RPA-CRISPR-Cas13a-assisted detection method of transmissible gastroenteritis virus.

Background and aim: Transmissible gastroenteritis virus (TGEV) is a highly contagious gastrointestinal virus that causes diarrhea, vomiting, anorexia, dehydration, and weight loss in piglets. In clinical practice, it often occurs in mixed infections with other pathogens, and is therefore difficult to diagnose and prevent. It mainly harms piglets of about 2 weeks old, causing huge losses on farms. The clinical confirmation of TGEV usually requires a laboratory diagnosis, but traditional PCR and immunofluorescence assays have some limitations. Moreover, most farms in China are ill-equipped to accurately diagnose the disease. Therefore, a new detection method with high sensitivity and specificity and less dependence on instrumentation is required. Methods: We used recombinase polymerase amplification (RPA), combined with the nuclease characteristics of the activated Cas13a protein to establish a visual CRISPR-Cas13a-assisted detection method for TGEV by adding a reporter RNA with fluorescent and quenching moieties to the system. Result: We selected the optimal RPA primer and best CRISPR RNA (crRNA). The reaction system was optimized and its repeatability, specificity, and sensitivity verified. The TGEV detection system did not cross-react with other common diarrhea viruses, and its detection limit was 101 copies, which is similar with the sensitivity of qPCR. We successfully established an RPA-CRISPR-Cas13a-assisted detection method, and used this detection system to analyze 123 pig blood samples. qPCR was used as the gold standard method. The sensitivity, specificity, positive coincidence rate, and negative coincidence rate of the new method were 100, 98.93, 96.66, and 100%, respectively.

Efficacy of Qingpeng ointment (a Tibetan medicine) for acute gouty arthritis: a multi-center, randomized, double-blind, placebo-controlled trial.

BACKGROUND: This study aims to assess the efficacy and safety of Qingpeng ointment (QPO), a Tibetan medicine for alleviating symptoms in individuals with acute gouty arthritis (AGA). METHODS: This study was a randomized, double-blind, placebo-controlled trial that involved individuals with AGA whose joint pain, as measured on a visual analog scale (VAS) from 0 to 10, was equal to or greater than 3. The participants were randomly assigned to either the QPO or the placebo group and received their respective treatments twice daily for seven consecutive days. In case of intolerable pain, the participants were allowed to use diclofenac sodium sustained-release tablets as a rescue medicine. The primary outcomes measured were joint pain and swelling, while the secondary outcomes included joint mobility, redness, serum uric acid levels, C-reactive protein levels, and the amount of remaining rescue medicine. Any adverse events that occurred during the trial were also recorded. RESULTS: A total of 203 cases were divided into two groups, with balanced baselines: 102 in the QPO group and 101 in the placebo group. For joint pain, differences between the groups were notable in the VAS scores [1.75 (0, 3.00) versus 2.00 (1.00, 3.50); P = 0.038], changes in VAS [5.00 (3.00, 6.00) versus 4.00 (2.00, 6.00); P = 0.036], and disappearance rate [26.47% compared to 15.84%; P = 0.046] after treatment. Concerning joint swelling, significant between-group differences were observed in the VAS scores [1.00 (0, 2.30) versus 2.00 (0.70, 3.00); P = 0.032] and disappearance rate [33.33% compared to 21.78%; P = 0.046] at treatment completion. The QPO group exhibited a statistically significant mobility improvement compared to the placebo group (P = 0.004). No significant differences were found in other secondary outcomes. Five patients, four from the QPO group and one from the other, encountered mild adverse events, primarily skin irritation. All of these cases were resolved after dosage reduction or discontinuation of the medication. CONCLUSIONS: Compared to the placebo, QPO exhibits positive effects on AGA by alleviating pain, reducing swelling, and enhancing joint mobility, without causing significant adverse effects. TRIAL REGISTRATION: ISRCTN34355813. Registered on 25/01/2021.

Effects of Continuous Blood Purification without Heparin on Strem-1, NSE, and IL-10 Levels In Patients With Sepsis.

