常用功率密度CO2激光治疗尖锐湿关疣时烟尘中HPV DAN的检测

目的：了解临床常用功率密度的ＣＯ２激光治疗尖锐湿疣术中烟尘是否含有ＨＰＶ ＤＮＡ。方法：选择皮损较多的尖锐湿疣，病程在６个月以内的患者作为研究对象，分别用５Ｗ、１０Ｗ、１５Ｗ对应功率密度下的ＣＯ２激光汽化在体疣体组织，离光斑５ｃｍ或５０ｃｍ处用吸尘器提吸烟尘于ＰＢＳ中，经固定后观察烟尘的细胞形态学，并用ＨＰＶ通用型引物作ＰＣＲ检测烟尘是否含有ＨＰＶ－ＤＮＡ。结果：三种功率密度的烟尘主要由炭化颗粒组     

He-Ne激光对大鼠离体颈上神经节后神经元膜电导的影响

应用细胞内生物电记录技术 ,观测不同功率、不同照射时间的 He- Ne激光 (脉冲频率 1Hz)对大鼠离体颈上神经节后神经元快兴奋性突触后电位 (f- EPSP)期间膜电导的影响。功率密度为 2 m W/ cm2 的 He- Ne激光在照射初期 (1min～ 2 min)引起快兴奋性突触后电位 (f- EPSP)幅值增大 ,同时膜电导增大 ;而在激光照射后期 (后 3m in～8m in)引起节后神经元膜电导减少。功率密度为 5 m W/ cm2 的 He- Ne激光照射期膜电导无明显变化 .结果表明 :功率密度为 2 m W/ cm2 的 He- Ne激光照射初期引起膜电导 (Gl=34.6± 5 .4 n S)较照射前 (Gf=2 6 .8± 6 .2 n S)有明显增大 (P<0 .0 5 ) ,照射后期膜电导减少。提示 :He- Ne激光照射可能是通过两时相效应改变节后神经元膜电导来影响交感神经节内兴奋传递过程。这可能是低功率激光对神经细胞的一种作用机制。     

倍频Nd：YAG激光对钝顶螺旋藻的诱变效应

利用倍频 Nd:YAG激光 (波长 53 2 nm,功率 50 0 m W,功率密度 1 60 m W/cm2 )诱变钝顶螺旋藻 ,辐照时间为 1 5min、1 0 min、5min通过测定藻丝形态参数、叶绿素 a、β 胡萝卜素、生长速度 ,比较倍频 Nd:YAG激光对钝顶螺旋藻生长的影响。实验结果表明 :与出发株相比 ,经倍频 Nd:YAG激光辐照后 ,藻丝形态发生变化 ,藻丝长、螺旋数、螺旋长变小 ;1 5min,1 0 min辐照组出现螺旋变松驰 ;1 0 min,5min辐照组促进藻的生长和叶绿素 a含量提高 ,使生长速度提高。三个诱变时间剂量都有利于 β 胡萝卡素积累 ,含量增幅最高达 2 2 .3 %。     

尖锐湿疣样本中HPV病毒的分子检测

调查男性和女性尖锐湿疣样本中人乳头瘤病毒（HPV）的检出率及病毒类型,为研发相关防治疫苗提供依据,以HPV 外壳蛋白DNA序列为模板设计特异引物,SSP－PCR扩增检测样本中HPV的感染率和病毒类型。收集了北京及邯郸市医院门诊尖锐湿疣样本22例,其中男性13例,女性9例。检测发现所有样本中存在着高浓度的HPV病毒DNA。男性样本中有5例感染HPV6型,6例感染HPV11型,2例为HPV6＋HPV11混合感染。女性样本中有3例感染HPV6型,2例感染HPV11型,4例为 HPV6＋HPV11混合感染。被诊断为宫颈湿疣的4位女性还在其含宫颈粘膜脱落细胞的样本中检出了HPV16、HPV18、HPV33、HPV35、HPV45、HPV54、HPV56或HPV58等高危险型病毒类型。所有检测到的HPV病毒DNA片段均TA克隆并将测定的DNA序列存入了国际基因数据库GenBank（DQ003066－DQ003079）。调查没有在单纯的男女尖锐湿疣组织块中检测到除HPV6和HPV11以外的其他HPV类型。该研究建立了灵敏可靠的HPV分子检测及分型方法,尖锐湿疣中HPV的检出率达100％。 本研究初步结果显示导致男女尖锐湿疣的HPV病毒类型没有显著差异,主要为HPV6及HPV11型。     

