Objective To investigate the mechanism of 2,6-dimethoxy-1,4-benzoquinone(DMQ),an active ingredients in fermented wheat germ extract,for inhibiting NLRP3 inflammasome activation and alleviating septic shock in mice.Methods Cultured murine bone marrow-derived macrophages(BMDM)stimulated with lipopolysaccharide(LPS)were treated with DMQ,followed by treatment with Nigericin,ATP,and MSU for activating the canonical NLRP3 inflammasome;the non-canonical NLRP3 inflammasome was activated by intracellular transfection of LPS,and AIM2 inflammasome was activated using Poly A:T.In human monocytic THP-1 cells,the effect of Nigericin on inflammasome activation products was examined using Western blotting and ELISA.Co-immunoprecipitation was performed to explore the mechanism of DMQ-induced blocking of NLRP3 inflammasome activation.In a male C57BL/6J mouse model of LPS-induced septic shock treated with 20 and 40 mg/kg DMQ,the levels of IL-1β and TNF-α in the serum and peritoneal lavage fluid were determined using ELISA,and the survival time of the mice within 36 h was observed.Results Treatment with DMQ effectively inhibited LPS-induced activation of canonical NLRP3 inflammasome in mouse BMDM and human THP-1 cells and also inhibited non-canonical NLRP3 inflammasome activation in mouse BMDM,but produced no significant effect on AIM2 inflammasome activation.DMQ significantly blocked the binding between ASC and NLRP3.In the mouse models of septic shock,DMQ treatment significantly reduced the levels of IL-1β in the serum and peritoneal fluid and obviously prolonged survival time of the mice.Conclusion DMQ can effectively block ASC-NLRP3 interaction to inhibit NLRP3 inflammasome activation and alleviate LPS-induced septic shock in mice.

Objective To investigate the mechanism of 2,6-dimethoxy-1,4-benzoquinone(DMQ),an active ingredients in fermented wheat germ extract,for inhibiting NLRP3 inflammasome activation and alleviating septic shock in mice.Methods Cultured murine bone marrow-derived macrophages(BMDM)stimulated with lipopolysaccharide(LPS)were treated with DMQ,followed by treatment with Nigericin,ATP,and MSU for activating the canonical NLRP3 inflammasome;the non-canonical NLRP3 inflammasome was activated by intracellular transfection of LPS,and AIM2 inflammasome was activated using Poly A:T.In human monocytic THP-1 cells,the effect of Nigericin on inflammasome activation products was examined using Western blotting and ELISA.Co-immunoprecipitation was performed to explore the mechanism of DMQ-induced blocking of NLRP3 inflammasome activation.In a male C57BL/6J mouse model of LPS-induced septic shock treated with 20 and 40 mg/kg DMQ,the levels of IL-1β and TNF-α in the serum and peritoneal lavage fluid were determined using ELISA,and the survival time of the mice within 36 h was observed.Results Treatment with DMQ effectively inhibited LPS-induced activation of canonical NLRP3 inflammasome in mouse BMDM and human THP-1 cells and also inhibited non-canonical NLRP3 inflammasome activation in mouse BMDM,but produced no significant effect on AIM2 inflammasome activation.DMQ significantly blocked the binding between ASC and NLRP3.In the mouse models of septic shock,DMQ treatment significantly reduced the levels of IL-1β in the serum and peritoneal fluid and obviously prolonged survival time of the mice.Conclusion DMQ can effectively block ASC-NLRP3 interaction to inhibit NLRP3 inflammasome activation and alleviate LPS-induced septic shock in mice.

Objective To investigate the effect of serum containing Huangqi decoction on the proliferation, migration, and tubulogenesis of rat liver sinusoidal endothelial cells (LSECs) induced by vascular endothelial growth factor (VEGF) and its mechanism of action based on the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway. Methods The serum containing Huangqi decoction was prepared, and rat LSECs were isolated and cultured in vitro . Then rat LSECs were randomly divided into blank group, VEGF group, serum control group, and low-, middle-, and high-dose serum containing Huangqi decoction groups. MTT colorimetry, Transwell assay, and tubulogenesis assay were used to measure the proliferation, migration, and tubulogenesis abilities of LSECs in each group, and Western Blot was used to measure the protein expression levels of platelet endothelial cell adhesion molecule-1 (CD31), endothelin-1 (ET-1), and endothelial nitric oxide synthase (eNOS), as well as AKT, phosphorylated-AKT (p-AKT), mTOR, and phosphorylated mTOR (p-mTOR) in the AKT/mTOR signaling pathway. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the Tukey's test was used for further comparison between two groups. Results Compared with the blank group, the VEGF group and the serum control group had significantly promoted proliferation, migration, and angiogenesis of rat LSECs (all P < 0.05), and Western Blot showed significant increases in the expression levels of CD31, ET-1, eNOS, and AKT/mTOR signaling pathway-related proteins (all P < 0.05). There were no significant differences in the above indices between the VEGF group and the serum control group (all P > 0.05). Compared with the serum group, the middle- and high-dose serum containing Huangqi decoction groups had significantly inhibited proliferation, migration, and angiogenesis of rat LSECs induced by VEGF (all P < 0.05), and Western Blot showed significant reductions in the expression levels of CD31, ET-1, eNOS, and AKT/mTOR signaling pathway-related proteins (all P < 0.01). Conclusion A relatively high dose of serum containing Huangqi decoction can significantly inhibit the proliferation, migration, and tubulogenesis of rat LSECs induced by VEGF, possibly by regulating the AKT/mTOR signaling pathway.

