Characterisation and Specificity of Glomerular Basement Membrane Antigens Identified by Sera of Patients with Anti-GBM Nephritis

The sera of 21 patients positive for antibodies against GBMin indirect immunofluorescence tests were examined by immunoblotting.We demonstrated antibodies against 50, 48, 43 and 29 kD molecularweight peptides in 20 of 21 sera using collagenase-digestedGBM, in 19 of 21 using trypsin-digested GBM, and in.10 of 21using elastase-digested GBM. Although the spectrum of molecularweights of the antigenic proteins was similar in all three digests,they differed with respect to preservation of antigenicity uponreduction with mercaptoethanol. Many of the sera of patientsand controls reacted with proteins unrelated to GBM, e.g. albuminand prealbumin. Furthermore, some control sera reacted withone single peptide of the above-mentioned specific GBM peptides.Our results suggest that the highly purified 29 kD peptide ofthe collagenase digest or the 50 kD peptide of the trypsin digestprovide the best antigens to develop a screening test for antibodiesagainst GBM. However, serum antibodies against these antigenswill not be absolutely specific for anti-GBM antibody-mediatednephritis, as shown by the immunoblot experiments.     

The Sperm Acrosome: Immunological Analysis using Specific Polyclonal and Monoclonal Antibodies Directed Against the Outer Acrosomal Membrane of Boar Spermatozoa

An antiserum to the purified porcine outer acrosomal membrane (OAM) was raised in female Balb/c mice and was characterized by means of an indirect ELISA. The hyperimmune serum reacted selectively with the acrosomal cap of the sperm head and showed an extremely good cross reactivity with bull and human spermatozoa when assayed by indirect immunofluorescence. Immunoelectron microscopy using the protein A-gold method further confirmed the specificity of the anti-OAM-antiserum for the OAM. In an effort to identify the OAM antigens recognized by the hyperimmune serum and to analyse the extent of cross reactivity on a molecular level, the SDS-extractable proteins were separated by SDS-PAGE, transblotted and immunoprinted using an 125J-conjugated anti-mouse-antibody. To facilitate functional and structural analysis of distinct OAM-proteins monoclonal antibodies were generated by hybridization of mouse myeloma cells with the splenocytes of female Balb/c mice immunized with the purified OAM. One fusion resulted in about 100 anti-OAM-antibodies secreting hybridoma cultures, of which about 30% showed cross reaction with human and bull spermatozoa. Four stable cell lines were selected for this study secreting antibodies directed against the outer acrosomal membrane of boar spermatozoa. Whereas the polyclonal immune mouse serum stained the entire acrosomal cap, the four hybridoma antibodies generated a patch-work-like immunofluorescence pattern over the acrosome. HPLC-ELISA of the solubilized OAM revealed first information on the nature of the corresponding membrane antigen.     

Venovenous versus venoarterial extracorporeal membrane oxygenation in congenital diaphragmatic hernia

Background: Extracorporeal membrane oxygenation (ECMO) has a significant role as a final rescue modality in severe respiratory failure of the newborn with congenital diaphragmatic hernia (CDH). The objective of this study was to compare the efficiency of venovenous (VV) versus venoarterial (VA) ECMO in newborns with CDH.Methods: A retrospective report of 11 years experience (1990 through 2001) of a single center, comparing VV and VA ECMO is given. VV ECMO was the preferred rescue modality for respiratory failure unresponsive to maximal medical therapy. Only when the placement of a VV ECMO 14F catheter was not possible, VA ECMO was used. Forty-six patients met ECMO criteria; 26 were treated with VV ECMO and 19 with VA ECMO. One patient underwent conversion from VV to VA ECMO.Results: Before ECMO, there was no difference between VV and VA ECMO patients in mean oxygenation index (83 v 83), mean airway pressure (18.4 v 18.9 cm H₂O), ECMO cannulation age (28 v 20 hours), or in the percentage of patients who needed dopamine and dobutamine (100% v 100%). From November 1994, nitric oxide (NO) was available; before ECMO, 11 of 14 (79%) VV ECMO patients received NO versus 9 of 10 (90%) patients in the VA group. VV ECMO patients were larger (3.34 v 2.77 kg; P < .05) and of advanced gestational age (39.0 v 36.9 wk; P < .05) compared with VA ECMO patients. There was no significant difference between VV and VA ECMO patients in survival rate (18 of 26, 69% v 13 of 19, 68%), ECMO duration (152 v 150 hours), time of extubation (32.0 v 33.5 days), age at discharge (73 v 81 days), or incidence of short-term intracranial complications (3.8% v 10.5%) or myocardial stun (3.8% v 15.8%).Conclusions: The authors conclude that VV ECMO is as reliable as VA ECMO in newborns with CDH in severe respiratory failure who need ECMO support and who can accommodate the VV double-lumen catheter. Because of its potential advantages, VV ECMO may be the preferred ECMO method in these infants.     

