Identified Small Open Reading Frame-Encoded Peptides in Human Serum with Nanoparticle Protein Coronas.

The low-molecular-weight proteins (LMWP) in serum and plasma are related to various human diseases and can be valuable biomarkers. A small open reading frame-encoded peptide (SEP) is one kind of LMWP, which has been found to function in many bioprocesses and has also been found in human blood, making it a potential biomarker. The detection of LMWP by a mass spectrometry (MS)-based proteomic assay is often inhibited by the wide dynamic range of serum/plasma protein abundance. Nanoparticle protein coronas are a newly emerging protein enrichment method. To analyze SEPs in human serum, we have developed a protocol integrated with nanoparticle protein coronas and liquid chromatography (LC)/MS/MS. With three nanoparticles, TiO2, Fe3O4@SiO2, and Fe3O4@SiO2@TiO2, we identified 164 new SEPs in the human serum sample. Fe3O4@SiO2 and a nanoparticle mixture obtained the maximum number and the largest proportion of identified SEPs, respectively. Compared with acetonitrile-based extraction, nanoparticle protein coronas can cover more small proteins and SEPs. The magnetic nanoparticle is also fit for high-throughput parallel protein separation before LC/MS. This method is fast, efficient, reproducible, and easy to operate in 96-well plates and centrifuge tubes, which will benefit the research on SEPs and biomarkers.

Self-Delivery Nanoplatform Based on Amphiphilic Apoptosis Peptide for Precise Mitochondria-Targeting Photothermal Therapy.

Mitochondria-targeting photothermal therapy could significantly enhance the tumor cell killing effect. However, since therapeutic reagents need to overcome a series of physiological obstacles to arrive at mitochondria accurately, precise mitochondria-targeting photothermal therapy still faces great challenges. In this study, we developed a self-delivery nanoplatform that specifically targeted the mitochondria of tumor cells for precise photothermal therapy. Photothermal agent IR780 was encapsulated by amphiphilic apoptotic peptide KLA with mitochondria-targeting ability to form nanomicelle KI by self-assembly through hydrophilic and hydrophobic interactions. Subsequently, negatively charged tumor-targeting polymer HA was coated on the surface of KI through electrostatic interactions, to obtain tumor mitochondria-targeting self-delivery nanoplatform HKI. Through CD44 receptor-mediated recognition, HKI was internalizated by tumor cells and then disassembled in an acidic environment with hyaluronidase in endosomes, resulting in the release of apoptotic peptide KLA and photothermal agent IR780 with mitochondria anchoring capacity, which achieved precise mitochondria guidance and destruction. This tumor mitochondria-targeting self-delivery nanoplatform was able to effectively deliver photothermal agents and apoptotic peptides to tumor cell mitochondria, resulting in precise destruction to mitochondria and enhancing tumor cell inhibition at the subcellular organelle level.

Adequate dietary magnesium intake may protect females but not males older than 55 years from cognitive impairment.

BACKGROUND: Magnesium is an essential nutrient required to maintain brain health throughout life, and adequate magnesium intake is positively associated with cognitive performance in older adults. However, sex differences in magnesium metabolism have not been adequately assessed in humans. OBJECTIVES: We investigated sex differences in the effect of dietary magnesium intake and the risk of different types of cognitive impairment in older Chinese adults. METHODS: We collected and assessed dietary data and cognitive function status in people aged 55 years and older in northern China who participated in the Community Cohort Study of Nervous System Diseases from 2018 to 2019 to explore the relationship between dietary magnesium intake and the risk of each type of mild cognitive impairment (MCI) in sex-specific cohorts of older adults. RESULTS: The study included 612 people: 260 (42.5%) men and 352 (57.5%) women. Logistic regression results showed that for the total sample and women's sample, high dietary magnesium intake reduced the risk of amnestic MCI (ORtotal = 0.300; ORwomen = 0.190) and multidomain amnestic MCI (ORtotal = 0.225; ORwomen = 0.145). The results of restricted cubic spline analysis showed that the risk of amnestic MCI (ptotal = 0.0193; pwomen = 0.0351) and multidomain amnestic MCI (ptotal = 0.0089; pwomen = 0.0096) in the total sample and women's sample gradually decreased with increasing dietary magnesium intake. CONCLUSIONS: The results suggest that adequate magnesium intake may have a preventive effect against the risk of MCI in older women.

[Association between vitamin E intake and mild cognitive impairment in people aged 55 years and older in Hebei Province].

