Microdissection genotyping of archival fixative treated tissue for Gaucher disease

The genetic diagnosis of Gaucher disease by molecular methods is complicated by the existence of a highly homologous transcribed pseudogene (96% identity) that is found in close proximity to the true gene on chromosome 1q21. In addition, the pseudogene sequence can mimic disease-causing mutations in the true gene. Selective polymerase chain reaction (PCR) amplification of the true gene can be accomplished in extracted DNA from fresh-frozen samples by designing oligonucleotide primers to hybridize to defined regions that are not present in the pseudogene. This standard molecular approach, which entails amplification of relatively long segments of intact DNA, is not feasible in archival, paraffin-embedded, solid-tissue specimens in which the negative effects of chemical fixation result in DNA strand scission and breakdown of nucleic acid. A novel approach, specifically created for use with archival, fixative-treated tissue specimens, was developed for detection and characterization of common mutations of Gaucher disease. Three separate robust PCR reactions were formulated, 2 for selective amplification of portions of only the true gene exons 2 and 9, with a third reaction targeting exon 10, wherein both the true and pseudogene were coamplified. In the latter, DNA sequencing was used to determine the presence of true and pseudogene allele content in addition to identification of base sequence alterations. This method, requiring a single, 4-microm-thick histologic section, was successfully applied to archival paraffin block tissue specimens that had been in storage for up to 75 years. It was capable of accurately genotyping common Gaucher disease mutations as well as discovering a novel mutation and genetic polymorphism. We recommend our approach when only fixative-treated tis sue is available for molecular genotyping.     

The AMeX method: a multipurpose tissue-processing and paraffin-embedding method. III. Extraction and purification of RNA and application to slot-blot hybridization analysis.

RNA was extracted from tissues processed by a new fixation and paraffin-embedding method (the AMeX method) and examined by Northern blot analysis and slot-blot analysis. The RNA extraction method for AMeX-processed tissue sections after the deparaffinization step was the same as that for fresh materials. The total amount of cellular RNA extracted from AMeX-processed mouse liver tissue was slightly less than that extracted from fresh tissue. In tissues of malignant lymphoma, the total amount of cellular RNA extracted from 25 sections each 20 microns thick was about 1.6-1.8 micrograms/mm2, regardless of the histological subtype and period of storage. The extracted RNA was moderately degraded, and usually could not be used for Northern blot hybridization analysis. The intensity of ethidium bromide staining and the hybridization signals of RNA extracted from AMeX-processed tissues were usually reduced in comparison with RNA from fresh material, but specific signals could be detected by slot-blot hybridization analysis. We have demonstrated previously that the AMeX method preserves high-molecular-weight DNA and various antigens. Since the present study showed that information on mRNA can be obtained from AMeX-processed tissue, the versatility and usefulness of this method were further proven.     

环境物理因子的最佳条件的选择

目的：利用黄金数比例规律找出环境物理因子对人体的最佳条件：方法：人体的相当一部分内容与黄金数有着密切的联系，为此把复杂的人体可以简化为比例加破缺的比例实体；环境物理因子：如环境温度、湿度、环境光波和声波以及环境气压和氧分压，当它们处在黄金数比例附近时，会在人体产生和谐合柏作用。结果：环境物理因子选择在黄金数比例附近时，在心理上产生美和舒适感。已经发现，当精神愉快时，人脑电波频率下限(8Hz)和上限(12．9Hz)之比恰为黄金数。结论：环境物理因子的最佳条件是可以选择的。     

Use and abuse of frozen section in the diagnosis of follicular thyroid lesions

The role of intraoperative frozen section in certain organ systems such as the thyroid continues to be problematic. In many cases, diagnoses are deferred or nonhelpful—“follicular lesion.” In the modern era, the widespread use of preoperative aspiration biopsy has allowed for more careful selection of patients who undergo thyroid surgery. In many cases, the fine-needle-aspiration (FNA) biopsy diagnosis can be definitive or can guide the specific surgical procedure. The literature supports our approach, which is summarized as follows: Intraoperative consultation is not needed on the intrathyroidal nodule if a preoperative FNA was definitive for papillary carcinoma. Frozen section is of no value in the intraoperative diagnosis of lesions diagnosed on FNA as “follicular neoplasm” or “Hürthle cell neoplasm” because the characterization of these lesions requires detailed analysis of the tumor capsule for the demonstration of capsular and/or vascular invasion—an analysis that is not practical in the intraoperative setting. Finally, intraoperative consultation including frozen section and intraoperative cytologic examination is most useful in those cases that are diagnosed as suspicious for papillary carcinoma by FNA, because the assessment of nuclear features needed for the definitive diagnosis is possible with intraoperative techniques in a significant number of cases.     

