角质细胞生长因子2对细胞生长、移行及创伤愈合的作用

角质细胞生长因子2(KGF-2)是成纤维细胞生长因子超家族的一员,由间质细胞合成并分泌,能特异促进上皮细胞增殖、分化与迁移,对脊椎动物多种组织和器官的发育起重要调控作用．通过PCR从人的肾组织cDNA文库中克隆分离获得了KGF-2的cDNA,表明该因子在成人肾中有表达．采用大肠杆菌表达并纯化重组蛋白用于生物学功能研究的结果显示：KGF-2在体外不仅能够促进角质细胞的生长和增殖,而且对其凋亡具有抑制作用,还对细胞的移行具有影响．在动物实验中,KGF-2能促进皮肤切除产生的伤口愈合,提示该蛋白质可以作为创伤治疗或辅助用药的候选分子．     

转基因红花中角质细胞生长因子KGF-1的表达

通过构建重组表达质粒载体p139035S-KGF1和根癌农杆菌介导在红花（Carthamus tinctorius）中表达角质细胞生长因子（KGF-1）。从侵染到诱导生根共需要14周,转化率达0.1%。红花子叶在潮霉素筛选培养基上培养4-5周后便可获得丛生芽,再生芽移入含潮霉素的伸长生根培养基,培养4-8周可诱导生根。通过PCR、Southern blot、RT-PCR及Western blot检测证明目的基因KGF-1已经整合到红花细胞的染色体中,实现了KGF-1外源蛋白在红花中的成功表达,为开发KGF-1蛋白新的生产途径奠定了基础。     

人角质细胞生长因子2真核表达载体的构建及功能验证

目的:构建人角质细胞生长因子2(KGF-2)基因真核表达载体,探索KGF-2的上皮细胞迁移功能。方法:以人乳腺cDNA文库为模板,PCR扩增KGF-2基因片段,将其插入pXJ-40-myc,经双酶切和测序验证后,将重组质粒转染HEK-293T细胞,采用Western印迹检测重组蛋白;转染人肠上皮细胞FHC,采用细胞划痕实验检测细胞迁移能力。结果:PCR扩增获得627 bp的DNA产物,插入pXJ-40-myc载体,双酶切及测序结果证明重组质粒含有目的序列;转染HEK-293T细胞,Western印迹检测到相对分子质量约23×103的目的蛋白;细胞划痕实验显示,转染Myc-KGF-2的FHC细胞较未转染或转染空载体的细胞迁移能力强。结论:构建了人KGF-2基因真核表达载体,并验证了其促进细胞迁移的功能。     

转基因红花中角质细胞生长因子KGF-1的表达

通过构建重组表达质粒载体p139035S-KGF1和根癌农杆菌介导在红花(Carthamus tinctorius)中表达角质细胞生长因子(KGF-1)。从侵染到诱导生根共需要14周, 转化率达0.1%。红花子叶在潮霉素筛选培养基上培养4–5周后便可获得丛生芽, 再生芽移入含潮霉素的伸长生根培养基, 培养4–8周可诱导生根。通过PCR、Southern blot、RT-PCR及Western blot检测证明目的基因KGF-1已经整合到红花细胞的染色体中, 实现了KGF-1外源蛋白在红花中的成功表达, 为开发KGF-1蛋白新的生产途径奠定了基础。     

皮肤局部应用维甲酸引致的表皮增殖机制

维甲酸（retinoic acid,学名：视黄酸）在皮肤局部给药时,常刺激皮肤基底层角质细胞的异常增殖。证据表明,位于皮肤表皮基底层上层角质细胞中的维甲酸受体二聚体对维甲酸引起的基底层细胞增殖是必需的。可能的机制是,位于表皮基底层上层的维甲酸受体首先诱导位于同一细胞层中的肝素结合表皮生长因子（HB—EGF）表达上调,该生长因子通过旁分泌途径激活位于基底层的受体（erbB1和erbB2）,刺激基底层细胞增殖。     

肺部铜绿假单胞菌感染前后肺组织角质细胞生长因子的变化研究

目的观察与分析肺部铜绿假单胞菌（PAE）感染前后患者肺组织角质细胞生长因子（KGF）的变化情况,并研究其临床意义。方法将2010年12月到2013年12月于沈阳军区总医院接受治疗的60例肺炎患者作为研究对象,分别于其感染前后收集肺组织标本,检测感染前后患者肺组织角质细胞生长因子的表达水平。结果在感染PAE后,患者肺组织角质细胞生长因子蛋白表达水平呈明显上升的态势,并于感染3 d后到达峰值。相较感染铜绿假单胞菌前,患者肺组织角质细胞生长因子蛋白表达水平明显提升,感染前后对比差异有统计学意义（P〈0.05）。结论在急性肺损伤的诊断与治疗中,重视对患者肺组织KGF蛋白表达水平的研究,及时实施KGF辅助干预,对抑制感染,控制肺炎的进展有十分重要的作用。     