OBJECTIVE: To investigate the relationship between dietary vitamin E(VE) intake and mild cognitive impairment(MCI). METHODS: Based on the data of Hebei Province in the 2018 National Key Research and Development Program Nervous system disease CCSNSD, 612 subjects were included in this study. All study participants were surveyed for dietary VE intake by the Dietary Frequency Questionnaire and assessed for cognitive function by the Montreal Cognitive Assessment Scale. The relationship between dietary VE intake and MCI and its subtypes was analyzed using logistic regression and restricted cubic splines. RESULTS: The study subjects included 260(42.5%) males and 352(57.5%) females, with an average age of(66.8±7.4) years and an average dietary VE intake of(12.17±4.91) mg/d. The prevalence of cognitive impairment in the study population was 41.3%. After adjusting the covariates of age, gender, energy intake, residence, education level, employment status, BMI, smoking, drinking, physical activity, hypertension, diabetes, VE intake was not associated with the risk of non-amnesic(naMCI), single-domain amnesic(aMCI-SD), but relate to the risk of multi-domain amnesic(aMCI-MD). What's more, compared to dietary vitamin E intake Q1 group, the OR(95%CI) for multi-domain amnesic mild cognitive impairment in Q2, Q3 and Q4 groups were 1.628(0.836-3.170), 0.313(0.124-0.791) and 0.727(0.330-1.602), respectively. Using vitamin E intake as a continuous-type variable, a non-linear dose-response relationship was found between VE intake and mild cognitive dysfunction of the multi-domain amnesic type(P=0.02). When VE intake was about 17 mg/d, the risk of aMCI-MD was the lowest. CONCLUSION: There is an approximate "U" shaped dose-response relationship between dietary VE intake and the risk of mild cognitive impairment with multi-domain amnesia type. Research suggests that moderate VE intake has a preventive effect on the development of mild cognitive impairment.

The association between low carbohydrate diet scores and cardiometabolic risk factors in Chinese adults.

Epidemiological studies on the association between the low carbohydrate diet (LCD) score and cardiovascular disease risk factors have limited and inconsistent results. Data are from the baseline survey of Community-based Cohort Study on Nervous System Diseases. A total of 4609 adults aged ≥18 years were included in the study. Dietary data were assessed using a validated semi-quantitative Food Frequency Questionnaire. Multivariable logistic regression analyses were used to estimate relationships of three LCD scores with low high-density lipoprotein cholesterol (HDL-C), high low-density lipoprotein cholesterol (LDL-C), hypercholesterolemia, hypertriglyceridemia, impaired fasting glucose (IFG), high blood pressure, and hyperuricemia after adjusting for potential confounders. A higher LCD score was negatively associated with low HDL-C [OR (95%CI): 0.65 (0.50, 0.83), P=0.0001] and IFG [OR (95%CI): 0.65 (0.51, 0.81), P=0.001] after the final adjustment. However, there are gender differences in this result. Males in the highest quintile of the animal-based or plant-based LCD scores showed a decreased risk of low HDL-C, and females in the highest quintile of the animal-based or plant-based LCD scores showed a decreased risk of IFG than those in the lowest quintile of the LCD scores. These results suggest that gender differences should be considered when using LCD to treat dyslipidemia and reduce fasting blood glucose.

Lipopolysaccharide-induced proliferation and glycolysis in airway smooth muscle cells via activation of Drp1.

Abnormal airway smooth muscle cells (ASMCs) proliferation is an important pathological process in airway remodeling contributes to increased mortality in asthma. Mitochondrial dynamics and metabolism have a central role in the maintenance of the cell function. In this study, lipopolysaccharide (LPS)-induced ASMCs proliferative model was used to investigate the effect of mitochondria on the proliferation of ASMCs and the possible mechanism. We used cell and molecular biology to determine the effect of dynamin-related protein 1 (Drp1) on LPS-mediated ASMCs cell cycle progression and glycolysis. The major findings of the current study are as follows: LPS promoted an increased mitochondrial fission and phosphorylation of Drp1 at Ser616 (p-Drp1 Ser616). LPS-induced ASMCs proliferation and cell cycle progression, which was significantly inhibited application of Drp1 RNA interfering. Glycolysis inhibitor 2-deoxyglucose (2-DG) depressed ASMCs proliferative process induced by LPS stimulation. LPS caused mitochondrial metabolism disorders and aerobic glycolysis in a dependent on Drp1 activation. These results indicated that Drp1 may function as a key factor in asthma airway remodeling by mediating ASMC proliferation and cell cycle acceleration through an effect on mitochondrial metabolic disturbance.

