肝脏及肢体缺血预处理抗大鼠肝缺血再灌注损伤的实验研究

目的研究肝脏缺血预处理(经典缺血预处理IPC)的第一保护窗(FW)与肢体缺血预处理(远端缺血预处理RPC)的第二保护窗(SW)及两者联合应用对大鼠肝脏缺血再灌注(I/R)损伤的保护作用及可能机制。方法大鼠随机分成5组:I/R组不行预处理;IPC组以肝缺血5 min行预处理;RPC组以双后肢缺血5 min,反复3次行预处理;RPC+IPC组先行RPC,24 h后行IPC作预处理;S组仅行开腹,不行其他处理。3个预处理组及I/R组均行肝缺血1 h再灌注3 h。取血用于血清谷丙转氨酶(ALT)与血清谷草转氨酶(AST)检测。切取肝组织用于测定肿瘤坏死因子α(TNF-α)和热休克蛋白70(HSP70)的表达、湿干比(W/D)及观察显微、超微结构的变化。结果与I/R组比较,IPC组,RPC组及RPC+IPC组ALT,AST,W/D及TNF-α阳性表达均明显降低(P0.01),HSP70表达量明显增加(P0.01),肝脏的显微及超微结构损伤减轻;IPC,RPC,RPC+IPC组3组间各项指标差异无统计学意义(P0.05)。结论IPC的FW,RPC的SW及两者联合应用对大鼠肝脏I/R损伤均有明显的保护作用,三者在保护强度上无明显差异。其机制可能与抑制TNF-α的产生、诱导保护性蛋白HSP70的表达、减轻肝脏水肿、改善肝组织微循环有关。     

In vivo and in vitro inhibitory effects of a traditional Chinese formulation on LPS-stimulated leukocyte-endothelial cell adhesion and VCAM-1 gene expression

Aim of the study

A traditional Chinese formulation Huang-Lian-Jie-Du-Tang (HLJDT) exerts anti-inflammatory effects. The present study aimed to investigate the effect of HLJDT on the LPS-stimulated leukocyte–endothelial cell adhesion and VCAM-1 gene expression both in vivo and in vitro.

Materials and methods

HLJDT was extracted from rhizoma coptidis, radix scutellariae, cortex phellodendri and fructus gardeniae in a weight rario of 1:1:1:1. In vivo leukocyte–endothelial cell adhesion was observed in rat lung after LPS stimulation (5 mg/kg, i.p.) with or without HLJDT (350 or 700 mg/kg, i.g.) pretreatment. The protein expression of vascular cell adhesion molecule 1 (VCAM-1) was analyzed by immunohistochemical method. In vitro leukocyte–endothelial cell adhesion was performed by examining the adhesion of THP-1 cells to LPS-stimulated human vascular endothelial cells with or without HLJDT pretreatment. The VCAM-1 expression at the RNA and protein levels was investigated by RT-PCR and western blot analysis, respectively. The activation of NF-κB was examined by the nuclear translocation of NF-κB by immunocytochemical method.

Results

In vivo, HLJDT dose-dependently reduced the number of leukocytes adhered to endothelium and VCAM-1 protein expression in lung venules of LPS-challenged rats. In vitro, HLJDT dose-dependently decreased the number of THP-1 cells adhered to LPS-stimulated endothelial cells and the expression of VCAM-1 both at the RNA and protein levels. The LPS-induced nuclear translocation of NF-kappa B in endothelial cells was also dose-dependently inhibited by HLJDT.

Conclusions

The present study demonstrated an additional mechanism underlying the anti-inflmmatory effect of HLJDT by inhibiting the leukocyte–endothelial cell adhesion and VCAM-1 gene expression. The inhibition of NF-kappa B activation by HLJDT might suggest a profound anti-inflammatory consequences.     

Treatment of retroperitoneal echinococcosis: the experience of a single center

Retroperitoneal echinococcosis (RE) is a rare condition that is associated with a high mortality and disability rate. It is associated with a high rate of misdiagnosis, a high risk of surgery, and is extremely difficult to manage. There is no uniform standard for determining the exact form of surgical method and the timing of surgery.This was a retrospective analysis of the characteristics and surgical management of patients diagnosed with RE in our hospital between 2012 and 2019.Between 2012 and 2019, 1257 cases of echinococcosis and 121 cases of RE were diagnosed in our hospital. Of these, 68 cases involved surgical treatment, 53 involved non-surgical treatment, and 12 cases were lost to follow-up (4 cases in the surgical group and 8 cases in the non-surgical group). Thus, 109 cases were followed-up. RE cases were divided according to different treatment methods into a radical resection group (Group A, 31 cases), a non-radical resection group (Group B, 37 cases), and a non-surgical group (Group C, 53 cases). We carried out a detailed analysis of the 109 cases experiencing surgical intervention with effective follow-up.Our analysis found that radical resection is the first line of treatment of RE, although non-radical surgery can benefit most patients. It is important to emphasize the importance of the first round of surgery, particularly in cases involving hepatic echinococcosis. If the lesion can be removed radically during the first round of surgery, then radical surgery should be performed.     

