Cryopreservation of human platelets with polyvinylpyrrolidone

Fourteen polymeric compounds were assessed for their abilities to protect human platelets from injury during freezing and thawing. The effect of polymer concentration and molecular weight, cooling rate, platelet concentration, and short-term storage on platelet recovery were studied. Polyvinylpyrrolidone, average molecular weight 10,000, was found to provide the greatest degree of protection to platelets. Recoveries of platelets of from 60 to 80 per cent were achieved using a 5 per cent final concentration of polyvinylpyrrolidone and a cooling rate of 8 C per minute. Methods of freezing small aliquots of platelets and platelet concentrates were developed. The frozen-thawed platelets were viable in vitro as shown by osmotic reversal, clot retraction, and aggregation by collagen and adenosine diphosphate. The techniques described could be developed readily to facilitate the long-term storage of platelets.     

Unusual human poisoning with Delphinium peregrinum

Abstract

Background: This is the 30^th Annual Report of the American Association of Poison Control Centers’ (AAPCC) National Poison Data System (NPDS). As of July 1, 2012, 57 of the nation's poison centers (PCs) uploaded case data automatically to NPDS. The upload interval was 7.58 [6.30, 11.22] (median [25%, 75%]) min, creating a near real-time national exposure and information database and surveillance system.

Methodology: We analyzed the case data tabulating specific indices from NPDS. The methodology was similar to that of previous years. Where changes were introduced, the differences are identified. Poison center cases with medical outcomes of death were evaluated by a team of 34 medical and clinical toxicologist reviewers using an ordinal scale of 1–6 to assess the Relative Contribution to Fatality (RCF) of the exposure to the death.

Results: In 2012, 3,373,025 closed encounters were logged by NPDS: 2,275,141 human exposures, 66,440 animal exposures, 1,025,547 information calls, 5,679 human confirmed nonexposures, and 218 animal confirmed nonexposures. Total encounters showed a 6.9% decline from 2011, while healthcare facility (HCF) exposure calls increased by 1.2%. All information calls decreased by 14.8% and HCF information calls decreased by 1.7%, medication identification requests (Drug ID) decreased by 22.0%, and human exposures reported to US PCs decreased by 2.5%. Human exposures with less serious outcomes have decreased by 3.7% per year since 2008, while those with more serious outcomes (moderate, major, or death) have increased by 4.6% per year since 2000.

The top five substance classes most frequently involved in all human exposures were analgesics (11.6%), cosmetics/personal care products (7.9%), household cleaning substances (7.2%), sedatives/hypnotics/antipsychotics (6.1%), and foreign bodies/toys/miscellaneous (4.1%). Analgesic exposures as a class increased the most rapidly (8,780 calls/year) over the last 12 years. The top five most common exposures in children aged 5 years or less were cosmetics/ personal care products (13.9%), analgesics (9.9%), household cleaning substances (9.7%), foreign bodies/toys/ miscellaneous (7.0%), and topical preparations (6.3%). Drug identification requests comprised 54.4% of all information calls. NPDS documented 2,937 human exposures resulting in death with 2,576 human fatalities judged related (RCF of 1-Undoubtedly responsible, 2-Probably responsible, or 3-Contributory).

Conclusions: These data support the continued value of PC expertise and need for specialized medical toxicology information to manage the more severe exposures, despite a decrease in calls involving less severe exposures. Unintentional and intentional exposures continue to be a significant cause of morbidity and mortality in the US. The near real-time, always current status of NPDS represents a national public health resource to collect and monitor US exposure cases and information calls. The continuing mission of NPDS is to provide a nationwide infrastructure for public health surveillance for all types of exposures, public health event identification, resilience response, and situational awareness tracking. NPDS is a model system for the nation and global public health.     

