The ctnF gene is involved in citrinin and pigment synthesis in Monascus aurantiacus

The application of Monascus is restricted by citrinin. So, it is important to explore the synthetic pathway of citrinin to completely inhibit the production of citrinin. In our previous study, we found that the protein encoded by the ctnF gene has a significant similarity to fructose-2,6-bisphosphatase (F26BPase). It is generally known that the bifunctional enzyme F26BPase regulates the glycolytic flux. So, we speculated that the CtnF protein strengthens carbon flux towards acetyl-CoA and malonyl-CoA which are precursor compounds in citrinin and pigment synthesis. In this study, the ctnF gene-targeting vector pctnF-HPH was constructed and transformed into Monascus aurantiacus. A ctnF-deficient strain was selected by four sets of primers and polymerase chain reaction amplification. Compared with the wild-type strain, citrinin content in the deficient strain was reduced by 34%, and the pigment production was decreased by 72%. These results indicate that the ctnF gene is involved in the common synthesis of citrinin and pigment, which is consistent with previous speculations.     

组织玻璃化冻存技术：优势与尚未完全解决的问题

文题释义：玻璃化冷冻保存：是利用这些高浓度的低温保护剂组合成玻璃化冻存液,通过与水分子发生强烈的水合作用,增加溶液黏性,降低冰晶形成速度,从而使细胞在快速降温或复温过程中得以保护。 玻璃化：对于非晶高分子,当高分子通过降温从高弹态转变为玻璃态,或者通过升温从玻璃态转变为高弹态的过程称之为玻璃化转变,发生玻璃化转变的温度叫玻璃化转变温度。对于结晶高分子,玻璃化转变是指其非晶部分所发生的由高弹态向玻璃态(或者玻璃态向高弹态)的转变。因此,玻璃化转变是高分子中普遍存在的现象。但是玻璃化转变现象并不局限于高分子,一些小分子化合物也存在玻璃化转变。 背景：玻璃化冷冻保存是一种应用前途广阔的低温冷冻方法,通过使用高浓度的玻璃化冻存试剂将生物材料进行玻璃态转变,从而实现活性保存。 目的：就玻璃化冻存的生物学原理及玻璃化冻存试剂的分类,卵巢、皮肤与角膜等医学组织标本的玻璃化冻存进行综述。 方法：以“tissue;vitrification;cryopreservation”为英文检索词;“组织;玻璃化;冷冻保存”为中文检索词,检索1994年1月至2019年10月 PubMed 数据库及万方医学网相关文献。按照纳入与排除标准筛选后,对最终纳入的45篇文献进行归纳总结。 结果与结论：玻璃化冻存可以防止细胞内外冰晶形成,避免了冰晶给细胞带来的多种损伤,有效保留了细胞的生物活性与基本功能。玻璃化冻存试剂主要分为渗透性和非渗透性2种,其操作简便、高效,唯一的缺点是高浓度的冻存试剂对细胞具有一定的毒性损伤。为了降低对组织整体损伤风险,可以混合使用多种低毒冻存试剂。目前玻璃化冻存技术已经成功应用于多种细胞,但组织冻存的技术难题尚未完全解决。 ORCID: 0000-0003-0140-9935(张源) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Haemorrhagic cystitis following the administration of voriconazole in the treatment of central nervous system aspergillosis: a case report

Central nervous system aspergillosis (CNS-A) is a rare and fatal fungal infection. Voriconazole is the recommended treatment for CNS-A. The therapeutic effect of voriconazole is good, but its use is limited due to adverse reactions. This case report describes a 37-year-old male patient that had previously been diagnosed with acute lymphoblastic leukaemia. He had received immunosuppressive agents for 1 year following a haematopoietic bone marrow transplant. He presented with a 1-month history of left limb weakness as well as recurrent fever. Brain magnetic resonance imaging showed that he had multiple cerebral infarctions. Subsequently, he was diagnosed with CNS-A by metagenomic next-generation sequencing. Voriconazole was added to his treatment regimen, but it resulted in severe haemorrhagic cystitis and possibly bladder rupture. The dose of voriconazole was adjusted and reparative bladder surgery was undertaken immediately. Eventually, the patient was successfully treated with voriconazole and there was no recurrence of symptoms after 1 year of follow-up. Haemorrhagic cystitis is a rare adverse drug reaction associated with voriconazole use. Based on the experience with this current case, physicians should be aware of urinary tract complications with voriconazole including haemorrhagic cystitis.     