Identification of phosphorylated small ORF-encoded peptides in Hep3B cells by LC/MS/MS.

Small ORF-encoded peptides (SEPs) are a class of low molecular weight proteins and peptides comprising <100 amino acids with important functions in various life activities. Although the sequence length is short, SEPs might also have post-translational modification (PTM). Phosphorylation is one of the most essential PTMs of proteins. In this work, we enriched phosphopeptides with IMAC and TiO2 materials and analyzed the phosphorylated SEPs in Hep3B cells. A total of 24 phosphorylated SEPs were identified, and 11 SEPs were coded by ncRNA. For the sequence analysis, we found that the general characteristics of phosphorylated SEPs are roughly the same as canonical proteins. Besides, two phosphorylation SEPs have the Stathmin family signature 2 motif, which can regulate the microtubule cytoskeleton. Some SEPs have domains or signal peptides, indicating their specific functions and subcellular locations. Kinase network analysis found a small number of kinases that may be a clue to the specific functions of some SEPs. However, only one-fifth of the predicted phosphorylation sites were identified by LC/MS/MS, indicating that many SEP PTMs are hidden in the dark, waiting to be uncovered and verified. This study helps expand our understanding of SEP and provides information for further SEP function investigation. SIGNIFICANCE: Small ORF-encoded peptides (SEPs) are important in various life activities. Although the sequence length is short (<100AA), SEPs might also have post-translational modification (PTM). Phosphorylation is one of the most essential PTMs of proteins. We enriched phosphopeptides and analyzed the phosphorylated SEPs in Hep3B cells. That is the first time to explore the PTM of SPEs systematically. Kinase network analysis found a small number of kinases that may be a clue to the specific functions of SEPs. More SEP PTMs are hidden in the dark and waiting to be uncovered and verified. This study helps expand our understanding of SEP and provides information for further SEP function investigation.

Comparative proteomic analysis of seed germination between allotetraploid cotton Gossypium hirsutum and Gossypium barbadense.

Seed germination, a key initial event in the plant life cycle, directly affects cotton yield and quality. Gossypium barbadense and Gossypium hirsutum gradually evolved through polyploidization, resulting in different characteristics, and this interspecific variation lacks genetic and molecular explanation. This work aimed to compare the proteomes between G. barbadense and G. hirsutum during seed germination. Here, we identified 2740 proteins for G. barbadense and 3758 for G. hirsutum. In the initial state, proteins in two cotton involved similar bioprocess, such as sugar metabolism, DNA repairing, and ABA signaling pathway. However, in the post-germination stage, G. hirsutum expressed more protein related to redox homeostasis, peroxidase activity, and pathogen interactions. Analyzing the different expression patterns of 915 single-copy orthogroups between the two kinds of cotton indicated that most of the differentially expressed proteins in G. barbadense were related to carbon metabolism. In contrast, most proteins in G. hirsutum were associated with stress response. Besides that, by proteogenomic analysis, we found 349 putative non-canonical peptides, which may be involved in plant development. These results will help to understand the different characteristics of these two kinds of cotton, such as fiber quality, yield, and adaptability. SIGNIFICANCE STATEMENT: Cotton is the predominant natural fiber crop worldwide; Gossypium barbadense and Gossypium hirsutum have evolved through polyploidization to produce differing traits. However, given their specific features, the divergence of mechanisms underlying seed germination between G. hirsutum and G. barbadense has not been discussed. Here, we explore what protein contributes to interspecific differences between G. barbadense and G. hirsutum during the seed germination period. This study helps to elucidate the evolution and domestication history of cotton polyploids and may allow breeders to understand their domestication history better and improve fiber quality and adaptability.

Interfacial modification of recombinant protein for immunoglobulin G adsorption with spindle-shaped MOF as nano molecular containers.

