The expression of Bcl-2 family proteins (Bcl-2, Bcl-x, Bax, Bak and Bim) in human lymphocytes

Bcl-2 family proteins regulate programmed cell death, and may play an important role in the selection of lymphocytes. We investigated the expression of Bcl-2, Bcl-x, Bax, Bak and Bim in human lymphocytes using flow-cytometry. Bcl-2 was down-regulated in CD4(+)8(+) (DP) thymocytes and CD19(+)38(+) tonsillar lymphocytes (GC B cells). Among DP thymocytes, cells co-expressing CD69 up-regulated Bcl-2, suggesting that the role of Bcl-2 is promoting survival of positively selected DP cells. Unexpectedly, the expression level of Bcl-x was higher in DP cells than in Single Positive (SP) cells and in CD69(+) DP thymocytes it was lower than in CD69(+) DP thymocytes. Expression of Bim was low in DP thymocytes but high in a subset of GC B cells. Bim and Bax were expressed more highly in SP than in DP thymocytes. Among peripheral blood lymphocytes (PBL), CD8(+) T cells expressed an approximately ten-fold higher level of Bcl-x than CD4(+) T cells while both subsets expressed similar levels of Bcl-2. Bak expression was low and Bim expression was absent in PBL. These results suggest that not only Bcl-2 but other members of the Bcl-2 family are involved in T cell development in the thymus and affinity maturation of B cells in the germinal center.     

Apparent genotype–phenotype correlation in childhood,adolescent, and adult Chediak‐Higashi syndrome

Chediak‐Higashi syndrome (CHS) is a rare autosomal recessive disorder characterized by severe immunologic defects, reduced pigmentation, bleeding tendency, and progressive neurological dysfunction. Most patients present in early childhood and die unless treated by bone marrow transplantation. About 10–15% of patients exhibit a much milder clinical phenotype and survive to adulthood, but develop progressive and often fatal neurological dysfunction. Very rare patients exhibit an intermediate adolescent CHS phenotype, presenting with severe infections in early childhood, but a milder course by adolescence, with no accelerated phase. Here, we describe the organization and genomic DNA sequence of the CHS1 gene and mutation analysis of 21 unrelated patients with the childhood, adolescent, and adult forms of CHS. In patients with severe childhood CHS, we found only functionally null mutant CHS1 alleles, whereas in patients with the adolescent and adult forms of CHS we also found missense mutant alleles that likely encode CHS1 polypeptides with partial function. Together, these results suggest an allelic genotype–phenotype relationship among the various clinical forms of CHS. © 2002 Wiley‐Liss, Inc.     

Crystal structure of human DAAM1 formin homology 2 domain

Reorganization of the actin filament is an essential process for cell motility, cell-cell attachment and intracellular transport. Formin proteins promote nucleation and elongation of the actin filament, and thus are key regulators for this process. The formin homology 2 (FH2) domain forms a head-to-tail ring-shaped dimer, and processively moves towards the barbed end. Dishevelled-associated activator of morphogenesis (DAAM) is a Rho-regulated formin implicated in neuronal development. Here, we present the crystal structure of human DAAM1 FH2 dimer at 2.8 A resolution. This is the first dimeric structure of the mammalian formin. The core structure of human DAAM1 is similar to those of mouse mDia1 and yeast Bni1p, whereas the orientations of the FH2 dimeric rings are different between human DAAM1 and yeast Bni1p, despite their similar dimer interactions. This difference supports the previous prediction that the dimer architecture of the formin is highly flexible in the actin-free state. The results of the actin assembly assays using the DAAM1 mutants demonstrated that the length of the linker connecting the N-terminal domain and the core region is crucial for the activity.     

A Study on the Inhibitory Effect of Matrine on Gastric Cancer SGC-7901 Cells

The objective of this paper was to investigate the antitumour mechanism of action of matrine by studying its inhibitory effect on gastric cancer SGC-7901 cells. SGC-7901 cells were chosen, and cell-killing capacity of matrine on gastric cancer SGC-7901 cells was determined using MTT assay and single PI staining assay. The results showed that matrine had an inhibitory effect on gastric cancer SGC-7901 cells, which was somewhat dose-dependent. The study concluded that matrine has a significant in-vitro inhibitory effect on SGC-7901 tumour cells, influences cell cycle of SGC-7901 cells, and induces their apoptosis.     

New CR system with pixel size of 50 microm for digital mammography: physical imaging properties and detection of subtle microcalcifications

PURPOSE: To investigate the physical imaging properties and detection of simulated microcalcifications of a new computed radiography (CR) system with a pixel size of 50 microm for digital mammography. MATERIALS AND METHODS: New and conventional CR were employed in this study. The new CR system included a high-resolution imaging plate coupled with the FCR5000MA (50 microm pixel pitch) including transparent support and a dual-sided reader. The conventional CR system was coupled with the FCR9000 (100 microm pixel pitch). Modulation transfer functions (MTFs) and Wiener spectra (WS) of the new and conventional CR systems were measured. Observer performance tests were conducted to compare the effects of pixel size (50 microm vs. 100 microm) on the diagnostic accuracy of CR systems in the detection of simulated microcalcifications. RESULTS: The presampling MTF of the new CR system was higher at high frequencies than the conventional CR system. The WS of the new CR system was comparable to that of the conventional CR system at all frequencies. The area under the receiver operating characteristic (ROC) curve (Az) obtained with the new CR and the conventional CR systems were 0.84 and 0.79, respectively. Results showed that the detection of simulated clustered microcalcifications was significantly improved by use of the new CR system compared with the conventional CR system (p<0.05). CONCLUSION: The new CR mammography system improved physical imaging properties and detection of simulated microcalcifications over conventional CR mammography.     

