用复合PCR检测淋球菌、沙眼衣原体和解脲支原体的感染研究

在性传播疾病中,泌尿生殖道感染的主要病原体是淋球菌、沙眼衣原体和解脲支原体,目前临床上常有多种病原体混合感染[1],其临床症状相似或无明显症状,给确诊和治疗带来不便。因此我们应用多重引物ＰＣＲ(ｍｕｌｔｉｐｌｅｘＰＣＲ),一次实验能从临床标本中检测出三种病原体...     

应用PCR-SSCP快速鉴定结核分枝杆菌复合群

通过聚合酶链反应（ＰＣＲ）－单链构象多态性（ＳＳＣＰ）技术分析结核分枝杆菌和非结核分枝杆菌临床株的１６ＳｒＤＮＡ基因。６０例临床标本中,２０例为阳性,与传统方法比较无差异。其中分型：１８例为结核分枝杆菌,２例为结核分枝杆菌和非结核分枝杆菌双重感染。２０例阴性标本中,ＰＣＲ－ＳＳＣＰ又检查出５例阳性。２０例对照标本中,３种传统方法与ＰＣＲ－ＳＳＣＰ法均检测为阴性。全部试验３ｄ报告结果。结核分枝杆菌复合群和卡介苗的图形相同以外,其它分枝杆菌的ＰＣＲ－ＳＳＣＰ电泳图谱均有差异。所以,应用ＰＣＲ－ＳＳＣＰ技术快速     

PCR技术检测性病病原体在男性泌尿生殖系感染中的意义

用ＰＣＲ技术对我院５８８例男性生殖泌尿系感染患者进行淋病Ｋ球菌（ＮＧＦ）沙眼衣原体（ＣＴ）及解脲支原体（ＵＵ）检测,结果淋球菌感染者７５例（１２．７６％）;沙眼衣原体感染者８４例（１４．２９％）;解脲支原体感染者５１例（８．６７％）。淋球菌与沙眼衣原体混和感染者２５例（４．２５％）;淋球菌与解脲支原体混合感染者９例（１．５３％）;沙眼支原体与解脲支原体混合感染者１２例（２．０４％）;未发现有三种病原体同时感染者,并对ＰＣＲ检测性病病原体的意义及性病病原体传播趋势进行讨论。     

家庭内性传播疾病感染研究

为了解性传播疾病（ＳｅｘｕａｌｌｙＴｒａｎｓｍｉｔｔｅｄＤｉｓｅａｓｅｓＳＴＤ）对家庭内传播的影响,用多聚酶链反应（ＰＣＲ）等方法检测了９４对夫妻和其中１５户家庭中１６名子女的ＳＴＤ病原体ＮＧ、ＣＴ、ＵＵ、ＨＰＶ、梅毒螺旋体和白色念珠菌。结果表明：男性不洁性史为６８．１５％而女性为４．２６％,两者间有显著差异（Ｐ＜０．０１）,但其发病率无统计上显著差异（男７６．６０％、女７３．４０％）。ＳＴＤ感染以ＣＴ感染最高,达４４．６８％,ＮＧ感染为３１．９１％,ＨＰＶ感染是１５．９６％,未检出梅毒和ＨＳＶ感染。患者中两种及以上病原感染过３５．１１％。５６．２５％的子女ＳＴＤ感染系其患ＳＴＤ父母传染的。这提示如果夫妻任何一方检出有ＳＴＤ感染,另一方须做系统监测与咨询,以便及时采取治疗和预防措施并避免子女受到感染。     

血沉标本中人巨细胞病毒DNA的检出

用聚合酶链反应（ＰＣＲ）和地高辛标记探针（ｄｉｇ－ｐｒｏｂｅ）检测临床血沉标本中人巨细胞病毒（ＨＣＭＶ）ＤＮＡ。结果表明,人群血标本中ＨＣＭＶ－ＤＮＡ携带率较高;ＰＣＲ技术较ｄｉｇ－ｐｒｏｂｅ更敏感、快速、简便,二者检出ＨＣＭＶ－ＤＮＡ阳性率分别为８３．３％和６０．３％;ＨＣＭＶ－ＤＮＡ检出率、ＨＣＭＶ－ＩｇＭ检出率与血沉值高低之间无相关性。     

