Human norovirus is the leading cause of viral gastroenteritis worldwide. Rapid detection facilitates management of disease outbreaks, but field diagnosis is difficult to achieve due to the lack of ...reliable and portable methods. Recombinase polymerase amplification (RPA) is a robust isothermal amplification method that is capable of rapidly amplifying and detecting nucleic acids using simple equipment. In this study, RPA combined with lateral flow (LF) strips specific for human genogroup II (GII) noroviruses was established and evaluated. The assay specifically detects purified GII noroviruses as well as RNA in boiled human stool samples, with a sensitivity of 50 norovirus genome copies per reaction. The whole detection procedure of the one-step RT-RPA-LF is completed within 20 min, which is eight times faster than that of the standard real-time RT-PCR. The RT-RPA-LF method described here is suitable for rapid field diagnosis of all GII noroviruses in human stool samples.
Oysters are major transmission vectors of noroviruses (NoVs) in the environment. Outbreaks of NoVs are often associated with the consumption of NoV-contaminated oysters. Laboratory confirmation of ...suspected oyster samples is a critical step in the surveillance and control of NoVs. Because of non-specific amplification, false-positive results are frequently obtained by semi-nested RT-PCR with the presently widely used primer set (G2SKF/G2SKR). Here, a novel universal PCR primer set N (NG2OF/NG2OR) specific for genogroup II (GII) NoVs was designed based on all GII NoV sequences available in public databases. Specific products were obtained with the primer set N when the NoV-positive oysters, spiked with each of five representative genotypes of GII NoVs (GII.17, GII.13, GII.4, GII.3, and GII.12), were subjected to analyzing. No products were detected with the primer set N for the NoV-negative oysters, while the primer set C gave various non-specific bands. Twenty-three out of 156 fresh oyster samples were NoV-positive with both the primer set N and the classic primer set, while eight were NoV-positive solely with the primer set N. Compared with the classic primer set, the newly designed primer set N had a higher detection rate and improved specificity for GII NoVs in oyster samples. These results show that the novel PCR primer pair is specific and applicable for the detection of GII NoVs in oysters.
Paeniclostridium sordellii lethal toxin (TcsL) is a potent exotoxin that causes lethal toxic shock syndrome associated with fulminant bacterial infections. TcsL belongs to the large clostridial toxin ...(LCT) family. Here, we report that TcsL with varied lengths of combined repetitive oligopeptides (CROPs) deleted show increased autoproteolysis as well as higher cytotoxicity. We next present cryo-EM structures of full-length TcsL, at neutral (pH 7.4) and acidic (pH 5.0) conditions. The TcsL at neutral pH exhibits in the open conformation, which resembles reported TcdB structures. Low pH induces the conformational change of partial TcsL to the closed form. Two intracellular interfaces are observed in the closed conformation, which possibly locks the cysteine protease domain and hinders the binding of the host receptor. Our findings provide insights into the structure and function of TcsL and reveal mechanisms for CROPs-mediated modulation of autoproteolysis and cytotoxicity, which could be common across the LCT family.
For patients with diabetes mellitus, excessive and long-lasting inflammatory reactions at the wound site commonly lead to the delayed refractory wound healing. The polarization of macrophages in ...terms of M1 and M2 phenotypes is closely related to the production of inflammatory cytokines. Quercetin is traditionally recognized to have anti-inflammatory effect; however, whether quercetin modulates macrophage polarization from M1 to M2 and thus promotes diabetic wound healing remain unknown.
Wounded male diabetic rats were equally divided into five groups: model group, solvent control group (10% DMSO), and three drug groups treated with quercetin (Q) at concentrations of 10 mg/mL (Q-LD low dose), 20 mg/mL (Q-MD medium dose), and 40 mg/mL (Q-HD high dose), respectively. The anti-inflammatory effect of quercetin on diabetic wounds was observed. Immunohistochemistry and quantificational real-time polymerase chain reaction were applied to test the changes in macrophage polarization and inflammatory responses.
