The HEAT repeat protein HPO-27 is a lysosome fission factor.

Lysosomes are degradation and signalling centres crucial for homeostasis, development and ageing1. To meet diverse cellular demands, lysosomes remodel their morphology and function through constant fusion and fission2,3. Little is known about the molecular basis of fission. Here we identify HPO-27, a conserved HEAT repeat protein, as a lysosome scission factor in Caenorhabditis elegans. Loss of HPO-27 impairs lysosome fission and leads to an excessive tubular network that ultimately collapses. HPO-27 and its human homologue MROH1 are recruited to lysosomes by RAB-7 and enriched at scission sites. Super-resolution imaging, negative-staining electron microscopy and in vitro reconstitution assays reveal that HPO-27 and MROH1 self-assemble to mediate the constriction and scission of lysosomal tubules in worms and mammalian cells, respectively, and assemble to sever supported membrane tubes in vitro. Loss of HPO-27 affects lysosomal morphology, integrity and degradation activity, which impairs animal development and longevity. Thus, HPO-27 and MROH1 act as self-assembling scission factors to maintain lysosomal homeostasis and function.

Tanshinone IIA, a component of the self-made Xiao-Yin decoction, ameliorates psoriasis by inhibiting IL-17/IL-23 and PTGS2/NF-κB/AP-1 pathways.

BACKGROUND: Psoriasis is a persistent inflammatory dermatological disorder. Tanshinone IIA (tan-IIA) is a biologically active compound in the self-made Xiao-Yin decoction (SMXYD) and exhibits diverse biological properties, such as anti-proliferative and anti-inflammatory effects. The objective of this investigation was to assess the potential of tan-IIA as a therapeutic agent against psoriasis. METHODS: Network pharmacology was employed to ascertain the active constituents and potential pathways associated with SMXYD and psoriasis. We conducted CCK-8, qRT-PCR, and western blotting to assess the proliferation of HaCaT keratinocytes and the expression of IL-17/IL-23 and PTGS2/NF-κB/AP-1 pathways. Additionally, we used H&E staining, western blotting, and ELISA to evaluate the therapeutic effects and signaling pathways of tan-IIA in psoriasis-like mice induced by imiquimod (IMQ). RESULTS: Network pharmacology analysis identified eight hub compounds. The Th17/IL-17 signaling was found to be a potential therapeutic pathway of SMXYD against psoriasis, with JUN (AP-1) as the core molecule. Next, PTGS2 was selected as the target of tan-IIA against psoriasis using network pharmacology analysis. Molecular docking showed a high affinity between PTGS2 and tan-IIA. Tan-IIA treatment attenuated M-5-induced hyperproliferation and inflammation in HaCaT keratinocytes. Additionally, Tan-IIA downregulated the PTGS2/NF-κB/AP-1 pathway in HaCaT keratinocytes. In the IMQ-induced psoriasis-like mouse, tan-IIA significantly reduced the severity of skin lesions and downregulated the PTGS2/NF-κB/AP-1 pathway. Moreover, the combination of methotrexate (MTX) and tan-IIA further inhibited the IL-17/IL-23 and PTGS2/NF-κB/AP-1 pathways. CONCLUSION: The administration of tan-IIA has shown a positive effect on psoriasis by inhibiting the IL-17/IL-23 and PTGS2/NF-κB/AP-1 pathways. The findings suggest that it has promising qualities that make it a potential candidate for the development of future anti-psoriatic agents.

The hippo pathway effector Yes-associated protein promotes epithelial proliferation and remodeling in chronic rhinosinusitis with nasal polyps.

BACKGROUND: Hippo-Yes-associated protein (YAP) pathway plays an important role in epithelial cell proliferation and development. However, its possible role in chronic rhinosinusitis with nasal polyps (CRSwNP) remains unknown. We aim to investigate it on nasal epithelial proliferation and remodeling in CRSwNP. METHODS: The expressions of hippo pathway components as well as Ki-67 and E-cadherin in the sinonasal mucosa and nasal epithelial cells were analyzed in 14 controls, 14 eosinophilic CRSwNP, and 14 noneosinophilic CRSwNP. Nasal epithelial cells from 6 controls, 6 eosinophilic CRSwNP, and 6 noneosinophilic CRSwNP were cultured and treated with lipopolysaccharide (LPS), Poly(I:C), or a selective YAP inhibitor verteporfin (VP). RESULTS: The hippo pathway components MST1, LATS1/2, YAP, and TEAD1 were increased in both eosinophilic and noneosinophilic CRSwNP, particularly in nasal epithelial cells, along with upregulation of Ki-67 and downregulation of E-cadherin. The mRNA levels of YAP positively correlated with the Ki-67 mRNA levels, and negatively associated with the E-cadherin mRNA levels in polyp tissues and epithelial cells from nasal polyps (NPECs). LPS and Poly(I:C) upregulated the YAP expression in nasal epithelial cells accompanied by increased TEAD1 and Ki-67 expression. Conversely, YAP inhibition by VP decreased TEAD1 and Ki-67 expression in NPECs. CONCLUSIONS: Hippo pathway components are abnormally upregulated in NPECs, and its effector YAP promotes nasal epithelial cells proliferation and remodeling in CRSwNP. It provides a rationale to explore inhibition of YAP as a novel therapeutic strategy for reducing the epithelial proliferation and remodeling in CRSwNP.

