泛素特异性蛋白酶25在颞叶癫痫大鼠颞叶皮质中表达变化的实验研究

目的探讨海人酸点燃杏仁核大鼠颞叶癫痫模型颞叶皮质中泛素特异性蛋白酶25(USP25)的表达情况。方法将52只SD雄性大鼠按随机数字表法分为癫痫模型组(共39只)和假手术对照组(简称对照组,13只)。癫痫模型组建立海人酸点燃杏仁核大鼠颞叶癫痫模型,再按构建成功的时间分为3组(构建成功1 d为急性期组、构建成功7 d为潜伏期组、构建成功30 d为慢性期组,每组各13只),在观察结束时取材。对照组在杏仁核注入生理盐水,与癫痫模型组伴随取材。免疫组织化学染色及免疫荧光双标法检测USP25在颞叶皮质中的表达及与神经元(NeuN)和星形胶质细胞(GFAP)的共定位情况;实时荧光定量PCR法和蛋白免疫印迹法检测USP25在颞叶皮质中的表达变化。结果在注药侧颞叶皮质中,癫痫模型组USP25与NeuN的共定位阳性细胞在癫痫后期增加,USP25 mRNA及蛋白水平在癫痫发作不同时期的表达差异均有统计学意义(F值分别为25.48、7.68,均P<0.05)。与对照组(mRNA:1.00±0.36;蛋白:1.00±0.46)比较,潜伏期组(mRNA:10.80±4.82;蛋白:1.88±0.32)和慢性期组(mRNA:12.97±4.48;蛋白:1.92±0.26)的表达水平显著增加,差异均有统计学意义(均P<0.05)。在未注药侧皮质中,USP25 mRNA及蛋白水平在癫痫发作不同时期的表达差异也均有统计学意义(F值分别为86.86、6.65,均P<0.05)。结论颞叶皮质中USP25的表达在癫痫大鼠潜伏期后增加,提示去泛素化通路参与颞叶癫痫的慢性病理过程。     

Inhaled corticosteroid use in HIV‐positive individuals taking protease inhibitors: a review of pharmacokinetics,case reports and clinical management

As a consequence of inhibition of the hepatic cytochrome P450 3A4 isozyme, treatment with HIV protease inhibitors can result in significant drug?drug interactions. One noteworthy interaction is between protease inhibitors and inhaled or intranasal corticosteroids. This interaction can result in adrenal insufficiency and iatrogenic Cushing's syndrome (with symptoms such as rapid weight gain, obesity, facial hirsutism and swelling), as well as hypertension, osteoporosis and decreased CD4 cell count. In this paper, we review and unite pharmacokinetic data, case reports and current research regarding this drug?drug interaction in order to suggest options for the clinical management of HIV‐positive patients requiring treatment with protease inhibitors and inhaled or intranasal corticosteroids.     

A critical review on serine protease: Key immune manipulator and pathology mediator

Proteolytic activity is fundamental to survival, so it is not surprising that all living organisms have proteases, especially seine protease. This enzyme in its numerous isoforms and homologues, constitutes the quintessential offence and defence factors, in the form of surface proteins, secreted molecules, gut digestive enzymes, venom in specialised glands or plant latex, among other manifestations. Occurring as trypsin, chymotrypsin, elastase, collagenase, thrombin, subtilisin etc., it mediates a diverse array of functions, including pathological roles as inflammatory, coagulatory to haemorrhagic. This review emphasizes that despite the superficial differences in mechanisms, most health issues, be they infectious, allergic, metabolic, or neural have a common conduit. This enzyme, in its various glycosylated forms leads to signal misinterpretations, wreaking havoc. However, organisms are endowed with serine protease inhibitors which might restrain this ubiquitous yet deleterious enzyme. Hence, serine proteases-driven pathogenesis and antagonising role of inhibitors is the focal point of this critical review.     

Management of protozoa-related diarrhea in HIV infection

Protozoa infections are an important cause of chronic diarrhea in patients infected with HIV. The introduction of highly active antiretroviral treatment to the management of HIV in the mid1990s has led to a dramatic reduction in the incidence of these opportunistic infections in Europe and America. In contrast, in the developing world where such treatments are not readily affordable, protozoa-related diarrhea remains a major cause of morbidity and mortality in HIV-infected individuals. In this review, the optimum investigations required to diagnose these pathogens in HIV-related diarrhea, as well as current treatment options, will be discussed.     