成年人肾小球基底膜厚度及基底膜变薄标准研究

目的 了解成年人肾小球基底膜（GBM）厚度及拟建议薄基底膜肾病（TBMN）的GBM弥漫变薄的标准。 方法 选取肾癌根治性切除患者29例,分析性别、年龄、尿常规、Scr以及既往史、家族史等临床资料。选取远离病灶的肾皮质组织,进行光镜、免疫荧光及透射电镜检查,并进行GBM厚度测量和Ⅳ型胶原α3、α5链免疫荧光检查。 结果 29例中,男15例、女14例,年龄（55.9±14.9）岁（20~80岁）,所有病例均无肾脏病家族史。肾组织GBM厚度为（363.6±46.8） nm。GBM厚度与性别相关,男性为（384.0±41.7） nm,女性为（335.0±39.2） nm,差异有统计学意义（P = 0.008）。建议以均数减去两倍标准差作为GBM变薄的标准,即GBM厚度＜270 nm。 结论 成年人肾组织的GBM厚度为（363.6±46.8） nm。GBM厚度和性别相关,男性GBM厚度大于女性,差异有统计学意义。TBMN的GBM弥漫变薄的诊断标准建议为GBM厚度＜270 nm,也建议今后制定TBMN标准中应考虑男女的差异。     

Overtime expression of plasma membrane and mitochondrial function markers associated with cell death in human spermatozoa exposed to nonphysiological levels of reactive oxygen species

In many cell types, the potential of reactive oxygen species to induce death processes has been largely demonstrated. Studies in spermatozoa have associated the imbalance of reactive oxygen species and phosphatidylserine externalisation as an apoptosis marker. However, the lack of consensus about time effect in the joint expression of these and other death markers has made it difficult to understand the set of mechanisms influenced beyond the concentration effect of reactive oxygen species to stimulate cell death. Here, the plasma membrane permeability and integrity, phosphatidylserine externalisation and mitochondrial membrane potential were jointly evaluated as death markers in human spermatozoa stimulated with H₂O₂. The results showed a profound and sustained effect of dissipation in the mitochondrial membrane potential and an increased phosphatidylserine externalisation in human spermatozoa exposed to 3 mmol⁻¹ of H₂O₂ at 30 min. This was followed by an increased membrane permeability after 45 min. The last observed event was the loss of cell membrane integrity at 60 min. In conclusion, mitochondria are rapidly affected in human spermatozoa exposed to reactive oxygen species, with the barely detectable mitochondrial membrane potential coexisting with the high phosphatidylserine externalisation in cells with normal membrane permeability.     

