ObjectiveTo investigate the effects of Huangqi Jianzhongtang （HQJZ） on macrophage polarization and fat consumption in cancer cachexia （CC） mice. MethodsUltra-performance liquid chromatography-quadrupole/electrostatic field Orbitrap high-resolution mass spectrometry （UPLC-Q-Orbitrap HRMS） was used to control the quality of HQJZ. （1） In vitro experiment： HQJZ-containing serum was prepared， and the optimal concentration was determined by cytotoxicity assay. Mouse monocyte-derived macrophages （RAW264.7） were cultured and randomly divided into six groups， including a blank group， a classically activated macrophages （M1） group， an alternatively activated macrophages （M2） group， a HQJZ + blank group， a HQJZ+M1 group， and a HQJZ + M2 group. The relative expression of macrophage marker genes CD86， inducible nitric oxide synthase （iNOS）， CD206， and arginase-1 （Arg1） was detected by real-time quantitative polymerase chain reaction （Real-time PCR ）. （2） In vivo experiment： Thirty-two BALB/c mice were randomly divided into a control group， a model group， a medroxyprogesterone acetate （MPA） group， and a HQJZ group. Except for the control group， the other mice were injected with CT-26 colon cancer cells to establish a CC model. Mice in the MPA and HQJZ groups were given MPA （0.13 g·kg^-1·d^-1） or HQJZ （13.13 g·kg^-1·d^-1） by gavage， respectively， while mice in the control and model groups were given an equal volume of saline by gavage， with interventions continued for 10 d. Real-time PCR was used to detect the expression of macrophage markers （iNOS， Arg1， CD86， CD206） and fat browning-related genes uncoupling protein 1 （UCP1） and peroxisome proliferator-activated receptor γ （PPARγ） in epididymal adipose tissue. Western blot （WB） was used to detect protein expression levels of UCP1 and PPARγ. Micro-computed tomography （micro-CT） was used to measure residual fat volume， and hematoxylin-eosin （HE） staining was used to assess fat browning and calculate pathological scores. ResultsIn vitro， the dominant effective concentration of HQJZ-containing serum was 12.5%. Real-time PCR results showed that， compared with the blank group， Arg1 expression decreased in the HQJZ+blank group （P<0.05）， CD206 showed a downward trend without statistical significance， while iNOS and CD86 expression were significantly increased （P<0.05）. Compared with the M1 group， Arg1 and CD206 expression decreased in the HQJZ+M1 group （P<0.05）. Compared with the M2 group， CD206 expression decreased in the HQJZ+M2 group （P<0.05）， CD86 expression increased significantly （P<0.01）. In vivo， Real-time PCR results showed that， compared with the control group， CD86 and CD206 expression levels were significantly increased in the model group （P<0.01）. Compared with the model group， CD206 expression in the MPA group was significantly decreased （P<0.01）. In the HQJZ group， CD206 was significantly decreased （P<0.01）. WB results showed that， compared with the model group， protein expression of UCP1 and PPARγ was significantly reduced in the HQJZ group （P<0.05， P<0.01）. micro-CT results showed that the total white fat volume in the HQJZ group was greater than that in the model group （P<0.05）. HE staining results showed that pathological scores in the HQJZ group were lower than those in the model group （P<0.05）. ConclusionHQJZ may inhibit white adipose tissue browning by promoting macrophage M1 polarization and suppressing M2 polarization， thereby delaying fat consumption in CC mice.