The wound contraction was fastest in Q-HD group. Hematoxylin and eosin (H&E) and Masson's trichrome staining revealed that fibroblast distribution and collagen deposition in quercetin-treated groups were significantly higher than those in the model group. Immunohistochemistry tests showed more CD206-positive cells and less iNOS-positive cells in quercetin-treated groups. Furthermore, the levels of proinflammatory factors in quercetin-treated groups were lower than those in the model group, whereas the levels of the anti-inflammatory factors and angiogenesis-related factors were relatively higher.
In short, quercetin inhibits inflammatory reactions via modulating macrophage polarization switching from M1 to M2 phenotype, thereby accelerating the diabetic wound repair.
The COVID-19 pandemic now affects the entire world and has many major effects on the global economy, environment, health, and society. Focusing on the harm COVID-19 poses for human health and ...society, this study used system dynamics to establish a prevention and control model that combines material supply, public opinion dissemination, public awareness, scientific and technological research, staggered work shifts, and the warning effect (of law/policy). Causal loop analysis was used to identify interactions between subsystems and explore the key factors affecting social benefit. Further, different scenarios were dynamically simulated to explore optimal combination modes. The main findings were as follows: (1) The low supervision mode will produce a lag effect and superimposed effect on material supply and impede social benefit. (2) The strong supervision mode has multiple performances; it can reduce online public opinion dissemination and the rate of concealment and false declaration and improve government credibility and social benefit. However, a fading effect will appear in the middle and late periods, and over time, the effect of strong supervision will gradually weaken (but occasionally rebound) and thus require adjustment. These findings can provide a theoretical basis for improving epidemic prevention and control measures.
The mechanism of early blood–brain barrier (BBB) disruption after stroke has been intensively studied but still not fully understood. Here, we report that microRNA-30a (miR-30a) could mediate BBB ...damage using both cellular and animal models of ischemic stroke. In the experiments in vitro, inhibition of miR-30a decreased BBB permeability, prevented the degradation of tight junction proteins, and reduced intracellular free zinc in endothelial cells. We found that the zinc transporter ZnT4 was a direct target of negative regulation by miR-30a, and ZnT4/zinc signaling pathway contributed significantly to miR-30a-mediated BBB damage. Consistent with these in vitro findings, treatment with miR-30a inhibitor reduced zinc accumulation, increased the expression of ZnT4, and prevented the loss of tight junction proteins in microvessels of ischemic animals. Furthermore, inhibition of miR-30a, even at 90 min post onset of middle cerebral artery occlusion, prevented BBB damage, reduced infarct volume, and ameliorated neurological deficits. Together, our findings provide novel insights into the mechanisms of cerebral ischemia-induced BBB disruption and indicate miR-30a as a regulator of BBB function that can be an effective therapeutic target for ischemic stroke.
The impairment in diabetic wound healing represents a significant clinical problem, with no efficient targeted treatments for these wound disorders. Curcumin is well confirmed to improve diabetic ...wound healing, however, its low bioavailability and poor solubility severely limit its clinical application. This study aims to provide the pharmacological basis for the use of (2
,6
)-2,6-bis(2-(trifluoromethyl)benzylidene)cyclohexanone (C66). The results showed that topically applied C66 improved cutaneous wound healing in vivo. Further studies showed that C66 treatment increased the level of microRNA-146a (miR-146a) in the wounds in streptozotocin (STZ)-induced diabetic mice, downregulated the expression of interleukin-1 receptor-associated kinase 1 (IRAK1) and phosphorylated nuclear factor-κB (NF-κB) p65 subunit (p-p65) (both
< 0.05), and suppressed the mRNA expression of inflammation-related cytokines, tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8), and interleukin-6 (IL-6). The in vitro data obtained in human umbilical vein endothelial cells (HUVECs) showed that C66 could reverse high glucose (HG)-induced NF-κB activation due to upregulation of miR-146a expression, which matched the in vivo findings. In conclusion, the present study indicates that C66 exerts anti-inflammation activity and accelerates skin wound healing of diabetic mice, probably via increasing miR-146a and inhibiting the NF-κB-mediated inflammation pathway. Therefore, C66 may be a promising alternative for the treatment of diabetic wounds.