Bacterial genome reduction using the progressive clustering of deletions via yeast sexual cycling

The availability of genetically tractable organisms with simple genomes is critical for the rapid, systems-level understanding of basic biological processes. Mycoplasma bacteria, with the smallest known genomes among free-living cellular organisms, are ideal models for this purpose, but the natural versions of these cells have genome complexities still too great to offer a comprehensive view of a fundamental life form. Here we describe an efficient method for reducing genomes from these organisms by identifying individually deletable regions using transposon mutagenesis and progressively clustering deleted genomic segments using meiotic recombination between the bacterial genomes harbored in yeast. Mycoplasmal genomes subjected to this process and transplanted into recipient cells yielded two mycoplasma strains. The first simultaneously lacked eight singly deletable regions of the genome, representing a total of 91 genes and ∼10% of the original genome. The second strain lacked seven of the eight regions, representing 84 genes. Growth assay data revealed an absence of genetic interactions among the 91 genes under tested conditions. Despite predicted effects of the deletions on sugar metabolism and the proteome, growth rates were unaffected by the gene deletions in the seven-deletion strain. These results support the feasibility of using single-gene disruption data to design and construct viable genomes lacking multiple genes, paving the way toward genome minimization. The progressive clustering method is expected to be effective for the reorganization of any mega-sized DNA molecules cloned in yeast, facilitating the construction of designer genomes in microbes as well as genomic fragments for genetic engineering of higher eukaryotes.Complexities of natural biological systems make it difficult to understand and define precisely the roles of individual genes and their integrated functions. The use of model organisms with a relatively small number of genes enables the isolation of core biological processes from their complex regulatory networks for extensive characterization. However, even the simplest natural microbes contain many genes of unknown function, as well as genes that can be singly or simultaneously deleted without any noticeable effect on growth rate in a laboratory setting (Hutchison et al. 1999; Glass et al. 2006; Posfai et al. 2006). Ill-defined genes and those mediating functional redundancies both compound the challenge of understanding even the simplest life forms.Toward generating a minimal cell where every gene is essential for the axenic viability of the organism, we are pursuing strategies to reduce the 1-Mb genome of Mycoplasma mycoides JCVI-syn1.0 (Gibson et al. 2010). Because we can (1) introduce this genome into yeast and maintain it as a plasmid (Benders et al. 2010; Karas et al. 2013a); and (2) “transplant” the genome from yeast into mycoplasma recipient cells (Lartigue et al. 2009), genetic tools in yeast are available for reducing this bacterial genome. Several systems offer advanced tools for bacterial genome engineering. Here we further exploit distinctive features of yeast for this purpose.Methods for serially replacing genomic regions with selectable markers are limited by the number of available markers. One effective approach is to reuse the same marker after precise and scarless marker excision (Storici et al. 2001). We have previously used a self-excising marker (Noskov et al. 2010) six times in yeast to generate a JCVI-syn1.0 genome lacking all six restriction systems (JCVI-syn1.0 ∆1-6) (Karas et al. 2013a). Despite the advantages of scarless engineering, sequential procedures are time-consuming. When applied to poorly characterized genes with the potential to interact with other genes, some paths for multigene knockout may lead to dead ends that result from synergistic mutant phenotypes. When a dead end is reached, sequentially returning to a previous genome in an effort to find a detour to a viable higher-order multimutant may be prohibitively time-consuming.An alternative approach to multigene engineering, available in yeast, is to prepare a set of single mutants and combine the deletions into a single strain via cycles of mating and meiotic recombination (Fig. 1A; Pinel et al. 2011; Suzuki et al. 2011, 2012). With a green fluorescent protein (GFP) reporter gene inserted in each deletion locus, the enrichment of higher-order yeast deletion strains in the meiotic population can be accomplished using flow cytometry. Here we apply this method to the JCVI-syn1.0 ∆1-6 exogenous, bacterial genome harbored in yeast to nonsequentially assemble deletions for genes predicted to be individually deletable based on biological knowledge or transposon-mediated disruption data. The functional identification of simultaneously deletable regions is expected to accelerate the effort to construct a minimal genome.Open in a separate windowFigure 1.Progressive clustering of deleted genomic segments. (A) Scheme of the method. Light blue oval represents a bacterial cell. Black ring or horizontal line denotes a bacterial genome, with the orange box indicating the yeast vector used as a site for linearization and recircularization. Gray shape denotes a yeast cell. Green dot in the genome indicates a deletion replaced with a GFP marker. (B) Map of deleted regions. Orange box indicates the yeast vector sequence used for genome linearization and recircularization. Green boxes indicate regions deleted in multimutant mycoplasma strains. Blue boxes denote restriction modification (RM) systems that are also deleted in the strains. (C) Pulsed-gel electrophoresis result for deleted genomes. The starting strain was the JCVI-syn1.0 ∆1–6 strain (1062 kb). Two strains were analyzed for each design of simultaneous deletion (962 kb for eight-deletion or 974 kb for seven-deletion genome). Ladder is a set of yeast chromosomes (New England BioLabs). (D) GFP-RFP ratio sorting result. Standard sorting was compared with sorting based on a GFP-RFP ratio (Methods).     