肛管内尖锐湿疣的治疗及HPV基因分型检测

目的:探讨降低肛管内尖锐湿疣复发率的治疗措施。方法:采用导流杂交基因芯片技术检测HPV基因分型。108例肛管内尖锐湿疣患者初次治疗为CO_2激光治疗去除疣体后,给予5-氨基酮戊酸光动力疗法(ALA-PDT),每周1次,共3次。末次治疗后随访6个月,复发患者再次CO_2激光治疗去除疣体后,给予ALA-PDT治疗,每周1次,共10次,结束后外用咪喹莫特乳膏,每周3次,共12周,随访6个月,观察临床疗效和复发率。结果:108例尖锐湿疣患者中HPV阳性者81例,阴性者27例,HPV感染率75.0%。一重HPV感染51例,占47.2%,一重感染中HPV 6型为30例,阳性检出率为27.8%,是主要的感染型别,HPV 11型21例,阳性检出率为19.4%。多重HPV感染30例,阳性检出率27.8%。所有患者经初次治疗后痊愈率为79.6%,有效率为100.0%,复发率为22.1%;对复发患者进行再次治疗后,痊愈率为52.7%,有效率为100.0%,复发率为20.0%,两次治疗之后总的复发率为2.6%(2/77)。结论:肛管内尖锐湿疣组织感染的HPV基因型别主要为HPV6、HPV11及HPV6+HPV11。CO_2激光联合ALA-PPT及外用咪喹莫特软膏治疗肛管内尖锐湿疣,可显著降低肛管内尖锐湿疣的复发率。     

应用树状DNA杂交(DDH)对生殖道尖锐湿疣中HPV DNA的分型检测

从手术切除的50例生殖道尖锐湿疣新鲜标本中,以及15例正常人血清中,提取基因组DNA,同时用树状DNA杂交(dendrimer DNA hybridizalion,DDH)技术和PCR进行HPV DNA的分型检测.结果50例尖锐湿疣中,以DDH方法检测,感染HPV6型者20例,感染11型者24例,6/11型混合感染者3例,阴性3例,总检测率达94%;以PCR方法检测,HPV6型感染者21例,11型感染者24例,6/11型混合感染者3例,阴性2例,总检测率为96%.15例正常人血清中,以DDH方法检测,HPV感染的假阳性率为0%;以PCR检测,假阳性率为6.67%.还以HPV阳性标本对DDH方法做了敏感度的测定,结果阳性病例DNA检测最低浓度为97.28pg/ml.研究表明,DDH技术具有较高敏感性和高特异性,且成本较低,操作安全简便,可适用于基层中小医院较大样本量筛查.     

UVB照射后小鼠脾淋巴细胞DNA损伤程度的检测

用强度为30 μW/cm2、60 μW/cm2、90 μW/cm2、120 μW/cm2、150 μW/cm2的中波红斑效应紫外线(UVB)分别照射小鼠脾淋巴细胞5 min、15 min、30 min,采用单细胞凝胶电泳法(SCGE)检测UVB照射对细胞DNA的损伤,结果照射5 min和15 min时,DNA损伤程度与照射强度呈正相关;而照射30 min时,30 μW/cm2组DNA损伤程度最高,60 μW/cm2、90 μW/cm2、120 μW/cm2组损伤程度有所降低,而150 μW/cm2组又出现了高DNA损伤.认为通过SCGE来检测UVB照射对小鼠脾淋巴细胞的损伤程度,可广泛应用于环境毒理学中对DNA具有损伤作用的分析,为环境生物学研究辐射对生物体的影响提供借鉴.     