One of the most common causes of mortality and disability across the world is sepsis, a condition marked by an abnormal immunological reaction in the body. The prognosis of patients with sepsis is much improved when they are diagnosed early and provided with proper therapy. Soluble triggering receptors expressed on myeloid cells (STREM-1), Interleukin-10 (IL-10), neuronal serum enolase (NSE), and so on are possible biomarkers for the diagnosis of sepsis based on the pathophysiology. Blood purification treatment to control the cytokine storm induced by sepsis was regarded to be promising. Recently, the treatment of sepsis is likely to shift toward a multimodal approach. Hence in this paper, we investigated the effect of a continuous blood purification technique (hemofiltration without heparin followed by hemoadsorption) on STREM-1, NSE, and IL-10 levels in patients suffering from sepsis. A sample of hundred patients suffering from sepsis was randomly allocated to one of the two groups (study and control groups). The control group got standard sepsis care, whereas the research group got continuous blood purification treatment. To compare the differences between the two groups, we used t-statistical analysis. Blood STREM-1, IL-10, and NSE concentrations of the study group were significantly lesser than that of the control group after therapy. As a result, continuous blood purification can significantly minimize the dysregulated immune response in patients with sepsis, promote immune function and improve the survival rate.

Interleukin 35 contributes to immunosuppression by regulating inflammatory cytokines and T cell populations in the acute phase of sepsis.

Cytokines interact closely with each other and play a crucial role in the progression of sepsis. We focused on the associations of a cytokine network with IL-35 in sepsis. First, the retrospective study included 42 patients with sepsis and 23 healthy controls. Blood samples were collected from patients on days 1, 2, 4. Levels of IL-35, IL-1ß, IL-4, IL-6, IL-10, IL-17A, TNF-α and IFN-γ were measured. They all increased to various extend on days 1, 2, 4, and strongly associated with markers of disease severity. Network analysis revealed a network formed by IL-35, with IL-6, IL-10, IL-17A, TNF-α and IFN-γ throughout the acute phase of sepsis(days 1, 2 and4). Then, the CLP-induced septic rats were used. The recombinant human IL-35(rIL-35) upregulated the levels of IL-10, but downregulated IL-4, IL-6, IL-17A, TNF-α and IFN-γ, while it had no significant effect on IL-1ß, and upregulated the percentages of CD4+CD25+Tregs, and iTR35, but downregulated Teff cells in the peripheral blood. The rIL-35 reduced inflammation damage and improved prognosis of the septic rats. IL-35 forms a network with other cytokines and plays a major role in the immunopathogenesis of sepsis.

Hierarchical-pore UiO-66 modified with Ag+ for π-complexation adsorption desulfurization.

Adsorption desulfurization represents an alternative technology for the effective removal of thiophenic compounds from fuels. Metal-organic frameworks have been the ideal candidates for the adsorptive desulfurization of fuel due to the high surface areas. Pristine UiO-66 is thought to be appropriate for the removal of small thiophenic compounds. This work developed a new type of hierarchical-pore (micro and mesopores) UiO-66 with a higher specific surface area and porosity for the removal of larger adsorbates using MOF-5 as the template. To enhance adsorption desulfurization performance, the Ag+-exchanged hierarchical-pore UiO-66 (HP-UiO-66-SO3Ag) with π-complexation was synthesized by the ion-exchange method. The HP-UiO-66-SO3Ag samples were characterized by FTIR, XRD, SEM, TEM, and N2 adsorption-desorption isotherms. Compared with the original UiO-66, the HP-UiO-66-SO3Ag has a higher specific surface area, pore volume, and pore size. The static adsorption experiments showed that HP-UiO-66-SO3Ag had a high adsorption capacity for thiophene and benzothiophene. Moreover, the HP-UiO-66-SO3Ag sample still exhibits high adsorptive performance in the presence of toluene. The regeneration results show that about 90% of the initial adsorption capacity of HP-UiO-66-SO3Ag can be regenerated after four cycles.

[Hsa-let-7b-5p Inhibits Proliferation of Human Leukemia THP-1 Cells via FTO/m6A/MYC Signaling Pathway].