Objective To specifically extract and analyze nascent proteins synthesized by bone marrow mesenchymal stem cells(BMSCs)after transplantation into ischemic hearts using a technique employing mutant methionyl-tRNA synthetase(MetRSL247G)for nascent protein labeling,in order to explore the potential mechanisms of action in BMSCs post-transplantation.Methods Point mutation at position 274 of the MetRS gene in BMSCs was induced via lentiviral infection to enable azidonorleucine(ANL)-mediated labeling of nascent proteins in BMSCs.The labeling efficiency was verified by means of fluorescent non-canonical amino-acid tagging(FUNCAT).Thirty healthy female C57BL/6J mice(8-10 weeks old)were divided into control and experimental groups,with 15 mice in each group.The acute myocardial infarction model was constructed by ligating the left anterior descending coronary artery in experimental group,while control mice underwent only thoracotomy without coronary ligation.After modeling,both groups received intramyocardial injections of MetRSL247G-modified BMSCs(MetRSL247G-BMSCs)at 3 different sites in the peri-infarct ischemic region.Mice were intraperitoneally injected with ANL every 6 hours for 4 times on postoperative days 0,2,and 6(n=5 for each time point)respectively,euthanized 24 h after the last injection,and cardiac tissues were isolated.The newly synthesized and labeled proteins produced by BMSCs after transplantation into the myocardium of experimental and control groups were collected,using an enrichment technique for ANL-tagged proteins and liquid chromatography-tandem mass spectrometry(LC-MS)analysis.Gene ontology(GO)analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis,protein-protein interaction(PPI)analysis,and heatmap visualization analysis were performed to identify differentially expressed proteins at the 3 time points and screen key pathways and genes.Results Under fluorescence microscopy,the MetRSL247G lentivirus-infected BMSCs were observed to be labelled with mCherry signals,confirming the successful construction of the MetRSL247G-BMSCs cell line.Green fluorescent signals were detected only in nascent proteins in culture medium containing both MetRSL247G-BMSCs and ANL,validating the sensitivity and specificity of the labeling method.GO analysis revealed that differentially expressed proteins were primarily involved in basic cellular biological processes such as extracellular exosome formation,extracellular matrix organization,and focal adhesion.KEGG and PPI analyses indicated that the differential proteins were mainly involved in complement and coagulation cascade pathway,actin cytoskeleton regulation pathway,and apoptosis pathway.Heatmap analysis showed significantly upregulated expression of anti-apoptosis and cell adhesion-related factors in experimental group on day 1(P＜0.05),upregulated anti-apoptotic factors,pro-apoptotic factors,and cell adhesion-related factors on day 3(P＜0.05),and upregulated anti-apoptotic factors,cell differentiation-related factors,and cell adhesion-related factors on day 7(P＜0.05)compared with control group.Expression of apoptosis-inducing factor 1 was significantly downregulated on days 1 and 7(P＜0.05).On day 3,most differentially expressed proteins,including anti-apoptosis factors(Protein S100-A11,Clusterin,Gelsolin),pro-apoptosis factor(Cathepsin B),cell differentiation-related factor(Transgelin-2),and cell adhesion-related factors(Cofilin-1,Periostin,Fibronectin)were significantly upregulated(P＜0.05).Conclusions The MetRSL247G mutation enables BMSCs to incorporate ANL and synthesize labeled proteins,confirming the feasibility of this nascent protein labeling technique.Nascent proteins of BMSCs in ischemic myocardium primarily contribute to extracellular exosome secretion and extracellular matrix organization.BMSCs may adapt to and respond to ischemic and hypoxic environments by influencing complement and coagulation cascades,activating inflammatory factors,regulating actin cytoskeleton structure,and modulating apoptosis,thereby maintaining the survival of BMSCs.

The efficacy mechanism of the alcoholic extract of Gnaphalium affine was investigated by observing its influence on oxidative stress and intestinal flora in rats modeled for chronic obstructive pulmonary disease(COPD). UPLC-MS was used to evaluate the quality of the alcoholic extract of G. affine, and 72 rats were randomly divided into six groups, with COPD models established in five groups by cigarette smoke combined with airway drip lipopolysaccharide, and the rats were given the positive drug of Danlong Oral Solution, as well as low-, medium-, and high-doses alcoholic extract of G. affine, respectively. After two weeks of continuous gastric gavage, the body weights and general morphology observations were performed; HE staining and Masson staining were used to verify the effects of the alcoholic extract of G. affine on alveolar inflammation and collagen deposition area in COPD rats; the oxidative stress indexes CAT and GSH in serum and SOD and MDA in lung tissue of the rats were measured, and the mRNA expression of HO-1, Nrf2, and NQO1 were determined by qRT-PCR. The protein expressions of HO-1, Nrf2, and NQO1 were determined by the Western blot method, and the mechanism by which the alcoholic extract of G. affine affected oxidative stress in COPD rats was explored. Finally, the influence of G. affine on the changes in intestinal flora caused by COPD was studied by 16S rRNA sequencing. The results showed that a total of 121 chemical components were identified by UPLC-MS, including 70 positive and 51 negative ion modes. In animal experiments, it was found that the alcoholic extracts of G. affine were able to reduce the percentage of collagen deposition, affect the oxidative stress indexes such as CAT, GSH, SOD, MDA, as well as the mRNA and protein expression of Nrf2, HO-1, and NQO1. The 16S rRNA sequencing results showed an increase in the level of Lactobacillales and a decrease in the level of Desulfovibrio and Desulfovibrionales, suggesting that the alcoholic extracts of G. affine could reverse the changes in intestinal flora caused by COPD. In conclusion, the alcoholic extracts of G. affine may exert anti-COPD effects by affecting the oxidative stress pathway and modulating the changes in intestinal flora.
Animals ; Oxidative Stress/drug effects* ; Pulmonary Disease, Chronic Obstructive/genetics* ; Rats ; Male ; Gastrointestinal Microbiome/drug effects* ; Rats, Sprague-Dawley ; Drugs, Chinese Herbal/administration & dosage* ; NF-E2-Related Factor 2/metabolism* ; Humans ; Lung/metabolism*

