Urinary nuclear magnetic resonance spectroscopy of a Bangladeshi cohort with hepatitis-B hepatocellular carcinoma: A biomarker corroboration study

AIM: To establish if a distinct urinary metabolic profile could be identified in Bangladeshi hepatitis-B hepatocellular carcinoma(HCC) patients compared to cirrhosis patients and controls. METHODS: Urine samples from 42 Bangladeshi patients with HCC(39 patients with hepatitis-B HCC), 47 with cirrhosis on a background of hepatitis B, 46 with chronic hepatitis B, and seven ethnically-matched healthy controls were analyzed using nuclear magnetic resonance(NMR) spectroscopy. A full dietary and medication history was recorded for each subject. The urinary NMR data were analyzed using principal component analysis(PCA) and orthogonal partial leastsquared discriminant analysis(OPLS-DA) techniques. Differences in relative signal levels of the most discriminatory metabolites identified by PCA and OPLSDA were compared between subject groups using an independent samples Kruskal-Wallis one-way analysis of variance(ANOVA) test with all pairwise multiple comparisons. Within the patient subgroups, the MannWhitney U test was used to compare metabolite levels depending on hepatitis B e-antigen(HBe Ag) status and treatment with anti-viral therapy. A BenjaminiHochberg adjustment was applied to acquire the level of significance for multiple testing, with a declared level of statistical significance of P 0.05.RESULTS: There were significant differences in age(P 0.001), weight(P 0.001), and body mass index(P 0.001) across the four clinical subgroups. Serum alanine aminotransferase(ALT) was significantly higher in the HCC group compared to controls(P 0.001); serum α-fetoprotein was generally markedly elevated in HCC compared to controls; and serum creatinine levels were significantly reduced in the HCC group compared to the cirrhosis group(P = 0.004). A threefactor PCA scores plot showed clustering of the urinary NMR spectra from the four subgroups. Metabolites that contributed to the discrimination between the subgroups included acetate, creatine, creatinine, dimethyamine(DMA), formate, glycine, hippurate, and trimethylamine-N-oxide(TMAO). A comparison of relative metabolite levels confirmed that carnitine was significantly increased in HCC; and creatinine, hippurate, and TMAO were significantly reduced in HCC compared to the other subgroups. HBe Ag negative patients showed a significant increase in creatinine(P = 0.001) compared to HBe Ag positive patients in the chronic hepatitis B subgroup, whilst HBe Ag negative patients showed a significant decrease in DMA(P = 0.004) in the cirrhosis subgroup compared to HBe Ag positive patients. There were no differences in metabolite levels in HCC patients who did or did not receive antiviral treatment. CONCLUSION: Urinary NMR changes in Bangladeshi HCC were identified, corroborating previous findings from Egypt and West Africa. These findings could form the basis for the development of a cost-effective HCC dipstick screening test.     

克隆并应用计算机软件分析小鼠LASS1基因启动子区的特征

目的:克隆并应用计算机分析小鼠LASS1基因启动子,为进一步研究在细胞衰老过程中LAG1基因的转录调控奠定基础。方法:实验于2006-03/07在汕头大学医学院细胞衰老实验室完成。培养EC109细胞株,预测小鼠LASS1基因启动子所在区域,用PCR技术扩增启动子区序列,构建重组质粒pGEM-T-501、pGEM-T-181、pGEM-T-26,并转化JM109感受态菌,并分别克隆入荧光素酶报告基因载体pGL3-Basic及增强型绿色荧光蛋白报告基因载体pEGFP-1,酶切pGEM-T-501、181、26重组质粒及pGL3-Basic、pEGFP-1载体,制备插入片段和载体,构建重组表达质粒pGL3-501、pGL3-181和pGL3-26及pEGFP-501、pEGFP-181和pEGFP-26,转化JM109感受态菌。用脂质体介导的方法瞬时转染EC109细胞,测定荧光素酶表达活性及观察绿色荧光蛋白的表达情况。结果:①在预测启动子区构建了3种荧光素酶报告基因表达体系及3种增强型绿色荧光蛋白报告基因表达体系,分别为pGL3-501(-501bp～ 106bp)、pGL3-181(-181bp～ 106bp)、pGL3-26( 26～ 106)、pEGFP-501、pEGFP-181和pEGFP-26。②pGL3-501表达载体与pGL3-181表达载体荧光素酶表达活性相近,pGL3-26表达载体荧光素酶表达活性极低,与阴性对照活性相近, 26bp～ 106bp无启动子活性。绿色荧光蛋白的表达情况也类似,pEGFP-501和pEGFP-181有明显的绿色荧光蛋白表达,pEGFP-26和空载体pEGFP-1无表达活性。结论:-181bp～ 26bp区域含有小鼠LASS1基因转录所必需的基本启动子序列。其中2个SP1保守序列是小鼠LASS1基因启动子所必需的。     

