胰腺癌B7-H4与相关血清肿瘤标志物的预后价值研究

目的:分析胰腺癌患者肿瘤组织B7-H4表达水平与术前血清中癌胚抗原(CEA)、糖类抗原(CA)199及CA242等肿瘤标志物表达水平的相关性,同时探讨其在胰腺癌中的临床应用价值。方法:采用抗人B7-H4单克隆抗体免疫组化染色方法测定188例胰腺癌患者手术切除组织中B7-H4表达水平。同时利用电化学发光法检测术前血清中CEA、CA199及CA242浓度。结果:胰腺癌患者术前血清CEA水平与胰腺癌原发灶、淋巴结及远端转移分期(TNM)及远端转移相关(P<0.05)。胰腺癌患者术前血清CA199及CA242水平与胰腺癌患者年龄及远端转移相关(P<0.05)。CA199与术后生存期相关(P<0.05)。胰腺癌患者肿瘤组织B7-H4表达水平与血清CEA表达存在显著正相关(P<0.05),与其他肿瘤标志物之间无显著相关性(P>0.05)。在胰腺癌患者术后6个月生存的预测分析中,B7-H4染色积分的曲线下面积(AUC)明显高于CEA和CA242(P<0.05);B7-H4染色积分联合术前血清CA199的AUC明显高于术前血清CA199(P<0.05),而与B...     

血清p185蛋白及VEGF-C水平在乳腺癌诊断中的应用及生存质量影响分析

目的:探讨免疫PCR检测的预测应用,检测血管内皮生长因子-C(VEGF-C)的表达水平及意义,以及乳腺癌患者生存质量的主要影响因素并总结研究方法。方法:本院2007-05/2010-04收治的136例乳腺癌患者(实验组)的临床资料进行了回溯式分析并总结研究,并与同期收治的132例乳腺良性肿瘤患者(对照组)的生存质量进行比较。结果:两组患者血清p185蛋白的免疫检测发现,p185蛋白阳性表达对乳腺癌的临床诊断及预后存在统计学意义。血清p185阳性患者80.5%可发生复发或转移,显著高于阴性组的12.5%(P<0.05)。实验组患者术前血清VEGF-C的水平显著高于对照组患者(P<0.05),但治疗1个月后对照组VEGF-C的水平较治疗前有显著降低(P<0.05)。结论:检测血清P185蛋白以及VEGF-C的水平对于乳腺癌患者的诊断、治疗及预后判断均有显著的重要意义。p185蛋白阳性患者易复发,且VEGF-C的水平术后降低越多者治疗效果越显著,生存质量越好。     

联合检测HIF-1α、ET-1、VEGF、CEA、CA153在乳腺癌转移及预后评估中的作用

目的研究联合检测HIF-1α、ET-1、VEGF、CEA、CA153在乳腺癌转移及预后评估中的作用。方法选取2013年11月至2015年10月在我院收治的84例乳腺癌患者为试验组,以及84例乳腺良性肿瘤患者作为对照组,对比分析两组患者的HIF-1α、ET-1、VEGF、CEA、CA153水平,以及与肿瘤转移和复发的关系,并分析联合检测乳腺癌患者HIF-1α、ET-1、VEGF水平对肿瘤是否转移和复发的诊断效能。结果乳腺癌患者的HIF-1α、ET-1、VEGF、CEA、CA153水平均显著高于乳腺良性肿瘤患者(P<0.05),肿瘤最大径>3cm、乳腺癌转移、复发的患者HIF-1α、ET-1、VEGF水平显著高于非转移、复发的患者(P<0.05),Logistic多因素回归模型表明乳腺癌患者的HIF-1α、ET-1、VEGF水平等均与肿瘤是否转移和复发存在正相关性,HIF-1α、ET-1、VEGF水平及联合检测诊断是否转移和复发均大于0.6,具有较好的检测价值,并且联合检测的ROC曲线下面积高于各单项检测,差异具有统计学意义(P<0.05)。结论乳腺癌患者的HIF-1α、ET-1、VEGF水平与肿瘤的转移和复发存在显著的正相关性,联合检测诊断肿瘤的转移和复发具有较好的检测效能,对临床的针对性治疗具有参考作用。     