人乳头瘤病毒11型L1主要衣壳蛋白的B细胞表位多肽作为疫苗的研究

分析了人乳头瘤病毒11型(HPV11)L1主要衣壳蛋白的B细胞优势表位,并以此为基础研制表位多肽疫苗。研究中采用Goldkey和．PC／Gene软件系统,分析HPV11的L1主要衣壳蛋白的二级结构、抗原性、B细胞表位,并引人氨基酸抗原性指数,综合评估其B细胞优势表位。Fmoc固相合成表位多肽,高效液相层析方法纯化,毛细管电泳分析其纯度。与0．2ml佐剂完全乳化后,按50μg／只的剂量免疫小鼠,进行动物水平的免疫效果评价。取免疫小鼠血清,与HPV11 DNA阳性的尖锐湿疣患者的疣体上清液结合,鉴定免疫后小鼠所产生抗体的特异性。发现HPV11的L1主要衣壳蛋白的第426～439位和第487～501位具有较高的免疫原性,可明显诱导小鼠血清抗体滴度升高,且该抗体与人尖锐湿疣的疣体组织上清液呈阳性反应。说明所选这两个肽段为HPV11的L1主要衣壳蛋白的B细胞优势表位,但是否具有功能特异性,尚需进一步研究。     

激光辐照花生种子的细胞学效应

本试验考察了用Ar (488 nm)激光辐照对花生种子胚轴细胞形态结构以及根尖细胞染色体的影响。研究结果表明:在辐照功率密度为5.12 W/cm2,时间为10 s、25 s、55 s情况下,胚轴细胞均有不同程度的损伤;且均能诱发根尖细胞染色体畸变。处理时间延长,细胞破坏加剧,主要表现在细胞中膜系统、脂体、蛋白体和细胞核,出现了异形、合并或结构混乱等现象。染色体畸变类型也随之增加。各激光处理剂量均对花生种子萌发产生抑制作用。     

用光敏生物素标记探针检测宫颈癌、尖锐湿疣及假性湿疣组织中HPV DNA

人乳头瘤病毒(简称HPV)能引起人皮肤及粘膜的多种良恶性肿瘤,特别多见于女性生殖系统如宫颈癌、尖锐湿疣和假性湿疣。但由于HPV尚不能在体外细胞培养中增殖,因此目前对其研究主要用核酸杂交和PCR方法,检测HPV DNA。本文采用光敏生物素标记HPV16、HPV18 DNA探针,分别对宫颈癌组织进行斑点杂交,检测宫颈癌组织中HPV16、18DNA并以正常宫颈组织做对照。以同样方法用HPV6b DNA探针检测尖锐湿疣及假性湿疣组织中HPV6b DNA。结果如下:18例宫颈癌组织中仅有1例与HPV16 DNA探针杂交阳性(5.7%);10例与HPV18杂交刚性(55.6%);4例为HPV16+18混合杂交阳性(22.2%)。12例尖锐湿疣组织中8例与HPV6b DNA杂交阳性(66.7%);12例假性湿疣组织中有6例与HPV6b探针杂交阳性(50%)。10例正常宫颈组织对照均为阴性。     

Characteristics of the membranes of the pellicle of the ciliatePseudomicrothorax dubius