Development of fresh solid phase extractant is critical for selective separation and purification of special proteins. Herein, we demonstrated a recombinant Staphylococcal Protein G (rSPG) with a His-tag modified the novel single-metal organic framework (rSPG@Ni-MOF-74). The proposed solid-phase extraction material possessed a uniform spindle-shaped structure, large surface area (709.60 m2 g-1) and pore volume (0.08 m3 g-1), high metal content (22.57 wt%), which facilitated the interaction between host and guest. As results, the composite displayed outstanding selective recognition and adsorption of IgG, due to synergistic effect of the binding ability of rSPG with the Fc region of IgG, maintained through hydrogen bonding and electrostatic attraction, as well as hydrophobic interaction. The adsorption performance and mechanism of rSPG@Ni-MOF-74 have been fully investigated. Additionally, the rSPG@Ni-MOF-74 composite could effectively separate IgG from serum obtained from healthy humans, with the purity of the separated IgG verified through SDS-PAGE analysis. Furthermore, LC-MS/MS analysis identified a high content of IgG (55.3 %) in the eluate from rSPG@Ni-MOF-74, suggesting the great potential of rSPG@Ni-MOF-74 in IgG separation and enrichment from complex matrix.

Peroxiredoxin 4 deficiency induces accelerated ovarian aging through destroyed proteostasis in granulosa cells.

Ovarian aging, a complex and challenging concern within the realm of reproductive medicine, is associated with reduced fertility, menopausal symptoms and long-term health risks. Our previous investigation revealed a correlation between Peroxiredoxin 4 (PRDX4) and human ovarian aging. The purpose of this research was to substantiate the protective role of PRDX4 against ovarian aging and elucidate the underlying molecular mechanism in mice. In this study, a Prdx4-/- mouse model was established and it was observed that the deficiency of PRDX4 led to only an accelerated decline of ovarian function in comparison to wild-type (WT) mice. The impaired ovarian function observed in this study can be attributed to an imbalance in protein homeostasis, an exacerbation of endoplasmic reticulum stress (ER stress), and ultimately an increase in apoptosis of granulosa cells. Furthermore, our research reveals a noteworthy decline in the expression of Follicle-stimulating hormone receptor (FSHR) in aging Prdx4-/- mice, especially the functional trimer, due to impaired disulfide bond formation. Contrarily, the overexpression of PRDX4 facilitated the maintenance of protein homeostasis, mitigated ER stress, and consequently elevated E2 levels in a simulated KGN cell aging model. Additionally, the overexpression of PRDX4 restored the expression of the correct spatial conformation of FSHR, the functional trimer. In summary, our research reveals the significant contribution of PRDX4 in delaying ovarian aging, presenting a novel and promising therapeutic target for ovarian aging from the perspective of endoplasmic reticulum protein homeostasis.

Branched-chain amino acids supplementation has beneficial effects on the progression of liver cirrhosis: A meta-analysis.

BACKGROUND: Liver cirrhosis (LC) is currently the 11th most common cause of death and 15th cause of morbidity globally. The treatment of LC is mainly aimed at etiological intervention, lifestyle intervention, prevention and treatment of complications and nutritional treatment. Nutritional treatment of LC mainly includes increasing dietary intake, food intake time and branched-chain amino acids (BCAAs). Despite the recommendation of BCAAs in some guidelines, adverse effects have been reported in studies so the efficacy and safety of BCAAs remain controversial. Currently, BCAAs have been widely used in chronic liver disease, while the summary of the effect of BCAAs on long-term prognosis is rare. AIM: To determine the effects of BCAAs in patients with LC. METHODS: The PubMed, Cochrane Library, Embase and Web of Science databases were searched. The retrieval deadline was 1 October 2021 and there were no language restrictions set in the retrieval. The study was performed in strict accordance with the inclusion and exclusion criteria. Nine studies were finally included. The primary outcome was complications of LC. The secondary outcomes were nutritional status and liver function. This meta-analysis used the Review Manager, version 5 statistical package (Cochrane Collaboration, Oxford, England) for analysis. RESULTS: The analysis included nine studies that consisted of 1080 patients (554 in the BCAA groups and 526 in the control groups). The nine studies were randomized control trials (RCTs). The quality of the studies was assessed using the risk of bias method recommended by the Cochrane Collaboration. BCAAs reduced the rate of complications in LC patients [Risk ratio: 0.70, 95% confidence interval (CI): 0.56-0.88, P = 0.002] and improved patients' albumin levels [std mean difference SMD: 0.26, 95%CI: 0.12-0.40, P = 0.0002]. Meanwhile, BCAAs significantly ameliorated the levels of alanine transaminase (SMD: -2.03, 95%CI: -2.52 to -1.53, P < 0.00001) and aspartate aminotransferase (SMD: -1.8, 95%CI: -2.14 to -1.46, P < 0.00001). Meanwhile, glucose in the LC was significantly increased in BCAA-treated patients (MD: 13.04, 95%CI: 6.81-19.89, P = 0.0002). CONCLUSION: BCAAs reduce the incidence of complications in patients with LC and ameliorate nutritional status.

