results of RCTs. Thus, the investigators of RCTs have to report these items in a complete, accurate and clear manner. Since 1998, we have conducted several evaluations on the reporting quality of RCTs published in Chinese journals on traditional Chinese medicine （TCM） and results have shown that there is an urgent need for higher quality RCTs on TCM.     

TCM Treatment of Ankylosing Spondylitis by Tonifying the Kidney and Strengthening the Governor Vessel

Ankylosing spondylitis is a chronic and progressive disorder with inflammation mainly involving the central axis joints. It mainly affects the cervical spine and the lumbosacral area, with the pathogenesis closely related to the kidney and the Governor Vessel (GV). TCM holds that the syndrome is deficiency in origin and excess in superficiality, which is due to insufficiency of the kidney, deficiency of GV, and blocking of the channels with the invasion of exogenous evil, leading to poor circulation of qi and blood and malnutrition of the bones, muscles and joints. The TCM method of tonifying the kidney and strengthening GV to regulate circulation of qi and blood and check the arthralgia pain should be adopted, with the Kidney-Tonifying and GV Strengthening Decoction (益肾强督汤) prescribed.     

IMPROVING P-gp EXPRESSION IN HUMAN MONONUCLEAR CELLS IN VITRO TRANSFECTED BY MULTIDRUG RESISTANCE-1 mRNA

CHEMOTHERAPY playsa greatrolein the treat- ment of malignanttumors,especiallyingynecolo- gicalones.But inanticancerchemotherapy,leuko-cytopeniaisfrequentlytheprimarydose-limitingsideeffect factor.Moreover,cancersarefrequentlychemoresistantbe-causeof overexpressionof P-glycoprotein(P-gp), which isencodedby multidrugresistancegene (MDR1 ) and detectableinup to50% ofhuman cancersand renderscellsresistancetoanticancerdrugs.The safetyand potentialtherapeuticbenefitof mdr1 gene transferredto h…