Human papillomavirus typing with a polymerase chain reaction-based genotyping array compared with type-specific PCR

BACKGROUND: Type-specific persistence of human papillomavirus (HPV) infection can cause invasive cervical cancer. OBJECTIVES: To evaluate the efficacy of HPV detection and typing with a general polymerase chain reaction (PCR)-based genotyping array and to compare it with a type-specific PCR assay. STUDY DESIGN: Four hundred and thirty-three cervical samples were tested with a modified MY11/GP6+ PCR-based reverse-blot assay (EasyChip HPV Blot; King Car, Taiwan [hereafter HPV Blot]) and with 20 genotypes of L1-type-specific PCR (HPV-6, -11, -16, -18, -31, -33, -35, -39, -45, -51, -52, -53, -56, -58, -59, -62, -66, -68, -70, and -71 [CP8061]). RESULTS: The concordance of the two tests in determining HPV positivity was 96.8% (419/433), with a Cohen's kappa=0.93 (95% CI: 0.90-0.97) and McNemar's test of P=1.0, which indicates excellent agreement. The overall concordance of the two tests in the identification of type-specific HPV was 91.0% (394/433). Sensitivity (90-100%), specificity (99.2-100%), and accuracy (98.6-100%) rates of HPV Blot against the gold standard were satisfactory for HPV-16, -18, -58, -33, -52, -39, -45, -31, -51, -70 while HPV-71 (63.6%) had suboptimal sensitivity. Though the kappa values between the two tests for many individual genotypes could not be reliably calculated because of low positivity, the kappa values for HPV-16, -52, and -58 were excellent (0.93, 0.96, and 0.95, respectively). CONCLUSION: The modified MY11/GP6+ PCR-based HPV Blot assay is accurate and sensitive for detection and genotyping of HPV in cervical swab samples.     

Serum immunoglobulin levels in various skin diseases

Serum immunoglobulin (IgG, IgA, IgM) determinations by the single radial immunodiffusion technique were done in patients with various skin diseases. The geometric means of the IgG, IgA and IgM concentrations of each of the patient groups were compared with those of a control group comprising 156 individuals.

A significantly raised IgG level was found only in patients with pemphigus; in patients with dermatitis herpetiformis, other eczemas, and psoriasis vulgaris the IgG level was significantly lowered. The IgA level was significantly elevated in patients with discoid lupus erythematosus and (bullous) pemphigoid, and the IgM level was significantly depressed in patients with atopic dermatitis.

IgE determinations, done with the Ouchterlony technique in a selected group of patients, showed raised levels in some patients with malignant reticulosis and other eczemas but not in those with atopic dermatitis.

Comparison of the results of the serum immunoglobulin determinations with the immunoglobulin synthesis of lesional skin showed no correlation between the local synthesis of immunoglobulins in the skin and changes in the serum levels.

     

Recombinant hepatitis delta antigen from E. coli promotes hepatitis delta virus RNA replication only from the genomic strand but not the antigenomic strand

Hepatitis delta antigen (HDAg) of hepatitis delta virus (HDV) typically consists of two related protein species. The small HDAg (S-HDAg) is a 24-kDa protein of 195 amino acids and the large HDAg (L-HDAg) is a 27-kDa protein with an additional 19 amino acids at its C-terminus. These two proteins have distinct functions in the HDV life cycle. We have developed conditions for expressing S-HDAg and L-HDAg in E. coli as soluble proteins to facilitate large-scale purification. These proteins were purified to homogeneity and shown to be biologically active. Transfection of the purified recombinant S-HDAg together with HDV genomic RNA resulted in viral RNA replication. Surprisingly, the purified S-HDAg could not initiate replication from the antigenomic-sense HDV RNA, even though the latter led to RNA replication when transfected with an mRNA encoding the S-HDAg. These results suggest that initiation of HDV RNA synthesis from the antigenomic RNA may require a form of HDAg that is modified in mammalian cells; in contrast, RNA synthesis from the genomic RNA could be initiated by the recombinant S-HDAg from E. coli. Interestingly, the purified L-HDAg appeared as multiple protein species, including one corresponding to S-HDAg, probably as a result of degradation. The partially proteolyzed L-HDAg also initiated HDV RNA replication under the same conditions. These results add to the mounting evidence that genomic- and antigenomic-strand HDV RNA syntheses are carried out by different mechanisms.     