Randomized controlled trial for time-restricted eating in overweight and obese young adults.

Time-restricted eating (TRE) is known to improve metabolic health, whereas very few studies have compared the effects of early and late TRE (eTRE and lTRE) on metabolic health. Overweight and obese young adults were randomized to 6-h eTRE (eating from 7 a.m. to 1 p.m.) (n = 21), 6-h lTRE (eating from 12 p.m. to 6 p.m.) (n = 20), or a control group (ad libitum intake in a day) (n = 19). After 8 weeks, 6-h eTRE and lTRE produced comparable body weight loss compared with controls. Compared with control, 6-h eTRE reduced systolic blood pressure, mean glucose, fasting insulin, insulin resistance, leptin, and thyroid axis activity, whereas lTRE only reduced leptin. These findings shed light on the promise of 6-h eTRE and lTRE for weight loss. Larger studies are needed to assess the promise of eTRE to yield better thyroid axis modulation and overall cardiometabolic health improvement.

AMPK activation enhances PPARα activity to inhibit cardiac hypertrophy via ERK1/2 MAPK signaling pathway.

Activation of adenosine monophosphate-activated protein kinase (AMPK) has been shown to inhibit cardiac hypertrophy through peroxisome proliferators-activated receptor-α (PPARα) signaling pathway, but the detailed mechanism remains unclear. A rat model of cardiac hypertrophy created by transaortic constriction (TAC) was used to investigate the mechanism involved in regulation of PPARα activity by AMPK. It was observed that treatment with AICAR (5-aminoimidazole 1 carboxamide ribonucleoside), an AMPK activator, significantly inhibited cardiac hypertrophy in vivo and in vitro. Phosphorylated extracellular signal regulated protein kinase (phospho-ERK1/2) and phospho-p38 mitogen-activated protein kinase (MAPK) protein levels were significantly up-regulated, while PPARα protein level was down-regulated in TAC rats. AICAR treatment reversed the changes of PPARα and phospho-ERK1/2, but increased phospho-p38 MAPK protein level in TAC rats. Similar changes of PPARα and phospho-ERK1/2 protein levels were observed in the hypertrophied cardiomyocytes induced by phenylephrine treatment. Epidermal growth factor (EGF, ERK1/2 activator), but not SB203580 (p38 inhibitor) blocked the up-regulation of PPARα protein level induced by AICAR. Luciferase assay showed that AICAR increased PPARα transcriptional activity which was abrogated by EGF, but not by SB203580. These results demonstrate that AMPK activation enhances the activity of PPARα to inhibit cardiac hypertrophy through ERK1/2, but not p38 MAPK, signaling pathway.

Associations Between Dietary Inflammatory Index and Sex Hormones Among 6- to 19-Year-Old Children and Adolescents in NHANES 2015-2016.

Objectives: This study aimed to assess the relationship between dietary inflammatory index (DII) and sex steroids in children (6-11 years old) and adolescents (12-19 years old) in the U.S. National Health and Nutrition Examination Survey, 2015-2016. Methods: Participants between the ages of 6-19 have 24-hour dietary intake data, serum sex hormones [total testosterone (TT), estradiol (E2)], and sex hormone-binding globulin (SHBG) available data (n = 1382). The free androgen index (FAI) is calculated as TT divided by SHBG and the ratio of TT to E2 (TT/E2). The constructed puberty state is defined as high levels of steroid hormones (TT≥50 ng/dL in men, E2≥20 pg/ml in women) or onset of menarche. Multiple linear regression analysis was stratified by gender-age and gender-pubertal status groups to evaluate the association between DII and sex hormone levels. Results: After adjusting for covariates, the association between consecutive DII and sex hormone indicators by gender and age group. In male adolescents, DII was always negatively associated with TT (P-trend = 0.09), FAI (P-trend = 0.03) and E2 (P-trend = 0.01), and monotonically positively associated with SHBG (P-trend = 0.02).In female adolescents, with the increase of DII, a significant positive correlation with SHBG was observed (ß 0.017, 95%CI: 0.009,0.053) (Table 3). Among female adolescents, a significant negative association between DII and TT and a significant positive association between SHBG were observed in this group. Moreover, DII was positively associated with SHBG of prepubertal males and negatively associated with FAI of prepubertal females. Conclusions: DII was associated with decreased levels of certain sex steroid hormones (TT, FAI, and E2) and increased levels of SHBG in adolescents or pubertal individuals, with the associations presenting somewhat sex-dependent pattern. However, there is little evidence that there is a significant association in children or prepubertal children. Further research needs to be carried out to verify our results.

