Enhanced effect of soluble transforming growth factor-β receptor II and IFN-γ fusion protein in reversing hepatic fibrosis

Objective

To examine the in vivo anti-fibrotic effect of rat soluble transforming growth factor β receptor II (RsTβRII) and IFN-γ fusion protein (RsTβRII-IFN-γ) in rat hepatic fibrosis model.

Methods

Model rats were divided into five groups and treated i.m. for 8 weeks: 1) fibrotic model group (each rat, 100 μl of 0.9% NaCl day^-1); 2) RsTβRII-IFN-γ treatment group (each rat, 0.136 mg· day^-1); 3) IFN-γ treatment group (each rat, 7.5 MU· day^-1); 4) RsTβRII treatment group (each rat, 0.048 mg· day^-1); and 5) mixture of IFN-γ and RsTβRII treatment group (each rat, IFN-γ 7.5 MU· day^-1+ RsTβRII 0.048 mg· day^-1). After treatment, hepatic fibrogenesis was evaluated by histopathological analysis and measurement of collagen III, α-smooth muscle actin (α-SMA), TGF-β1, TGF-βRII and their mRNA.

Results

Immunohistochemistry, Western blot and real-time RT-PCR showed that RsTβRII-IFN-γ treatment significantly inhibited liver expression of collagen III, α-SMA, TGF-β1 and TGF-βRII at both protein and mRNA levels. Histopathological analysis also showed that the enhanced anti-fibrotic effects were achieved in model rats treated with RsTβRII-IFN-γ.

Conclusion

Our results confirmed that RsTβRII-IFN-γ has the enhanced effects in reversing hepatic fibrosis.     

Chlamydia pneumoniae infection in adults with chronic cough compared with healthy blood donors.

In a small uncontrolled study, persistent cough has recently been found to be associated with serological evidence of acute Chlamydia pneumoniae infection. In order to assess whether C. pneumoniae plays a role in chronic cough, the prevalence of C. pneumoniae infection in 201 adult patients with chronic cough was compared with the prevalence in 106 healthy blood donors without respiratory tract symptoms in the preceding 3 months. A microimmunofluorescence antibody test was used to determine C. pneumoniae antibodies in the immunoglobulin (Ig)M, IgG and IgA fractions. Further, nasopharyngeal aspirates from the 201 patients were examined for C. pneumoniae deoxyribonucleic acid by polymerase chain reaction (PCR). As judged by serology, nine patients (4%) and one control (1%) had acute C. pneumoniae infection, and 92 patients (46%) and 42 controls (40%) had previous or chronic C. pneumoniae infection. Of the nine patients with acute infection, three were C. pneumoniae PCR positive, and they all had an IgM antibody titre response. The remaining six patients had either an IgG antibody titre of > or =512 (five patients) or an IgA antibody titre of > or =512 (one patient). None of these six patients had detectable IgM antibodies. The mean cough period for the five IgG positive patients (10.8 weeks) was significantly longer than the mean cough period for the remaining patient population (6.4 weeks; p=0.004). It is concluded that Chlamydia pneumoniae infection was not statistically significantly more prevalent in patients with chronic cough than in healthy blood donors, and that Chlamydia pneumoniae appears to have a minor role in patients with chronic cough. Direct detection of Chlamydia pneumoniae by polymerase chain reaction on nasopharyngeal aspirates is highly correlated with detectable immunoglobulin M antibodies, but in the late stages of prolonged cough serological testing of immunoglobulin G and immunoglobulin A may be more beneficial for obtaining a microbiological diagnosis.     

Insulin priming effect on estradiol-induced breast cancer metabolism and growth

Diabetes is a risk factor for breast cancer development and is associated with poor prognosis for breast cancer patients. However, the molecular and biochemical mechanisms underlying the association between diabetes and breast cancer have not been fully elucidated. Here, we investigated estradiol response in MCF-7 breast cancer cells with or without chronic exposure to insulin. We found that insulin priming is necessary and specific for estradiol-induced cancer cell growth, and induces anaplerotic shunting of glucose into macromolecule biosynthesis in the estradiol treated cells. Treatment with ERK or Akt specific inhibitors, U0126 or {"type":"entrez-nucleotide","attrs":{"text":"LY294002","term_id":"1257998346","term_text":"LY294002"}}LY294002, respectively, suppressed estradiol-induced growth. Interestingly, molecular analysis revealed that estradiol treatment markedly increases expression of cyclin A and B, and decreases p21 and p27 in the insulin-primed cells. In addition, estradiol treatment activated metabolic genes in pentose phosphate (PPP) and serine biosynthesis pathways in the insulin-primed cells while insulin priming decreased metabolic gene expression associated with glucose catabolism in the breast cancer cells. Finally, we found that anti-diabetic drug metformin and AMPK ligand AICAR, but not thiazolidinediones (TZDs), specifically suppress the estradiol-induced cellular growth in the insulin-primed cells. These findings suggest that estrogen receptor (ER) activation under chronic hyperinsulinemic condition increases breast cancer growth through the modulation of cell cycle and apoptotic factors and nutrient metabolism, and further provide a mechanistic evidence for the clinical benefit of metformin use for ER-positive breast cancer patients with diabetes.     

