Objective To observe the immunohistochemical changes of ICAM-1,MMP-2 and MMP-9 in the lungs of immunocompromised rats with Pseudomonas aeruginosa pneumonia and their relationships with lung inflammation.Methods After the establishment of pseudomonas aeruginosa pneumonia infected immunocompromised rat mode,the pathological changes of lungs were observed, lung wet/dry ratios and total protein concentration in bronchial alveolar lavage fluid were tested,and imunnohistochemical study of ICAM-1,MMP-2 and MMP 9 in lung tissue were performed.Results 1.The staining intensity of ICAM-1 in alveolar epithelial cells turned stronger in rats with pulmonary infection than those without of both groups(P＜0.05);2.The staining intensity of MMP-2 in lung tissue was stronger in rats with pulmonary infection than those without infection in both groups,and reached peak at 6～9 h after inoculation.Immunohistochemical changes of MMP-9 exhibited a similar pattern,4.Immunohistochemical changes of ICAM-1,MMP-2 and MMP-9 showed some correlation with numbers of polymorphonuclears in lung tissue(P＜0.05);5.A correlation between the stai- ning intensity of MMP-9 in bronchial epithelial eells and total protein concentrations were observed(r_s =0.484,P＜0.05),similar association were found between the staining intensity of MMP-2 in alveolar epithelial cells,endothelium of arterioles and venules and tissues beneath endothelium and to- tal protein eoncentrations in bronchial alveolar lavage fluid(r_s were 0.457,0.492 and 0.429,respec- tively,P＜0.05).Conclusion In immunocompromised rats,the staining intensity of ICAM-1, MMP-2 and MMP-9 in lung tissue of those with pseudomonas aeruginosa pneumonia were stronger than those without infection,and the changes were demonstrated some correlation with the levels of polymorphonuclears infiltration or severity of lung injury.

Abstract: Objective　To analyze the detection of SARS‑CoV‑2 in household environment and public place environment of Huangpu District, and describe the feature of SARS‑CoV‑2 contamination in the environment exposure to the infected cases, so as to support the control strategies such as disinfection and health communication. Methods　The results of RT-PCR test for the environmental samples exposure to the cases infected by SARS‑CoV‑2 during February 1 to March 31 2022 in Huangpu District of Shanghai were collected as the research data. Pearson χ2 was used to test the significance of the differences between positive rates of SARS‑CoV‑2 contamination. Results　From February 1 to March 31, household environment samples had a higher positive rate (6.47%, 234/3 618) of SARS‑CoV‑2 contamination while the public place samples had a lower one (1.22%, 47/3 582) in Huangpu District of Shanghai (χ2=141.908, P<0.01). Among the household buildings, the lane houses of old style representing poorer living condition had the highest positive rates (8.31%, 96/1 155) of SARS‑CoV‑2 contamination while the apartments representing better living condition had the lowest (3.59%, 22/612) (F=5.25，P<0.05). Among the samples from household environment, samples regarding sewerage had the highest positive rates (13.30%,58/436) of SARS‑CoV‑2 contamination, while samples regarding the tool of cooking and sweeping had the lowest (3.10%,17/548) (F=9.84，P<0.01). Among the samples from public place environment, samples regarding entertainment tools had the highest positive rates (13.33%, 2/15) of SARS‑CoV‑2 contamination, while samples regarding the tool of cooking and sweeping had the lowest (0.62%, 4/646) (F=4.22，P<0.01). Conclusion　In the environment exposure to the SARS‑CoV‑2 infected cases, the disinfection, ventilation and cleaning should be intensified strictly. SARS‑CoV‑2's surviving in sewage environment should be evaluation dynamically. More health communication should be pushed to people of poorer living condition.