[Development of a radiomics signature to predict Ki-67 expression level in non-small cell lung cancer].

OBJECTIVE: To develop a radiomics signature based on CT image features to estimate the expression level of Ki-67 in non-small cell lung cancer (NSCLC).
 Methods: A total of 108 NSCLC patients, who underwent non-enhanced and contrast-enhanced CT scan in our hospital from January 2014 to November 2017, were retrospectively analyzed. They were confirmed by histopathological examination and undergone Ki-67 expression level test within 2 weeks after CT examination. The non-enhanced and contrast-enhanced CT three-dimensional structural images of the lesions were manually delineated by MaZda software, and the texture features of the region of interest were extracted. Combination of feature selection and classification methods were used to build radiomics signatures, and the classification were assessed using misclassification rates. The MaZda software provides texture feature selection methods including mutual information (MI), Fisher coefficients (Fisher), classification error probability combined with average correlation coefficients (POE+ACC), and Fisher+POE+ACC+MI (FPM), and texture feature analysis including raw data analysis (RDA), principal component analysis (PCA), linear classification analysis (LDA) and nonlinear classification analysis (NDA).
 Results: Among the 108 patients, 50 cases were at high levels of Ki-67 expression and 58 cases were at low levels of Ki-67 expression, respectively. The differences of gender, age and pathological type between the two groups were statistically significant (P<0.05). The radiomics signature built by FPM feature selection combined with NDA feature analysis based on non-enhanced CT images achieved the best performance for predicting the level of Ki-67 with a misclassification rate of 14.81%. However, radiomics signature based on contrast-enhanced CT images did not reduce the misclassification rate.
 Conclusion: The radiomics signature based on conventional CT image texture features is helpful to predict the expression of Ki-67 in NSCLC lesions, which can provide a non-invasive technique for assessing the invasiveness and prognosis for NSCLC.

Diagnostic value of gadobutrol versus gadopentetate dimeglumine in enhanced MRI of brain metastases.

