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Peroxynitrite anion(ONOO?)is one of the most active species of reactive oxygen species(ROS)and reactive nitrogen species(RNS).As an important physiologically active molecule,the abnormal expression of ONOO?will damage the protein and DNA in cells,leading to inflammation and other serious diseases in vivo.In recent years,fluorescence detection technology has been used to realize rapid and sensitive monitoring of bioactive molecules,and imaging tools have been used to conduct high-resolution tracing.Therefore,many organic small molecule fluorescent probes have been developed to detect ONOO?.In this paper,the recent research progresses of ONOO? fluorescence detection methods based on intramolecular charge transfer(ICT),photoinduced electron transfer(PET),fluorescence resonance energy transfer(FRET),excited state intramolecular charge transfer(ESIPT)and other fluorescence response mechanisms were reviewed.
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Previous studies have shown that high blood glucose-induced chronic microinflammation can cause inflammatory podocyte injury in patients with diabetic kidney disease(DKD). Therein, necroptosis is a new form of podocyte death that is closely associated with renal fibrosis(RF). To explore the effects and mechanisms in vivo of total flavones of Abelmoschus manihot(TFA), an extract from traditional Chinese herbal medicine Abelmoschus manihot for treating kidney diseases, on podocyte necroptosis and RF in DKD, and to further reveal its scientific connotation with multi-pathway and multi-target, the authors randomly divided all rats into four groups: a namely normal group, a model group, a TFA group and a rapamycin(RAP) group. After the modified DKD rat models were successfully established, four group rats were given double-distilled water, TFA suspension and RAP suspension, respectively by gavage every day. At the end of the 4th week of drug treatment, all rats were sacrificed, and the samples of their urine, blood and kidneys were collected. And then, the various indicators related to podocyte necroptosis and RF in the DKD model rats were observed, detected and analyzed, respectively. The results indicated that, general condition, body weight(BW), serum creatinine(Scr), urinary albumin(UAlb), and kidney hypertrophy index(KHI) in these modified DKD model rats were both improved by TFA and RAP. Indicators of RF, including glomerular histomorphological characteristics, fibronectin(FN) and collagen type Ⅰ(collagen Ⅰ) staining extent in glomeruli, as well as the protein expression levels of FN, collagen Ⅰ, transforming growth factor-β1(TGF-β1) and Smad2/3 in the kidneys were improved respectively by TFA and RAP. Podocyte damage, including foot process form and the protein expression levels of podocin and CD2AP in the kidneys was improved by TFA and RAP. In addition, tumor necrosis factor-α(TNF-α)-mediated podocyte necroptosis in the kidneys, including the morphological characteristics of podocyte necroptosis, the extent and levels of the protein expression of TNF-α and phosphorylated mixed lineage kinase domain like pseudokinase(p-MLKL) was improved respectively by TFA and RAP. Among them, RAP had the better effect on p-MLKL. More importantly, the activation of the receptor interacting serine/threonine protein kinase 1(RIPK1)/RIPK3/MLKL signaling axis in the kidneys, including the expression levels of its key signaling molecules, such as phosphorylated receptor interacting serine/threonine protein kinase 1(p-RIPK1), p-RIPK3, p-MLKL and cysteinyl aspartate specific proteinase-8(caspase-8) was improved respectively by TFA and RAP. Among them, the effect of TFA on p-RIPK1 was superior. On the whole, in this study, the authors demonstrated that TFA alleviates podocyte necroptosis and RF in DKD through inhibiting the activation of the TNF-α-mediated RIPK1/RIPK3/MLKL signaling axis in diabetic kidneys. The authors' findings provide new pharmacological evidence to reveal the scientific connotation of TFA in treating RF in DKD in more depth.