Morphometric study of glomerular basement membrane and proximal tubular basement membrane in adult thin basement membrane disease

Background. Thin basement membrane disease (TBMD) is a benign hereditary glomerulopathy with a diffuse attenuation of glomerular basement membrane (GBM). Whether the development of renal basement membranes other than GBM is normal in TBMD has not yet been resolved. Methods. We performed a morphometric study to measure the thickness of GBM and proximal tubular basement membrane (P-TBM) in 44 adult patients with TBMD and in 10 adult diseased controls confirmed to have minor glomerular abnormalities. Results. There was a significant difference between the patients with TBMD and the diseased controls in the thickness of the GBM; however, there was no significant difference between the two groups in the thickness of the P-TBM. In the patients with TBMD, the thickness of the GBM was unchanged with age, but the thickness of the P-TBM increased with age, as did that in the diseased controls. Conclusion. Our morphometric study clarified that the development of P-TBM was normal in the patients with TBMD. Received: October 7, 1998 / Accepted: May 27, 1999     

几丁糖联合聚乳酸薄膜预防硬膜外粘连的研究

目的 观察几丁糖加聚乳酸薄膜联合运用对椎板切除术后预防硬膜外瘢痕粘连的效果.方法 80只成年家兔[体质量(2.0±0.2)kg]制作椎板切除模型,在椎板缺损处分别覆盖等渗盐水(A组)、聚乳酸薄膜(B组)、几丁糖(C组)及几丁糖加聚乳酸薄膜(D组),术后12周对椎板切除部位进行大体观察、组织学观察及透射电镜比较各组间瘢痕形成和粘连情况.结果 B、C、D组的Rydell-Balazs粘连度评分、Nussvaum组织学评分均优于A组(P<0.01),D组优于B组和C组(P<0.01),B组与C组间差异无统计学意义(P>0.05).结论 联合应用几丁糖加聚乳酸薄膜能有效预防硬膜外瘢痕粘连,比单独应用几丁糖或聚乳酸薄膜效果好.     

几丁糖联合聚乳酸薄膜预防硬膜外粘连的研究

目的 观察几丁糖加聚乳酸薄膜联合运用对椎板切除术后预防硬膜外瘢痕粘连的效果.方法 80只成年家兔[体质量(2.0±0.2)kg]制作椎板切除模型,在椎板缺损处分别覆盖等渗盐水(A组)、聚乳酸薄膜(B组)、几丁糖(C组)及几丁糖加聚乳酸薄膜(D组),术后12周对椎板切除部位进行大体观察、组织学观察及透射电镜比较各组间瘢痕形成和粘连情况.结果 B、C、D组的Rydell-Balazs粘连度评分、Nussvaum组织学评分均优于A组(P<0.01),D组优于B组和C组(P<0.01),B组与C组间差异无统计学意义(P>0.05).结论 联合应用几丁糖加聚乳酸薄膜能有效预防硬膜外瘢痕粘连,比单独应用几丁糖或聚乳酸薄膜效果好.     

几丁糖联合聚乳酸薄膜预防硬膜外粘连的研究

目的 观察几丁糖加聚乳酸薄膜联合运用对椎板切除术后预防硬膜外瘢痕粘连的效果.方法 80只成年家兔[体质量(2.0±0.2)kg]制作椎板切除模型,在椎板缺损处分别覆盖等渗盐水(A组)、聚乳酸薄膜(B组)、几丁糖(C组)及几丁糖加聚乳酸薄膜(D组),术后12周对椎板切除部位进行大体观察、组织学观察及透射电镜比较各组间瘢痕形成和粘连情况.结果 B、C、D组的Rydell-Balazs粘连度评分、Nussvaum组织学评分均优于A组(P<0.01),D组优于B组和C组(P<0.01),B组与C组间差异无统计学意义(P>0.05).结论 联合应用几丁糖加聚乳酸薄膜能有效预防硬膜外瘢痕粘连,比单独应用几丁糖或聚乳酸薄膜效果好.     