PURPOSE: To compare gadobutrol and gadopentetate dimeglumine (Gd-DTPA) contrast-enhanced magnetic resonance imaging (MRI) at 3T for visualizing brain metastases. MATERIALS AND METHODS: The present randomized study included 60 consecutive patients with known or suspected brain metastases from systemic malignancies. Two enhanced cerebral MR scans were performed in each patient within an interval of 2-5 days using different contrast agents (gadobutrol or Gd-DTPA) at 3T. The dose of the contrast agents (0.1 mmol/kg Gd) was also identical. The axial T1 FLAIR images at 3, 7, and 10 minutes after the injection of the contrast agent were obtained for evaluation. Two experienced radiologists performed subjective evaluation of the image quality, made the choice of the optimal images, and performed an objective evaluation including: signal-to-noise ratio (SNR) of the brain metastases, contrast-to-noise ratio (CNR), contrast enhancement (CE), contrast-to-brain ratio (CBR), and contrast enhancement ratio (CER) of the brain metastases. RESULTS: Subjective evaluation showed that at 3, 7, and 10 minutes gadobutrol elicited higher scores (margin score: 3.56 ± 0.74 vs. 3.33 ± 0.93, 3.68 ± 0.57 vs. 3.45 ± 0.81, 3.58 ± 0.71 vs. 3.43 ± 0.76; interior score: 2.83 ± 0.42 vs. 2.63 ± 0.61, 2.86 ± 0.38 vs. 2.73 ± 0.52, 2.80 ± 0.42 vs. 2.69 ± 0.53; and overall score: 4.42 ± 0.98 vs. 4.09 ± 1.19, 4.57 ± 0.75 vs. 4.26 ± 1.05, 4.48 ± 0.83 vs. 4.21 ± 1.03, respectively) in displaying the details and overall lesions than Gd-DTPA (repeated measures analysis of variance [ANOVA], margin score: P = 0.001, < 0.0001, 0.006; interior score: P < 0.0001, 0.004, 0.009; and overall score: P = 0.001, < 0.0001, < 0.0001, respectively). Subjective optimal image evaluation showed that the percentage of image assessed as "gadobutrol was better than Gd-DTPA (41.2-44.1%)" was greater than that assessed as "Gd-DTPA was better than gadobutrol (5.9-26.5%)." Objective evaluation showed that at 3, 7, and 10 minutes the SNR (214.17 ± 85.70 vs. 199.57 ± 85.08, 214.80 ± 86.03 vs. 199.19 ± 84.74, and 213.83 ± 82.46 vs. 193.68 ± 79.59, respectively), CNR (68.64 ± 50.18 vs. 57.88 ± 51.06, 75.42 ± 53.19 vs. 63.74 ± 53.91, and 77.13 ± 51.86 vs. 63.21 ± 51.71, respectively), CE (101.76 ± 63.31 vs. 87.61 ± 64.85, 99.85 ± 61.56 vs. 85.08 ± 64.98, and 100.33 ± 58.63 vs. 82.73 ± 61.73, respectively), CBR (0.48 ± 0.32 vs. 0.40 ± 0.33, 0.54 ± 0.34 vs. 0.46 ± 0.35, and 0.56 ± 0.34 vs. 0.47 ± 0.34, respectively), and CER (0.99 ± 0.69 vs. 0.88 ± 0.81, 0.97 ± 0.68 vs. 0.86 ± 0.84, and 0.98 ± 0.65 vs. 0.85 ± 0.80, respectively) were all higher when using gadobutrol compared with Gd-DTPA in the enhanced MR (repeated measures ANOVA, all P < 0.0001). On Gd-DTPA enhanced images, 289, 292, and 292 lesions at 3, 7, and 10 minutes were detected by the two radiologists, while 295, 301, and 301 lesions were detected on gadobutrol-enhanced images, respectively. CONCLUSION: Using a 3T T1 FLAIR sequence, gadobutrol (0.1 mmol/kg body weight)-enhanced MR resulted in more conspicuous brain metastases, and more metastases compared with the same dose of Gd-DTPA. A delay time of 7 minutes for postcontrast MRI in patients with brain metastases is suggested in clinical practice. LEVEL OF EVIDENCE: 2 J. MAGN. RESON. IMAGING 2017;45:1827-1834.

Neuroprotective effects of Eucommia ulmoides Oliv. and its bioactive constituent work via ameliorating the ubiquitin-proteasome system.

BACKGROUND: Parkinson's disease (PD) is a chronic neurodegenerative disorder characterized by a loss of dopaminergic neurons in the substantia nigra, decreased striatal dopamine levels, and consequent extrapyramidal motor dysfunction. The purpose of this study was to investigate potential in vivo protective effects of Duzhong against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), as well as the bioactive constituents against 1-methyl-4-phenylpyridinium (MPP(+)) toxicity in vitro. METHODS: Male C57BL/6 mice were intraperitoneally administrated five consecutive injections of MPTP every 24 h at a dose of 30 mg/kg to induce an in vivo PD model. Pole and traction tests were performed in mice to evaluate motor deficits and bradykinesia after the final MPTP administration. The striatal levels of dopamine and its metabolites, 3,4-dihydroxyphenylacetic acid and homovanilic acid, were measured using a High-performance liquid chromatography-electrical conductivity detector. To further explore the bioactive constituents and protective mechanisms of Duzhong, seven compounds from Duzhong were tested on MPP(+)-treated SH-SY5Y cell lines in vitro. A proteasome enzymatic assay and Cell Counting Kit-8 were performed to examine proteasomal activity and cell viability of Duzhong-treated cells, respectively, after exposure to MPP(+) and proteasome inhibitor MG132. RESULTS: Duzhong antagonized the loss of striatal neurotransmitters and relieved the associated anomaly in ambulatory locomotor activity in PD mice after a 3-day pre-treatment of Duzhong crude extract. The five Duzhong compounds attenuated MPP(+)-induced dysfunction of protease activity and reduced MG132-induced cytotoxicity. CONCLUSION: Duzhong could serve as a potential candidate for PD treatment, and its mechanism involves the amelioration of the ubiquitin-proteasome system.

Danhong injection attenuates ischemia/reperfusion-induced brain damage which is associating with Nrf2 levels in vivo and in vitro.