穿膜肽KGF-2的构建及其对HaCaT增殖作用的影响

目的:构建穿膜肽KGF-2(TAT-KGF-2),研究其对人角质形成细胞HaCaT的增殖作用。方法:采用普通PCR技术和特殊引物从人胚胎肺成纤维细胞cDNA中扩增出目的基因TAT-KGF-2,并将其插入pET-28a(+)载体中构建pET28a-TAT-KGF-2重组质粒;将该质粒转化大肠杆菌Rosetta(DE3),IPTG诱导表达重组蛋白TAT-KGF-2;通过Ni-NTA柱纯化获得目标重组蛋白并利用Western blot进行鉴定;MTT法研究TAT-KGF-2对HaCa的增殖作用。结果:成功构建pET28a-TAT-KGF-2重组质粒,经序列比对,与GenBank上公布的人KGF-2基因序列同源性达到100%;获得穿膜肽KGF-2蛋白,分子量约为19 kDa,纯度达到95%以上;TAT-KGF-2对HaCaT在12.5ng/ml时具有增殖作用,并呈浓度依赖性,而且与不具有穿膜功能的KGF-2相比有更强的增殖效果。结论:成功纯化出TAT-KGF-2蛋白,并验证其对HaCaT具有增殖作用,为进一步研究该蛋白的结构和功能奠定了基础。     

人的原代上皮角质形成细胞和永生化的上皮角质形成细胞HaCaT的增殖能力的比较

目的：筛选适合充当组织工程皮肤的种子细胞,比较原代培养的人原代上皮角质形成细胞和永生化的上皮角质形成细胞HaCaT的增值能力。方法：将两种细胞（小儿包皮环切术后的组织培养表皮角质形成细胞,永生化的上皮角质形成细胞HaCaT）,分别接种于96孔板,通过MTT检测细胞1,3,5,7,9,11天的生长情况;当两种细胞融合至60%时分别取1×106个细胞,通过流式细胞检测细胞的周期。结果：永生化的上皮角质形成细胞HaCaT每隔一天即可传代一次,原代上皮角质形成细胞每3天传代一次;细胞周期：永生化上皮角质形成细胞HaCaT的G1期和S期的比例高于原代上皮角质形成细胞。生长曲线：MTT检测两种细胞1,3,5,7,9,11的生长情况,永生化的上皮角质形成细胞HaCaT的生长速度明显高于原代上皮角质形成细胞。结论：永生化上皮角质形成细胞HaCaT的增殖能力要高于原代培养的上皮角质形成细胞。     

壳聚糖复合KGF 2突变体温敏敷料联合简易负压吸引法对压力性损伤的疗效

目的探究壳聚糖复合角质细胞生长因子(keratinocyte growth factor-2,KGF-2)突变体温敏敷料联合简易负压吸引法对长期卧床老年患者大面积压力性损伤的治疗效果。方法选择2016年1月至2018年12月我院普外科收治的90名大面积压力性损伤患者,随机分为对照组与观察组,各45例。对照组患者采用简易负压吸引联合传统泡沫敷料治疗,观察组患者在此基础上联合壳聚糖复合KGF-2突变体温敏敷料填充。比较两组患者的伤口愈合效果以及创面细菌检出率。结果观察组患者的伤口愈合总有效率(93.3%)明显高于对照组(75.6%),差异有统计学意义(χ~2=5.41,P=0.003)。观察组患者细菌检出率(8.89%)显著低于对照组(35.56%),差异有统计学意义(χ~2=3.02,P=0.003)。结论壳聚糖复合KGF-2突变体温敏敷料联合简易负压吸引法能显著促进长期卧床老年患者大面积压力性损伤创面愈合,降低交叉感染的风险,具有较好的临床应用效果。     

角质细胞生长因子-2与核糖体蛋白L22相互作用的发现及在哺乳动物细胞中的验证

通过聚合酶链式反应从人胎肝cDNA文库中钓取KGF-2 cDNA,构建诱饵蛋白载体pAS2-1-KGF-2并对其自身转录激活活性进行鉴定,利用酵母双杂交系统筛选人胎肝cDNA文库,挑选双阳性克隆.DNA序列分析和同源检索显示,所获侯选蛋白为人核糖体蛋白L22(RPL22).将KGF-2和侯选蛋白分别克隆至哺乳动物细胞双杂交的BD、AD质粒中,共同转染COS-7细胞,通过CAT分析验证了KGF-2和侯选蛋白之间的相互作用.为阐明KGF-2作用的分子机制提供有益线索.     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.