几丁糖联合聚乳酸薄膜预防硬膜外粘连的研究

目的 观察几丁糖加聚乳酸薄膜联合运用对椎板切除术后预防硬膜外瘢痕粘连的效果.方法 80只成年家兔[体质量(2.0±0.2)kg]制作椎板切除模型,在椎板缺损处分别覆盖等渗盐水(A组)、聚乳酸薄膜(B组)、几丁糖(C组)及几丁糖加聚乳酸薄膜(D组),术后12周对椎板切除部位进行大体观察、组织学观察及透射电镜比较各组间瘢痕形成和粘连情况.结果 B、C、D组的Rydell-Balazs粘连度评分、Nussvaum组织学评分均优于A组(P<0.01),D组优于B组和C组(P<0.01),B组与C组间差异无统计学意义(P>0.05).结论 联合应用几丁糖加聚乳酸薄膜能有效预防硬膜外瘢痕粘连,比单独应用几丁糖或聚乳酸薄膜效果好.     

Blastocystis hominis infection in a post-cardiotomy patient on extracorporeal membrane oxygenation support: A case report and literature review

INTRODUCTIONOpportunistic pathogens can cause severe damage leading to irreversible complications in immune-compromised patients. Here we describe a patient who sustained Blastocystis hominis infection resulting in severe sepsis while on extracorporeal membrane oxygenation (ECMO) support, and the course of treatment taken to treat him.PRESENTATION OF CASEOur case, a 34-year-old Filipino man, was hospitalized for valvular disease and received valve replacements. ECMO and an intra-aortic balloon pump (IABP) were implemented when the patient developed progressive heart failure after cardiac surgery. Unfortunately, the patient suffered from sepsis with persistent fever and diarrhea, and subsequent examinations indicated the patient was infected by B. hominis. After adequate administration of the antibiotic metronidazole, the patient's symptoms subsided and he was discharged.DISCUSSIONBlastocystis hominis is a unicellular protozoa commonly found in the intestinal tract, and the prevalence of B. hominis is 1.5–10% in developed countries and 30–50% in developing countries. The patient needed the support of ECMO and IABP, was immunocompromised to a certain extent; B. hominis can be a harmful opportunistic pathogen for them and lead to severe irreversible complications such as death.CONCLUSIONThis is the first published article showing that the opportunistic pathogen, B. hominis, can cause severe infection in patients on ECMO support, a result that should be kept in mind when patients come from a place with a high prevalence of B. hominis. The prophylactic medication should be administered routinely when patients live in the region and extracorporeal life-support is used.     

Development of a New Hollow Fiber Silicone Membrane Oxygenator: In Vitro Study

An experimental silicone hollow fiber membrane oxygenator for long-term extracorporeal membrane oxygenation (ECMO) was developed in our laboratory using an ultrathin silicone hollow fiber. However, the marginal gas transfer performances and a high-pressure drop in some cases were demonstrated in the initial models. In order to improve performance the following features were incorporated in the most recent oxygenator model: increasing the fiber length and total surface area, decreasing the packing density, and modifying the flow distributor. The aim of this study was to evaluate the gas transfer performances and biocompatibility of this newly improved model with in vitro experiments. According to the established method in our laboratory, in vitro studies were performed using fresh bovine blood. Gas transfer performance tests were performed at a blood flow rate of 0.5 to 6 L/min and a V/Q ratio (V = gas flow rate, Q = blood flow rate) of 2 and 3. Hemolysis tests were performed at a blood flow rate of 1 and 5 L/min. Blood pressure drop was also measured. At a blood flow rate of 1 L/min and V/Q = 3, the O2 and CO2 gas transfer rates were 72.45 +/- 1.24 and 39.87 +/- 2.92 ml/min, respectively. At a blood flow rate of 2 L/min and V/Q = 3, the O2 and CO2 gas transfer rates were 128.83 +/- 1.09 and 47.49 +/- 5.11 ml/min. Clearly, these data were superior to those obtained with previous models. As for the pressure drop and hemolytic performance, remarkable improvements were also demonstrated. These data indicate that this newly improved oxygenator is superior to the previous model and may be clinically acceptable for long-term ECMO application.     