肠毒素大肠杆菌攻毒小鼠模型的建立以及疫苗候选株保护效果的评价

目的:建立肠毒素大肠杆菌攻毒小鼠模型以及应用模型对疫苗候选株免疫效果进行评价．方法:通过鼻饲半数致死量(LD50)肠毒素大肠杆菌E44813,E44815和E11881A观察小鼠肺病理学变化、肺部细菌清除情况变化,建立肠毒素大肠杆菌鼻饲小鼠模型;应用鼻饲小鼠模型观察疫苗候选株FE1,FE3,FE6保护效果．结果:鼻饲LD50剂量肠毒素大肠杆菌小鼠的病理特征是肺组织存在大量的淋巴细胞、巨噬细胞、中性粒细胞和浆细胞,为多病灶支气管肺炎,肺部细菌清除缓慢,至第7天仍能检测到105数量级的细菌．应用疫苗候选株免疫后进行攻毒,小鼠没有发病和死亡,病理特征主要是淋巴细胞少量增多,肺部细菌清除迅速,至第7天已检测不到细菌,与对照有显著性差异(0 CFU／g vs 6．2×105,5．4×105,2．3×105 CFU／g,P<0．05)．结论:肠毒素大肠杆菌鼻饲小鼠模型能够为疫苗筛选和评价提供了有效途径,同时也证实了疫苗候选株FE1,FE3,FE6具有良好的免疫保护效果．     

Increased mortality in Klinefelter syndrome

Klinefelter syndrome (KS) is the most prevalent sex chromosome disorder in man and is a common cause of hypogonadism. To describe mortality in KS, we conducted an epidemiological study, using Danish registers covering the entire nation. We constructed a cohort of 781 Danish boys and men diagnosed with KS (from the Danish Cytogenetic Central Register) and a control group of 3803 men, matched by month and year of birth. Vital status was obtained from the Centralized Civil Register, and causes of death were obtained from the National Register of Causes of Death. We used Cox regression with stratification on groups of diagnoses according to International Classification of Diseases, 10th version. Where significant results were found, subsequent analyses were performed on subdivisions of diagnoses. We found that Klinefelter syndrome was associated with a significant increase in mortality risk of 40% (hazard ratio, 1.40; 95% confidence interval, 1.13-1.74), corresponding to a significantly reduced median survival of 2.1 yr. The increased mortality was mainly due to increased mortality from infectious, neurological, circulatory, pulmonary, and urinary tract diseases. Whether this increase is caused by the syndrome per se (i.e. hypogonadism) or other factors, e.g. socioeconomic, are involved is presently unknown.     

An in vitro limiting-dilution assay of long-term repopulating hematopoietic stem cells in the mouse

We have developed a limiting-dilution assay of long-term repopulating hematopoietic stem cells in the mouse using a miniturized stroma- dependent bone marrow culture assay in vitro. The cells were overlaid on irradiated stromal layers in microtiter wells in a range of concentrations, and frequencies of cobblestone area-forming cells (CAFC) were calculated by employing Poisson statistics. The production of secondary granulocyte/macrophage colony-forming units (CFU-G/M) in the adherent layer of individual wells was correlated with the presence of such cobblestone areas. CAFC frequencies were determined in bone marrow cell suspensions that were either enriched for marrow repopulating ability (MRA) in vivo, while depleted for spleen colony- forming units (CFU-S), or vice versa. The separation of bone marrow cells (BMC) was either based on centrifugal elutriation, or monoclonal antibody-mediated magnetic depletion of cells carrying cell surface differentiation antigens, and subsequent sorting on the basis of light scatter and rhodamine-123 retention as a measure of mitochondrial activity. In addition, 5-fluorouracil-resistant BMC were studied. Our investigations show that a time-dependent cobblestone area formation exists that reflects the turnover time and primitiveness of CAFC. The frequency of precursors forming cobblestone areas on day 28 after overlay is proposed to be a measure for MRA, whereas the day-7 CAFC frequency closely corresponds with day-12 CFU-S numbers in the suspensions tested.