血清TSH、TSGF与p185蛋白联合诊断乳腺癌的临床意义

目的探讨血清促甲状腺激素(TSH)、肿瘤特异性生长因子(TSGF)水平、p185蛋白阳性率与乳腺癌疾病的相关性。方法选择2016年8月至2018年10月应城市人民医院收治的60例乳腺癌患者作为研究对象,按照TNM分期分为Ⅰ-Ⅱ组(Ⅰ-Ⅱ期患者)36例和Ⅲ-Ⅳ组(Ⅲ-Ⅳ期患者)24例,另选取我院同期健康体检人群60例作为对照组。对比分析3组患者血清TSH、TSGF水平及p185蛋白阳性率及不同病理特征下患者TSH、TSGF水平及p185蛋白阳性率之间的差异,分析联合检测和单独检测之间的差异。结果3组间TSH、TSGF水平及p185蛋白阳性率比较,差异有统计学意义(P<0.05)。与对照组相比,Ⅰ-Ⅱ组和Ⅲ-Ⅳ组患者血清中TSH、TSGF水平及p185蛋白阳性率更高,差异有统计学意义(P<0.05)。不同组织分级、TNM分期以及淋巴结转移情况的乳腺癌患者的TSH、TSGF水平及p185蛋白阳性率比较,差异有统计学意义(P<0.05);而不同肿瘤直径和分子分型的乳腺癌患者的TSH、TSGF水平及p185蛋白阳性率比较,差异无统计学意义(P>0.05)。Spearman相关性分析显示,组织分级、TNM分期、淋巴结转移、病理状态与乳腺癌患者的TSH、TSGF水平及p185蛋白阳性率呈显著相关性(P<0.05)。Logistic多因素分析结果显示,血清TSH(OR=1.019,P=0.001)、TSGF(OR=1.632,P=0.001)、p185蛋白阳性率(OR=1.089,P=0.000)是乳腺癌发生的独立危险因素。ROC诊断曲线结果显示,联合血清TSH、TSGF水平及p185蛋白阳性率的诊断效能显著高于单独指标的诊断效能(P<0.05)。结论乳腺癌患者的血清TSH、TSGF水平及p185蛋白阳性率与病理分期显著相关,且联合检测诊断效能更高,可作为诊断的依据。     

乳腺癌手术治疗后血清E-cad、CA153和VEGF水平检测的临床意义

目的探讨了乳腺癌患者手术治疗前后血清E-cad、CA153和VEGF水平的变化及临床意义.方法分别应用放射免疫分析法和酶免法对36例乳腺癌患者进行了手术治疗前后血清E-cad,CA153和VEGF水平检测,并与35名正常健康人作比较.结果手术前乳腺癌患者血清E-cad、CA153和VEGF水平均非常显著地的高于正常人组(P<0.01),手术治疗后1年复发组有明显地高于无复发组(P<0.01).结论检测乳腺癌患者手术治疗前后血清E-cad、CA153和VEGF水平的变化与患者的病情和预后密切相关,具有重要的临床价值.     

血清CA153、CEA和TPA联检在术前乳腺肿瘤鉴别诊断中的价值

目的:本文探讨血清CA153、CEA和TPA联检在术前乳腺肿瘤鉴别诊断中价值,以期为乳腺癌患者即时手术提供帮助.方法:采用放射免疫分析,检测乳腺肿瘤患者和健康人群血清CA153、CEA和TPA含量.结果:在CA153、CEA和TPA血清学检测中,术前乳腺癌组与乳腺良性肿瘤和健康人群组表达水平存在明显差异.术前乳腺癌组阳性率:CA153为63.8%,CEA为22.4%,TPA为62.1%;术前乳腺癌组联检阳性率:CA153和CEA为69.8%,CA153和TPA为87.1%,三者联检为90.5%;术前乳腺癌组检测特异性:CA153为77.9%,CA153和CEA联检为77.9%,CA153和TPA联检为71.9%,三者联检为73.4%.结论:当CA153、CEA和TPA三者联检时,术前乳腺癌组诊断阳性率显著提高,阳性组特异性无明显下降,值得临床推广应用.     