Danhong injection, a Chinese Materia Medica standardized product extracted from Radix Salviae miltiorrhizae and Flos Carthami tinctorii, is used extensively for the treatment of cerebrovascular diseases such as acutely cerebral infarction in clinic. In this study, we further investigated the mechanisms of Danhong injection on cerebral ischemia/reperfusion (I/R) damage relating to Nrf2/ARE signalling pathway in vivo and in vitro. For in vivo experiment, cerebral I/R injury was induced through middle cerebral artery occlusion. Rats were randomly divided into five groups: sham-operated group, I/R injury group, 6 mg/kg edaravone injection (positive control drug) group, 0.9 ml/kg Danhong injection (DHI-L) group, 1.8 ml/kg Danhong injection (DHI-H) group. The neurological score, cerebral infarction and brain edema were assessed while levels of superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) in brain tissue were also evaluated. Transcription levels of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), NAD(P)H: quinone oxidoreductase 1 (NQO1) were analysed by real-time polymerase chain reaction. For in vitro experiment, mouse Neuro-2A cells were wounded with H2O2 then cell viability and mRNA transcriptions levels of Nrf2, HO-1, NQO1 were detected. Protein expression level of Nrf2 was assayed by western blotting. The results showed that Danhong injection could ameliorate neurological score, cerebral infarction and brain edema. Also it can increase levels of SOD, GSH and decrease of MDA and upregulate expressions of Nrf2, HO-1, NQO1 in ischemic brain tissue in vivo. What's more, it increased the mRNA transcription of Nrf2 and HO-1 and upregulated protein expression of Nrf2 in vitro. These findings suggested that Danhong injection could prevent I/R-induced brain damage through activating Nrf2/ARE signaling pathway.

Hawthorn Proanthocyanidin Extract Inhibits Colorectal Carcinoma Metastasis by Targeting the Epithelial-Mesenchymal Transition Process and Wnt/ß-Catenin Signaling Pathway.

Colorectal carcinoma (CRC) is a major global health concern, with cancer metastasis being the main cause of patient mortality, and current CRC treatments are challenged by drug resistance. Although natural compounds, especially in foods like hawthorn proanthocyanidin extract (HPOE), have good anticancer activity, their effects on CRC metastasis remain unknown. Therefore, our objective was to investigate the impact and potential mechanisms of HPOE on the movement and infiltration of cells in the HCT116 CRC cells. Firstly, scratch-healing experiments confirmed the anti-migratory and anti-invasive capabilities of HPOE. Then, network pharmacology identified 16 possible targets, including MMP-9. Subsequently, RT-qPCR and Western blotting experiments confirmed that HPOE downregulated epithelial-mesenchymal transition-related factors (N-cadherin and MMP-9) and inhibited Wnt/ß-catenin pathway activation. Finally, these results were experimentally validated using the Wnt pathway activator Licl and inhibitor XAV939. It was confirmed that HPOE had a certain inhibitory effect on the activation of the Wnt signaling pathway caused by the activator Licl and could enhance the inhibitory effect of the inhibitor XAV939. Our findings provide a basis for developing functional foods or dietary supplements, especially positioning HPOE as a functional food raw material for adjuvant treatment of CRC, given its ability to inhibit metastasis through the Wnt/ß-catenin pathway.

Metabolomic insights into the profile, bioaccessibility, and transepithelial transport of polyphenols from germinated quinoa during in vitro gastrointestinal digestion/Caco-2 cell transport, and their prebiotic effects during colonic fermentation.

The health-promoting activities of polyphenols and their metabolites originating from germinated quinoa (GQ) are closely related to their digestive behavior, absorption, and colonic fermentation; however, limited knowledge regarding these properties hinder further development. The aim of this study was to provide metabolomic insights into the profile, bioaccessibility, and transepithelial transport of polyphenols from germinated quinoa during in vitro gastrointestinal digestion and Caco-2 cell transport, whilst also investigating the changes in the major polyphenol metabolites and the effects of prebiotics during colonic fermentation. It was found that germination treatment increased the polyphenol content of quinoa by 21.91%. Compared with RQ group, 23 phenolic differential metabolites were upregulated and 47 phenolic differential metabolites were downregulated in GQ group. Compared with RQ group after simulated digestion, 7 kinds of phenolic differential metabolites were upregulated and 17 kinds of phenolic differential metabolites were downregulated in GQ group. Compared with RQ group after cell transport, 7 kinds of phenolic differential metabolites were upregulated and 9 kinds of phenolic differential metabolites were downregulated in GQ group. In addition, GQ improved the bioaccessibilities and transport rates of various polyphenol metabolites. During colonic fermentation, GQ group can also increase the content of SCFAs, reduce pH value, and adjust gut microbial populations by increasing the abundance of Actinobacteria, Bacteroidetes, Verrucomicrobiota, and Spirochaeota at the phylum level, as well as Bifidobacterium, Megamonas, Bifidobacterium, Brevundimonas, and Bacteroides at the genus level. Furthermore, the GQ have significantly inhibited the activity of α-amylase and α-glucosidase. Based on these results, it was possible to elucidate the underlying mechanisms of polyphenol metabolism in GQ and highlight its beneficial effects on the gut microbiota.