The ubiquitin system is crucial for the development and fitness of higher plants. De-etiolation, during which green plants initiate photomorphogenesis and establish autotrophy, is a dramatic and complicated process that is tightly regulated by a massive number of ubiquitylation/de-ubiquitylation events. Here we present site-specific quantitative proteomic data for theubiquitylomes of de-etiolating seedling leaves of Zea mays L. (exposed to light for 1, 6, or 12 h) achieved through immunoprecipitation-based high-resolution mass spectrometry (MS). Through the integrated analysis of multiple ubiquitylomes, we identified and quantified 1926 unique ubiquity-lation sites corresponding to 1053 proteins. We analyzed these sites and found five potential ubiqui-tylation motifs, KA, AXK, KXG, AK, and TK. Time-course studies revealed that the ubiquitylation levels of 214 sites corresponding to 173 proteins were highly correlated across two replicate MS exper-iments, and significant alterations in the ubiquitylation levels of 78 sites (fold change >1.5) were detected after de-etiolation for 12 h. The majority of the ubiquitylated sites we identified corre-sponded to substrates involved in protein and DNA metabolism, such as ribosomes and histones. Meanwhile, multiple ubiquitylation sites were detected in proteins whose functions reflect the major physiological changes that occur during plant de-etiolation, such as hormone synthesis/signaling pro-teins, key C4 photosynthetic enzymes, and light signaling proteins. This study on the ubiquitylome of the maize seedling leaf is the first attempt ever to study the ubiquitylome of a C4 plant and provides the proteomic basis for elucidating the role of ubiquitylation during plant de-etiolation.

Nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are the major enzymatic source of reactive oxygen species (ROS). NOX2 and NOX4 are expressed in the heart but its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion is unclear. This study investigated the effect of NOX on ANP secretion induced by hypoxia in isolated beating rat atria. The results showed that hypoxia significantly upregulated NOX4 but not NOX2 expression, which was completely abolished by endothelin-1 (ET-1) type A and B receptor antagonists BQ123 (0.3 µM) and BQ788 (0.3 µM). ET-1-upregulated NOX4 expression was also blocked by antagonists of secreted phospholipase A2 (sPLA2; varespladib, 5.0 µM) and cytosolic PLA2 (cPLA2; CAY10650, 120.0 nM), and ET-1-induced cPLA2 expression was inhibited by varespladib under normoxia. Moreover, hypoxia-increased ANP secretion was evidently attenuated by the NOX4 antagonist GLX351322 (35.0 µM) and inhibitor of ROS N-Acetyl-D-cysteine (NAC, 15.0 mM), and hypoxia-increased production of ROS was blocked by GLX351322. In addition, hypoxia markedly upregulated Src expression, which was blocked by ET receptors, NOX4, and ROS antagonists. ET-1-increased Src expression was also inhibited by NAC under normoxia. Furthermore, hypoxiaactivated extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) were completely abolished by Src inhibitor 1 (1.0 µM), and hypoxia-increased GATA4 was inhibited by the ERK1/2 and Akt antagonists PD98059 (10.0 µM) and LY294002 (10.0 µM), respectively. However, hypoxia-induced ANP secretion was substantially inhibited by Src inhibitor. These results indicate that NOX4/Src modulated by ET-1 regulates ANP secretion by activating ERK1/2 and Akt/GATA4 signaling in isolated beating rat hypoxic atria.

Nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are the major enzymatic source of reactive oxygen species (ROS). NOX2 and NOX4 are expressed in the heart but its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion is unclear. This study investigated the effect of NOX on ANP secretion induced by hypoxia in isolated beating rat atria. The results showed that hypoxia significantly upregulated NOX4 but not NOX2 expression, which was completely abolished by endothelin-1 (ET-1) type A and B receptor antagonists BQ123 (0.3 µM) and BQ788 (0.3 µM). ET-1-upregulated NOX4 expression was also blocked by antagonists of secreted phospholipase A2 (sPLA2; varespladib, 5.0 µM) and cytosolic PLA2 (cPLA2; CAY10650, 120.0 nM), and ET-1-induced cPLA2 expression was inhibited by varespladib under normoxia. Moreover, hypoxia-increased ANP secretion was evidently attenuated by the NOX4 antagonist GLX351322 (35.0 µM) and inhibitor of ROS N-Acetyl-D-cysteine (NAC, 15.0 mM), and hypoxia-increased production of ROS was blocked by GLX351322. In addition, hypoxia markedly upregulated Src expression, which was blocked by ET receptors, NOX4, and ROS antagonists. ET-1-increased Src expression was also inhibited by NAC under normoxia. Furthermore, hypoxiaactivated extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) were completely abolished by Src inhibitor 1 (1.0 µM), and hypoxia-increased GATA4 was inhibited by the ERK1/2 and Akt antagonists PD98059 (10.0 µM) and LY294002 (10.0 µM), respectively. However, hypoxia-induced ANP secretion was substantially inhibited by Src inhibitor. These results indicate that NOX4/Src modulated by ET-1 regulates ANP secretion by activating ERK1/2 and Akt/GATA4 signaling in isolated beating rat hypoxic atria.