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Humanos , Ratas , Animales , Nefropatías Diabéticas/tratamiento farmacológico , Abelmoschus , Flavonas/farmacología , Podocitos , Factor de Necrosis Tumoral alfa/metabolismo , Necroptosis , Proteína Serina-Treonina Quinasas de Interacción con Receptores/metabolismo , Fibrosis , Treonina/farmacología , Colágeno/metabolismo , Serina/farmacología , Diabetes Mellitus/tratamiento farmacológicoRESUMEN
Non-alcoholic fatty liver disease(NAFLD) is a chronic metabolic condition with rapidly increasing incidence, becoming a public health issue of worldwide concern. Studies have shown that farnesoid X receptor(FXR)-based modulation of downstream targets can improve liver function and metabolic status in the patients with NAFLD and may be a potential drug target for treating this di-sease. Great progress has been achieved in the development of drugs targeting FXR for the treatment of NAFLD. A number of studies have explored the traditional Chinese medicine and their active ingredients for the treatment of NAFLD via FXR considering the high safety and efficacy and mild side effects. This paper systematically describes the mechanism of traditional Chinese medicines in the treatment of NAFLD via FXR and the downstream targets, aiming to provide precise targets for the drug development and clinical treatment of NAFLD.
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Humanos , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Hígado , Medicina Tradicional China/efectos adversos , Receptores Citoplasmáticos y Nucleares/metabolismoRESUMEN
Tanshinone IIa is a key ingredient extracted from the traditional Chinese medicine Salvia miltiorrhiza (Danshen), and is widely used to treat various cardiovascular diseases. Vascular calcification is a common pathological change of cardiovascular tissues in patients with chronic kidney disease, diabetes, hypertension and atherosclerosis. However, whether Tanshinone IIa inhibits vascular calcification and the underlying mechanisms remain largely unknown. This study aims to investigate whether Tanshinone IIa can inhibit vascular calcification using high phosphate-induced vascular smooth muscle cell and aortic ring calcification model, and high dose vitamin D3 (vD3)-induced mouse models of vascular calcification. Alizarin red staining and calcium quantitative assay showed that Tanshinone IIa significantly inhibited high phosphate-induced vascular smooth muscle cell and aortic ring calcification. qPCR and Western blot showed that Tanshinone IIa attenuated the osteogenic transition of vascular smooth muscle cells. In addition, Tanshinone IIa also significantly inhibited high dose vD3-induced mouse aortic calcification and aortic osteogenic transition. Mechanistically, Tanshinone IIa inhibited the activation of NF-κB and β-catenin signaling in normal vascular smooth muscle cells. Similar to Tanshinone IIa, inhibition of NF-κB and β-catenin signaling using the chemical inhibitors SC75741 and LF3 attenuated high phosphate-induced vascular smooth muscle cell calcification. These results suggest that Tanshinone IIa attenuates vascular calcification at least in part through inhibition of NF-κB and β-catenin signaling, and Tanshinone IIa may be a potential drug for the treatment of vascular calcification.
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Animales , Ratones , FN-kappa B/metabolismo , beta Catenina/metabolismo , Transducción de Señal , Miocitos del Músculo Liso/metabolismo , Calcificación Vascular/metabolismo , Fosfatos/metabolismoRESUMEN
BACKGROUND: Autophagy is believed to participate in embryonic development, but whether the expression of autophagy-associated genes undergoes changes during the development of human embryonic kidneys remains unknown. METHODS: In this work, we identified 36,151 human renal cells from embryonic kidneys of 9-18 gestational weeks in 16 major clusters by single-cell RNA sequencing (scRNA-seq), and detected 1350 autophagy-related genes in all fetal renal cells. The abundance of each cell cluster in Wilms tumor samples from scRNA-seq and GDC TARGET WT datasets was detected by CIBERSORTx. R package Monocle 3 was used to determine differentiation trajectories. Cyclone tool of R package scran was applied to calculate the cell cycle scores. R package SCENIC was used to investigate the transcriptional regulons. The FindMarkers tool from Seurat was used to calculate DEGs. GSVA was used to perform gene set enrichment analyses. CellphoneDB was utilized to analyze intercellular communication. RESULTS: It was found that cells in the 13th gestational week showed the lowest transcriptional level in each cluster in all stages. Nephron progenitors could be divided into four subgroups with diverse levels of autophagy corresponding to different SIX2 expressions. SSBpod (podocyte precursors) could differentiate into four types of podocytes (Pod), and autophagy-related regulation was involved in this process. Pseudotime analysis showed that interstitial progenitor cells (IPCs) potentially possessed two primitive directions of differentiation to interstitial cells with different expressions of autophagy. It was found that NPCs, pretubular aggregates and interstitial cell clusters had high abundance in Wilms tumor as compared with para-tumor samples with active intercellular communication. CONCLUSIONS: All these findings suggest that autophagy may be involved in the development and cellular heterogeneity of early human fetal kidneys. In addition, part of Wilms tumor cancer cells possess the characteristics of some fetal renal cell clusters.