Artificial intelligence for segmentation and classification of lobar, lobular, and interstitial pneumonia using case-specific CT information.

Background: Pneumonia can be anatomically classified into lobar, lobular, and interstitial types, with each type associated with different pathogens. Utilizing artificial intelligence (AI) to determine the anatomical classifications of pneumonia and assist in refining the differential diagnosis may offer a more viable and clinically relevant solution. This study aimed to develop a multi-classification model capable of identifying the occurrence of pneumonia in patients by utilizing case-specific computed tomography (CT) information, categorizing the pneumonia type (lobar, lobular, and interstitial pneumonia), and performing segmentation of the associated lesions. Methods: A total of 61 lobar pneumonia patients, 60 lobular pneumonia patients, and 60 interstitial pneumonia patients were consecutively enrolled at our local hospital from June 2020 and May 2022. All selected cases were divided into a training cohort (n=135) and an independent testing cohort (n=46). To generate the ground truth labels for the training process, manual segmentation and labeling were performed by three junior radiologists. Subsequently, the segmentations were manually reviewed and edited by a senior radiologist. AI models were developed to automatically segment the infected lung regions and classify the pneumonia. The accuracy of pneumonia lesion segmentation was analyzed and evaluated using the Dice coefficient. Receiver operating characteristic curves were plotted, and the area under the curve (AUC), accuracy, precision, sensitivity, and specificity were calculated to assess the efficacy of pneumonia classification. Results: Our AI model achieved a Dice coefficient of 0.743 [95% confidence interval (CI): 0.657-0.826] for lesion segmentation in the training set and 0.723 (95% CI: 0.602-0.845) in the test set. In the test set, our model achieved an accuracy of 0.927 (95% CI: 0.876-0.978), precision of 0.889 (95% CI: 0.827-0.951), sensitivity of 0.889 (95% CI: 0.827-0.951), specificity of 0.946 (95% CI: 0.902-0.990), and AUC of 0.989 (95% CI: 0.969-1.000) for pneumonia classification. We trained the model using labels annotated by senior physicians and compared it to a model trained using labels annotated by junior physicians. The Dice coefficient of the model's segmentation improved by 0.014, increasing from 0.709 (95% CI: 0.589-0.830) to 0.723 (95% CI: 0.602-0.845), and the AUC improved by 0.042, rising from 0.947 to 0.989. Conclusions: Our study presents a robust multi-task learning model with substantial promise in enhancing the segmentation and classification of pneumonia in medical imaging.

Construction and Evaluation of Nomogram for Hematological Indicators to Predict Pathological Response after Neoadjuvant Chemoradiotherapy in Locally Advanced Rectal Cancer.

OBJECTIVE: A retrospective study was conducted by developing prediction models to evaluate the association between hematological indexes, their changes during neoadjuvant chemoradiotherapy (NCRT), and tumor pathological response in patients with locally advanced rectal cancer. METHODS: The clinical data of 202 patients who received NCRT and radical surgery in Sichuan Cancer Hospital were retrospectively analyzed. Univariate and logistic multivariate regression analyses were used to identify hematological indexes with predictive significance. The independent risk factors were imported into the R software, and a nomogram prediction model was developed. The bootstrap method and ROC curve were used to evaluate the discriminative degree of the model. RESULTS: Univariate analysis demonstrated age, tumor diameter, preoperative T, distance from tumor to the anal verge, CEA before NCRT, preoperative CEA, lymphocyte changes, platelet changes, and pathology of rectal cancer after NCRT were associated. Multivariate analysis demonstrated that age, tumor distance from the anus, preoperative CEA, lymphocyte changes, and platelet changes were independent risk factors. The independent risk factors were imported into the R software to construct a nomogram model. The area under the ROC was 0.76, and the slope of the calibration curve of the nomogram was close to 1. CONCLUSION: A low preoperative CEA level, a young age, a high tumor from the anal verge, the maintenance of circulating lymphocyte level, and a decreased platelet level after NCRT are important factors for favorable outcomes after NCRT. Developing a nomogram prediction model with good discrimination and consistency can provide some guidance for predicting pathological responses after NCRT.