Objective To identify the characteristics of the subtype of PMA-induced THP-1 macrophages by flow cytometry analysis. Methods THP-1 monocytic cells differentiate into macrophages promoted by PMA, then induced into M1 and M2 by adding different cytokines, such as LPS,IL-6 and IFN-γ for THP-1-M1, IL-4,IL-13 and IL-6 for THP-1-M2. Morphology of cells were observed under a microscope and the expression of CD14, CD68, CD16, CD80, CD86, CD163, CD206, CD209, CD83, CD1a, CD11c, HLA-DR were detected by flow cytometry. Results The macrophages stimulated by PMA became adherent;THP-1-M1 and THP-1-M2 lost their spherical morphology, appeared more irregular with many obvious projections. The expression of CD83, CD1a, CD11c, HLA-DR which had the function of antigen presenting on the surface of THP-1-Mφwere very low, and most of them were negative, but those of THP-1-M1 and THP-1-M2 were very high. Conclusions The macrophages differentiated from THP-1 stimulated by PMA are weak in antigen presenting function.

Objective To investigate the preparation and evaluation of animal models of depression associated with autoimmune thyroiditis,and to verify the NLRP3/Caspase-1/GSDMD pathway based on this condition.Methods 32 NOD.H-2H4 mice were randomly divided into a normal(N)group,depression(DP)group,autoimmune thyroiditis with depression(AIT+DP)group,autoimmune thyroiditis(AIT)group,with 8 animals in each group.The N group was fed normally,the DP group was subjected to chronic unpredictable mild stress(CUMS)for 5 weeks,the AIT group was given 0.05％sodium iodide water to establish an autoimmune thyroiditis model,and the AIT+DP group was subjected to 5 weeks of CUMS to establish the AIT animal model.We evaluated whether the mouse autoimmune thyroiditis model had been successfully prepared by observing the thyroid tissue structure,lymphocyte infiltration,and serum TGAb and TPOAb levels.Changes in body weight,sugar water preference rate,open field behavior(central quadrant time,central quadrant time proportion,standing rate,frequency of defecation,and hair grooming time),and hippocampal pathological changes were used to evaluate the depression status of the mice.When the model mice met the above-mentioned indicators related to depression and autoimmune thyroiditis,the AIT+DP animal model was considered successfully prepared.Results Compared with the levels in the N group,the AIT group's and AIT+DP group's serum TGAb and TPOAb levels were significantly increased(P＜0.01),and a large number of inflammatory cells had infiltrated the thyroid gland.The central quadrant time and central quadrant time proportion,standing rate,frequency of defecation,and hair grooming time were reduced to varying degrees in the DP group and AIT+DP group.In addition,the numbers of glial cells in the cerebral cortex increased and neuronal cells decreased,accompanied by some nuclear atrophy,and the expression levels of NLRP3,IL-1β,Caspase-1,and GSDMD-N significantly increased,especially in the AIT+DP group(P＜0.01).Conclusions 0.05％sodium iodide water and CUMS create autoimmune thyroiditis with depression model animals that better simulate the external performance and internal index changes of the diseases.These mice can provide an animal model reference for research into autoimmune thyroiditis with depression.