OBJECTIVE: To investigate the down-regulation effect of let-7b-5p on the expression of FTO in acute myeloid leukemia cell line THP-1 and inhibitory effect on THP-1 proliferation via m6A/MYC signaling pathway. METHODS: The acute myeloid leukemia cell line THP-1 and the normal human peripheral blood mononuclear cells (PBMNC) were selected as subjects. The expression of let-7b-5p and FTO mRNA in those cells was detected by qPCR, further the expression of FTO protein in those cells was detected by Western blot. And, the luciferase reporter gene assay was used to verify the targeting effect of let-7b-5p on FTO. Finally, THP-1 cells were transfected respectively with let-7b-5p mimic, and PBMNC with let-7b-5p inhibitor, there after the C-MYC mRNA m6A enrichment level in transfected cells was analyzed by dot blot, the expression levels of let-7b-5p, FTO and c-MYC were assayed by RT-PCR, the expressions of FTO and c-MYC protein were verified by Western blot, and the proliferation level of cells after transfection was detected by MTT assay. RESULTS: Compared with PBMNC, the expression of let-7b-5p in THP-1 significantly decreased, while the expression of FTO was significantly increased (P<0.05). After transfection with let-7b-5p mimic combined with FTO 3'-UTR, the luciferase activity of transfected THP-1 cells significantly decreased, but the luciferase activity significantly increased after transfection with mutant 3'-UTR, which was significantly different from the negative control group(blank vector) (P<0.05). Let-7b-5p inhibitor down-regulated c-MYC mRNA m6A enrichment, and then up-regulated the expression of FTO in transfected PBMNC cells, the effects of which were significant (P<0.05). However, let-7b-5p mimic up-regulated c-MYC mRNA m6A enrichment level and down-regulated the expression of FTO in the transfected THP-1 cells, and the cell proliferation rate was significantly lower than that in the negative control group (blank vector) (P<0.05). CONCLUSION: Human acute myeloid leukemia cell line THP-1 low expresses the let-7b-5p, which regulates c-MYC expression through let-7b-5p-/FTO-/m6A axis and promotes the proliferation of leukemia cell line THP-1.

IL-10 Promotes Neurite Outgrowth and Synapse Formation in Cultured Cortical Neurons after the Oxygen-Glucose Deprivation via JAK1/STAT3 Pathway.

As a classic immunoregulatory and anti-inflammatory cytokine, interleukin-10 (IL-10) provides neuroprotection in cerebral ischemia in vivo or oxygen-glucose deprivation (OGD)-induced injury in vitro. However, it remains blurred whether IL-10 promotes neurite outgrowth and synapse formation in cultured primary cortical neurons after OGD injury. In order to evaluate its effect on neuronal apoptosis, neurite outgrowth and synapse formation, we administered IL-10 or IL-10 neutralizing antibody (IL-10NA) to cultured rat primary cortical neurons after OGD injury. We found that IL-10 treatment activated the Janus kinase 1 (JAK1)/signal transducers and activators of transcription 3 (STAT3) signaling pathway. Moreover, IL-10 attenuated OGD-induced neuronal apoptosis by down-regulating the Bax expression and up-regulating the Bcl-2 expression, facilitated neurite outgrowth by increasing the expression of Netrin-1, and promoted synapse formation in cultured primary cortical neurons after OGD injury. These effects were partly abolished by JAK1 inhibitor GLPG0634. Contrarily, IL-10NA produced opposite effects on the cultured cortical neurons after OGD injury. Taken together, our findings suggest that IL-10 not only attenuates neuronal apoptosis, but also promotes neurite outgrowth and synapse formation via the JAK1/STAT3 signaling pathway in cultured primary cortical neurons after OGD injury.

IL-10 Protects Neurites in Oxygen-Glucose-Deprived Cortical Neurons through the PI3K/Akt Pathway.

IL-10, as a cytokine, has an anti-inflammatory cascade following various injuries, but it remains blurred whether IL-10 protects neurites of cortical neurons after oxygen-glucose deprivation injury. Here, we reported that IL-10, in a concentration-dependent manner, reduced neuronal apoptosis and increased neuronal survival in oxygen-glucose-deprived primary cortical neurons, producing an optimal protective effect at 20ng/ml. After staining NF-H and GAP-43, we found that IL-10 significantly protected neurites in terms of axon length and dendrite number by confocal microscopy. Furthermore, it induced the phosphorylation of AKT, suppressed the activation of caspase-3, and up-regulated the protein expression of GAP-43. In contrast, LY294002, a specific inhibitor of PI3K/AKT, reduced the level of AKT phosphorylation and GAP-43 expression, increased active caspase-3 expression and thus significantly weakened IL-10-mediated protective effect in the OGD-induced injury model. IL-10NA, the IL-10 neutralizing antibody, reduced the level of p-PI3K phosphorylation and increased the expression of active caspase-3. These findings suggest that IL-10 provides neuroprotective effects by protecting neurites through PI3K/AKT signaling pathway in oxygen-glucose-deprived primary cortical neurons.