多聚酶链反应在耐新青霉素Ⅰ金黄色葡萄球菌感染诊断中的应用

多聚酶链反应（ＰＣＲ）是８０年代后期发展起来的先进的体外基因复制技术,用ＰＣＲ技术检测各种病原体具有高度灵敏、特异,且简便、快速的特点。本文就ＰＣＲ在耐新青霉素Ⅰ金黄色葡萄球菌（ＭＲＳＡ）感染诊断中的应用作一综述。     

PCR检测淋球菌的探讨

多聚酶链反应（ＰＣＲ）具有灵敏度高,特异性强,快速高效的特点,在病原微生物检测领域有广阔的前景。本文对４０８例疑似淋病患者的泌尿生殖道分泌物作了ＰＣＲ检测淋球菌的探索,结果１７０例阳性,占４１．７％,而培养阳性（２１．６％）,ＰＣＲ法显著高于培养法（Ｐ＜０．０１）。     

PCR技术在三种男性性病检测中的应用

本文用聚合酶链反应ＰＣＲ技术对３６９例男性泌尿生殖道炎患者进行淋球菌（ＮＧ）,解脲支原体（ＭＰＵ）和沙眼衣原体（ＣＴ）的检测,阳性率分别为３５．５％,３０．５％和１１．６％,同时,我们对高度怀疑为淋病患者７４例及６３例非淋病性尿道炎患者进行细菌培养和ＰＣＲ的同时检测,淋病患者细菌培养阳性检测率为３１．１％,与９２-９４年间所进行的培养检测结果基本一致,均低于ＰＣＲ检测结果（７７．１％）,解脲支原体培养阳性率为１７．５％,亦明显低于ＰＣＲ结果（３０．５％）,此外,对同一患者进行ＮＧ＋ＭＰＵ和ＭＰＵ＋ＣＴＰＣＲ检测,发现约４０％的淋病患者并发ＭＰＵ感染,而ＭＰＵ和ＣＴ复合感染亦在１０％左右。上述结果说明ＰＣＲ方法在性病病原体的诊断中比细菌培养法更为可靠,特别是在多种病原体复合感染情况下更为有效。     

PCR检测HPV－18DNA

采用聚合酶链式反应（ＰＣＲ）技术,对ＨＰＶ－１８序列中引物ＨＰ＿１、ＨＰ＿２之间的片段（Ｆ）进行扩增,通过两组阴、阳性对照实验证明扩增片段的特异性。用不同Ｍｇ浓度的缓冲系统进行ＰＣＲ反应发现,缓冲系统中Ｍｇ浓度高低是影响ＨＰＶ－１８／ＨＰ＿１、ＨＰ＿２特异扩增的重要因素,高浓度Ｍｇ导致扩增特异性降低。对１７例宫颈癌组织ＤＮＡ进行ＰＣＲ检测,有９例检出Ｆ片段,其检出率是５３％,为ＨＰＶ－１８与宫颈癌的相关性提供证据。     

甲状腺良、恶性病变中增殖细胞核抗原、P_(53)蛋白与AgNOR的检测及相关性研究

本文应用ＬＳＡＢ免疫组织化学方法及ＡｇＮＯＲ技术对７３例甲状腺良、恶性病变ＰＣ－ＮＡ、Ｐ５３蛋白及ＡｇＮＯＲ进行了检测并对其相关性进行了研究。结果显示①ＰＣＮＡ表达在良、恶性病变中有显著差异（Ｐ＜０．０１）;②ＡｇＮＯＲ颗粒均数及其形态在良、恶性病变中有较显著差异;③ＰＣＮＡ表达与ＡｇＮＯＲ计数呈显著正相关性（Ｐ＜０．０１）;④Ｐ５３蛋白在良、恶性病变中的表达均呈阴性,只是在淋巴结转移灶内呈阳性表达。根据研究我们认为检测甲状腺良、恶性病变中ＰＣＮＡ及ＡｇＮＯＲ颗粒对甲状腺良、恶性病变的鉴别、预后及对临床治疗均有意义     

Synthesis and luminescence properties of a blue-emitting Sr(3.5) Y(6.5) O(2) (PO(4) )(1.5) (SiO(4) )(4.5) :Eu(2+) phosphor