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Objective:To determine the disinfection effect in child care institutions in Qingpu District of Shanghai, and provide evidence for improving the disinfection quality and preventing and controlling infectious diseases. Methods:According to the monitoring program of disinfection quality in child care institutions in Shanghai, disinfection quality was monitored for 3 years. Results:A total of 979 samples were monitored in the consecutive three years, in which 895 samples were qualified with a total qualification rate of 91.42%. The qualification rate of air samples was 100.00%, while the rate of staff hand samples was the lowest, with an average of 79.60%. During 2016-2018, the difference among the annual qualification rates was statistically significant(χ2=23.809,P<0.05), whereas it was not significant in the rates between public institutions and private ones (χ2=0.141,P>0.05). Conclusion:The quality of preventive disinfection in child care institutions in Qingpu District is generally good, but the disinfection of tableware, surface of objects and staff needs to be further improved.
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As the body ages, the immune system will undergo a series of changes, which are termed "immunosenescence" and are embodied in immune cells. Previous studies have shown that the immune cells involved in the regulation of immunosenescence include intrinsic immune cells and adaptive immune cells. Intrinsic immune cells are neutrophils, monocytes/macrophages, myeloid-derived suppressor cells, dendritic cells, natural killer cells, etc., and the underlying mechanisms involve the regulation of cell number, phagocytosis, chemotaxis, adhesion, the function of toll-like receptor (TLR), antigen presentation, macrophage polarization, cytotoxicity, migration, etc. The adaptive immune cells include T-lymphocytes and B-lymphocytes, and the underlying mechanisms involve the regulation of cell development, proliferation, differentiation, cell number, telomerase activity, self-reactive antibodies, etc. Immunosenescence is the manifestation of aging in the human body and is also an important target for delaying aging by Chinese medicine and western medicine. In recent years, scholars have found some classical prescriptions and their active components (such as Dushentang and total saponins in Panax ginseng leaves, and Shengmaiyin and anwulignan and total saponins in P. ginseng stems and leaves) can regulate immunosenescence by targeting the immune cells and interfering with their molecular regulatory mechanisms. In addition, the mechanisms of the classical prescriptions in regulating immunosenescence are closely related to autophagy. The representative prescription embodying the therapeutic principles of resolving blood stasis and promoting regeneration, Dahuang Zhechongwan, can delay D-galactose-induced renal aging in mice, and its underlying mechanisms are related to the regulation of the number and activity of thymic immune cells and improvement of the protein expression of autophagy-related markers and inflammatory cytokines in the kidney. Therefore, exploring the effects of the classical prescriptions and their active components by targeting the mechanisms of immunosenescence will become a new direction for investigating and developing anti-aging drugs.
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This study aimed to investigate apoptosis induction of ginsenoside compound K (ginsenoside CK) in human liver cancer SMMC-7721 cells and the involvement of TGF-β1/Smads signaling pathway. MTT assay was used to detect cell viability following ginsenoside CK treatment in SMMC-7721 cells. Annexin V-FITC/PI assay was used to detect apoptosis. After ginsenoside CK, or TGF-β1/Smads pathway activator TGFβ1 and inhibitor LY2109761 treatment, the TGF-β1/Smads pathway proteins and apoptosis proteins were detected by Western blot. The results showed that ginsenoside CK inhibited the proliferation of SMMC-7721 cells in a dose- and time-dependent manner. Annexin V-FITC/PI showed that ginsenoside CK induced apoptosis in SMMC-7721 cells. Meanwhile, ginsenoside CK inhibited the expression of Smad2/3, p-Smad2/3, Smad4, but promoted Smad7 expression, cleavage of caspase-3 and down-regulated Bcl-2/Bax. Compared with TGFβ1 treatment alone, levels of Smad2/3, p-Smad2/3, Smad4 and the ratio of Bcl-2/Bax were down-regulated, whereas Smad7 or cleaved caspase-3 was up-regulated in the ginsenoside CK+TGF-β1 group. In addition, Smad2/3, p-Smad2/3 and Smad4 expression were decreased in LY2109761 group. Compared with LY2109761 group, cleaved caspase-3 expression and Bcl-2/Bax have no significant change in ginsenoside CK+LY2109761 group. Taken together, our results showed that ginsenoside CK induced apoptosis in SMMC-7721 cells, and such induction is related to inhibiting TGF-β1/Smads signaling pathway.