Nanoparticle-directed sub-cellular localization of doxorubicin and the sensitization breast cancer cells by circumventing GST-Mediated drug resistance

Resistance to single or multiple chemotherapeutic drugs is a major complication in clinical oncology and is one of the most common treatment limitations in patients with reoccurring cancers. Nanoparticle (NP)-based drug delivery systems (DDS's) have been shown to overcome drug resistance in cancer cells mainly by avoiding the activation of efflux pumps in these cells. We demonstrate in this work that polyester-based hyperbranched dendritic-linear (HBDL)-based NPs carrying doxorubicin (Dox) can effectively overcome microsomal glutathione transferase 1 (MGST1)-mediated drug resistance in breast cancer cells. Our DDS was much more effective at considerably lower intracellular Dox concentrations (IC₅₀ 6.3 μm vs. 36.3 μm) and achieved significantly greater reductions in viability and induced higher degrees of apoptosis (31% vs. 14%) compared to the free drug in the resistant cells. Dox-loaded HBDL NPs were found to translocate across the membranes of resistant cells via active endocytic pathways and to be transported to lysosomes, mitochondria, and the endoplasmic reticulum. A significantly lower amount of Dox accumulated in these cytoplasmic compartments in resistant cells treated with free Dox. Moreover, we found that Dox-HBDL significantly decreased the expression of MGST1 and enhanced mitochondria-mediated apoptotic cell death compared to free Dox. Dox-HBDL also markedly activated the JNK pathway that contributes to the apoptosis of drug-resistant cells. These results suggest that HBDL NPs can modulate subcellular drug distribution by specific endocytic and trafficking pathways and that this results in drug delivery that alters enzyme levels and cellular signaling pathways and, most importantly, increases the induction of apoptosis. Our findings suggest that by exploiting the cell transport machinery we can optimize the polymeric vehicles for controlled drug release to overcome drug resistance combat drug resistance with much higher efficacy.     

A recombinant adenovirus expressing CFP10, ESAT6, Ag85A and Ag85B of Mycobacterium tuberculosis elicits strong antigen-specific immune responses in mice

Tuberculosis (TB) is caused by an infection of Mycobacterium tuberculosis (Mtb) and remains an enormous and increasing health burden worldwide. To date, Mycobacterium bovis Bacillus Calmette Guerin (BCG) is the only licensed anti-TB vaccine worldwide, which provides an important but limited protection from the Mtb infection. The development of alternative anti-TB vaccines is therefore urgently needed. Here we report, the generation of Ad5-CEAB, a recombinant adenovirus expressing Mtb antigens of CFP10, ESAT6, Ag85A and Ag85B proteins in a form of mixture. In order to evaluate the immunogenicity of Ad5-CEAB, mice were immunized with Ad5-CEAB by intranasal instillation three times with 2-week intervals. The results demonstrated that Ad5-CEAB elicited a strong antigen-specific immune response, particularly of the Th1 immune responses that were characterized by an increased ratio of IgG2a/IgG1 and secretions of Th1 type cytokines, IFN-γ, TNF-α, IL-2 and IL-12. In addition, the Ad5-CEAB also showed an ability to enhance humoral responses with a dramatically augmented antigen-specific serum IgG. Furthermore, an elevated sIgA were also found in the bronchoalveolar lavage fluid of the immunized mice, suggesting the elicitation of mucosal immune responses. These data indicate that Ad5-CEAB can induce a broad range of antigen-specific immune responses in vivo, which provides a promising and novel route for developing anti-TB vaccines and warrants further investigation.     

Upregulated expression of human alpha-defensins 1, 2 and 3 in hypercholesteremia and its relationship with serum lipid levels

Human alpha-defensins are natural antimicrobial peptides of neutrophils evolved in host defense reactions and circulating nonstressed alpha-defensins may be associated with serum lipid levels. The aim of this work was to examine whether the expression of alpha-defensins 1, 2 and 3 genes are changed and whether this changes are reversed following treatment in patients with hypercholesteremia. A total of 40 individuals of hypercholesteremia group were studied, compared with 40 individuals of normal control group. Protein levels and gene expression levels of alpha-defensins 1, 2 and 3 were significantly higher in patients with hypercholesteremia compared with subjects in normal control group. In patients with hypercholesteremia, protein levels of alpha-defensins 1, 2 and 3 correlated positively with the levels of total cholesterol and low-density lipoprotein cholesterol. Protein levels and gene expression levels of alpha-defensins 1, 2 and 3 were decreased significantly after a treatment with atorvastatin calcium 20 mg daily compared with the patients before the treatment. Our results suggest that the expression of alpha-defensins 1, 2 and 3 genes is involved in dyslipidemia in patients with hypercholesteremia.