Changes in physicochemical and gut microbiota fermentation property induced by acetylation of polysaccharides from Cyperus esculentus.

In this study, the impact of acetylation on physicochemical, digestive behavior and fermentation characteristics of Cyperus esculentus polysaccharides (CEP) was investigated. Results indicated that the acetylation led the molecules to be more likely aggregated, followed by a higher crystallinity, a lower apparent viscosity and a higher ratio of G" to G' (tan δ). Importantly, the acetylated polysaccharides (ACEP) had a lower digestibility, but its molecular weight was lower than that of original polysaccharides (CEP) following a simulated saliva-gastrointestinal digestion. Gut microbiota fermentation indicated that both polysaccharides generated outstanding short-chain fatty acids (SCFAs), in which the acetylated polysaccharides had a faster fermentation kinetics than the original one, followed by a quicker reduction of pH and a more accumulation of SCFAs, particularly butyrate. Fermentation of both polysaccharides promoted Akkermansia, followed by a reduced richness of Klebsiella. Importantly, the current study revealed that the fermentation of acetylated polysaccharides enriched Parabacteroides, while fermentation of original ones promoted Bifidobacterium, for indicating their individual fermentation characteristics and gut environmental benefits.

Structurally manipulated antioxidant peptides derived from wheat bran: Preparation and identification.

Bioactive peptide's development is facing two challenges in terms of its lower yield and limited understanding of structurally orientated functionality. Therefore, peptides were prepared from wheat bran via a cocktail enzyme for achieving a higher level of hydrophobic amino acids than traditional method. The obtained peptides exhibited great antioxidant activities against H2O2-induced oxidative stress in HepG2 cells. Among them, 91 bioactive peptides were selected through the virtual screening, and their N-terminal and C-terminal contained many hydrophobic amino acids. Then the peptides with capacity to interact with Keap1 were identified by in silico simulation, because Keap1 acts as a sensor of redox insults. The results revealed that peptides DLDW and DLGL demonstrated the highest binding affinities, and a bridge was formed between Asp of DLGL and Arg415 of Klech domain, contributing to interfering Keap1-Nrf2 interaction. These findings implied a potential application of wheat bran peptides as nutraceuticals and health-promoting ingredients.

Structurally Orientated Rheological and Gut Microbiota Fermentation Property of Mannans Polysaccharides and Oligosaccharides.

Three mannan polysaccharides and their oligosaccharides were investigated in terms of physicochemical characteristics and effects on gut microbiota. Oligosaccharides from guar gum had the fastest fermentation kinetics for SCFAs generation at the initial stage, while the locust bean of both polymers and oligosaccharides demonstrated the lowest SCFAs through the whole fermentation process. In contrast, konjac gum steadily increased SCFAs and reached its maximum level at 24 h fermentation, indicating its fermentation character may be associated with its rheological properties. Compared to their corresponding polysaccharides, all the oligosaccharides demonstrated a faster fermentation kinetics, followed by an enriched abundance of propionate-producing bacterial Prevotella and a decreased abundance of Megamonas and Collinsella. Meanwhile, oligosaccharides reduced the Firmicutes/Bacteroidota ratio as well as the abundance of Bacteroidetes and Escherichia-Shigella. The fermentation of konjac substrate significantly promoted the abundance of butyrate-producing bacterial Faecalibacterium. In contrast, although the fermentation of locust bean and guar gum substrates benefited Bifidobacterium abundance due to their similar structure and monosaccharides composition, the fermentation of locust bean gum led to greater Bifidobacterium than the others, which may be associated with its higher mannose composition in the molecules. Interestingly, the partial hydrolysis of the three polysaccharides slightly reduced their prebiotic function.

Association between thyroid stimulating hormone levels and papillary thyroid cancer risk: A meta-analysis.

High thyroid stimulating hormone (TSH) levels may stimulate papillary thyroid cancer (PTC) cell proliferation; however, the relationship between TSH levels and PTC risk remains controversial. We aim to ascertain the association through a meta-analysis. Literature searches were conducted in PubMed, Embase, and Web of Science databases. After literature screening, the methodological quality was assessed using the Newcastle-Ottawa Scale and Agency for Healthcare Research and Quality methods. Cochran's Q and I 2 tests were used to evaluate heterogeneity in the meta-analysis. Egger's test was applied to assess publication bias. A total of 12 eligible studies were included in this meta-analysis; all were of moderate and high methodological quality. The pooled results suggested that increased TSH levels were significantly associated with PTC risk; however, the included studies were significantly heterogeneous. Stratification analysis indicated that the heterogeneity might be from the area or type of control. Although significant publication bias existed among the studies, the trim-and-fill method and sensitivity analysis revealed that the combined results were stable and robust. TSH levels are significantly associated with the PTC risk; however, more high-quality studies in large sample sizes are recommended to verify the extrapolation of these findings.