This study is to determine the preventive effect and mechanism of targeting IL-17A on pulmonary inflammation and fibrosis after acute lung injury. Mice were treated with anti-IL-17A antibody on the day 7 and sacrificed on the day 14 after bleomycin lung injury. The pulmonary inflammatory status and the deposition of collagen were measured by HE and Sirius stains staining. The contents of hydroxyproline and collagen were measured by using commercial kits. The survival rate of mice was calculated by Kaplan-Meier methods. The inflammatory cytokines in bronchoalveolar lavage fluid were measured by ELISA and the expressions of inflammation-related molecules were detected by Western blotting assay. Targeting of IL-17A could prevent the development of lung inflammation, decrease collagen deposition and the contents of hydroxyproline, and protect against the development of pulmonary fibrosis, which together led to an increase in the animal survival. Moreover, blocking IL-17A decreased the expression ofpro-fibrotic cytokines such as IL-17A, TGF-beta1 and IL-13; increased the expression of anti-fibrotic or anti-inflammatory factors such as IFN-gamma, COX-2, 5-LOX, 15-LOX. Indeed, IL-17A antagonism suppressed the activation of pro-inflammatory p65NF-kappaB but enhanced the activation of pro-resolving p50NF-kappaB. In conclusion, that blockade of IL-17A prevents the development of pulmonary fibrosis from acute lung injury, is because blocking IL-17A may prevent acute inflammation converting to chronic inflammation.
Acute Lung Injury ; chemically induced ; complications ; Animals ; Bleomycin ; Collagen ; metabolism ; Hydroxyproline ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-17 ; antagonists & inhibitors ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; NF-kappa B p50 Subunit ; metabolism ; Pneumonia ; etiology ; metabolism ; Pulmonary Fibrosis ; etiology ; metabolism ; prevention & control ; Random Allocation ; Transcription Factor RelA ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Up-Regulation

Objective To explore the feasibility and accuracy of neutrophil-to-lymphocyte ratio(NLR)in the prediction of testicular torsion(TT)in children with acute scrotal pain.Methods A retrospective case-control study was performed on 158 pediatric patients with ultrasound suspicion of TT who underwent surgical testicular examination during Jan.2017 and Jan.2024.The patients were divided into TT group and non-TT group.Clinical data and laboratory data at admission were analyzed.Sensitivity and specificity of NLR to TT were determined with the area under the curve(AUC)represented on the receiver operating characteristic(ROC)curves.Results There were with no statistically significant differences in clinical data between the two groups(P＞0.05).The NLR was significantly higher in the TT group than in the non-TT group[(4.82±2.37)vs.(2.85±0.75),P＜0.05].The optimal cut-off value of TT predicted by NLR was 2.07,the AUC was 0.809(95％CI:0.709-0.909),and the sensitivity and specificity were 97.9％and 93.3％,respectively,which were significantly higher than other factors.Conclusion For suspicious ultrasound diagnosis of pediatric acute scrotal pain cases,NLR can be used to predict the possibility of TT and may help to evaluate the urgent surgical treatment in these patients.

To explore the anti-HSV-1 effect of silencing gD gene expression by RNA interference, five 21-nucleotide duplex small interfering RNAs(siRNAs) targeting the HSV1 gD sequence were designed and the gD-EGFP fusion gene expression vector was constructed, then co-transfected into Vero cell, and screened the effective siRNA through analyzing the intensity of the EGFP fluorescence. Finally, the anti-HSV1 effect was confirmed by plaque reduction assay, real-time PCR and daughter virus titration of HSV1 infected Vero cells transfected with siRNAs. The study demonstrated that siRNAs could effectively and specifically inhibit gD gene expression in HSV1-infected cells, but only had a little effect on HSV1 infection, so taking gD as the target of siRNA against HSV1 needs further study.
Animals ; Cercopithecus aethiops ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Herpesvirus 1, Human ; genetics ; Humans ; Polymerase Chain Reaction ; RNA Interference ; RNA, Small Interfering ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Vero Cells ; Viral Envelope Proteins ; genetics ; metabolism