Horizontal transmission of foot‐and‐mouth disease virus O/JPN/2010 among different animal species by direct contact

Foot‐and‐mouth disease (FMD) is highly contagious and easily transmitted among species of cloven‐hoofed animals. To investigate the transmission of FMD virus (FMDV) among different animal species, experimental infections using the O/JPN/2010 strain were performed in cows, goats and pigs. One cow or two goats/pigs were housed with a different species of inoculated animals, and clinical observations, virus shedding and antibody responses were analysed daily. Whilst all cows and goats were infected horizontally by contact with inoculated pigs, transmission from cows to goats/pigs and from goats to cows/pigs was not observed in all in‐contact animals. In particular, no pigs were infected horizontally by contact with inoculated goats. Comparison with our previous study on experimental infections among animals of the same species indicates that horizontal transmission occurred more easily between animals of the same species than between those of the different species. These findings will be useful for establishing and performing species‐specific countermeasures in farms and regions where multiple species of animals coexist in potential future outbreaks.     

Strong cytoplasmic expression of NF-κB/p65 correlates with a good prognosis in patients with triple-negative breast cancer

Purpose

Recent studies have indicated that constitutive NF-κB activity could be involved in the proliferation of triple-negative breast cancer.

Methods

The NF-κB/p65 expression and the effects of a NF-κB inhibitor, (?)-DHMEQ, were examined in triple-negative MDA-MB-231 breast cancer cells. Women with triple-negative breast cancer treated with neoadjuvant chemotherapy between 2002 and 2012 were retrospectively analyzed for their expression of NF-κB/p65, Bcl2 and Ki67 by immunohistochemistry in pre- and post-treatment specimens. The factors predicting the response to neoadjuvant chemotherapy and the prognosis were analyzed.

Results

NF-κB/p65 was predominantly expressed in the cytoplasm of MDA-MB-231 cells. Of 34 triple-negative breast cancer patients, positive staining for NF-κB/p65 expression was detected in the nuclei of a few cells in seven tumors before neoadjuvant chemotherapy, while the expression of NF-κB/p65 in the cytoplasm was detected in almost all tumor cells of 33 tumors. The expression levels of NF-κB/p65 were not associated with the response to neoadjuvant chemotherapy, although the cytoplasmic NF-κB/p65 staining intensity was significantly decreased in the post-treatment tumor samples compared with the pretreatment samples. All patients whose tumors showed strong cytoplasmic NF-κB/p65 expression before neoadjuvant chemotherapy are currently disease free.

Conclusion

Our results suggest that strong cytoplasmic NF-κB/p65 expression could be a prognostic marker for patients with triple-negative breast cancer.

     

Liver abscess complicating intratumoral ethanol injection therapy for HCC

We report a patient who developed multiple liver abscesses and sepsis caused by lactobacilli after the percutaneous intratumoral injection of ethanol for hepatocellular carcinoma. We diagnosed the liver abscess at an early stage because of a the finding of gas on ultrasound and computed tomography. Blood cultures grew Gram-positive rods, which were of the Lactobacillus species. The patient responded to the administration of antibiotics, and his hepatic tumors have not recurred in the 7 months since treatment. This is the first report of liver abscess following percutaneous ethanol injection therapy.     

当归多糖对高糖状态下大鼠骨髓间充质干细胞成骨向分化的影响

目的 探讨当归多糖（ASP）对高糖状态下大鼠骨髓间充质干细胞（BMSCs）成骨向分化的影响。方法 培养并收集第3代BMSCs进行成骨成脂分化诱导鉴定。将BMSCs分为3组进行培养：正常对照组（葡萄糖浓度5.5 mmol·L ^-1）、高糖组（葡萄糖浓度25.5 mmol·L ^-1）、ASP+高糖组（葡萄糖浓度25.5 mmol·L ^-1+40 mg·L ^-1 ASP）。CCK8检测各组BMSCs的增殖活性,茜素红染色和碱性磷酸酶活性检测成骨活性。实时荧光定量聚合酶链反应检测成骨标记基因Runt相关转录因子2（Runx-2）、锌指结构转录因子（Osx）、骨钙蛋白（OCN）、Ⅰ型胶原酶（COL-Ⅰ）mRNA及Wnt/β-catenin信号通路关键因子CyclinD1及β-catenin的mRNA表达。建立2型糖尿病大鼠模型,将大鼠分为3组：正常对照组（正常大鼠）、糖尿病组（糖尿病大鼠）、糖尿病+ASP组（糖尿病大鼠,ASP喂养）,制备大鼠胫骨骨缺损,进行组织学检测,观察骨缺损修复情况。结果 高糖组、ASP+高糖组的BMSCs增殖高于正常对照组（P<0.05）,高糖组和ASP+高糖组二者之间无统计学差异（P>0.05）。高糖组中BMSCs钙结节数量、碱性磷酸酶活性及Runx-2、OCN、Osx、COL-Ⅰ、CyclinD1、β-catenin的mRNA表达均低于正常对照组和ASP+高糖组（P<0.05）,正常对照组和ASP+高糖组之间无统计学差异（P<0.05）。组织学检测结果显示,糖尿病组骨小梁数量少于正常对照组和糖尿病+ASP组（P<0.05）,而正常对照组和糖尿病+ASP组二者之间无统计学差异（P>0.05）。结论 ASP可促进高糖状态下大鼠BMSCs的成骨分化及2型糖尿病大鼠的骨缺损修复,这种促进作用可能与Wnt/β-catenin信号通路的激活有关。