Obstructive Sleep Apnea-induced Endothelial Dysfunction Is Mediated by miR-210.

Rationale: Obstructive sleep apnea (OSA)-induced endothelial cell (EC) dysfunction contributes to OSA-related cardiovascular sequelae. The mechanistic basis of endothelial impairment by OSA is unclear. Objectives: The goals of this study were to identify the mechanism of OSA-induced EC dysfunction and explore the potential therapies for OSA-accelerated cardiovascular disease. Methods: The experimental methods include data mining, bioinformatics, EC functional analyses, OSA mouse models, and assessment of OSA human subjects. Measurements and Main Results: Using mined microRNA sequencing data, we found that microRNA 210 (miR-210) conferred the greatest induction by intermittent hypoxia in ECs. Consistently, the serum concentration of miR-210 was higher in individuals with OSA from two independent cohorts. Importantly, miR-210 concentration was positively correlated with the apnea-hypopnea index. RNA sequencing data collected from ECs transfected with miR-210 or treated with OSA serum showed a set of genes commonly altered by miR-210 and OSA serum, which are largely involved in mitochondrion-related pathways. ECs transfected with miR-210 or treated with OSA serum showed reduced [Formula: see text]o2 rate, mitochondrial membrane potential, and DNA abundance. Mechanistically, intermittent hypoxia-induced SREBP2 (sterol regulatory element-binding protein 2) bound to the promoter region of miR-210, which in turn inhibited the iron-sulfur cluster assembly enzyme and led to mitochondrial dysfunction. Moreover, the SREBP2 inhibitor betulin alleviated intermittent hypoxia-increased systolic blood pressure in the OSA mouse model. Conclusions: These results identify an axis involving SREBP2, miR-210, and mitochondrial dysfunction, representing a new mechanistic link between OSA and EC dysfunction that may have important implications for treating and preventing OSA-related cardiovascular sequelae.

Low-temperature adaptation and preservation revealed by changes in physiological-biochemical characteristics and proteome expression patterns in post-harvest Hami melon during cold storage.

MAIN CONCLUSION: The proteome and its time-dependent effects reveal the importance of stress response (including expression regulation of heat-shock proteins) and fatty acid metabolism in cold adaptation and preservation of Hami melon. To better understand the molecular mechanism of how Hami melons respond to low-temperature stress, this study investigated the relevant physiological characteristics, catalytic antibody activity, and quantitative proteomics of Hami melon (Jiashi muskmelon) during low-temperature storage. Jiashi muskmelon was stored inside two refrigerators set at 21 °C (control group) and 3 °C, respectively, for 24 days. Low-temperature storage led to a significantly reduced decay rate, weight loss rate, and loss of relative conductivity. It also maintained fruit firmness, inhibited the production rate of malondialdehyde and H2O2, and induced over-expression of antioxidant enzyme and ATPase. A total of 1064 differentially expressed proteins (DEPs) were identified during low-temperature storage. Stimulation response was the main process in response to low-temperature. To further verify the proteome data, we selected four heat-shock proteins (HSP) displaying relatively high expression levels. Real-time fluorescence PCR results confirmed that HmHSP90 I, HmHSP90 II, HmHSP70, and HmsHSP were significantly up-regulated upon low-temperature induction. These proteins may protect the Hami melon from physiological and cellular damage due to the low-temperature stress by acting alone or synergistically. Additionally, the main enrichment term of the fatty acid metabolism-related DEPs was fatty acid beta oxidation at 21 °C in contrast to fatty acid biosynthesis processes at 3 °C. It is speculated that Hami melon enhances low-temperature adaptability by slowing down the oxidative degradation of fatty acids and synthesizing new fatty acids at low temperatures. This study provides new insights into the mechanism of low-temperature adaptation and preservation in post-harvest Hami melon during cold storage.