肿瘤标志物CEA,CA50和CA19-9在大肠癌诊断中的意义

目的:观察三种肿瘤标志物CEA、CA50、CA19-9在大肠癌患者血清中的表达水平,探讨其在大肠 癌诊断中的临床意义,并评价其联合检测对大肠癌检测的价值。方法:采用化学发光法测定血清CEA、CA19- 9水平,免疫放射分析法测定血清CA50水平,收集病例包括大肠癌患者94例,大肠良性疾病患者20例及正常 对照者37例。结果:三种血清肿瘤标志物水平在大肠癌组明显高于大肠良性疾病组及正常对照组(P< 0.05),而后两组之间没有明显差异。其中以CEA的灵敏度(57.4%)和特异度(85.9%)最高。采用三项联检 可提高大肠癌诊断的敏感度(62.7%)和特异度(96.5%)。三项肿瘤标志物在结肠癌中的表达水平高于直肠 癌,其表达水平与肿瘤大小有关,并在淋巴结阳性组及远处转移组中表达水平显著高于其相应对照组(P< 0.05)。各项肿瘤标志物的表达水平与大肠癌病程呈正相关(P<0.05)。结论:血清CEA、CA50、CA19-9对大 肠癌诊断、病变范围程度和生物学行为的评估有一定临床意义,联检可提高肿瘤标志物对大肠癌的诊断准确 性。     

钙离子结合蛋白S100A9在乳腺癌中表达及临床意义

目的:探讨钙离子结合蛋白S100A9在乳腺癌患者中的表达及临床意义。方法:选取39例乳腺癌患者术前、15例术后血清及10例健康女性血清,12例手术切除肿瘤组织标本,应用ELISA检测乳腺癌患者血清S100A9水平,采用免疫组织化学方法检测乳腺癌组织中S100A9的表达,分析S100A9表达与临床病理特征的关系。结果:S100A9在乳腺癌组患者血清中表达水平高于健康组(P<0.01),有淋巴结转移乳腺癌组血清S100A9表达水平高于无淋巴结组(P<0.01),乳腺癌患者术后S100A9血清表达水平降低(P<0.01)。乳腺癌患者血清中S100A9水平与患者年龄、肿瘤最大径、组织学分级和病理类型有关(P<0.01)。S100A9在乳腺癌组织中表达明显高于癌旁正常组织。结论:S100A9表达水平在乳腺癌血清和组织中明显升高,表明S100A9可作为乳腺癌患者诊断和疗效观察指标。     

乳腺癌患者手术治疗前后血清CA153、CA125、TSGF检测的临床意义

目的:探讨乳腺癌患者手术治疗前后血清CA153、CA125和TSGF水平的变化及意义.方法:应用放射免疫分析和生化法对36例乳腺癌患者进行了血清CA153、CA125和TSGF检测,并与35名正常健康人作比较.结果:手术治疗前血清CA153、CA125和TSGF水平非常显著地高于正常人组(P＜0.01),手术治疗后1个月皆已接近正常.手术治疗后6个月再测,未复发者仍属正常水平,而复发者又恢复至手术治疗前水平.结论:测定乳腺癌患者血清CA153、CA125和TSGF水平的变化对患者的治疗和预后均具有重要的临床价值.     

浸润性乳腺癌中OPN和MK蛋白的表达及临床意义

目的通过检测浸润性乳腺癌术后患者石蜡标本中骨桥蛋白(ostenopontin,OPN)和中期因子(minkine,MK)的表达,回顾性分析其与患者临床病理特征及预后的关系,为浸润性乳腺癌的治疗、预后提供有价值的生物学标志。方法应用免疫组化PV-9000两步法检测90例浸润性乳腺癌和30例乳腺增生症术后石蜡标本中OPN和MK蛋白的表达情况。结果90例浸润性乳腺癌组织中OPN和MK蛋白的阳性表达率分别为71.11%(64/90)和78.89%(71/90),明显高于乳腺增生症30.00%(9/30)和46.67%(14/30)(P0.05);OPN的表达与肿瘤的大小、腋窝淋巴结转移、术后复发及p185受体表达密切相关(P0.05),与患者TNM分期、组织学类型及分级、雌激素及孕激素受体状况无关(P0.05)。MK的表达与肿瘤的大小、腋窝淋巴结转移、术后复发、p185受体及TNM分期表达密切相关(P0.05),与患者的组织学类型及分级、雌激素及孕激素受体状况无关(P0.05)。OPN和MK蛋白的表达有相关性(r=0.274,P=0.009)。OPN和MK蛋白阳性表达组患者TTP、OS值均低于阴性表达组(P0.05)。结论OPN和MK蛋白的表达与浸润性乳腺癌的进展存在一定相关性,检测二者的表达对浸润性乳腺癌治疗及判断预后提供理论依据。     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