A novel blue‐emitting Sr_3.5Y_6.5O₂(PO₄)_1.5(SiO₄)_4.5:Eu²⁺ phosphor was synthesized via a solid‐state reaction. Powder X‐ray diffraction (XRD) analysis demonstrated that the Sr_3.5Y_6.5O₂(PO₄)_1.5(SiO₄)_4.5 host had a hexagonal crystal structure in the space group P6₃/m and unit cell parameters a = 9.418 Å, c = 6.900 Å. The as‐prepared phosphor showed a blue emission and all the main emission peaks were located at around 466 nm for different excitation wavelengths of 297, 333 and 391 nm. The temperature dependence of the photoluminescence property was investigated in the range 20–250 °C, and the emission intensity decreased to 71% of the initial value at room temperature on increasing the temperature to 150 °C. According to the classical theory of fluorescent thermal quenching, the activation energy (ΔE) for the thermal quenching luminescence of the as‐prepared Sr_3.45Y_6.5O₂(PO₄)_1.5(SiO₄)_4.5:0.05Eu²⁺ phosphor was determined to be 0.20 eV. Copyright © 2011 John Wiley & Sons, Ltd.     

Collagen-like triple helix formation of synthetic (Pro-Pro-Gly)10 analogues: (4(S)-hydroxyprolyl-4(R)-hydroxyprolyl-Gly)10, (4(R)-hydroxyprolyl-4(R)-hydroxyprolyl-Gly)10 and (4(S)-fluoroprolyl-4(R)-fluoroprolyl-Gly)10.

For the rational design of a stable collagen triple helix according to the conventional rule that the pyrrolidine puckerings of Pro, 4-hydroxyproline (Hyp) and 4-fluoroproline (fPro) should be down at the X-position and up at the Y-position in the X-Y-Gly repeated sequence for enhancing the triple helix propensities of collagen model peptides, a series of peptides were prepared in which X- and Y-positions were altogether occupied by Hyp(R), Hyp(S), fPro(R) or fPro(S). Contrary to our presumption that inducing the X-Y residues to adopt a down-up conformation would result in an increase in the thermal stability of peptides, the triple helices of (Hyp(S)-Hyp(R)-Gly)(10) and (fPro(S)-fPro(R)-Gly)(10) were less stable than those of (Pro-Hyp(R)-Gly)(10) and (Pro-fPro(R)-Gly)(10), respectively. As reported by B?chinger's and Zagari's groups, (Hyp(R)-Hyp(R)-Gly)(10) which could have an up-up conformation unfavorable for the triple helix, formed a triple helix that has a high thermal stability close to that of (Pro-Hyp(R)-Gly)(10). These results clearly show that the empirical rule based on the conformational preference of pyrrolidine ring at each of X and Y residues should not be regarded as still valid, at least for predicting the stability of collagen models in which both X and Y residues have electronegative groups at the 4-position.     

Role of tfdC(I)D(I)E(I)F(I) and tfdD(II)C(II)E(II)F(II) gene modules in catabolism of 3-chlorobenzoate by Ralstonia eutropha JMP134(pJP4)

The enzymes chlorocatechol-1,2-dioxygenase, chloromuconate cycloisomerase, dienelactone hydrolase, and maleylacetate reductase allow Ralstonia eutropha JMP134(pJP4) to degrade chlorocatechols formed during growth in 2,4-dichlorophenoxyacetate or 3-chlorobenzoate (3-CB). There are two gene modules located in plasmid pJP4, tfdC(I)D(I)E(I)F(I) (module I) and tfdD(II)C(II)E(II)F(II) (module II), putatively encoding these enzymes. To assess the role of both tfd modules in the degradation of chloroaromatics, each module was cloned into the medium-copy-number plasmid vector pBBR1MCS-2 under the control of the tfdR regulatory gene. These constructs were introduced into R. eutropha JMP222 (a JMP134 derivative lacking pJP4) and Pseudomonas putida KT2442, two strains able to transform 3-CB into chlorocatechols. Specific activities in cell extracts of chlorocatechol-1,2-dioxygenase (tfdC), chloromuconate cycloisomerase (tfdD), and dienelactone hydrolase (tfdE) were 2 to 50 times higher for microorganisms containing module I compared to those containing module II. In contrast, a significantly (50-fold) higher activity of maleylacetate reductase (tfdF) was observed in cell extracts of microorganisms containing module II compared to module I. The R. eutropha JMP222 derivative containing tfdR-tfdC(I)D(I)E(I)F(I) grew four times faster in liquid cultures with 3-CB as a sole carbon and energy source than in cultures containing tfdR-tfdD(II)C(II)E(II)F(II). In the case of P. putida KT2442, only the derivative containing module I was able to grow in liquid cultures of 3-CB. These results indicate that efficient degradation of 3-CB by R. eutropha JMP134(pJP4) requires the two tfd modules such that TfdCDE is likely supplied primarily by module I, while TfdF is likely supplied by module II.     