Severe cerebral edema in substance-related cardiac arrest patients.

BACKGROUND: Studies of neurologic outcomes have found conflicting results regarding differences between patients with substance-related cardiac arrests (SRCA) and non-SRCA. We investigate the effects of SRCA on severe cerebral edema development, a neuroimaging intermediate endpoint for neurologic injury. METHODS: 327 out-of-hospital comatose cardiac arrest patients were retrospectively analyzed. Demographics and baseline clinical characteristics were examined. SRCA categorization was based on admission toxicology screens. Severe cerebral edema classification was based on radiology reports. Poor clinical outcomes were defined as discharge Cerebral Performance Category scores > 3. RESULTS: SRCA patients (N = 86) were younger (P < 0.001), and more likely to have non-shockable rhythms (P < 0.001), be unwitnessed (P < 0.001), lower Glasgow Coma Scale scores (P < 0.001), absent brainstem reflexes (P < 0.05) and develop severe cerebral edema (P < 0.001) than non-SRCA patients (N = 241). Multivariable analyses found younger age (P < 0.001), female sex (P = 0.008), non-shockable rhythm (P = 0.01) and SRCA (P = 0.05) to be predictors of severe cerebral edema development. Older age (P < 0.001), non-shockable rhythm (P = 0.02), severe cerebral edema (P < 0.001), and absent pupillary light reflexes (P = 0.004) were predictors of poor outcomes. SRCA patients had higher proportion of brain deaths (P < 0.001) compared to non-SRCA patients. CONCLUSIONS: SRCA results in higher rates of severe cerebral edema development and brain death. The absence of statistically significant differences in discharge outcomes or survival between SRCA and non-SRCA patients may be related to the higher rate of withdrawal of life-sustaining treatment (WLST) in the non-SRCA group. Future neuroprognostic studies may opt to include neuroimaging markers as intermediate measures of neurologic injury which are not influenced by WLST decisions.

Delayed rFGF21 Administration Improves Cerebrovascular Remodeling and White Matter Repair After Focal Stroke in Diabetic Mice.

Type 2 diabetes mellitus (T2DM) is a major comorbidity exacerbating ischemic brain injury and impairing post-stroke recovery. Our previous study suggested that recombinant human fibroblast growth factor (rFGF) 21 might be a potent therapeutic targeting multiple aspects of pathophysiology in T2DM stroke. This study aims to evaluate the potential effects of rFGF21 on cerebrovascular remodeling after T2DM stroke. Permanent distal middle cerebral artery occlusion was performed in heterozygous non-diabetic db/ + and homozygous diabetic db/db mice. Daily rFGF21 administration was initiated 1 week after stroke induction and maintained for up to 2 weeks thereafter. Multiple markers associated with post-stroke recovery, including angiogenesis, oligodendrogenesis, white matter integrity, and neurogenesis, were assessed up to 3 weeks after stroke. Our results showed an impairment in post-stroke vascular remodeling under T2DM condition, reflected by the decreased expression of trophic factors in brain microvessels and impairments of angiogenesis. The defected cerebrovascular remodeling was accompanied by the decreased oligodendrogenesis and neurogenesis. However, delayed rFGF21 administration normalized post-stroke hyperglycemia and improved neurological outcomes, which may partially be via the promotion of pro-angiogenic trophic factor expression in brain microvessels and cerebrovascular remodeling. The better cerebrovascular remodeling may also contribute to oligodendrogenesis, white matter integrity, and neurogenesis after T2DM stroke. Therefore, delayed rFGF21 administration may improve neurological outcomes in T2DM stroke mice, at least in part by normalizing the metabolic abnormalities and promoting cerebrovascular remodeling and white matter repair.

Chitosan treatment reduces softening and chilling injury in cold-stored Hami melon by regulating starch and sucrose metabolism.

Cold-stored Hami melon is susceptible to chilling injury, resulting in quality deterioration and reduced sales. Pre-storage treatment with chitosan reduces fruit softening and chilling injury in melon; however, the underlying mechanism remains unclear. In this study, Gold Queen Hami melons were treated with 1.5% chitosan solution for 10 min before cold storage at 3°C and then the effect of chitosan was examined on fruit firmness, weight loss, chilling injury, soluble solid content (SSC), pectin, and soluble sugar contents of melon fruit. Also, the enzyme activities and gene expressions related to fruit softening and starch and sucrose metabolism were investigated. Chitosan treatment reduced the fruit softening and chilling injury, maintained the high levels of starch and sucrose contents, and regulated the enzyme activities and gene expressions related to starch and sucrose metabolism. Fruit firmness was significantly positively correlated with sucrose and starch contents. Altogether, we uncovered the potential mechanism of chitosan coating mitigating melon softening and chilling injury through the regulation of starch and sucrose metabolism.