The universal muscular chain reaction,muscle spasm,Torsions, ruptures and extravasations. Chameleons of pathology and some manifestations of simple muscular disorders

Most bodily functions require the coordinated actions of complementary and supplementary paired muscle groups. Where this essential muscular cooperation is lacking, hollow organs may burst and others become literally screwed up, giving rise to many similar spastic diseases such as Torticollis, Twisted ovarian cyst, Torsion of the Testis, Volvulus of the intestines, Varicose Veins, Megacolon, Aortamegaly, Scoliosis, Erb's Palsy, Peyronie's Disease, Main-en-Griffe, Undescended Foot (Pes Cavus), Talipes, Strabismus. Spasm is “panenepidemic” and unclassified examples of Torsion Dystonia and Dyskinesia really are as common as debt and taxes.     

The dominant diseases of modernized societies as omega-3 essential fatty acid deficiency syndrome: Substrate beriberi

About 1900, modern food selection and processing caused widespread $\text{epidemics}$ of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a $\text{panh}$ypovitaminosis B, i.e., a mixture of $\text{substrate}$ beriberi and $\text{substrate}$ pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse $\text{endemic}$ disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for $\text{primates}$. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.     

Hypo- und Hypermagnesämie aus kardiologischer Sicht

Zusammenfassung Eine Reihe pathologischer Zustände bedingen Magnesiummangel. Zustände mit Hypermagnesämie sind ebenfalls bekannt, doch wesentlich seltener. Für den Kardiologen beachtenswert ist, daß unter Therapie mit bestimmten Diuretica bei Herzinsuffizienz, bei Herzinfarkt, Kardiomyopathie, Digitalisintoxikation und bestimmten Herzrhythmusstörungen Hypomagnesämie beobachtet wurde. Leider kann in der klinischen Routine nur ein extracelluläres Magnesiumdefizit durch Serumbestimmungen gemessen werden; über Magnesiummangel einzelner Organe kann nichts ausgesagt werden. Hinweise für Magnesiummangel geben aber neben der Messung des Serumspiegels Anamnese, klinischer Befund, bestimmte EKG-Veränderungen wie auch evtl. Hypokalämie, ein Zustand, bei dem sich oft — besonders bei Aldosteronismus — parallele Veränderungen zeigten.Tierexperimente deuten darauf hin, daß infarktähnliche Läsionen unter Magnesiummangel entstehen, doch ob Herzinfarkt beim Menschen durch Magnesiummangel ausgelöst werden kann, ist noch ungeklärt. In Leichenherzen zeigte sich im Infarktgebiet neben Calciumakkumulation signifikanter Magnesiumverlust, wobei unklar blieb, ob sich Ursache oder Folge des Infarktes widerspiegelten. Falls ein ursächlicher Zusammenhang besteht, ist er im Myokardstoffwechsel selbst zu suchen, wie bei der Alkoholkardiomyopathie, wo myokardialer Magnesiummangel zumindest als pathogenetischer Teilfaktor anerkannt wird. Andererseits versucht man aber auch Beziehungen zwischen Atherosklerose, Blutgerinnung und Hypomagnesämie herzustellen, in der Meinung, daß Magnesiummangel auch über den coronaren Pathomechanismus des Herzinfarktes wirken könnte. Sicher scheint, daß gewisse EKG-Veränderungen und Herzrhythmusstörungen durch einen irritierten Magnesiumhaushalt bedingt sein können, da sie bei Gabe bzw. Entzug von Magnesium verschwinden. Daß Magnesiummangel die Glykosidtoleranz verringert, wird tierexperimentell bestätigt. Unter Hypomagnesämie bewirkt Acetylstrophanthidin eher und länger Rhythmusstörungen als ohne, außerdem lassen diese sich durch Magnesiumgaben eliminieren. Da in gewissen Fällen spontane und digitalisinduzierte Herzrythmusstörungen durch Magnesiuminjektionen beseitigt wurden, scheint Magnesium als Therapeuticum angebracht. Einsatz verschiedener Magnesiumsalze bei Angina pectoris, degenerativen Herzerkrankungen und Herzinsuffizienz ohne geprüften und offensichtlich gestörten Magnesiumhaushalt ist fragwürdig, weil keine eindeutigen klinischen Erfolgsbeweise vorliegen. Immerhin mag es aber larvierte, durch Serumbestimmungen nicht erfaßbare Mangelzustände geben. Allgemein erscheint es aus kardiologischer Sicht ratsam, den Magnesiumhaushalt zu überwachen und in entsprechenden Fällen auszugleichen, um möglichen Myokardläsionen oder fatalen Herzrhythmusstörungen entgegenzuwirken.     