The discovery and development of boceprevir

Introduction: Boceprevir was the first direct acting agent developed for the treatment of hepatitis C virus infection. Boceprevir functions by targeting NS3 protease, a viral enzyme essential for replication. This peptidomimetic molecule was optimized from a peptide lead to provide a potent, selective and orally bioavailable drug that can be combined with ribavirin and peg interferon to achieve sustained viral response (undetectable HCV RNA levels for 24 weeks after completion of therapy) in patients infected with Genotype 1 of the virus.

Areas covered: This article provides a review of the pre-clinical and clinical discovery of boceprevir. This review includes the role and function of its molecular target, NS3 protease, as well as the assays used to measure in vitro efficacy, compound optimization and clinical studies to demonstrate safety and efficacy.

Expert opinion: As the first direct acting anti-HCV agent, boceprevir represents an important advance in therapy of this widespread chronic disease. Yet, while this therapy is a valuable approach, it does have limitations. Studies have suggested that 30% of patients do not achieve sustained viral response and 11% of patients have developed anemia and/or neutropenia. Current drug discovery and development efforts are underway to develop novel therapeutic options that address these issues.     

Clinical and Local Effects of Antacids and Physical Therapy as Shown Gastroscopically

All physicians should be alert to indications of suicidal intent on the part of depressed or mentally ill patients. Hospital personnel generally should also know the importance of immediately reporting unusual behavior on the part of patients. Patients who seem likely to attempt suicide should be hospitalized and carefully supervised. Treatment should not be undertaken at home. Some of the newer antidepressant drugs are effective in treating potentially suicidal patients.     

Does lopinavir/ritonavir alter the primary gingival epithelium?

Evaluation of: Israr M, Mitchell D, Alam S, Dinello D, Kishel JJ, Meyers C. The HIV protease inhibitor lopinavir/ritonavir (Kaletra) alters the growth, differentiation and proliferation of primary gingival epithelium. HIV Med. DOI: 10.1111/j.1468-1293.2010.00863.x (2010) (Epub ahead of print).

In clinical practice, a significant proportion of HIV-infected patients receive lopinavir/ritonavir (LPV/r) as a component of highly active antiretroviral therapy (HAART). The study by Israr et al. was designed to evaluate the effects of the HIV protease inhibitor LPV/r on gingival epithelium growth and differentiation. The authors isolated gingival keratinocytes from human gingival tissue from patients undergoing dental surgery. Raft cultures of gingival keratinocytes were established and treated with a range of LPV/r concentrations. LPV/r inhibited the growth of gingival epithelium when the drug was present throughout the growth period of the tissue. When LPV/r was added on day 8 of tissue growth, it compromised tissue integrity and altered the proliferation and differentiation of gingival keratinocytes. The expression pattern of cytokeratin-5, -14, -10 and -6, proliferating cell nuclear antigen and cyclin A were altered in treated rafts. These findings suggest that LPV/r compromised tissue integrity and deregulated the cell cycle/proliferation and differentiation pathways, resulting in abnormal epithelial repair and proliferation. However, these findings do not correlate with those observed in clinical practice. In clinical trials, the adverse effects in the oral cavity have been uncommon in patients receiving HAART containing standard doses of LPV/r. This disparity may be due to a few limitations in the study design. The investigators assumed that the blood levels of LPV/r would be the same as in the saliva, and thus chose the peak concentration of LPV/r in blood serum as the baseline concentration in the study. However, the correlation between LPV/r levels in blood serum and in oral tissues has not been widely studied. Lopinavir and ritonavir are highly (98–99%) bound to serum proteins. Thus, the concentrations of these drugs in saliva would be expected to be 50–100-times lower than total plasma concentrations, rather than equivalent, as the authors assumed. Thus, the concentrations assessed in this in vitro study are very likely to be much higher than the actual concentrations of LPV/r encountered in the saliva of patients.     

Atazanavir: improving the HIV protease inhibitor class

Protease inhibitors are potent agents against HIV but their use is constrained by poor pharmacokinetics, cross-resistance and metabolic toxicities. Atazanavir [Reyataz?] is a new protease inhibitors with once-daily dosing and minimal lipid and glycemic effects. Resistance studies of clinical isolates reveal a mutational pattern distinctive from that of other protease inhibitors. Atazanavir selects for the I50L mutation in HIV protease that confers increased susceptibility to other protease inhibitors in vitro. Clinical trials have shown comparable efficacy to nelfinavir (Viracept^®) and efavirenz (Sustiva^®) in treatment-naive patients, and in preliminary studies, ritonavir-boosted atazanavir is effective in patients failing previous protease inhibitor-containing regimens. Reversible elevations in bilirubin occur in some patients but are not associated with hepatic injury. Atazanavir improves upon aspects of currently-available protease inhibitors and appears useful for initial and possibly subsequent HIV therapy.