Effects of heavy metals on health risk and characteristic in surrounding atmosphere of tire manufacturing plant,Taiwan

The health and environmental effects of metal-containing carbon black (CB) particles emitted from a CB feeding area near a tire manufacturing plant were investigated. The mass ratios of PM₁ and PM_0.1 (UFPs) relative to TSP were 13.84% ± 4.88% and 50.84% ± 4.29%, respectively. The most abundant elements in all fractions were Fe, Al, and Zn. The mean percentage contributions of Al, Fe, Zn, Cu, and Co to the coarse particles ranged from 49.1% to 69.1%, thus indicating that the Al, Fe, and Zn contents in the CB particles were affected by workplace emissions. The ratios of the total mean deposition fluxes of atmospheric particle-bound heavy metals in the human respiratory tracts of workers/adults, workers/children, and adults/children were approximately 5.5, 11.0, and 2.0, respectively. The integrated risks of five elements via two exposure pathways to adults and children were 1.1 × 10⁻⁴ and 1.7 × 10⁻⁵, respectively; these numbers reflect the high cumulative carcinogenic risk posed by these toxic metals to local residents (both adults and children; limit, 10⁻⁶). These results demonstrate the potential health risk presented by particle-bound heavy metals to humans residing near tire manufacturing plants via inhalation and dermal contact exposure.

The health and environmental effects of metal-containing carbon black (CB) particles emitted from a CB feeding area near a tire manufacturing plant were investigated.     

An In‐House Prevocational Training Program for Newly Discharged Psychiatric Inpatients: Exploring Its Employment Outcomes and the Predictive Factors

Individuals with severe mental disorders continue to experience low employment rates. Occupational therapists play an important role in helping these individuals develop the skills and obtain the supports necessary for productive living. This retrospective cohort study aimed to explore employment outcomes and identify factors predictive of the outcomes of an in‐house prevocational training program designed for newly discharged psychiatric inpatients. Data retrieved from the files of 58 participants including demographics, diagnostic history, physical fitness, functional assessment results, the use of vocational counselling service and employment status were analyzed. The overall employment rates among the participants were high (67.2–79.3%) within the 6 months following the prevocational training program. No significant differences were found in the employment rates across the 1, 3 and 6‐month time periods post‐training. Vocational counselling service post‐training and hand function were two factors predictive of participants' employment outcomes. Occupational therapists should attend to the clients' need for continuous vocational support and carefully identify the personal, functional and environmental factors contributing to successful employment. Caution in interpreting the results is warranted because of the lack of control and randomization in this retrospective study. Additional longitudinal cohort or experimental studies would add further certainty to the current findings. Copyright © 2015 John Wiley & Sons, Ltd.     

Lower risk of primary Sjogren’s syndrome in patients with dengue virus infection: a nationwide cohort study in Taiwan

Clinical Rheumatology - The data concerning the association between dengue viruses (DV) infection and autoimmune diseases (ADs) remain unclear and are scarce. This nationwide population-based...     

An adult case of Chryseobacterium meningosepticum meningitis

Chryseobacterium meningosepticum is an uncommon pathogen causing adult bacterial meningitis. Herein, we report the case history of one 21-year-old woman with this uncommon central nervous system infection. A diagnosis of adult C. meningosepticum meningitis can only be confirmed by a positive cerebrospinal fluid (CSF) culture. The patient had insulin-dependent diabetes mellitus as the underlying condition associated with this infection. The clinical presentations were fever, headache, consciousness disturbance, and seizure. CSF analysis revealed a purulent inflammatory reaction. After a 21-day course of intravenous cefepime (6 g/day) treatment, this patient was discharged in a state of complete recovery.