Association between the Lymphotoxin-α A252g Gene Polymorphism and the Risk of Sepsis and Mortality: A Meta-Analysis.

BACKGROUND: The association between the lymphotoxin-α (LTA) A252G polymorphism and sepsis risk has been extensively studied, but the results have been controversial. This study is aimed at investigating the overall association between the LTA A252G polymorphism and the risk of sepsis/septic shock and sepsis-related mortality. METHODS: We searched the PubMed and EMBASE databases to identify studies that investigated the association between the LTA A252G polymorphism and risks of sepsis, septic shock, and mortality. The relevant data were extracted, and statistical analyses were performed using the Revman 5.0 and STATA 12 software. RESULTS: A total of 32 publications were included in the meta-analysis. The results demonstrated that the LTA A252G polymorphism showed no significant association with sepsis risk (GG+GA vs. AA: OR = 0.92, 95%CI = 0.79-1.07, p = 0.27) or with sepsis shock risk (GG+GA vs. AA: OR = 1.01, 95%CI = 0.84-1.22, p = 0.91). However, in the subgroup analyzed by ethnicity, the LTA A252G polymorphism significantly decreased sepsis risk in the Asian population for the recessive model [GG vs. GA+AA: OR = 0.82, 95%CI = 0.68-0.99, p = 0.04] but not in the Caucasian population. Moreover, comparisons between sepsis patients who survived and those who did not suggested that the LTA A252G polymorphism decreases the risk of mortality [GG+GA vs. AA: OR = 0.57, 95%CI = 0.41-0.80, p < 0.01]. CONCLUSION: Our results suggested that the A252G polymorphism in the LTA gene decreased the risk of sepsis in Asians and may reduce mortality in septic individuals.

The association study of Apolipoprotein E polymorphisms and chronic obstructive pulmonary disease in the Chinese population: A case-control study.

Chronic obstructive pulmonary disease (COPD) patients have increased cardiovascular morbidity and mortality. Apolipoprotein E (ApoE) is involved in chronic inflammation which is the common characteristic of emphysema and cardiovascular disease. ApoE polymorphisms are associated with cardiovascular disorders and atherosclerosis. There is no report about the association between ApoE polymorphism and COPD.A total of 480 COPD patients and 322 controls who were unrelated Chinese Han individuals were enrolled. Rs429358 and rs7412 were genotyped and the associations between ApoE polymorphisms and COPD risk were analyzed by logistic regression analysis. Online software SHEsis were applied to perform linkage disequilibrium (LD) and haplotypes analysis. The interactions of ApoE and environmental factor on COPD susceptibility was analyzed by software MDR3.0.2.No significant association was found between rs429358, rs7412 and COPD under different genetic models. Rs429358 and smoking formed the best model in the MDR analysis. The frequency of E2/E2 phenotype was the lowest in 2 groups. E3/E3 was the most common phenotype, accounting for 69.8% of COPD patients and 68.9% of controls. No statistically difference was identified between the cases and controls under different phenotypes.This was the first genetic association study between ApoE and COPD. No positive association was found in the Chinese Han population. Rs429358 and smoking status existed significant interaction, indicating that both of ApoE and smoking may be involved in the development of COPD disease.

COVID-19 Is Distinct From SARS-CoV-2-Negative Community-Acquired Pneumonia.

Background: Corona virus disease (COVID-19) is an infectious respiratory disease that has spread rapidly across the world. Many studies have already evaluated the clinical features of COVID-19, but how it compares with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-negative community-acquired pneumonia (SN-CAP) is still unclear. Moreover, COVID-19 mortality is correlated with disease severity, but indicators for severity grading have not been specified. We aimed to analyze the clinical characteristics of COVID-19 in comparison with SN-CAP and find indicators for disease severity in COVID-19. Methods: Patients diagnosed with COVID-19 and SN-CAP were enrolled. Clinical, radiological, and laboratory data were analyzed. Results: The numbers of COVID-19 and SN-CAP patients enrolled were 304 and 138, respectively. The age of the patients was not significantly different between the groups. Compared with SN-CAP, COVID-19 patients had more symptoms of fever and dyspnea; and showed significant difference in blood count results. Computed tomography (CT) imaging of COVID-19 patients showed patchy ground-glass opacities that correlated with disease severity, whereas the CT imaging of SN-CAP patients showed patchy high-density shadows. COVID-19 patients were classified into moderate, severe, and critically severe groups. The severe and critically severe groups had elevated levels of white blood cells (WBC), neutrophils, platelets, C-reaction protein (CRP), lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), troponin-I, creatinine, and blood urea nitrogen (BUN). However, they had decreased levels of lymphocytes, lymphocyte ratio, and albumin. Compared with the younger patients, the older COVID-19 individuals had more chronic diseases and significantly elevated levels of WBC, neutrophil, and CRP levels. Conclusion: SN-CAP showed more inflammatory reaction than COVID-19. Old people with chronic diseases are more susceptible to COVID-19 and have a high likelihood of developing severe and critically severe infection. Levels of WBC, lymphocytes, neutrophils, CRP, NLR, PLR, troponin-I, creatinine, and BUN are important indicators for severity grading in COVID-19.