Induction of in vitro flowering in Dendrobium Madame Thong-In (Orchidaceae) seedlings is associated with increase in endogenous N(6)-(Delta (2)-isopentenyl)-adenine (iP) and N (6)-(Delta (2)-isopentenyl)-adenosine (iPA) levels

We analysed the endogenous cytokinin levels of Dendrobium Madame Thong-In seedlings grown in vitro during vegetative and flowering-inductive periods. HPLC was used to fractionate the extracts and radioimmunoassay (RIA) was used for assay of zeatin (Z), dihydrozeatin (DZ), N(6)-(Delta(2)-isopentenyl)-adenine (iP) and their derivatives. Coconut water used in experiments was found to contain high level (>136 pmol ml(-1)) of zeatin riboside (ZR). Protocorms and seedlings cultured in medium with coconut water were found to contain 0.5-3.9 pmol g(-1) FW of the cytokinins analysed. Seedlings (1.0-1.5 cm) cultured in flowering-inductive liquid medium containing 6-benzyladenine (BA, 4.4 muM) and coconut water (CW, 15%) contained up to 200 and 133 pmol g(-1) FW of iP and iPA, respectively. These levels were significantly higher than all other cytokinins analysed in seedlings of the same stage and were about 80- to 150-folds higher than seedlings cultured in non-inductive medium. During the transitional (vegetative to reproductive) stage, the endogenous levels of iP (178 pmol g(-1) FW) and iPA (63 pmol g(-1) FW) were also significantly higher than cytokinins in the zeatine (Z) and dihydrozeatin (DZ) families in the same seedlings. Seedlings that grew on inductive medium but remained vegetative contained lower levels of iPA. The importance of the profiles of iP and its derivatives in induction of in vitro flowering of D. Madame Thong-In is discussed.     

Biology of Bactrocera (Zeugodacus) tau (Walker) (Diptera: Tephritidae)

The biology of the fruit fly Bactrocera tau, an important horticultural pest, was studied under laboratory conditions at 25°C and 60–70% relative humidity on Cucurbita maxima. The duration of mating averaged 408.03 ± 235.93 min. After mating, the female fly had a preoviposition period of 11.7 ± 4.49 days. The oviposition rate was 9.9 ± 8.50 eggs and fecundity was 464.6 ± 67.98 eggs/female. Eggs were elliptical, smooth and shiny white, turning darker as hatching approached, and measured 1.30 ± 0.07 mm × 0.24 ± 0.04 mm. The chorion has polygonal microsculpturing and is species-specific with polygonal walls. The egg period lasts for 1.3 ± 0.41 days. The duration of the larval period is 1.2 ± 0.42, 1.7 ± 0.48 and 4.0 ± 0.94 days for first, second and third instars, respectively. Pupation occurs in the sand or soil and pupal periods are 7.0 ± 0.47 days. The life cycle from egg to adult was completed in 14.2 ± 1.69 days; the longevity of mated females and males was 130.33 ± 14.18 and 104.66 ± 31.21 days, respectively. At least two to three generations were observed from June 2008 to June 2009.     

The helix-coil transitions of poly (dA)-poly (dT) and poly (dA-dT)-poly (dA-dT)

Helix–coil transition curves are calculated for poly (dA) poly(dT) and poly (dA-dT) poly (dA-dT) using the integral equation approach of Goel and Montroll.⁵ The transitions are described by the loop entropy model with the exponent of the loop entropy factor, k, remaining an arbitrary constant. The theoretical calculations are compared with experimental transition curves of the two polymers. Results indicate that the stacking energies for these two polymers differ by about 1 kcal/mole of base pairs. Also, a fit between theory and experiment was not possible for k > 1.70.     

Spectroscopic investigations of Er(3+) :CdO-Bi(2) O(3) -B(2) O(3) glasses

This article reports on the optical properties of Er³⁺ ions doped CdO–Bi₂O₃–B₂O₃ (CdBiB) glasses. The materials were characterized by optical absorption and emission spectra. By using Judd–Ofelt theory, the intensity parameters Ω_λ (λ = 2, 4, 6) and also oscillatory strengths were calculated from the absorption spectra. The results were used to compute the radiative properties of Er³⁺:CdBiB glasses. The concentration quenching and energy transfer from Yb³⁺–Er³⁺ were explained. The stimulated emission cross‐section, full width at half maximum (FWHM) and FWHM × values are also calculated for all the Er³⁺:CdBiB glasses. Copyright © 2011 John Wiley & Sons, Ltd.