人端粒酶反转录酶基因转染人胚胎大脑皮质神经元的凋亡

背景：人端粒酶反转录酶重组腺病毒转染原代培养的人胚胎大脑皮质神经元,可以促进细胞生存,抑制凋亡。目的：观察人端粒酶反转录酶对β淀粉样蛋白（amyloidβ-protein,Aβ）诱导人胚胎大脑皮质神经元凋亡的影响。方法：人胚胎大脑皮质神经元原代培养后,分为3组：对照组、Aβ25-35组和人端粒酶反转录酶组。Aβ25-35组和人端粒酶反转录酶组细胞在培养144 h后,用Aβ25-35 5μmol/L干预24 h。人端粒酶反转录酶组在Aβ25-35 5μmol/L干预72 h进行人端粒酶反转录酶基因转染。结果与结论：原代培养的人胚胎大脑皮质神经元培养7 d后,出现凋亡细胞,而Aβ25-35干预后使凋亡细胞数量增加,而人端粒酶反转录酶可防止Aβ25-35诱导的人胚胎大脑皮质神经元的凋亡。     

儿童特发性血小板减少性紫癜与人类巨细胞病毒感染

目的为探讨特发性血小板减少性紫癜（ITP）与人类巨细胞病毒（HCMV）感染之间的关系，采用非抗凝血清用ELISA方法检测CMV—IgM及清洁晨尿用聚合酶链反应（PCR）方法测定HCMV—DNA含量。结果显示36例ITP患者中，HCMV—DNA阳性者17例，阳性率47．2％，儿童保健门诊正常体检儿童对照组28例，HCMV—DNA阳性者4例，阳性率14．3％，P〈0．01，差异有显著性。ITP患者CMV—IgM阳性者8例，阳性率22．2％，儿童保健门诊正常体检儿童对照组中CMV—IgM阳性者0例。P（0．01，差异有显著性。结论HCMV感染与ITP密切相关。     

Studies of human sera with cytotoxic activity

42 human sera showing in vitro cytotoxic activity of restricted or broad HL-A specificities with test human lymphocytes were studied for the molecular and immunoglobulin class of cytotoxic antibody using sucrose gradient separations, DEAE-cellulose chromatography, and Sephadex G-200 gel filtration. Sera originated from patients with previous multiple pregnancies (19), multiply transfused patients (8), subacute bacterial endocarditis (4), systemic lupus (2), and human umbilical cord sera (9). In 32 of 42 instances, predominant cytotoxic activity was found in high molecular weight gradient fractions; however, DEAE chromatographic separations revealed cytotoxic activity in initial buffer fractions containing primarily gammaG globulin. Gradient separations of cytotoxic activity within initial gammaG DEAE fractions showed localization of cytotoxicity only in high molecular weight materials. Confirmation of high molecular weight gammaG cytotoxic activity was obtained by resistance to mercaptoethanol treatment, abolition of activity after absorption only with specific anti-gammaG antisera, and by the finding that high molecular weight cytotoxic activity in gradients or gel filtration run at neutral pH 7.4 became 7S when separations were rerun at an acidic pH of 4.0. Such 7S activity again became rapidly sedimenting when the same fractions were again rerun in gradients at neutral pH.19S gammaM cytotoxic activity was documented in a panel of 15 human sera containing anti-"I" cold agglutinins. In this instance the cytotoxic activity appeared to be related to the cold agglutinin antibody since it was mercaptoethanol sensitive and could be demonstrated in monoclonal antibody eluates containing primarily gammaM.No in vitro protection again cytotoxic effect was demonstrated when isolated human 19S anti-gamma-globulins were aded to the lymphocytotoxic assay system.With the exception of cold agglutinins, human cytotoxic antibodies appear to be primarily gammaG producing in vitro lymphocyte killing either as 7S gammaG globulin or as rapidly sedimenting aggregates or complexes of gammaG molecules.     