Environmental risk factors and their role in the management of atopic dermatitis

Introduction: The etiology of atopic dermatitis (AD) is multifactorial with interaction between genetics, immune and environmental factors.

Areas covered: We review the role of prenatal exposures, irritants and pruritogens, pathogens, climate factors, including temperature, humidity, ultraviolet radiation, outdoor and indoor air pollutants, tobacco smoke exposure, water hardness, urban vs. rural living, diet, breastfeeding, probiotics and prebiotics on AD.

Expert commentary: The increased global prevalence of AD cannot be attributed to genetics alone, suggesting that evolving environmental exposures may trigger and/or flare disease in predisposed individuals. There is a complex interplay between different environmental factors, including individual use of personal care products and exposure to climate, pollution, food and other exogenous factors. Understanding these complex risk factors is crucial to developing targeted interventions to prevent the disease in millions. Moreover, patients require counseling on optimal regimens for minimization of exposure to irritants and pruritogens and other harmful exposures.     

HLA genes in Amerindians from Mexico San Vicente Tancuayalab Teenek/Huastecos

Huastecos or Teenek Amerindians are presently living at North East Mexico (San Luis Potosi State). They have probably one of the most ancient culture of Mexico and Central America together with Mayas and Olmec groups with which also show close relationships. Proximity to Atlantic Ocean/Mexican Gulf originated that Spaniards had very early contact with them at about 1519 CE or before. In the present paper we have aimed to study HLA gene profile which may be useful for HLA and disease epidemiology and transplant programs in Teeneks. HLA-DRB1*04:07, -DRB1*14:06 and -DRB1*04:11 have been found in high frequency like in other Amerindian groups. High frequency typical Amerindians HLA extended haplotypes have been found, such as A*02-B*35-DRB1*04:07-DQB1*03:02; A*68-B*39-DRB1*04:07-DQB1*03:02 and A*02-B*39-DRB1*04:07-DQB1*03:02; also new haplotypes have been described, like A*02-B*52-DRB1*04:11-DQB1*03:02, A*68-B*35-DRB1*14:02-DQB1*03:01 and A*68-B*40-DRB1*16:02-DQB1*03:01. Genetic proximity is observed not only to linguistically close Mayans, but also to Mazatecans, Mixtecans and Zapotecans, who speak an altogether different languages; it shows once more that genes and languages do not correlate. This population was greatly diminished after European contact between 1500 and 1600 years CE; in fact, North and South America First Inhabitants population was brought from 80 down to 8 million people because of diseases (i.e.: measles, smallpox or influenza), slavery and war.     

'Very sore nights and days': the child's experience of illness in early modern England, c.1580-1720

Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.     

Guidelines for the Validation and Use of Immunoassays for Determination of Introduced Proteins in Biotechnology Enhanced Crops and Derived Food Ingredients

Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.     

Ultracryotomy: development and application (with examples from the yeast cytology) (author's transl)

The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.