Structural Properties and Dynamics of Thiophene in Sub/Supercritical Carbon Dioxide from Car-Parrinello Molecular Dynamics Simulations.

Structrual and dynamic properties of thiophene (C4H4S) in supercritical carbon dioxide were studied using Car-Parrinello molecular dynamics simulations. The geometries and energies optimized for the thiophene-CO2 complex show a stable C-H···O hydrogen bond interactions both in gas phase and in supercritical CO2. The radial distribution functions of CO2 around thiophene in the supercritical phase state show a correlation suggesting C-H···O hydrogen bond and S···C interaction. Local structural properties of the mixtures were investigated by angular-radial distributions and spatial distribution functions. The results show a mutually parallel arrangement between the thiophene plane and CO2 molecules at short distances and a high probability of the thiophene being located in the radial directions of the CO2 molecules. The decay of orientational correlations at 318.15 K shows slower relaxation compared to those of 298.15 K for first and second rank correlations. The vibrations of CO2 and thiophene molecules have been examined through an analysis of the velocity autocorrelation functions of the atoms. The C-H stretching modes of thiophene in the isolated configuration are less red-shifted and have a much narrower frequency range than that in the mixtures.

Computer simulation of the adsorption of thiophene in all-silica Y and Na-Y.

A grand canonical ensemble Monte Carlo simulation is performed to investigate the adsorption, heat of adsorption, and distributions of thiophene in all-silica Y and Na-Y zeolites. Biased particle insertions and deletions were implemented to allow the computation of equilibrium adsorption isotherms of such molecules. The calculated number of absorbed thiophene molecules in these zeolites is in good agreement with the experimental data. The calculated results show that the number absorbed of thiophene molecules in Na-Y is much greater than that in all-silica Y over the range of pressure. The calculated heat of adsorption is in good agreement with experimental results. The Na-Y zeolite, rather than all-silica Y, preferentially adsorbs the thiophene. A distribution analysis of the adsorbed phase structure reveals a different adsorption site in the zeolites.

[Distribution and sources of polycyclic aromatic hydrocarbons in sediments from rivers of Pearl River Delta and its nearby South China Sea].

Polycyclic aromatic hydrocarbons (PAHs) are measured in surface sediments from rivers and estuary of Pearl River Delta and its nearby South China Sea. Total PAH concentration varied from 255.9 - 16 670.3 ng/g and a moderate to low level compare to relevant areas worldwide. The order of PAHs concentration in sediments was: rivers of Pearl River Delta > estuary > South China Sea, and the most significant PAH contamination was at Guangzhou channel of Zhujiang river. A decrease trend for PAHs concentration with distance from estuary to open sea can be sees in South China Sea. Coal and biomass combustion is the major source of PAHs in nearshore of South China Sea, and petroleum combustion is the main source of pyrolytic PAHs in rivers and estuary of Pearl River Delta according to PAHs diagnostic ratios. Petroleum PAHs are revealed have a high contribution to PAHs in Xijiang River, estuary and some stations in Zhujiang River. A comparison of data from study in 1997 with data from present study indicates that there is no clear change in the PAH concentration over time but the source of PAHs in Pearl River Delta have been change from a main coal combustion to petroleum combustion and being reflect in the sediments in rivers and estuary of Pearl River Delta where there have high sedimentation rate.

Computer simulation of the adsorption of ethanethiol in silicalite of MFI and MOR.

The method of configurational-bias Monte Carlo simulation is used for investigating the adsorption and distributions of ethanethiol in MFI and MOR type zeolites. The calculated adsorption quantity of ethanethiol in MFI is close agreement with the experimental data by means of IR spectroscopy and thermogravimetry. The calculated results show the adsorption quantity of ethanethiol in MFI is much more than that in MOR under coequal conditions. The observation of distribution of MFI is attributed to preferential adsorption of the ethanethiol molecules in the straight and zig-zag channels. In the intersection of straight channels and zig-zag channels, it accommodates the ethanethiol molecules like a reservoir. The straight channels for MOR accommodate all ethanethiol molecules almost, however, in the other channels, the ethanethiol molecules cannot be found.