Summary The membranes of the pellicle of the ciliatePseudomicrothorax dubius are investigated using thin section electron microscopy and freeze-fracture replicas. The plasma membrane is covered by a surface coat and is connected to the outer alveolar membrane by short, sometimes branched, bridges. The inner alveolar membrane is coated on both sides. The epiplasm lies in intimate contact with the cytoplasmic surface of this membrane, and there is a corresponding deposit on the other surface. This deposit is regularly striated.The epiplasmic layer and the alveoli are interrupted at sites of cytotic activity,e.g., the attachment sites of trichocysts, the cytoproct, and the parasomal sacs. The striated deposit ends where the epiplasm ends, indicating a direct relationship between these two epimembranous layers.There is a deposit along the sides of the first part of the tip of the trichocysts, and in this region the trichocyst membrane is free of intramembranous particles.The membrane of the parasomal sacs has a coat on both surfaces. That on the extraplasmic surface is similar to the surface coat of the plasma membrane. The origin of the cytoplasmic coat is unknown. The cytotic activity of these sacs is indicated by their highly irregular profiles.     

The early development of the tail and the transformation of the shape of the nucleus of the spermatid of the domestic fowl,Gallus gallus

Summary The differentiation of the spermatid, especially in reference to the formation of the flagellum, and transformation of the shape of the nucleus was investigated in the domestic fowl.In the early stage of the spermatid, a prominent Golgi apparatus appears around the centrioles. The Golgi vesicles then surround the axial-filament complex which develops from the distal centriole. These vesicles fuse to form continuous membrane at the earliest stage of flagellar formation, and in the succeeding stage Golgi lamellae are attached to the plasma membrane of the developing flagellum. From these observations, it is assumed that Golgi apparatus may be a source of the membrane system of the flagellum.The microtubules distributed around the nucleus form the circular manchette. The anterior region of the nucleus with the manchette is cylindrical in shape and the posterior region without it remains irregular in shape. When the circular manchette has been completed, the whole nucleus acquires a slender cylindrical shape. The circular manchette then changes into the longitudinal manchette. The nuclei of spermatids without a longitudinal manchette are abnormal in shape. In view of these observations it is assumed that the nuclear shaping of the spermatid may be accomplished by circular manchette and the maintenance of shape of the elongated nucleus by longitudinal manchette.The authors wish to thank Mr. Takayuki Mori for his helpful suggestions and technical advices     

Characterization of the composition of the aqueous humor and the vitreous body of the eye of the frog Rana temporaria L

This study aimed to analyze the aqueous humor (AH) and the vitreous body (VB) of the eye of the adult frog Rana temporaria L. as a representative species of amphibians, which lead a semi-terrestrial life. The presence of collagen, albumin, uric acid and electron donors was shown in both media; however, there are slight differences in their concentrations. To determine collagen, a spectral-fluorescent probe, cyanine dye, was used. The presence of collagen in AH of the frog was found at the first time. The total content of electron donors (ascorbic and uric acids, tryptophan, and tyrosine) in VB and HA was roughly estimated at ~ 1.5 × 10^− 4 mol/L. Both VB and AH absorb light in similar UV regions. The total protein and albumin contents in AH were found to be somewhat higher than those in VB. The uric acid content was at an equally low level in both intraocular media. It is supposed that the similarity of VB and AH compositions shown in this work is due to some exchange between VB and AH contents in the course of accommodation. The role of intraocular fluids in physiological functions of the eye and in protecting the retina against UV light is discussed.     

Observations on the permeability of the choriocapillaris of the eye

Summary The choriocapillaris is a fenestrated capillary bed located posterior to the retinal pigment epithelium. It serves as the main source of supply to the photoreceptors, retinal pigment epithelium, and other cells of the outer retina. The permeability of these capillaries to intravenously injected ferritin (MW — approx. 480,000; mol. diam. 11 nm) was examined in the mouse, rabbit, and guinea pig, each of which is characterized by a different type of retinal vascularization. In all three species, the bulk of the ferritin remained in the capillary lumina, where it appeared to be blocked at the level of the diaphragmed fenestrae. Some ferritin was present in endothelial cell vacuoles. The results confirm previous work on the rat choriocapillaris and indicate that the barrier function of the choriocapillary endothelium is present even among species in which the retinal circulation differs significantly.Supported by NIH grant EY03418