Conformation and gas permeability of poly(α-amino acid) membranes

The permeability of some poly(?-amino acid) (PAA) membranes like such of poly(N,δ-carbobenzoxy-L -lysine) (PCBL), poly(N,δ-carbobenzoxy-L -ornithine) (PCBO) as well as of copolymers of L -lysine and carbobenzoxy-L -lysine to oxygen and carbon dioxide were determined at different water content especially with regard to their use for artificial lungs. The conformations of these polymer membranes were studied by means of IR-spectra and X-ray diffraction. The results on the diffusion coefficient obtained for PCBL and PCBO were analysed in terms of Eyring's theory. They suggest that the microvoids in the interstices between the α-helices, assumed on the basis of X-ray analyses, play an important role, especially below the glass transition temperature T_g of the side-chains. This seems also to contribute partly to the diffusion of gas through the membrane even above T_g and results in an abnormally high “apparent activation energy” of diffusion in the wet state below T_g. It was found that the permeability of the membrane of partially decarbobenzoxylated PCBL for oxygen in a certain temperature range is higher than that for carbon dioxide. This has never been observed for any other synthetic polymer, because usually carbon dioxide has a higher solubility in these materials than oxygen. In the case of the partially decarbobenzoxylated PCBL membranes, however, the diffusion coefficient of carbon dioxide, and therefore the permeation of this gas, decreases much more than that of oxygen with increasing decarbobenzoxylation accompanied by the formation of β-structure and concomitantly the formation of intermolecular hydrogen bonds. Obviously, this phenomenon may be responsible for the permselectivity of these membranes for oxygen compared with carbon dioxide.     

Effects of low-level microwave irradiation on amphetamine hyperthermia are blockable by naloxone and classically conditionable

In a series of experiments, we investigated the effects of pulsed low-level microwave irradiation on amphetamine-induced hyperthermia in the rat. Rats were irradiated in a 2,450-MHz cylindrical waveguide exposure system at 1 mW/cm², 2 s pulses, 500 pps, average SAR of 0.6 W/kg. Acute (45 min) exposure to microwaves attenuated amphetamine-induced hyperthermia. This effect was blocked by pretreatment of the animals with the narcotic antagonist naloxone. In another experiment, rats were subjected to ten daily sessions of microwave exposure (45 min/session). On day 11, amphetamine-induced hyperthermia was studied in the animals immediately after a session of either microwave or sham exposure. Similar to the acute effect, amphetamine-induced hyperthermia was attenuated in rats irradiated with microwaves (unconditioned effect). In the sham-irradiated animals we observed a potentiation of the amphetamine-induced hyperthermia, which was a conditioned effect of microwaves. Thus, the conditioned effect (potentiation) was opposite in direction to the unconditioned effect (attenuation). No tolerance developed to the unconditioned effect after subchronic exposure. Furthermore, both conditioned and unconditioned effects of microwaves on amphetamine-induced hyperthermia could be blocked by treatment with naloxone. These data suggest that (1) microwave irradiation may activate endogenous opioids, which in turn alter the actions of psychoactive drugs, and (2) the effect of microwaves on drug action can be classically conditioned. Offprint requests to: H. Lai     

Reduction in Maternal Energy Intake during Lactation Decreased Maternal Body Weight and Concentrations of Leptin,Insulin and Adiponectin in Human Milk without Affecting Milk Production,Milk Macronutrient Composition or Infant Growth

