人表皮细胞与角膜上皮细胞蛋白质组的差异表达初步研究

目的：应用蛋白质组技术比较人表皮细胞与角膜上皮细胞之间的蛋白质表达差异。方法：实验于2005-11，2006-10在中山大学中山眼科中心国家眼科学重点实验室完成。分别培养人表皮细胞与角膜上皮细胞，裂解原代细胞抽提总蛋白，精确定量后进行双向电泳，扫描电泳凝胶获得二维总蛋白图谱。图谱使用软件辅助比较分析，找出差异表达点，从凝胶中抠取差异点，胶内酶解后用基质辅助激光解吸附，电离飞行时间串级质谱法鉴定差异蛋白。结果：人表皮细胞与角膜上皮细胞蛋白质凝胶分析获得600余个清晰的蛋白质斑点，在比较分析出的26个差异点中初步鉴定出4个差异表达蛋白，分别是细胞内氯离子通道1、Psodasin、乳酸脱氢酶B和丝氨酸蛋白酶抑制因子。结论：人表皮细胞与角膜上皮细胞在蛋白质表达上有较高的相似性以及较好的可比性，为探索表皮细胞横向分化成角膜上皮细胞的分子机制做出了有益的尝试，并提供了具有一定可操作性的实验方法。     

腰椎定点牵压与硬膜外隙注药疗法单独或联合应用治疗腰椎间盘突出症

目的:对比观察硬膜外隙注药与腰椎定点牵压疗法及其联合应用治疗腰椎间盘突出症的疗效。方法:①选择2004-01/2006-01解放军总医院康复医学科门诊诊治的腰椎间盘突出症患者180例,男125例,女55例,年龄20～65岁。患者对治疗方案知情同意。按随机数字表法将患者分为3组:硬膜外隙注药组、腰椎定点牵压疗法组、联合治疗组,每组60例。硬膜外隙注药组:骶管注射利多卡因注射液、胞二磷胆碱、维生素B12、地塞米松混合液,每5d注射1次,共4次。腰椎定点牵压疗法组:采用胸带与下肢固定带牵引,待牵引床启动逐渐使患者腰脊柱拉伸时,术者双手拇指关节突关节连线,由上腰段向腰骶段滑行推压,当拇指推压到病变间隙时,牵引力须达到患者体质量1.5倍左右,迅速向脊柱前方施压。共2次。联合治疗组为两种疗法联合应用。每2次硬膜外隙注药后施行腰椎定点牵压疗法疗法1次,共2次。②于治疗前和治疗后3,6个月采用疼痛强度评分评估疼痛程度(0～10分,0分为无痛,10分为最痛),治疗前和治疗后3个月观察临床体征和评估疗效,疗效评估依据胡有谷的腰椎间盘突出症和国家中医药管理局(1994年)制定的中医病症诊断疗效标准。③计量和计数资料差异比较分别采用t检验和χ2检验。结果:腰椎间盘突出症患者180例均进入结果分析。①疼痛强度变化:治疗后3个月3组疼痛强度评分均较治疗前降低,其中联合治疗组与治疗前比较,差异明显(χ2=2.13,P<0.01)。治疗后6个月,各组病例的疼痛症状大多数获得控制,其中联合治疗组疼痛强度评分与治疗前比较,差异明显(χ2=4.03,P<0.01),联合治疗组和硬膜外隙注药组疼痛强度评分明显低于腰椎定点牵压疗法组(χ2=5.62,6.16,P<0.05)。②临床体征变化:各组治疗后3个月4项体征均较治疗前改善,其中联合治疗组直腿抬高试验阳性患者数明显少于硬膜外隙注药组和腰椎定点牵压疗法组(7,19,14例,χ2=9.24,9.14,P<0.01)。③疗效:联合治疗组治疗有效率明显高于其他硬膜外隙注药组和腰椎定点牵压疗法组[100%(60/60),88%(53/60),92%(55/60),χ2=6.26,6.04,P<0.01],而其他2组间比较,差异不明显(χ2=8.63,P>0.05)。结论:硬脊膜外注药及腰椎定点牵压疗法均是治疗腰椎间盘突出症的有效疗法,联合应用疗效更好。     

Blood usage in lung transplantation

BACKGROUND: Few published data are available regarding perioperative blood usage in lung transplantation. STUDY DESIGN AND METHODS: The medical records of all patients undergoing lung transplantation at a university medical center in 1994 and 1995 were reviewed. RESULTS: Ninety patients underwent lung transplantation during this period. Six patients were excluded: two received a living related-donor lung, three underwent retransplantation and one underwent concomitant repair of a tetralogy of Fallot. Of the 84 evaluable patients, 59 underwent single lung transplantation and 25 double lung transplantation. Double-lung recipients used more red cells (6.4 vs. 1.7 units, p = 0.0002) and were more likely to receive red cells, platelets, plasma, or any component (92 vs. 32%, p< or =0.0001) than were single-lung recipients. Double- lung recipients were more likely to require cardiopulmonary bypass (40 vs. 12%, p = 0.003), and cardiopulmonary bypass was associated with greater transfusion requirements (p< or =0.0001). However, among patients requiring cardiopulmonary bypass, blood use did not differ between those undergoing double lung transplantation and those undergoing single lung transplantation. In the subset of patients not requiring cardiopulmonary bypass, double-lung recipients received more red cells (4.5 vs. 0.7 units, p< or =0.0001) and more plasma (2.0 vs. 0.2 units, p = 0.006). CONCLUSION: Double-lung recipients require more perioperative transfusions than single-lung recipients. The greater transfusion requirement is due to the more frequent need for cardiopulmonary bypass as well as the greater complexity of the procedure. These data are useful for developing surgical blood ordering guidelines for lung transplantation.     