Genital infection with human papillomavirus in adolescents

Human papillomavirus (HPV) infection is one of the most common sexually transmitted infections in adolescents. Such infection is associated with substantial health risks and is unpredictable in its resolution. Genital warts are the most common clinical manifestations of genital HPV infections. Most genital warts are caused by low-risk HPV types (notably HPV-6 and -11). The majority of genital HPV infections are latent or subclinical. Although the outcome of a genital HPV infection is variable, the infection is usually transient and complete resolution is common. However, persistent infections with high-risk HPV types (notably HPV-16 and -18) are associated with the development of cervical intraepithelial dysplasia (CIN), which may progress to cervical cancer. Genital warts, generally diagnosed in the clinic, can be treated with imiquimod, podofilox, podophyllin, liquid nitrogen, bichloroacetic or trichloroacetic acid, or surgery. Cervical cytology screening is an ideal screening test for subclinical HPV infection with resultant CIN. Annual cervical cytology screening should begin approximately 3 years after initiation of sexual intercourse but no later than age 21 years. Because of the high rate of regression of low-grade squamous intraepithelial lesions (LSIL) in adolescents, the cytologic study should be repeated within 6 to 12 months. Colposcopy should be reserved for persistent LSIL. Patients with high-grade squamous intraepithelial lesions (HSIL) should be referred for colposcopy and biopsy. Confirmed HSIL should be treated with cryotherapy, laser therapy, or loop electrosurgical excisional procedure. The use of therapeutic vaccines is still experimental. HPV infection can be prevented, to a certain extent, by delaying the initiation of sexual activity, limiting the number of sexual partners, and using latex condoms. Several viruslike particle candidate vaccines are under development for the prevention of HPV. These vaccines have been proven safe, well tolerated, highly immunogenic, and highly efficacious. Such vaccines are urgently needed and ultimately may be an important preventive measure.     

Initial experiences with subcutaneous recombinant human hyaluronidase

We report here our retrospective observations on the use of recombinant human hyaluronidase (rHuPH20) for the facilitation of subcutaneous hydration and drug infusion. Thirty-two patients were treated with rHuPH20 in a hospice setting over a 6-month period. Of these, 26 received this agent to enhance hypodermoclysis with standard hydration fluids for symptom control of delirium, myoclonus and mild to moderate dehydration. Flow rates up to 500 mL/hr were attained without difficulty. Electrolyte replacement in hydration fluid was achieved without incident in 5 patients receiving potassium and in 1 patient receiving both potassium and magnesium. In addition to use for hydration, 6 patients received recombinant human hyaluronidase to enhance subcutaneous infusion of 9 medications, primarily because the medication dosage required subcutaneous flow rates greater than the standard 3 mL/hr. There were no significant adverse events. Induration at the infusion site occurred in 1 patient receiving hydration and higher than expected serum lidocaine concentration was observed in another patient. Based on our positive initial experience with recombinant human hyaluronidase, there is interest in expanding its use in our facility in both the inpatient and outpatient settings.     

Interaction of imatinib mesilate with human P-glycoprotein

The interaction of imatinib mesilate with P-glycoprotein (P-gp) was examined using pig kidney epithelial LLC-PK1 cells versus L-MDR1 cells, which overexpress human P-gp on the apical membrane. The basal-to-apical transport of imatinib mesilate in L-MDR1 cells significantly exceeded that in the parental LLCPK1 cells. The intracellular accumulation of imatinib mesilate after its basal application to LLC-PK1 and L-MDR1 cells was 35% and 15%, respectively. A P-gp modulator, cyclosporin A, inhibited the basal-to-apical transport in L-MDR1 cells. The intracellular accumulation of imatinib mesilate in L-MDR1 cells was also increased by cyclosporin A. The rhodamine 123 efflux assay showed that the efflux of rhodamine 123 in K562/DXR cells, which overexpress human P-gp, could be blocked markedly by imatinib mesilate in a dose-dependent fashion. The Ki values for the inhibition of P-gp function by cyclosporin A and imatinib mesilate were estimated to be 6.1 and 18.3 muM, respectively, using a calcein-AM efflux assay. These observations demonstrate that imatinib mesilate is a substrate as well as a modulator of human P-gp, suggesting that imatinib mesilate drug interactions may occur via P-gp. It is necessary to consider the pharmacokinetic and pharmacodynamic interactions of imatinib mesilate with other drugs via P-gp.     

人感染猪链球菌病患者的心理干预

目的探讨人感染猪链球菌病患者的心理干预方法及体会。方法总结分析我院49例人感染猪链球菌病患者的心理反应及心理干预效果。结果49例中,否认患者在1天内接受患病事实,孤独患者在2天内减轻孤独感,焦虑患者在3天内焦虑症状减轻,依赖患者2周内正确转换角色,除7例病人因病情过重死亡外,其余患者身心康复,痊愈出院。结论对人感染猪链球菌病患者在实施准确及时的治疗和护理措施的同时,重视病人心理护理,给予有针对性心理干预措施,可有效的促进病人身心健康的恢复,促进疾病治愈。