Identification of Glutathione S-Transferase Genes in Hami Melon (Cucumis melo var. saccharinus) and Their Expression Analysis Under Cold Stress.

As a group of multifunctional enzymes, glutathione S-transferases (GSTs) participate in oxidative stress resistance and cellular detoxification. Here, we identified 39 CmGST genes with typical binding sites from the Hami melon genome, and they can be classified into seven subfamilies. Their molecular information, chromosomal locations, phylogenetic relationships, synteny relationships, gene structures, protein-protein interactions, structure of 3-D models, and expression levels under cold stress were analyzed. Expression analysis indicates that cold-tolerant Jia Shi-310 (JS) had higher GST enzyme activities and expression levels of 28 stress-related genes under cold stress. Some CmGSTs belonging to Tau, Phi, and DHAR classes play significant roles under cold stress, and they could be regarded as candidate genes for further studies. The present study systematically investigated the characterization of the Hami melon GST gene family, extending our understanding of Hami melon GST mediated stress-response mechanisms in this worldwide fruit.

Diabetes Mellitus/Poststroke Hyperglycemia: a Detrimental Factor for tPA Thrombolytic Stroke Therapy.

Intravenous administration of tissue-type plasminogen activator (IV tPA) therapy has long been considered a mainstay in ischemic stroke management. However, patients respond to IV tPA therapy unequally with some subsets of patients having worsened outcomes after treatment. In particular, diabetes mellitus (DM) is recognized as a clinically important vascular comorbidity that leads to lower recanalization rates and increased risks of hemorrhagic transformation (HT). In this short-review, we summarize the recent advances in understanding of the underlying mechanisms involved in post-IV tPA worsening of outcome in diabetic stroke. Potential pathologic factors that are related to the suboptimal tPA recanalization in diabetic stroke include higher plasma plasminogen activator inhibitor (PAI)-1 level, diabetic atherogenic vascular damage, glycation of the tPA receptor annexin A2, and alterations in fibrin clot density. While factors contributing to the exacerbation of HT in diabetic stroke include hyperglycemia, vascular oxidative stress, and inflammation, tPA neurovascular toxicity and imbalance in extracellular proteolysis are discussed. Besides, impaired collaterals in DM also compromise the efficacy of IV tPA therapy. Additionally, several tPA combination approaches developed from experimental studies that may help to optimize IV tPA therapy are also briefly summarized. In summary, more research efforts are needed to improve the safety and efficacy of IV tPA therapy in ischemic stroke patients with DM/poststroke hyperglycemia.

Dual antiplatelet therapy reduced stroke risk in transient ischemic attack with positive diffusion weighted imaging.

Dual antiplatelet therapy (DAPT) reduced stroke risk in high-risk transient ischemic attack (TIA) patients assessed by ABCD2 score. Patients with positive diffusion-weighted imaging (DWI) were identified as imaging-based high-risk. The present study aims to investigate whether DAPT could reduce stroke risk in TIA with DWI positive. The study enrolled TIA patients within 72 h of onset from the prospective TIA database of the First Affiliated Hospital of Zhengzhou University. The predictive outcome was ischemic stroke at 90-day. The relationship between DAPT and stroke was analyzed in a cox proportional hazards model. The Kaplan-Meier curves of TIA patients with DAPT and monotherapy were plotted. Total of 661 TIA patients were enrolled, 279 of whom were DWI positive and 281 used DAPT. The 90-day stroke risk was higher in patients used monotherapy than those used DAPT in TIA with positive DWI (23.7% vs. 13.4%, p = 0.029). DAPT was associated with reduced stroke risk in TIA patients with positive DWI (hazard ratio [HR] = 0.54; 95% confidence interval [CI], 0.30-0.97; p = 0.037). However, the benefit didn't exist in TIA patients with negative DWI (HR = 0.43; 95% CI, 0.14-1.33; p = 0.142). Early use of DAPT reduced stroke risk in TIA patients with positive DWI.

iTRAQ-based quantitative proteomics analysis of cantaloupe (Cucumis melo var. saccharinus) after cold storage.