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Objective:To establish a method for rapidly screening human antibodies recognizing HEV capsids proteins from pe-ripheral blood.The antibodies recognizing HEV capsids proteins were screened from the peripheral blood of vaccinator and the properties of the antibodies were analyzed.Methods:The HEV capsids proteins specific memory B cells in peripheral blood were obtained by flow cytometry sorting.Then antibody variable genes were acquired through single-cell RT-PCR and recombined to express in eukaryocyte.Finally,the properties analysis of recombinant expressed human monoclonal antibodies were carried out.Results:Six hu-manized monoclonal antibodies recognizing HEV capsids proteins were successfully obtained,and most of them had binding activity and neutralizing activity.Conclusion:The sequence of human monoclonal antibodies recognizing HEV capsid proteins is successfully screened and successfully expressed in the eukaryocyte.The properties of the antibodies are identified,which lay the foundation for studying antibody evolution in the human body after vaccination.
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To investigate the antiviral effect of thymopolypeptides combined with 4 kinds of matrine type alkaloids on HepG2.2.15 cells, oxymatrine, sophocarpidine, sophocarpine, and sophoridine (at concentration of 0.2 mmol•L⁻¹ respectively) were respectively combined with thymopolypeptides (0.025, 0.1 g•L⁻¹), and after 48 h and 72 h treatment on HepG2.2.15 cells, the cells and supernatants were collected. The cells activity in various groups was determined by CCK-8 method to evaluate the toxic effects of the drugs on HepG2.2.15 cells. Enzyme linked immunosorbent assay (ELISA) was used to determine HBeAg and HBsAg levels in cellular supernatants. HBV DNA levels in cellular supernatants andcells were quantified with fluorogenic quantitative PCR method; and the expression level of IFN-α in supernatants was detected with CBA method. The results indicated that single thymopolypeptides at 0.025-0.4 g•L⁻¹ had no toxicity to cells. Thymopolypeptides in this concentration range combined with 0.2 mmol•L⁻¹ matrine type alkaloids also had no toxicity to cells. Anti-HBV activity of drug combination was better than that of alkali or thymopolypeptides alone. Thymopolypeptides at 0.025 g•L⁻¹ had better inhibitory effect than thymopolypeptides at 0.1 g•L⁻¹ on intracellular HBV DNA expression, but the inhibitory effect on supernatant HBeAg level was on the contrary. Anti-HBV activity was similar between alkaloids combined with 0.1 g•L⁻¹ and alkaloids combined with 0.025 g•L⁻¹. There was no statistical difference in anti-HBV effect between various combined groups (P<0.05). In general, 72 h anti-HBV effect was better than 48 h anti-HBV effect (P<0.05). The expression of IFN-α was increased after drug combination, with positive correlation to the changes of other four indicators (P<0.05). In conclusion, oxymatrine, sophocarpidine, sophocarpine and sophoridine combined with thymopolypeptides could inhibit HBsAg and HBeAg secretion in HepG2.2.15 cells and HBV DNA replication, and further promote the antiviral effect by promoting the expression of IFN-α.
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5-10 years old were 33 cases;febrile children were 55 cases,persistent cough in 65 cases,among of those cases,the wheezing were 8 cases,nonproductive cough 45 cases,nasal discharge 7 cases in early right lung pneumonia in 38 cases,left lung pneumonia in 23 cases,pulmones involved at same time were 8 cases.Among of those children who senerated plenritis were 9 cases,pleural effussion 1 case;complication besides lung appeared 32 children.Complication of digestive and urinary system had 8 cases(11.6%),respectively.Complication of cardiovascular system had 3 cases(4.3%).Complication of nervous system and skin had 4 cases(5.8%),respectively.Complication of hematological system had 5 cases(7.2%).Eight cases(11.6%) showed damage of no more one organ,Fifty-nine cases first treated by erythromycin in vein,then by azithromycin in oral,10 cases were treated by azithromycin oral medication.Thirty-three cases fully recovered,36 cases improved.Conclusions SMPP are more founded in school children,the complication of besides lung are more serious than that of non-segmental pneumonia,it is security and relatively satisfaction to treat with erythromycin and azithromycin in sequential therapy.