A new variant of concern for SARS-CoV-2,Omicron(B.1.1.529),was designated by the World Health Organization on November 26,2021.This study analyzed the viral genome sequenc-ing data of 108 samples collected from patients infected with Omicron.First,we found that the enrichment efficiency of viral nucleic acids was reduced due to mutations in the region where the primers anneal to.Second,the Omicron variant possesses an excessive number of mutations compared to other variants circulating at the same time(median:62 vs.45),especially in the Spike gene.Mutations in the Spike gene confer alterations in 32 amino acid residues,more than those observed in other SARS-CoV-2 variants.Moreover,a large number of nonsynonymous mutations occur in the codons for the amino acid residues located on the surface of the Spike protein,which could potentially affect the replication,infectivity,and antigenicity of SARS-CoV-2.Third,there are 53 mutations between the Omicron variant and its closest sequences available in public databases.Many of these mutations were rarely observed in public databases and had a low muta-tion rate.In addition,the linkage disequilibrium between these mutations was low,with a limited number of mutations concurrently observed in the same genome,suggesting that the Omicron vari-ant would be in a different evolutionary branch from the currently prevalent variants.To improve our ability to detect and track the source of new variants rapidly,it is imperative to further strengthen genomic surveillance and data sharing globally in a timely manner.

Objective:To evaluate the feasibility of brain injury after cardiopulmonary resuscitation (CPR) in rats based on T2WI image texture analysis.Methods:Eighteen SD rats were randomly divided into the sham group ( n=8) and model group ( n=10). The rats in the model group underwent MRI scanning at 6 h after return of spontaneous circulation (ROSC), and the rats in the sham group received MRI scanning at 6 h after the operation. The differences in the texture features of T2WI images and the expressions of AQP4 and NSE between the two groups were analyzed. The receiver operating characteristic curve (ROC) was used to evaluate the diagnostic efficacy of statistically different texture features between the two groups for brain injury. The associations between texture features and AQP4 and NSE expressions in the sham group and model group were analyzed using Spearman correlation coefficients. Results:The minimum intensity, standard deviation, and inverse difference moment of the whole brain T2WI texture features of the model group were significantly lower than those of the sham group ( P<0.05), while the difference entropy and characteristics of high gray in homogeneity were significantly higher than those of the sham group ( P<0.05). The difference entropy was the best with an area under curve (AUC) of 0.922, a sensitivity of 100% and a specificity of 75%. The AQP4 and NSE expressions in the model group were significantly higher than those in the sham group ( P<0.05). The minimum intensity value was positively correlated with AQP4 and NSE expressions ( r=0.501, 0.568, P=0.048, 0.022). The standard deviation was positively correlated with AQP4 and NSE expressions ( r=0.620, 0.530, P=0.010, 0.035). The difference entropy was negatively correlated with AQP4 expression ( r=-0.535, P=0.033). Conclusions:Texture analysis on T2WI images can evaluate the degree of brain edema and neuronal damage. The minimum intensity, standard deviation, and difference entropy are sensitive indicators to evaluate brain injury after CPR, and difference entropy has the highest sensitivity and specificity.

Animals ; CRISPR-Cas Systems ; Gene Editing ; Humans

Objective:To investigate the pathological changes of JNET classification of colorectal tumors.Methods:Data of patients with colorectal neoplasms who underwent narrow-band imaging combined with magnifying endoscopy, and whose postoperative pathological specimens were obtained at the endoscopy center of the Affiliated Hospital of Guizhou Medical University from January 2015 to June 2018 were analyzed retrospectively. The endoscopic JNET diagnosis and pathological features (surface pit epithelial exfoliation, surface mucosal necrosis structure, surface mucosal ethmoid reticular structure, fibrous tissue reactive hyperplasia, mucosa muscle without residual and carcinomatous interstitial reaction) were analyzed.Results:A total of 81 patients with colorectal neoplasms (diameter >1.5 cm) were enrolled with 74 cases receiving endoscopic treatment and seven cases receiving surgical treatment. The frequency of surface mucosal sieve reticular structure in type 3 of JNET (20.00%) was lower than that in type 2B (42.86%), which was in turn higher than that in type 2A (2.78%), with significant differences ( P<0.05). The frequencies of fibrous tissue reactive proliferation, no residual mucosal muscles, and carcinomatous interstitial reaction in type 2A and 2B were lower than that of type 3, with significant differences (all P<0.05). There were significant differences in the consistency group and inconsistency group of JNET classification and pathological diagnosis in surface pit epithelial exfoliation and surface mucosal necrosis (all P<0.05). Conclusion:Type 2B of JNET classification indicates that the pathological features may be the surface mucosal reticular structure; type 3 indicates reactive hyperplasia of fibrous tissue, no residual mucosal muscles and cancerous interstitial reaction. The exfoliation of the surface pit epithelium and the necrosis of the surface mucosa may be the pathological interference factors affecting the accuracy of JNET classification diagnosis.

Animals ; Archaeal Proteins ; genetics ; metabolism ; Deoxyribonuclease I ; genetics ; metabolism ; Gene Editing ; methods ; Humans ; Natronobacterium ; enzymology ; genetics

This paper mainly discussed the present situation of the pension mode transformation from traditional family pension to social pension. The existing ethical issues in pension institution include: old filial piety thought deeply rooted;institutional care services cannot be carried out caused by unfair distribution balance; humanistic thought of human ethical care implementation does not do well; some agencies have occasional bad behavior. Therefore, this paper proposed some advices based bioethical perspective:for old people and their children, advo-cate the new filial piety;pension institutions not only improving the quality of the hardware but also should strength-en humanistic ethics concern;establish and perfect the laws and regulations, combining the rule of law with moral and ethical;the relevant government departments should insist on and strengthen the principle of fair justice.