OBJECTIVE: To investigate the clinical efficacy of moxibustion (Mox) combined with low-dose tadalafil (TAD) in the treatment of diabetes mellitus-induced erectile dysfunction (DMED) with the syndrome of Qi deficiency and blood stasis. METHODS: According to the inclusion and exclusion criteria, we selected 90 patients with DMED for this trial and equally randomized them into a Mox, a TAD, and a Mox combined with TAD (Mox+TAD) group to be treated by mild Mox applied to the acupoints Zusanli, Sanyinjiao and Yinlingquan qd alt, oral medication with low-dose TAD at 5 mg per dose qd, and combination of the above two therapies, respectively, all for 4 weeks. We obtained from the patients their IIEF-5 scores, traditional Chinese medicine (TCM) symptoms scores, Erectile Hardness Scale (EHS) scores, corpus cavernosal hemodynamic indexes, and the peak systolic velocity (PSV), end diastolic velocity (EDV) and resistance index (RI) of the corpus cavernosal arteries before and after treatment, and compared them among the three groups. RESULTS: The total effectiveness rate was significantly higher in the Mox+TAD (90.0%) than in the Mox (46.7%) and TAD groups (60.0%) (P< 0.05). Compared with the baseline, the IIEF-5 and EHS scores were increased, while the TCM symptoms scores decreased in all the three groups after treatment, more significantly in the Mox+TAD group than in the other two (P< 0.05). And the PSV and RI were remarkably increased, while the EDV decreased (P< 0.05) in all the three groups (P< 0.05) after treatment, with PSV even higher in the Mox+TAD than in the Mox and TAD groups (P< 0.05). CONCLUSION Moxibustion combined with tadalafil has a definite efficacy and safety for the treatment of DMED, which can effectively improve the erectile function of the patients by increasing penile blood supply, benefiting qi and activating blood circulation.
Humans ; Male ; Tadalafil ; Erectile Dysfunction/etiology* ; Moxibustion ; Middle Aged ; Prospective Studies ; Adult ; Carbolines/administration & dosage* ; Diabetes Complications/therapy* ; Aged ; Treatment Outcome ; Combined Modality Therapy

This consensus will introduce the characteristics of fillers used in the surgical cavities of domestic nasal surgery patients based on relevant literature and expert opinions. It will also provide recommendations for the selection of cavity fillers for different nasal diseases, with chronic sinusitis as a representative example.
Humans ; Nasal Cavity/surgery* ; Nasal Surgical Procedures ; China ; Consensus ; Sinusitis/surgery* ; Dermal Fillers

Rapid and ultrasensitive detection of pathogen-associated biomarkers is vital for the early diagnosis and therapy of bacterial infections. Herein, we developed a close-packed and ordered Au@AgPt array coupled with a cascade triggering strategy for surface-enhanced Raman scattering (SERS) and colorimetric identification of the Staphylococcus aureus biomarker micrococcal nuclease (MNase) in serum samples. The trimetallic Au@AgPt nanozymes can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) molecules to SERS-enhanced oxidized TMB (oxTMB), accompanied by the color change from colorless to blue. In the presence of S. aureus, the secreted MNase preferentially cut the nucleobase AT-rich regions of DNA sequences on magnetic beads (MBs) to release alkaline phosphatase (ALP), which subsequently mediated the oxTMB reduction for inducing the colorimetric/SERS signal fade away. Using this "on-to-off" triggering strategy, the target S. aureus can be recorded in a wide linear range with a limit of detection of 38 CFU/mL in the colorimetric mode and 6 CFU/mL in the SERS mode. Meanwhile, the MNase-mediated strategy characterized by high specificity and sensitivity successfully discriminated between patients with sepsis (n = 7) and healthy participants (n = 3), as well as monitored the prognostic progression of the disease (n = 2). Overall, benefiting from highly active and dense "hot spot" substrate, MNase-mediated cascade response strategy, and colorimetric/SERS dual-signal output, this methodology will offer a promising avenue for the early diagnosis of S. aureus infection.