Maternal diet has the potential to affect human milk (HM) composition, but very few studies have directly assessed the effect of maternal diets on HM composition. The primary aim of this study was to assess the effect of improving dietary quality in lactating women over 2 weeks on the concentrations of macronutrients and metabolic hormones in HM. The secondary aims were to assess the impact of the dietary intervention on 24 h milk production, maternal body composition and infant growth. Fifteen women completed a 1-week baseline period followed by a 2-week dietary intervention phase targeted towards reducing fat and sugar intake. Maternal anthropometric and body composition and infant growth measurements were performed weekly. Total 24 h milk production was measured before and after the dietary intervention, and HM samples were collected daily. Maternal intakes of energy (−33%), carbohydrate (−22%), sugar (−29%), fat (−54%) and saturated fat (−63%) were significantly reduced during the dietary intervention. HM insulin, leptin and adiponectin concentrations were 10–25% lower at the end of the dietary intervention, but HM concentrations of macronutrients were unaffected. Maternal body weight (−1.8%) and fat mass (−6.3%) were significantly reduced at the end of the dietary intervention, but there were no effects on 24 h milk production or infant growth. These results suggest that reducing maternal energy, carbohydrate, fat and sugar intake over a 2-week period is associated with significant reductions in HM insulin, leptin and adiponectin concentrations. These changes may be secondary to decreases in maternal weight and fat mass. The limited studies to date that have investigated the association between metabolic hormone concentrations in HM and infant growth raise the possibility that the changes in HM composition observed in the current study could impact infant growth and adiposity, but further studies are required to confirm this hypothesis.     

中孕期胎儿染色体异常高危孕妇羊水细胞胎儿染色体核型及介入性产前诊断指征的大样本分析

目的探讨中孕期胎儿染色体异常(FCA)高危孕妇羊水细胞胎儿染色体核型(FK)及其介入性产前诊断(IPD)指征。方法选择2014—2018年,在四川大学华西第二医院产前诊断中心进行羊膜腔穿刺术羊水细胞FK检测的63581例FCA高危孕妇(孕龄为19~27+6孕周)为研究对象。回顾性分析孕妇不同IPD指征及其胎儿染色体核型异常(FKA)检测结果。本研究遵循的程序符合四川大学华西第二医院伦理委员会所制定的伦理学标准,并通过该伦理委员会批准[审批文号:医学科研2019伦审批第(077)号]。结果①本组63581例中孕期孕妇的FKA检出率为3.13%(1989/63581)。这63581例孕妇的IPD指征分别为高龄(预产期年龄≥35岁)(25648例)、血清学筛查高风险(29009例)、无创产前筛查(NIPT)高风险(333例)、夫妇一方为染色体异常携带者(603例)、超声筛查发现胎儿结构异常及超声软指标异常(2647例)、异常儿生育史(1546例)、于外院进行羊水细胞FK分析结果异常者(7例)、孕妇智力低下及合并智力低下家族史(149例)、孕妇早孕期有害物质接触史(1400例)、其他(2239例),FKA检出率分别为3.26%(836/25648)、2.04%(593/29009)、65.17%(217/333)、34.33%(207/603)、3.51%(93/2647)、1.68%(26/1546)、85.71%(6/7)、4.03%(6/149)、0.14%(2/1400)、0.13%(3/2239)。②351例孕妇的IPD指征为超声筛查发现胎儿结构异常中,IPD指征为胎儿淋巴水囊瘤、皮肤水肿的FKA检出率最高,为26.09%(6/23)。2296例孕妇的IPD指征为胎儿超声软指标异常中,IPD指征为胎儿颈项透明层(NT)值≥2.5 mm的FKA检出率最高,为7.74%(66/853)。结论孕妇高龄、血清学筛查高风险、NIPT高风险、夫妇一方为染色体异常携带者、超声筛查发现胎儿结构异常及超声软指标异常等,均为中孕期孕妇FCA的IPD指征,孕妇IPD指征类型不同,FCA风险不同。羊膜腔穿刺术羊水细胞FK检测,可预测有IPD指征孕妇的妊娠结局,为降低FKA出生率提供参考。     

胃癌术中动脉区域灌注化疗与全身静脉化疗的临床疗效比较

目的　评价胃癌术中动脉区域灌注化疗药物的临床疗效。方法　对 40例经病理确诊的胃癌患者 (治疗组 )于术中经供应肿瘤的区域动脉行插管化疗 ,同时进行手术治疗 ,并与同期术中行全身静脉化疗的 2 4例胃癌患者 (对照组 )进行临床疗效比较。结果　术后病理检查显示 ,治疗组原发灶中的癌细胞均有不同程度地变性坏死 ,其近期有效率为 95 % ,对照组有效率为 41.6 7% ,两组比较差异显著 (P<0 .0 5 )。结论　胃癌患者术中经动脉灌注化疗 ,其近期抗癌效果好 ,毒副反应轻 ,对减少或防止术中癌细胞的医源性扩散与种植以及术后复发 ,均有重要的临床意义