A new estimation of carbon emissions from land use and land cover change in China over the past 300 years.

Scientific estimation of carbon emissions induced by historical land use and land cover change (LUCC) can improve the accuracy of terrestrial ecosystem carbon budget estimates and deepen understanding of the future carbon-sink potential of terrestrial ecosystems. The present study, using historical-document-based data for provincial cropland, forest, and grassland area in China, and experimental-data-based information for provincial vegetation and soil organic carbon density, re-estimates China's LUCC-induced carbon emissions for 1700-1980 using a bookkeeping model in which we updated tabulated functions for carbon losses and gains. The past 300 years have witnessed a dramatic LUCC in China. The cropland area has increased by 67.11 million ha, while the forest and grassland areas have decreased by 127.96 million ha and 16.72 million ha, respectively. Accordingly, the net carbon emissions for 1700-1980 are 6.17-12.35 Pg C, with 8.55 Pg C in the moderate scenario. Among the contributing factors, deforestation was the largest carbon source, accounting for over 90 % of the total carbon emissions. According to our estimates, over 70 % of carbon emissions were caused by harvesting wood, while <30 % were from converting forest and grassland to cropland. Spatially, for the whole period, carbon emissions in southwestern China (Chuan-Yu, Yunnan, and Guangxi), northeastern China (Liaoning, Jilin, and Heilongjiang), and parts of northwestern China (Gan-Ning, Qinghai, and Xinjiang) were as high as 6.03 Pg C, accounting for 70 % of the total carbon emissions. Extending previous studies, we updated the historical LUCC data, carbon density data, and tabulated functions for carbon losses and gains. The estimation results objectively reveal the historical spatiotemporal changes in LUCC-induced emissions.

Lysosomal chloride transporter CLH-6 protects lysosome membrane integrity via cathepsin activation.

Lysosomal integrity is vital for cell homeostasis, but the underlying mechanisms are poorly understood. Here, we identify CLH-6, the C. elegans ortholog of the lysosomal Cl-/H+ antiporter ClC-7, as an important factor for protecting lysosomal integrity. Loss of CLH-6 affects lysosomal degradation, causing cargo accumulation and membrane rupture. Reducing cargo delivery or increasing CPL-1/cathepsin L or CPR-2/cathepsin B expression suppresses these lysosomal defects. Inactivation of CPL-1 or CPR-2, like CLH-6 inactivation, affects cargo digestion and causes lysosomal membrane rupture. Thus, loss of CLH-6 impairs cargo degradation, leading to membrane damage of lysosomes. In clh-6(lf) mutants, lysosomes are acidified as in wild type but contain lower chloride levels, and cathepsin B and L activities are significantly reduced. Cl- binds to CPL-1 and CPR-2 in vitro, and Cl- supplementation increases lysosomal cathepsin B and L activities. Altogether, these findings suggest that CLH-6 maintains the luminal chloride levels required for cathepsin activity, thus facilitating substrate digestion to protect lysosomal membrane integrity.

Tracheobronchial Amyloidosis Accompanied with Asthma: A Case Report and a Mini-Review.

Tracheobronchial amyloidosis is a rare disease characterized by amyloid deposits on the tracheal and bronchial tissue. Patients with tracheobronchial amyloidosis are asymptomatic or exhibit symptoms, such as chronic wheezing, dyspnea, and cough, that are common manifestations of other disorders, including asthma. A bronchoscopic tissue biopsy using Congo red staining is the key standard for diagnosing tracheobronchial amyloidosis. Treatment strategies vary depending on the degree of airway obstruction. If the obstruction is significant and the patient is symptomatic, repeated bronchoscopic treatment, including local resection, laser therapy, stent placement, and radiation therapy, is considered a safer and better option. It is often misdiagnosed as asthma, but cases of tracheobronchial amyloidosis accompanied with asthma have not been reported. We report a case of intermittent wheezing, cough for 33 years, and shortness of breath on exertion for 7 years, which had aggravated in the previous 22 days. A pulmonary examination revealed diffuse wheezing. Pulmonary function testing revealed an obstructive ventilation dysfunction. Computerized tomography (CT) imaging revealed circumferential and irregular thickening of the tracheobronchial wall tissue with calcification and atelectasis of the right middle and lower lobe of the lung. Bronchoscopy revealed diffuse thickening of the mucosa of the trachea and bilateral main bronchi, with multiple nodular protuberances and relatively narrow lumens. The bronchial biopsies revealed massive amyloid deposits under the bronchial mucosa. The deposits exhibited a green birefringence under crossed polarized light after Congo red positive staining. The patient received standard treatment for asthma, and remains in good general condition without wheezing. It is not difficult to distinguish tracheobronchial amyloidosis through chest CT examination for patients with wheezing as long as this disease was considered. It was interesting that we present a rarer case of patient with tracheobronchial amyloidosis accompanied with asthma which both can cause symptoms such as wheezing.