几丁糖联合聚乳酸薄膜预防硬膜外粘连的研究

目的 观察几丁糖加聚乳酸薄膜联合运用对椎板切除术后预防硬膜外瘢痕粘连的效果.方法 80只成年家兔[体质量(2.0±0.2)kg]制作椎板切除模型,在椎板缺损处分别覆盖等渗盐水(A组)、聚乳酸薄膜(B组)、几丁糖(C组)及几丁糖加聚乳酸薄膜(D组),术后12周对椎板切除部位进行大体观察、组织学观察及透射电镜比较各组间瘢痕形成和粘连情况.结果 B、C、D组的Rydell-Balazs粘连度评分、Nussvaum组织学评分均优于A组(P<0.01),D组优于B组和C组(P<0.01),B组与C组间差异无统计学意义(P>0.05).结论 联合应用几丁糖加聚乳酸薄膜能有效预防硬膜外瘢痕粘连,比单独应用几丁糖或聚乳酸薄膜效果好.     

几丁糖联合聚乳酸薄膜预防硬膜外粘连的研究

目的 观察几丁糖加聚乳酸薄膜联合运用对椎板切除术后预防硬膜外瘢痕粘连的效果.方法 80只成年家兔[体质量(2.0±0.2)kg]制作椎板切除模型,在椎板缺损处分别覆盖等渗盐水(A组)、聚乳酸薄膜(B组)、几丁糖(C组)及几丁糖加聚乳酸薄膜(D组),术后12周对椎板切除部位进行大体观察、组织学观察及透射电镜比较各组间瘢痕形成和粘连情况.结果 B、C、D组的Rydell-Balazs粘连度评分、Nussvaum组织学评分均优于A组(P<0.01),D组优于B组和C组(P<0.01),B组与C组间差异无统计学意义(P>0.05).结论 联合应用几丁糖加聚乳酸薄膜能有效预防硬膜外瘢痕粘连,比单独应用几丁糖或聚乳酸薄膜效果好.     

几丁糖联合聚乳酸薄膜预防硬膜外粘连的研究

目的 观察几丁糖加聚乳酸薄膜联合运用对椎板切除术后预防硬膜外瘢痕粘连的效果.方法 80只成年家兔[体质量(2.0±0.2)kg]制作椎板切除模型,在椎板缺损处分别覆盖等渗盐水(A组)、聚乳酸薄膜(B组)、几丁糖(C组)及几丁糖加聚乳酸薄膜(D组),术后12周对椎板切除部位进行大体观察、组织学观察及透射电镜比较各组间瘢痕形成和粘连情况.结果 B、C、D组的Rydell-Balazs粘连度评分、Nussvaum组织学评分均优于A组(P<0.01),D组优于B组和C组(P<0.01),B组与C组间差异无统计学意义(P>0.05).结论 联合应用几丁糖加聚乳酸薄膜能有效预防硬膜外瘢痕粘连,比单独应用几丁糖或聚乳酸薄膜效果好.     

几丁糖联合聚乳酸薄膜预防硬膜外粘连的研究

目的 观察几丁糖加聚乳酸薄膜联合运用对椎板切除术后预防硬膜外瘢痕粘连的效果.方法 80只成年家兔[体质量(2.0±0.2)kg]制作椎板切除模型,在椎板缺损处分别覆盖等渗盐水(A组)、聚乳酸薄膜(B组)、几丁糖(C组)及几丁糖加聚乳酸薄膜(D组),术后12周对椎板切除部位进行大体观察、组织学观察及透射电镜比较各组间瘢痕形成和粘连情况.结果 B、C、D组的Rydell-Balazs粘连度评分、Nussvaum组织学评分均优于A组(P<0.01),D组优于B组和C组(P<0.01),B组与C组间差异无统计学意义(P>0.05).结论 联合应用几丁糖加聚乳酸薄膜能有效预防硬膜外瘢痕粘连,比单独应用几丁糖或聚乳酸薄膜效果好.     