BACKGROUND: Cantaloupe is susceptible to cold stress when it is stored at low temperatures, resulting in the loss of edible and commercial quality. To ascertain the molecular mechanisms of low temperatures resistance in cantaloupe, a cold-sensitive cultivar, Golden Empress-308 (GE) and a cold-tolerant cultivar, Jia Shi-310 (JS), were selected in parallel for iTRAQ quantitative proteomic analysis. RESULTS: The two kinds of commercial cultivars were exposed to a temperature of 0.5 °C for 0, 12 and 24 days. We found that the cold-sensitive cultivar (GE) suffered more severe damage as the length of the cold treatment increased. Proteomic analysis of both cultivars indicated that the number of differentially expressed proteins (DEPs) changed remarkably during the chilly treatment. JS expressed cold-responsive proteins more rapidly and mobilized more groups of proteins than GE. Furthermore, metabolic analysis revealed that more amino acids were up-regulated in JS during the early phases of low temperatures stress. The DEPs we found were mainly related to carbohydrate and energy metabolism, structural proteins, reactive oxygen species scavenging, amino acids metabolism and signal transduction. The consequences of phenotype assays, metabolic analysis and q-PCR validation confirm the findings of the iTRAQ analysis. CONCLUSION: We found that the prompt response and mobilization of proteins in JS allowed it to maintain a higher level of cold tolerance than GE, and that the slower cold responses in GE may be a vital reason for the severe chilling injury commonly found in this cultivar. The candidate proteins we identified will form the basis of future studies and may improve our understanding of the mechanisms of cold tolerance in cantaloupe.

Effects of pretreatment methods and leaching methods on jujube wine quality detected by electronic senses and HS-SPME-GC-MS.

Conventional analysis, electronic senses and HS-SPME-GC-MS were applied to evaluate the effects of pretreatment methods and leaching methods on jujube wine quality. Significant differences (p < 0.05) in the levels of alcohol content, color, taste and aroma were observed among all the jujube wine samples, in which the pulp and pectase fermented jujube wine was the best among all. Moreover, rather than taste, aroma is the most significantly (p < 0.05) affected. In regard to aroma, a total of 182 volatile compounds were identified by HS-SPME-GC-MS. It was found that the blended-into-pulp treatment and the leached-by-pectase treatment had notable positive effects on jujube wine. The pulp and pectase fermented jujube wines exhibited the highest concentration of total volatile compounds as well as alcohols, esters, acids and aldehydes. Thus, the optimal pretreatment method and leaching method for jujube wine fermentation are blended-into-pulp and leached-by-pectase respectively.

Soluble vascular endothelial-cadherin in CSF after subarachnoid hemorrhage.

OBJECTIVE: To determine if CSF and plasma levels of soluble vascular endothelial (sVE)-cadherin are associated with functional outcome after subarachnoid hemorrhage (SAH) and to investigate sVE-cadherin effects on microglia. METHODS: Serial CSF and plasma were collected from prospectively enrolled patients with nontraumatic SAH from a ruptured aneurysm in the anterior circulation and who required an external ventricular drain for clinical indications. Patients with normal-pressure hydrocephalus without SAH served as controls. For prospective assessment of long-term outcomes at 3 and 6 months after SAH, modified Rankin Scale scores (mRS) were obtained and dichotomized into good (mRS ≤ 2) vs poor (mRS > 2) outcome groups. For SAH severity, Hunt and Hess grade was assessed. Association of CSF sVE-cadherin levels with long-term outcomes, HH grade, and CSF tumor necrosis factor (TNF)-α levels were evaluated. sVE-cadherin effects on microglia were also studied. RESULTS: sVE-cadherin levels in CSF, but not in plasma, were higher in patients with SAH and were associated with higher clinical severity and higher CSF TNF-α levels. Patients with SAH with higher CSF sVE-cadherin levels over time were more likely to develop worse functional outcome at 3 months after SAH. Incubation of cultured microglia with sVE-cadherin resulted in increased inducible nitric oxide synthase, interleukin-1ß, reactive oxygen species, cell soma size, and metabolic activity, consistent with microglia activation. Microinjection of sVE-cadherin fragments into mouse brain results in an increased number of microglia surrounding the injection site, compared to injection of denatured vascular endothelial-cadherin fragments. CONCLUSIONS: These results support the existence of a novel pathway by which sVE-cadherin, released from injured endothelium after SAH, can shift microglia into a more proinflammatory phenotype and contribute to neuroinflammation and poor outcome in SAH.