Clinical and Radiological Features Between Patients with Stable COPD from Plateau and Flatlands: A Comparative Study.

Background: COPD patients living in Tibet are exposed to specific environments and different risk factors and probably have different characteristics of COPD from those living in flatlands. We aimed to describe the distinction between stable COPD patients permanently residing at the Tibet plateau and those in flatlands. Methods: We conducted an observational cross-sectional study that enrolled stable COPD patients from Tibet Autonomous Region People's Hospital (Plateau Group) and Peking University Third Hospital (Flatland Group), respectively. Their demographic information, clinical features, spirometry test, blood routine and high-resolution chest CT were collected and evaluated. Results: A total of 182 stable COPD patients (82 from plateau and 100 from flatland) were consecutively enrolled. Compared to those in flatlands, patients in plateau had a higher proportion of females, more biomass fuel use and less tobacco exposure. CAT score and frequency of exacerbation in the past year were higher in plateau patients. The blood eosinophil count was lower in plateau patients, with fewer patients having an eosinophil count ≥300/µL. On CT examination, the proportions of previous pulmonary tuberculosis and bronchiectasis were higher in plateau patients, but emphysema was less common and milder. The ratio of diameters of pulmonary artery to aorta ≥1 was more often in plateau patients. Conclusion: Patients with COPD living at Tibet Plateau had a heavier respiratory burden, lower blood eosinophil count, less emphysema but more bronchiectasis and pulmonary hypertension. Biomass exposure and previous tuberculosis were more common in these patients.

Analysis of Risk Factors for Anastomotic Leakage After Laparoscopic Anterior Resection of Rectal Cancer and Construction of a Nomogram Prediction Model.

Objective: To explore the risk factors of anastomotic leakage (AL) after laparoscopic anterior resection (AR) of rectal cancer and establish a nomogram prediction model. Methods: Clinical and surgical data of patients who underwent AR of rectal cancer at Sichuan Cancer Hospital from January 2017 to December 2020 were retrospectively collected. Univariate and multivariate logistic regression analyses were used to screen the independent risk factors of AL after AR. A nomogram risk prediction model was established based on the selected independent risk factors and the predictive performance of nomogram was evaluated. Results: A 1013 patients undergoing laparoscopic AR were included, of which 67 had AL, with an incidence of 6.6%. Univariate and multivariate logistic regression analyses showed that male gender, tumors distance from the anus verge of ≤ 5cm, tumors distance from the anus verge of 5-10cm, circumferential tumor growth, operation time ≥ 240min, and no diverting stoma were independent risk factors for AL after AR. A nomogram prediction model was established based on these results. The calibration curve showed that the predicted occurrence probability of the nomogram model was in good agreement with the actual occurrence probability. The area under the receiver operating characteristic (ROC) curve was 0.749. Conclusion: The nomogram prediction model based on the independent risk factors of patients undergoing AL after AR can effectively predict the probability of AL.

M05B5.4 (lysosomal phospholipase A2) promotes disintegration of autophagic vesicles to maintain C. elegans development.

The autophagosome has two lipid bilayer membranes. The outer membrane fuses with the lysosome, while the inner membrane is degraded to release autophagic contents for degradation. It remains unclear how the inner vesicle of the autophagosome (called the autophagic vesicle) is disintegrated after autophagosome-lysosome fusion. Here, we identified C. elegans LPLA-2/M05B5.4 as a key enzyme that degrades membranous material in lysosomes. LPLA-2 is homologous to human PLA2G15, a lysosomal phospholipase A2 family protein that catalyzes cleavage of membrane phospholipids. We found that loss of LPLA-2 causes accumulation of large membrane whor ls in enlarged lysosomes and both phenotypes are suppressed by blocking macroautophagy/autophagy. Moreover, autophagic vesicles persisted in enlarged lysosomes in PLA2G15 knockdown cells and lpla-2(lf) mutants, which suggests that the breakdown of the inner autophagosomal membrane in lysosomes is impaired. lpla-2(lf) mutants exhibit severe defects in both embryonic and larval development. Our data suggest that disintegration of the inner autophagosomal membrane by LPLA-2 promotes the release and subsequent degradation of autophagic contents in lysosomes, which is essential for C. elegans development.Abbreviations: ATG: autophagy related; EPG: ectopic PGL granules; GFP: green fluorescent protein; LGG-1: LC3, GABARAP and GATE-16 family; LPLA-2: lysosomal phospholipase A2; PGL: P granule abnormality protein; PLA2: phospholipase A2; SD: standard deviation; SEPA-1: suppressor of ectopic P granules in autophagy mutant; SQST-1: sequestosome related.