With ongoing urbanization, traffic congestion and the air pollution it induces are worsening. Using a system dynamics (SD) approach, this study constructed a driving-restriction policy model to ...explore the effects of different stages of policy implementation on variables such as traffic congestion, emissions, and parking demand. Medium- and long-term dynamic simulation showed that the effect of the policy was obvious in the initial stage but gradually weakened in the medium term, leading to a “fading” effect on traffic-congestion alleviation; a “rebound” effect was even observed at the end of the simulation. Thus, the policy will not effectively reduce traffic congestion in the long term and will induce a new demand for car purchases, resulting in paradoxical effects, which will aggravate parking demand, congestion, and pollution. Yet, it was also found that introducing penalty policies and an air pollution charging fee could weaken the paradoxical effects and compensate for some defects of the policy. Such strategies could help reduce emissions, traffic congestion, parking demand, the number of illegal trips, and the overall number of vehicle trips. These findings can provide not only a theoretical basis for further research but also practical guidance for policy improvement.
Graphical abstract
Delayed rockburst occurs after the stress adjustment of surrounding rocks with discontinuous microseismicity, which is difficult to predict accurately. To predict and prevent the occurrence of the ...delayed rockbursts, study on time behavior of the surrounding rock failure and the mechanism behind the delayed rockbursts is necessary. In this article, based on the rockburst data from Jinping II Hydropower Station headrace tunnels, China, the characteristics of the delayed rockbursts are summarized. Considering the microseismic monitoring results of two typical delayed rockbursts, the formation process of delayed rockbursts is analyzed. Subsequently, the time behavior of rock failure is revealed by a uniaxial constant loading test. The results from our study suggest the surrounding rocks are in a relatively stable state, where the delayed rockburst occurs. Geo-stress, geological conditions and excavation are the key factors affecting the delayed rockbursts. Within the rockburst-affected area, the microseismic activities were frequent during the early stage of excavation, then the stress adjustment of the surrounding rocks gradually stabilized. As a result, the surrounding rocks were in a dormant stage, under the long-term high stress. Supported by laboratory experiments, our results suggest an obvious time behavior in the rock failure which corresponds to the time behavior of the crack propagation, under the high stress. Under the condition of long-term high stress, when the damage is accumulated to a certain extent, the rock mass is destabilized and then the rockburst is triggered, accordingly. The results from this study can provide a reference for the prediction and prevention of the delayed rockbursts, in similar situations, e.g. deep-buried tunnels, around the world.
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a potentially curative treatment for acute myeloid leukemia (AML). However, most patients experience relapse after allo-HSCT, with a ...poor prognosis, and treatment options are limited. The lack of an ideal targetable antigen is a major obstacle for treating patients with relapsed AML. CD38 is known to be expressed on most AML and myeloma cells, and its lack of expression on hematopoietic stem cells (HSCs) renders it a potential therapeutic target for relapsed AML. To investigate the clinical therapeutic efficacy and safety of CD38-targeted chimeric antigen receptor T (CAR-T-38) cells, we enrolled 6 AML patients who experienced relapse post-allo-HSCT (clinicaltrials.gov: NCT04351022). Prior to CAR-T-38 treatment, the blasts in the bone marrow of these patients exhibited a median of 95% (92-99%) CD38 positivity. Four weeks after the initial infusion of CAR-T-38 cells, four of six (66.7%) patients achieved complete remission (CR) or CR with incomplete count recovery (CRi); the median CR or CRi time was 191 (range 117-261) days. The cumulative relapse rate at 6 months was 50%. The median overall survival (OS) and leukemia-free survival (LFS) times were 7.9 and 6.4 months, respectively. One case relapsed 117 days after the first CAR-T-38 cell infusion, with remission achieved after the second CAR-T-38 cell infusion. All six patients experienced clinically manageable side effects. In addition, multiparameter flow cytometry (FCM) revealed that CAR-T-38 cells eliminated CD38 positive blasts without off-target effects on monocytes and lymphocytes. Although this prospective study has a limited number of cases and a relatively short follow-up time, our preliminary data highlight the clinical utility and safety of CAR-T-38 cell therapy in treating relapsed AML post-allo-HSCT. Keywords: Chimeric antigen receptor T cells, CAR-T-38, Relapsed acute myeloid leukemia, Allogeneic hematopoietic stem cell transplantation, Cytokine release syndrome
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