自由体位分娩对自然分娩率和剖宫产率影响的临床观察

目的:观察自由体位分娩对不同分娩方式的影响。方法:选取94例来我院分娩的孕产妇为研究对象,按入院顺序将其分为两组,采取常规卧位分娩的为对照组,应用自由体位分娩的则为观察组。对比两组产妇的产程时间及分娩方式。结果:①产程时间方面,两组产妇第二、第三产程时间差异不明显(P>0.05),但观察组第一及总产程时间显著低于对照组(P<0.05)。②分娩方式方面,观察组自然分娩率要显著高于对照组,剖宫产率明显低于对照组,P<0.05。结论:通过采取自由体位的方式进行分娩,可缩短产程,降低剖宫产率。     

剖宫产术后阴道分娩产妇的临床分析

目的探讨剖宫产术后阴道分娩(VBAC)产妇的母儿结局。 方法选择2016年1月1日至2019年12月31日,四川大学华西第二医院收治的VBAC足月妊娠产妇397例为研究对象,纳入研究组。按照1∶1比例,采用随机数字表法随机抽取同期于本院足月经阴道分娩的非瘢痕子宫产妇397例,作为对照,纳入对照组。观察2组产妇年龄、身高、人体质量指数(BMI)、本次分娩距前次分娩间隔时间、分娩时孕龄,第一、第二、第三产程时间,术中出血量、产后出血(PPH)量(2 h)、PPH率、严重PPH率,以及新生儿出生体重与窒息发生率等指标,并采用成组t检验、Wilcoxon秩和检验、χ²检验和Fisher确切概率法对上述指标进行统计学比较。对2016—2019年本院产妇分娩情况进行分析。2组产妇年龄、身高、BMI、分娩时孕龄等一般临床资料比较,差异均无统计学意义(P>0.05)。本研究遵循的程序符合2013年新修订的《世界医学协会赫尔辛基宣言》要求。 结果①2016—2019年,本院剖宫产术分娩率为58.45%～62.37%,剖宫产术后瘢痕子宫(SUAC)患者再次妊娠的剖宫产术分娩率为93.06%～95.83%,平均为94.60%(10 942/11 567),非SUAC患者的剖宫产术分娩率为50.27%(18 586/36 972)。瘢痕子宫患者为13 963例(28.77%),2017年瘢痕子宫患者所占比例最高,为33.74%(3 361/9 961),此后呈下降趋势。②研究组产妇第一、第二产程时间及术中出血量、PPH量(2 h)、PPH率分别为363 min(212～450 min)、46 min(22～60 min)、312 mL(200～350 mL)、348 mL(230～380 mL)、10.8%(43/397),均显著长于、大于、高于对照组的294 min(180～380 min)、15 min(7～20 min)、231 mL(150～280 mL)、264 mL(180～310 mL)、4.3%(17/397),研究组新生儿出生体重为(3 216±359) g,则显著低于对照组的(3 369±369) g,并且差异均有统计学意义(Z＝4.797、16.065、8.028、8.122,χ²＝12.188,t＝-17.013,均为P<0.001)。③SUAC患者再次妊娠的子宫破裂发生率为1.73/10 000(2/11 567),占VBAC者的0.32%(2/625)。 结论VBAC是可行的,子宫破裂发生率低,但是其PPH风险增加。临床对于此类产妇,产程中应做好监护,积极预防PPH。     

产科急症子宫切除术16例分析

回顾分析近12a产科急症子宫切除术16例,16例均为子宫出血,其中剖宫产13例,阴道产3例,子宫出血的主要原因是各种胎盘异常,其发生与多次流产及宫腔操作史有关,阴道产后子宫切除失血量往往多于剖宫产者,提示在产后血失代偿期经导迅速治疗处理后,子宫出血仍不能有效控制,应当机立断行子宫切除术,以挽救产妇的生命。     

子宫腔填塞纱条治疗前置胎盘剖宫产术中大出血32例分析

目的 总结32例子宫腔填塞纱条治疗前置盘剖宫产术中大出血的临床经验,方法 对1991年2月至1999年10月,在我科用子宫腔填塞纱条治疗前置胎盘剖宫产术中大出血32例进行分析。结果 32例前胎盘剖宫产术中大出血用纱条填塞子宫腔后均成功止血,20－28小时后经阴道顺利取出,未发生阴道大出血,产后42天随访28例,子宫复旧良好。结论 子宫腔填塞纱条是治疗前置胎盘剖宫产术中大出血的有效方法。     

妊娠晚期羊水过少56例临床分析

目的：探讨妊娠晚期羊水过少的分娩方式,寻找正确的处理方法,降低围生儿死亡率。方法：对妊娠晚期羊水过少56例作回顾性分析。结果：56例中,30例以羊水过少作为指标行剖宫产,新生儿窒息率为0,19例在临产过程中12例并发胎儿窘迫,7例并发其它高危因素行剖宫产,新生儿窒息率为5．3％,7例经阴道分娩,新生儿均有不同程度窒息。结论：羊水过少一经确诊,应积极引产,估计短时间内不能分娩,及时剖宫产是处理羊水过少较为安全的重要措施。     

不缝合腹膜剖宫产术式的临床观察

目的　探讨不缝合腹膜剖宫产术式的临床效果。方法　选取剖宫产不缝合腹膜 80例产妇作为观察组 ,与同期分娩按传统方法缝合腹膜产妇 80例作为对照组 ,观察两组术后病率、肠功能恢复及切口愈合情况。结果　观察组术后病 6例 ,对照组 15例 ,两组比较 ,差异有显著性 (P <0 0 5 ) ;观察组术后肛门排气时间为 2 0 5± 5 33h ,对照组为 36 6 7± 6 17h ,两组比较 ,差别有高度统计学意义 (P <0 0 1) ;腹部切口愈合两组无差异。结论　不缝合腹膜剖宫产术式安全、有效、并发症少 ,值得在临床上推广应用。