Development of an Artificial Intelligence Model for Analyzing the Relationship between Imaging Features and Glucocorticoid Sensitivity in Idiopathic Interstitial Pneumonia.

High-resolution CT (HRCT) imaging features of idiopathic interstitial pneumonia (IIP) patients are related to glucocorticoid sensitivity. This study aimed to develop an artificial intelligence model to assess glucocorticoid efficacy according to the HRCT imaging features of IIP. The medical records and chest HRCT images of 150 patients with IIP were analyzed retrospectively. The U-net framework was used to create a model for recognizing different imaging features, including ground glass opacities, reticulations, honeycombing, and consolidations. Then, the area ratio of those imaging features was calculated automatically. Forty-five patients were treated with glucocorticoids, and according to the drug efficacy, they were divided into a glucocorticoid-sensitive group and a glucocorticoid-insensitive group. Models assessing the correlation between imaging features and glucocorticoid sensitivity were established using the k-nearest neighbor (KNN) algorithm. The total accuracy (ACC) and mean intersection over union (mIoU) of the U-net model were 0.9755 and 0.4296, respectively. Out of the 45 patients treated with glucocorticoids, 34 and 11 were placed in the glucocorticoid-sensitive and glucocorticoid-insensitive groups, respectively. The KNN-based model had an accuracy of 0.82. An artificial intelligence model was successfully developed for recognizing different imaging features of IIP and a preliminary model for assessing the correlation between imaging features and glucocorticoid sensitivity in IIP patients was established.

Filamin FLN-2 promotes MVB biogenesis by mediating vesicle docking on the actin cytoskeleton.

Multivesicular bodies (MVBs) contain intralumenal vesicles that are delivered to lysosomes for degradation or released extracellularly for intercellular signaling. Here, we identified Caenorhabditis elegans filamin FLN-2 as a novel regulator of MVB biogenesis. FLN-2 co-localizes with V-ATPase subunits on MVBs, and the loss of FLN-2 affects MVB biogenesis, reducing the number of MVBs in C. elegans hypodermis. FLN-2 associates with actin filaments and is required for F-actin organization. Like fln-2(lf) mutation, inactivation of the V0 or V1 sector of V-ATPase or inhibition of actin polymerization impairs MVB biogenesis. Super-resolution imaging shows that FLN-2 docks V-ATPase-decorated MVBs onto actin filaments. FLN-2 interacts via its calponin-homology domains with F-actin and the V1-E subunit, VHA-8. Our data suggest that FLN-2 mediates the docking of MVBs on the actin cytoskeleton, which is required for MVB biogenesis.

Hawthorn Juice Simulation System for Pectin and Polyphenol Adsorption Behavior: Kinetic Modeling Properties and Identification of the Interaction Mechanism.

The interaction between polyphenols and polysaccharides plays an important role in increasing the turbidity stability of fruit juice and improving unpleasant sensory experiences. The binding adsorption behavior between hawthorn pectin (HP) and polyphenols (epicatechin and chlorogenic acid) accorded with the monolayer adsorption behavior driven by chemical action and were better fitted by pseudo-second order dynamic equation and Langmuir model. The HP binding sites (Qm) and adsorption capacity (Qe) to epicatechin were estimated at 75.188 and 293.627 µg/mg HP, respectively, which was about nine and twelve times higher than that of chlorogenic acid. The interaction between HP and polyphenols exhibited higher turbidity characteristics, particle size and lower zeta potential than epicatechin and chlorogenic acid alone. Meanwhile, according to Fourier Transform Infrared Spectroscopy (FT-IR) analysis, it could be speculated that the interaction between HP and polyphenols resulted in chemical combination. Moreover, ΔH < 0 and TΔS < 0, which indicated that the interaction between HP and polyphenols was mainly driven by hydrogen bonds and van der Waals forces.