Comparative study on operative trauma between microwave ablation and surgical treatment for papillary thyroid microcarcinoma.

AIM: To compare the effect and postoperative trauma of ultrasound-guided percutaneous microwave ablation and surgical resection in the treatment of papillary thyroid microcarcinoma (PTMC). METHODS: Eighty-seven patients with PTMC treated at Fudan University affiliated Shanghai Fifth People's Hospital were enrolled as subjects. The patients were divided into a microwave ablation group (41 cases) and a surgical group (46 cases). The operative time, intraoperative blood loss, length of hospital stay, serum C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), thyroid-related hormonal changes, and complications 7 d and 30 d after surgery were observed. RESULTS: The operative time, intraoperative blood loss, and length of hospital stay in the surgical group were significantly higher than those in the microwave ablation group (P < 0.05). The levels of CRP, IL-6, and TNF-α in the surgical group were significantly higher than those in the microwave ablation group (P < 0.05). The free triiodothyronine (FT3) and free thyroxin (FT4) levels in the surgical group were significantly lower than those in the microwave ablation group (P < 0.05). However, the postoperative thyroid stimulating hormone (TSH) level was significantly higher than that in the microwave ablation group (P < 0.05). There were significant interactions between the FT3, FT4, and TSH 7 d and 30 d after operation and the treatment methods (P < 0.05). There was no significant difference in the complications between the two groups (P > 0.05). CONCLUSION: Microwave ablation for papillary microcarcinoma of the thyroid gland has less trauma to the body, quicker recovery, and no scars. It can effectively shorten the length of hospital stay and improve the quality of life of patients.

Evaluation of elastography combined with serological indexes for hepatic fibrosis in patients with chronic hepatitis B.

AIM: To investigate the value of ultrasound elastography combined with serological indexes in diagnosing liver fibrosis and assessing its severity. METHODS: A total of 338 chronic hepatitis B (CHB) patients were divided into a disease group (patients with hepatic fibrosis) and control group (subjects without hepatic fibrosis). The disease group was further divided into S1-S4 according to the degree of fibrosis. Independent risk factors for hepatic fibrosis were analyzed using multivariate logistic regression. The diagnostic values of hepatic fibrosis from different indicators were compared using receiver operating characteristic (ROC) curves. The combination of elastography and serological indexes was explored to assess the severity of hepatic fibrosis. RESULTS: The multivariate logistic regression analysis results revealed that shear wave velocity (SWV), hyaluronic acid (HA), type IV collagen (CIV) and aspartate aminotransferase-to-platelet ratio index (APRI) significantly affected the occurrence of hepatic fibrosis. The ROC curve revealed that the accuracy of the diagnosis of hepatic fibrosis for SWV and HA were 87.3% and 84.8%, respectively. The accuracy of SWV combined with HA was 88.9%. The multiple linear regression analysis revealed that SWV, aspartate aminotransferase (AST)/alanine aminotransferase (ALT), HA, CIV, APRI and fibrosis index based on the 4 factor (FIB-4) were screened as statistically significant independent factors. The established regression equation was: Fibrosis level = -4.046 + 1.024 × SWV + 1.170 × AST/ALT + 0.011 × HA + 0.020 × CIV + 0.719 × APRI + 0.379 × FIB-4. CONCLUSION: SWV combined with serological indexes can improve the accuracy of diagnosis for CHB hepatic fibrosis. Serum indexes can help diagnose the degree of hepatic fibrosis.

Interaction of heat shock protein 60 (HSP60) with microRNA in Chinese mitten crab during Spiroplasma eriocheiris infection.