[Effects of epidermal growth factor receptor on airway inflammation in a rat asthmatic model].

OBJECTIVE: To explore the possible roles of epidermal growth factor receptor (EGFR) in the process of acute and chronic airway inflammation in a rat asthmatic model. METHODS: Forty-five Sprague-Dawley (SD) rats were randomly divided into control groups (subgroups A1, A2, A4), asthmatic groups (subgroups B1, B2, B4) and treatment groups (subgroups C1, C2, C4), with 5 mice in each subgroup. Mice in the asthmatic and treatment groups were exposed to OVA challenge for 1 week, 2 weeks and 4 weeks. Rats in the treatment groups received intraperitoneal injection of a tyrosine kinase inhibitor Genistein (Rongli China) with the dose of 20 mg/kg 1 hour before OVA exposure. Total cell counts and cell differentials in bronchoalveolar lavage fluid (BALF) were performed. A semi-quantified method of airway inflammation score was used to evaluate airway inflammation by hematoxylin-eosin (HE) staining. Expression of EGFR and tyrosine phosphorylation (EGFR activation) in airway epithelium at different times of OVA exposure were evaluated by immunohistochemical and immunofluorescence. All data were expressed as mean +/- SD. One-way ANOVA was used for comparison between 2 groups and post-hoc multiple comparisons of means were performed by using Least Significant Difference. RESULTS: (1) The total cell counts and cell differentials in the BALF of subgroups B1, B2 and B4 were higher than those of subgroups A1, A2 and A4. The total cell counts and eosinophils (EOS) in the BALF of subgroups C1, C2, and C4 [Total cells (48 +/- 6) x 10(5), (51 +/- 9) x 10(5), (57 +/- 12) x 10(5); EOS (2.5 +/- 0.5) x 10(5), (2.7 +/- 0.6) x 10(5), (2.6 +/- 0.5) x 10(5), respectively] decreased significantly as compared to those of subgroups B1, B2 and B4 [Total cells (70 +/- 10) x 10(5), (88 +/- 8) x 10(5), (72 +/- 10) x 10(5); EOS (5.6 +/- 0.8) x 10(5), (6.6 +/- 0.6) x 10(5), (4.3 +/- 0.4) x 10(5)], all P < 0.05. There was no significant difference in the counts of neutrophils and lymphocytes in BALF between the treatment groups and the asthmatic groups. The count of epithelial cells in group C1 [(2.5 +/- 0.5) x 10(5)] was lower than that in group B1[(4.9 +/- 0.7) x 10(5)], q = 4.671, P < 0.05. But that in group C4[(5.7 +/- 1.2) x 10(5)] was higher than that in group B4 [(4.3 +/- 0.4) x 10(5)], q = 4.012, P < 0.05. (2) The airway inflammation score in group C4(3.6 +/- 0.6) was less than that in group B4(5.1 +/- 0.6), q = 4.923, P < 0.05. The scores of group C1 and C2 were less than those of group B1 and B2, but the differences did not reach statistical significance. (3) The expression of EGFR and tyrosine phosphorylation in airway epithelium of the OVA sensitized subgroups were increased statistically as compared to the control subgroups (all P < 0.05). Genistein decreased tyrosine phosphorylation of EGFR in subgroups C1, C2 and C4[(3.12 +/- 0.24), (3.00 +/- 0.28), (2.69 +/- 0.54)] as compared to subgroups B1, B2 and B4[(3.69 +/- 0.43), (3.57 +/- 0.29), (4.46 +/- 0.47), respectively] (all P < 0.05). (4) There were positive correlations between expression and activation of EGFR in airway epithelium and total cell counts, EOS counts, neutrophil and lymphocyte counts in BALF, and airway inflammation scores (all P < 0.05). CONCLUSIONS: EGFR is involved in airway inflammation of asthmatic rats. Tyrosine kinase inhibitor Genistein inhibits acute and chronic airway inflammation in the asthmatic model.