Dehydroepiandrosterone inhibits human vascular smooth muscle cell proliferation independent of ARs and ERs.

Dehyroepiandrosterone (DHEA), an adrenal-derived steroid, has been clinically implicated in protection against coronary artery disease and experimentally in inhibition of atherosclerosis and plaque progression. Because DHEA is enzymatically metabolized to androgens or estrogens, it is not clear whether DHEA exerts effects directly or after conversion to these hormones, both of which are associated with well-characterized pathways of action. We therefore examined the effects of DHEA on proliferation of human vascular smooth muscle cells (VSMCs) in culture in the presence or absence of the ER antagonist ICI 182,780 and the AR antagonist flutamide and compared them with the effects of 17beta-estradiol, androstenedione, and T. We also determined the affinity of DHEA for ERs and ARs in VSMC and its specific binding in intact cells. To explore a possible mechanism for DHEA action in these cells, we measured the phosphorylation of ERK-1, c-jun N-terminal protein kinase, and p38 (three members of the MAPK superfamily). Both DHEA and 17beta-estradiol significantly inhibited platelet derived growth factor (PDGF)-BB-induced increases in VSMC proliferation, whereas androstenedione and T increased proliferation. Although E2-induced inhibition of the PDGF effect was abolished by ICI 182,780 and T-induced stimulation was abolished by flutamide, neither receptor antagonist altered the inhibitory effect of DHEA. Binding studies confirmed the presence of both ERs and ARs; DHEA showed minimal affinity for either receptor but bound specifically and with high affinity to putative receptors in intact cells. Following 4-h incubation with DHEA (1-100 nM), ERK1 phosphorylation was significantly reduced in a dose-dependent manner, whereas neither c-jun N-terminal protein kinase nor p38 kinase activity was altered by either PDGF-BB or DHEA. DHEA inhibits human VSMC proliferation by a mechanism independent of either ARs or ERs, presumably via a DHEA-specific receptor that involves ERK1 signaling pathways.     

The 'Silver Book' on elderly care in hospitals and community settings

The number of people aged 85 and over is set to increase by two-thirds in the next 20 years, making it imperative that appropriate structures are in place and guidance is available for clinicians in every setting on best practice in caring for older people over the first 24 hours of an urgent care episode. This article discusses the launch of the Silver Book, which recommends ways in which emergency admissions can be reduced and the experience of those admitted improved.     

Validity of clinical case definitions for influenza surveillance among hospitalized patients: results from a rural community in North India