几丁糖联合聚乳酸薄膜预防硬膜外粘连的研究

目的 观察几丁糖加聚乳酸薄膜联合运用对椎板切除术后预防硬膜外瘢痕粘连的效果.方法 80只成年家兔[体质量(2.0±0.2)kg]制作椎板切除模型,在椎板缺损处分别覆盖等渗盐水(A组)、聚乳酸薄膜(B组)、几丁糖(C组)及几丁糖加聚乳酸薄膜(D组),术后12周对椎板切除部位进行大体观察、组织学观察及透射电镜比较各组间瘢痕形成和粘连情况.结果 B、C、D组的Rydell-Balazs粘连度评分、Nussvaum组织学评分均优于A组(P<0.01),D组优于B组和C组(P<0.01),B组与C组间差异无统计学意义(P>0.05).结论 联合应用几丁糖加聚乳酸薄膜能有效预防硬膜外瘢痕粘连,比单独应用几丁糖或聚乳酸薄膜效果好.     

Development of an Intravenous Membrane Oxygenator: Enhanced Intravenous Gas Exchange Through Convective Mixing of Blood around Hollow Fiber Membranes

Abstract: In vitro testing of a new prototype intravenous membrane oxygenator (IMO) is reported. The new IMO design consists of matted hollow fiber membranes arranged around a centrally positioned tripartite balloon. Short gas flow paths and consistent, reproducible fiber geometry after insertion of the device result in an augmented oxygen flux of up to 800% with balloon activation compared with the static mode (balloon off). Operation of the new IMO device with the balloon on versus the balloon off results in a 400% increase in carbon dioxide flux. Gas flow rates of up to 9. 5 L/min through the 14–cm–long hollow fibers have been achieved with vacuum pressures of 250 mm Hg. Gas exchange efficiency for intravenous membrane oxygenators can be increased by emphasizing the following design features: short gas flow paths, consistent and reproducible fiber geometry, and most importantly, an active means of enhancing convective mixing of blood around the hollow fiber membranes     

几丁糖联合聚乳酸薄膜预防硬膜外粘连的研究

目的 观察几丁糖加聚乳酸薄膜联合运用对椎板切除术后预防硬膜外瘢痕粘连的效果.方法 80只成年家兔[体质量(2.0±0.2)kg]制作椎板切除模型,在椎板缺损处分别覆盖等渗盐水(A组)、聚乳酸薄膜(B组)、几丁糖(C组)及几丁糖加聚乳酸薄膜(D组),术后12周对椎板切除部位进行大体观察、组织学观察及透射电镜比较各组间瘢痕形成和粘连情况.结果 B、C、D组的Rydell-Balazs粘连度评分、Nussvaum组织学评分均优于A组(P<0.01),D组优于B组和C组(P<0.01),B组与C组间差异无统计学意义(P>0.05).结论 联合应用几丁糖加聚乳酸薄膜能有效预防硬膜外瘢痕粘连,比单独应用几丁糖或聚乳酸薄膜效果好.     

Pediatric cardiac extracorporeal membrane oxygenation in congenital heart disease: the cardiologist's view

Abstract: There is no doubt that extracorporeal membrane oxygenation (ECMO) as a powerful therapeutic modality in critically ill newborn infants and older children with congenital heart disease has implications for the pediatric cardiologist. His responsibilities as consultant in the intensive care unit include screening for unsuspected cyanotic heart disease in neonatal candidates referred for ECMO and appraisal of surgical repair in postcardiotomy patients as well as assessment of postoperative hemodynamics and detection of complications during perfusion. A close cooperation between intensive care specialists and other appropriate specialists (pediatric cardiologists, cardiac surgeons, and anesthesiologists) is required for the process of decision making prior to initiation of postoperative ECMO in the individual patient with congenital heart disease. Long-term survival, morbidity, cerebrovascular complications, and neurodevelopmental sequelae of these near miss children remain a critical issue. Furthermore, there is a strong need for professional psychosocial support of affected parents, both in the hospital and after discharge.