[The role of 12-alkylated chitosans/plasmid-encoding antisense endothelin converting enzyme complex nanoparticles in immunomodulation of allergic airway inflammation].

OBJECTIVE: To explore a novel nonspecific immunomodulation for the treatment of allergic airway inflammation by RNA interference for endothelin converting enzyme (ECE) using 12-alkylated chitosans/plasmid-encoding antisense ECE complex nanoparticles. METHODS: Forty BALB/c mice were randomly divided into Group N (normal control), Group NM (OVA + 12-ACSs/antisense-ECE plasmid), Group As (OVA) and Group DNA (OVA + antisense-ECE plasmid), and sensitized by intraperitoneal injection of OVA at day 1 and day 14, followed by challenge with aerosol of 1% OVA at day 24, 25 and 26, but with saline as a control (N). Supernatants from cultured splenocytes and lung homogenates were subjected to detection of the levels of interleukin-4, 5, 10, 13 (IL-4, 5, 10, 13), interferon-gamma (IFN-gamma) and endothelin-1 (ET-1) by using ELISA. Lung tissues were embedded, sliced and HE stained for histopathologic examination. The cultured splenocytes were subjected to flow cytometry detection (IL-4, IL-10 and IFN-gamma). RESULTS: Mice in group NM showed a lower level of cell count than that in either group AS or group DNA. Compared with N group, the lung tissues taken from the mice in As and DNA groups displayed allergic inflammation with eosinophil infiltration, while the pulmonary inflammation was decreased significantly in group NM. The levels of ET-1, IL-4, IL-13 and IL-5 were down regulated in group NM compared to As and DNA groups (P < 0.05 or P < 0.01). After stimulation by OVA, the splenocytes from the mice in NM group produced a higher level of IL-10 than that from As and DNA groups (P < 0.05 or P < 0.01). The number of Th2 lymphocytes (CD(4)(+) T cells with IL-4 expression) was significantly elevated in the mice of As, DNA and NM groups respectively, while the number of Th2 lymphocytes was lower in the mice of group NM than in the mice of group As or group DNA. The number of CD(4)(+)CD(25)(+) cells with IL-10 expression was up-regulated in the mice of As, DNA and NM groups respectively compared to the control. The percentage of T regulating cells was higher in the mice of group NM compared with that in the mice of group As or group DNA. No detectable difference in the level of Th1 cells (CD(4)(+) T cells with IFN-gamma expression) was found among the 4 groups. CONCLUSION: 12-ACSs can encapsulate and deliver antisense-ECE expression plasmid into bronchial epithelial cells in vitro and 12-ACSs/antisense ECE plasmid complex nanoparticles have the capability to down regulate the synthesis of ET-1 and thus decrease the allergic airway inflammation in OVA-sensitized mice.

Paeonol regulates hypoxia-induced proliferation of pulmonary artery smooth muscle cells via EKR 1/2 signalling.

Pulmonary hypertension (PH) is a disease with a developmental origin characterized by obstructive vascular remodelling that is partially due to excessive pulmonary arterial smooth muscle cells (PASMCs) proliferation. Paeonol has important effects on vascular cell proliferation, migration, and inflammation, but researchers have not determined whether paeonol participates in the development and progression of pulmonary vascular remodelling. We explored the remarkable anti-proliferative effects of paeonol on hypoxic PASMCs, which are postulated to be mediated by the extracellular signal-regulated kinase 1 and 2 (ERK1/2) signalling pathway. In this study, hypoxic rodent PH models, Western blotting, flow cytometry, immunochemistry, and morphometric analyses of the lung vasculature and right ventricle (RV) vessels were performed. Paeonol reversed hypoxia-induced increases in right ventricular function, right ventricular systolic pressure and thickening of medial walls. Meanwhile, paeonol ameliorated the hypoxia-induced PASMCs proliferation. Furthermore, paeonol modulated cell proliferation and cell cycle transitions from G0/G1 phase to S phase and G2/M phase in an ERK1/2-dependent manner. Our findings emphasize the central function of paeonol in regulating PASMCs proliferation in subjects with PH. Therefore, paeonol represents a potential novel therapeutic approach for the treatment of PH.