CENTRAL NEUROGENIC NEUROPROTECTION: PROTECTION OF BRAIN FROM FOCAL ISCHEMIA BY CEREBELLAR STIMULATION

Electrical stimulation of the cerebellar fastigial nucleus (FN) elevates regional cerebral blood flow (rCBF) independently of cerebral metabolism (rCGU) throughout brain. One hour of FN stimulation also reduces, by up to 50%, the volume of the focal ischemic infarction produced by occlusion of the middle cerebral artery in rat. Protected areas correspond to the ischemic penumbra. Neuroprotection, while reversible, persists for weeks after 1h of stimulation. It cannot be attributed to increasing rCBF and/or reducing rCGU to improve matching of flow and metabolism. Conditional stimulation of FN initiates long-lived inhibition of expression of peri-infarction depolarizing waves, possibly by altering potassium-channel function and suppresses induction of inducible nitric oxide synthase (iNOS) and ICAM in cerebral microvessels. The brain contains intrinsic networks which may protect the brain from ischemic injury, possibly by producing widespread and longterm suppression of electrical excitability and/or and expression of proinflammatory molecules.     

A rapid method for harvesting autologous red cells from patients with hemoglobin S disease

Red cells from patients with sickle cell disease (HbSS and HbSC) are more resistant to lysis in hypotonic NaCl solutions than normal (HbAA) red cells. Taking advantage of this inherent resistance to osmotic stress, patient red cells (HbSS or HbSC) were rapidly isolated from donor red cells (HbAA or HbAS) by washing with hypotonic (0.3%) NaCl. The hypotonic method of washing provides a previously unavailable means for obtaining autologous HbSS or HbSC red cells from samples containing transfused donor red cells. Once isolated, these red cells can be used for phenotyping, autoadsorption, or evaluation of positive direct antiglobulin tests.     

Van der Woude syndrome: dentofacial features and implications for clinical practice

Background:  Van der Woude syndrome (VWS) is the most common clefting syndrome in humans. It is characterized by the association of congenital lower lip fistulae with cleft lip and/or cleft palate. VWS individuals have a high prevalence of hypodontia. Although caused by a single gene mutation, VWS has variable phenotypic expression. This study aimed to describe the range of clinical presentations in 22 individuals with VWS to facilitate its diagnosis.
Methods:  A retrospective study of 22 patients with a diagnosis of VWS was undertaken at the Australian Craniofacial Unit (ACFU) in Adelaide. Three extended families with affected members were included in the study cohort.
Results:  The overall prevalence of lip pits in this study cohort was 86%. Cleft phenotypes included bilateral cleft lip and palate (32%); unilateral cleft lip and palate (32%); submucous cleft palate (23%); and isolated cleft hard and soft palate (9%). Missing permanent teeth were reported in 86% of affected individuals.
Conclusions:  Submucous cleft palate in VWS may go undiagnosed if the lower lip pits are not detected. Associated hypodontia and resultant malocclusions will also require management by a dental team.     

The impact of cytokines on the expression of drug transporters,cytochrome P450 enzymes and chemokine receptors in human PBMC

Background and purpose:

The function of transporters in peripheral blood mononuclear cells (PBMC) has been characterized, but less is known about cytochrome P450 (CYP) enzyme function in these cells. Given that cytokines are dysregulated in many diseases, the purpose of this work was to assess the impact of cytokines on the expression of CYPs, transporters and chemokine receptors in PBMC.

Experimental approach:

Human PBMC were incubated with cytokines for 48 h. ATP-binding cassette (ABC)B1, ABCC1, ABCC2, CYP2B6, CYP3A4, CXCR4 and CCR5 expression were measured by quantitative polymerase chain reaction and flow cytometry at 0, 4, 8, 24 and 48 h. Enzyme activity was assessed using fluorescent probes.

Key results:

We show here functional activity of CYP3A4 and CYP2B6 in PBMC. Furthermore, cytokines had a significant impact on the mRNA and protein expression of all proteins. For example, interleukin-2 (IL-2) had a marked impact on ABCB1 mRNA (% control 4745 ± 11961) and protein (% control 200 ± 57). Increases in drug efflux transporter expression, in response to cytokines, resulted in reduced cellular accumulation of digoxin [decrease of 17% and 26% for IL-2 and interferon-γ (IFNγ) respectively] and saquinavir (decrease of 28% and 30% for IL-2 and IFNγ respectively). The degree to which drug transporter and chemokine receptor expression changed in response to cytokines was positively correlated (e.g. ABCB1 and CXCR4, r² = 0.545).

Conclusions and implications:

These data have important implications for diseases in which cytokines are dysregulated and for which pharmacological intervention targets immune cells.     

Stage T1 glottic carcinoma: results of radiation therapy or laser excision

A retrospective analysis was made of the data on 60 patients with stage T1 glottic carcinoma (43 T1a, 17 T1b) who received radiation therapy and 17 patients with T1a disease who underwent laser excision as the primary treatment modality. Patients who received radiation therapy achieved 3- and 5-year actuarial local control rates of 92% and 89% for T1a and 77% and 77% for T1b disease, respectively. In patients who underwent laser excision (all with T1a disease), the 3-year local control rate was 77%. Of the 42 evaluable irradiated T1a patients, 31 (74%) had a normal to near-normal voice, eight (19%) had mild or intermittent hoarseness, and three (7%) had persistent hoarseness. Of the 13 evaluable patients in the laser-excision group, four (31%) had a normal to near-normal voice, five (38%) had mild or intermittent hoarseness, and four (31%) had persistent hoarseness. The difference in the quality of voice between these two groups is statistically significant (P = .012), although the ultimate local control rate after salvage therapy for irradiated patients (97%) was similar to that for laser-excision patients (94%).