Heat shock protein 60 from the Chinese mitten crab Eriocheir sinensis (EsHSP60) was previously identified in relation to Spiroplasma eriocheiris infection by isobaric tags for relative and absolute quantitation labelling followed by liquid chromatography-tandem mass spectrometry. In the present study, to validate the immune function of this protein, the cDNA of the EsHSP60 gene was cloned. Various crab tissues were assessed using real-time PCR, which showed that EsHSP60 transcription occurred in all tissues examined. The expression profiles of EsHSP60 in haemolymph at transcription and protein levels when infected with S. eriocheiris were investigated by real-time PCR and Western blot analysis, respectively. A significant increase of EsHSP60 transcription and protein expression appeared post-injection in response to S. eriocheiris infection when compared to the control group. The double-luciferase reporter gene assay showed that the microRNA PC-533-3p interacted with the 3'-untranslated region of EsHSP60 and inhibited the translation of EsHSP60. The expression profiles of PC-533-3p during S. eriocheiris infection were also investigated by real-time PCR. However, the change tendency of PC-533-3p was opposite to that of the EsHSP60 after S. eriocheiris challenge. These data indicate that the EsHSP60 proteins may play an important role in mediating the immune responses of E. sinensis to an S. eriocheiris challenge.

Association between peripheral blood cells mitochondrial DNA content and severity of coronary heart disease.

BACKGROUND AND AIMS: Leukocyte mitochondrial DNA (mtDNA) content reflects the oxidant-induced cell damage, which has been observed in a wide range of cardiovascular diseases. However, whether it correlates with coronary heart disease (CHD), which closely relates to oxidative stress, has never been elucidated before. The aim of this study was to explore association between mtDNA content and the presence and severity of CHD. METHODS: The study population consisted of 400 individuals (290 with CHD and 110 controls). A quantitative real-time PCR was performed to measure the relative content of mtDNA in peripheral blood cells (PBCs). Gensini score was used to evaluate the severity of coronary stenotic lesions. An unconditional multivariate logistic regression was developed to estimate the association between CHD risk and mtDNA content by using odds ratio (OR). This study is registered with ClinicalTrials.gov, number NCT02500823. RESULTS: CHD patients, compared to controls, had lower mtDNA content (median, 0.78 vs. 0.83, p < 0.001), and mtDNA levels significantly decreased following an increasing Gensini score (p < 0.001). By using the first (highest mtDNA content) quartile of mtDNA content of controls as reference, the adjusted ORs (95% CIs) for individuals in the second, third and highest quartile of mtDNA content were 1.78 (95% CI, 1.15-3.51), 2.21 (95% CI, 1.65-3.74) and 4.83 (95% CI, 2.67-8.64), respectively (p for trend <0.001). CONCLUSIONS: These preliminary results suggest that expression of mtDNA may be associated with atherogenesis. The level of peripheral blood mtDNA in predicting the severity of coronary atherosclerosis may have a relatively certain value.

Survey of Clinical Extended-Spectrum Beta-Lactamase-Producing Enterobacter cloacae Isolates in a Chinese Tertiary Hospital, 2012-2014.

A total of 72 consecutive and nonduplicate clinical extended-spectrum ß-lactamase (ESBL)-producing Enterobacter cloacae isolates were collected from our hospital from 2012 to 2014 for analyzing the prevalence of plasmid-mediated quinolone resistance (PMQR) genes, 16S rRNA methyltransferase (16S-RMTase) encoding genes, and carbapenem-hydrolyzing ß-lactamase (CHßL) genes, as well as integrons. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were carried out to characterize the genetic relatedness. The isolates mainly harbored blaCTX-M (n = 51, 70.8%) and blaSHV (n = 46, 63.9%) genes. High prevalence of PMQR genes qnrA1 (n = 24, 33.3%), qnrB (n = 4, 5.6%), qnrS1 (n = 2, 2.8%), and aac(6')-Ib-cr (n = 21, 29.2%) was observed. Furthermore, CHßLs IMP-4 (n = 6, 8.3%) and IMP-8 (n = 4, 5.6%), as well as class I integrons (n = 29, 40.3%), were found in the ESBL-producing E. cloacae isolates. PFGE revealed 69 pulsotypes. MLST distinguished 44 sequence types (STs) with ST124 (n = 7, 9.7%), ST50 (n = 3, 4.2%), ST45 (n = 3, 4.2%), and ST93 (n = 3, 4.2%) being the predominant STs. The results indicate a possible clonal transmission of ST124 isolates in the hospital that needs further surveillance. The genetic diversity of the other numerous distinctive STs indicates that most of the ESBL-producing E. cloacae in our hospital might arise through stepwise accumulations of multiple drug-resistance determinants in different clones.