Objective: Clinical case definitions used for influenza surveillance among hospitalized patients vary and need systematic evaluation. Design, setting and sample: During July 2009–August 2011, we collected clinical data and specimens (nasal and throat swabs) from rural patients hospitalized for acute medical illnesses. Specimens were tested by rRT‐PCR for influenza viruses. Main outcome measures: Case definitions evaluated the following: influenza‐like illness (ILI: measured fever plus cough or sore throat); severe acute respiratory illness (SARI: ILI with difficulty breathing in ≥5 years, Integrated Management of Childhood Illness–defined pneumonia or severe pneumonia, or physician diagnosed lower respiratory infection in <5 years); acute respiratory infection (ARI: ≥1 of cough, nasal discharge, difficulty breathing or sore throat); febrile acute respiratory illness (FARI: fever plus either cough, sore throat, runny nose, difficulty breathing, or earache). Variants that included “reported fever” and additional sign–symptom combinations were also evaluated. Results: We enrolled 1043 hospitalized patients, including 257 children <5 years of age (range 1 day–86 years). Seventy‐four patients tested influenza virus positive (including 28 A(H1N1)pdm09). Sensitivity(95% CI) and specificity (95% CI) for influenza infection were 78% (67–87) and 60% (57–63) for ILI (measured/reported fever); 37% (26–49) and 78% (75–80) for SARI (measured/reported fever); 82% (72–90) and 57% (54–60) for FARI (measured/reported fever); 88% (78–94) and 45% (42–49) for ARI; and 74% (63–84) and 61% (58–64) for measured/reported fever plus cough. Case definitions including only measured fever had lower sensitivity. Conclusion: ILI and FARI with measured/reported fever provided good balance between sensitivity and specificity among hospitalized patients. The simpler case definition of measured/reported fever plus cough is suited for field surveillance.     

Brief Report: Attention Deficit and Hyperactivity Disorder Scores Are Elevated and Respond to N‐Acetylcysteine Treatment in Patients With Systemic Lupus Erythematosus

Objective

To investigate whether attention deficit hyperactivity disorder (ADHD) may serve as a marker of neuropsychiatric disease and as a target for N‐acetylcysteine (NAC) treatment in patients with systemic lupus erythematosus (SLE).

Methods

The ADHD Self‐Report Scale (ASRS) was used to assess 49 patients with SLE and 46 matched healthy control subjects. Twenty‐four of the patients with SLE were randomized to receive either placebo, NAC at a dosage of 2.4 gm/day, or NAC at a dosage of 4.8 gm/day. Disease activity was evaluated monthly using the British Isles Lupus Assessment Group (BILAG) index, the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI), the Fatigue Assessment Scale (FAS), and the ASRS, before and during the 3‐month treatment period and after a 1‐month washout period.

Results

The cognitive/inattentive (ASRS part A), hyperactivity/impulsive (ASRS part B), and combined (total) ASRS scores were increased in patients with SLE compared with control subjects (mean ± SEM 17.37 ± 1.03 [P = 3 × 10⁻⁷], 14.51 ± 0.89 [P = 2 × 10⁻⁴], and 31.92 ± 1.74 [P = 8 × 10⁻⁷], respectively, versus 10.41 ± 1.02, 9.61 ± 1.21, and 20.02 ± 1.98, respectively. ASRS part A scores correlated with SLEDAI (r = 0.53, P < 0.0001) and BILAG scores (r = 0.36, P = 0.011). ASRS total scores also correlated with SLEDAI (r = 0.45, P = 0.0009) and BILAG scores (r = 0.31, P = 0.025). ASRS part A (r = 0.73, P < 0.0001), ASRS part B (r = 0.47, P = 0.0006), and ASRS total scores (r = 0.67, P < 0.0001) correlated with the FAS score. Relative to the scores in placebo‐treated patients, ASRS total scores were reduced in SLE patients treated with NAC dosages of 2.4 gm/day and 4.8 gm/day combined (P = 0.037). ASRS part A scores were reduced by NAC dosages of 2.4 gm/day (P = 0.001) and 4.8 gm/day (P < 0.0001) as well as by NAC at dosages of 2.4 gm/day and 4.8 gm/day combined (P = 0.001).

Conclusion

In patients with SLE, elevated ASRS scores reveal previously unrecognized and clinically significant symptoms of ADHD that respond to NAC treatment.

     

Inflammatory myofibroblastic tumor of the spleen: a case report

Inflammatory myofibroblastic tumors (IMTs), otherwise known as the inflammatory pseudotumor, is a rare solid mesenchymal tumor, simulating malignant neoplasms, histologically characterized by the proliferation of spindle cells in a fibrous myxoid stroma containing inflammatory cells. CT and MR imaging are the most used tools in their assessment. Clinical features are nonspecific and depend on the localization of the tumor, radiologic findings are polymorphic and no-conclusive and present a diagnostic challenge to the radiologist. Although histology remains obligatory for the final diagnosis. Heren, we report a case of splenic IMT with histological correlation.