Objective The evaluation of lymph node properties before lung cancer surgery has a great impact with the choice of surgical methods.Although there are various examination methods,many methods have invasive or accuracy problems.In order to improve the accuracy of diagnosis,we mainly discuss the value of wide-body spectral CT in the differential diagnosis of mediastinal metastatic lymph nodes,non-metastatic lymph nodes in lung cancer patients and reactive hyperplastic lymph nodes.Methods The clinical and imaging data of 64 patients with lung cancer and 28 patients with pulmonary inflammatory lesions were retrospectively analyzed.All patients underwent plain scan and enhanced dual-phase spectral CT scan.The size,density,three-phase IC,NIC,and λHU of lymph nodes in metastatic,non-metastatic and inflammatory reactive hyperplasia groups were measured on 70 keV single-energy images and iodine-based images,respectively.The single-factor variance and Kruskal-Wallis H rank sum test were used to analyze and compare the differences.Results The short diameter of metastatic lymph nodes was larger than that of non-metastatic lymph nodes and reactive hyperplastic lymph nodes(P<0.001).The plain scan density of reactive hyperplastic lymph nodes was higher than that of metastatic lymph nodes(P<0.001),but there was no significant difference between non-metastatic lymph nodes(P=0.325).The CT values of reactive hyperplastic lymph nodes in arterial phase and venous phase were higher than those of metastatic and non-metastatic lymph nodes(P<0.05).Except for NIC in arterial phase,IC,NIC and λHU in plain scan,IC and λHU in arterial phase,IC,NIC and λHU in venous phase of reactive hyperplastic lymph nodes and metastatic lymph nodes were statistically significant(all P<0.05).There was no significant difference in IC,NIC and λHU between reactive hyperplastic lymph nodes and non-metastatic lymph nodes in plain scan,arterial phase and venous phase(all P>0.05).Conclusion The quantitative and spectral curve slope of iodine in mediastinal metastatic lymph nodes of lung cancer were basically lower than those in reactive hyperplastic lymph nodes.The quantitative parameters of spectral CT had certain diagnostic efficacy in differentiating metastatic lymph nodes and reactive hyperplastic lymph nodes,while the spectral parameters of non-metastatic lymph nodes and reactive hyperplastic lymph nodes were not statistically significant.

Objective To develop deep learning-based auxiliary diagnostic models for diverse pulmonary diffuse cystic diseases,and subsequently evaluate their classification performance to identify the optimal model for clinical diagnosis.Methods A total of 288 patients diagnosed with idiopathic pulmonary fibrosis(IPF),pulmonary lymphangioleiomyomatosis(PLAM),and pulmonary Langerhans cell histiocytosis(PLCH)were prospectively enrolled from the First Affiliated Hospital of Guangzhou Medical University between January 2010 and October 2022,comprising 76 cases of IPF,179 cases of PLAM,and 33 cases of PLCH.A total of 877 CT cases were collected,comprising 232 cases of IPF,557 cases of PLAM,and 88 cases of pulmonary PLCH.Based on the cutoff date of December 31,2019,the CT scans were divided into two datasets:dataset A consisted of 500 CT scans including 185 IPF cases,265 PLAM cases,and 50 PLCH cases;while dataset B comprised 377 CT scans with a distribution of 47 IPFcases,292 PLAMcases,and 38 PLCH cases.The Dataset A was randomly partitioned into training set,validation set,and test set in a ratio of 7∶1∶2.Subsequently,six distinct deep learning neural networks were employed for training after preprocessing and data augmentation.Receiver operating characteristic curves were generated to assess the model performance using metrics such as area under the curve(AUC),accuracy,sensitivity,specificity,and F1 score in order to identify the optimal model.Furthermore,a test set B comprising 30 randomly selected cases from dataset B for each disease type was utilized to evaluate the trained optimal model by employing the same aforementioned metrics.Results In test A,six well-established diagnostic models demonstrated superior classification performance for IPF and LAM,with an AUC greater than 0.9.For LCH,EfficientNet exhibited low classification efficiency with an AUC between 0.6 and 0.7,while Vgg11 showed an AUC between 0.8 and 0.9;the other four models displayed excellent classification efficiency with an AUC greater than 0.9.Except for Inception V3,the remaining five diagnostic models performed poorly in identifying and classifying LCH lesions.Considering multiple indicators,the InceptionV3 model showcased optimal comprehensive performance among the six models,achieving high evaluation parameters such as overall accuracy(94.90%),precision(93.49%),recall(90.84%),and specificity(96.91%).TestB was conducted using the trained InceptionV3 model resulting in an accuracy of 81%,precision of 82%,recall of 81%,and specificity of 90%.Conclusions Six recognition and classification models,developed using deep learning technology in conjunction with pulmonary CT images,demonstrate effective discrimination between LAM,LCH,and IPF.Notably,the model constructed utilizing the InceptionV3 neural network exhibits superior efficiency in accurately recognizing and classifying IPF and LAM.

Objective To investigate the related factors of early neurological deterioration in elderly patients with thrombolysis for acute occlusive stroke of large intracranial vessels. Methods Clinical materials of 480 elderly patients with thrombolysis for acute occlusive stroke of large intracranial vessels were retrospectively analyzed. The National Institutes of Health Stroke Scale (NIHSS) was used to evaluate the neurological function of patients within 72 hours after thrombectomy, and the patients were divided into deterioration group and non-deterioration group. The baseline materials were compared between the two groups, the factors of early neurological deterioration were analyzed, and the receiver operating characteristic (ROC) curve was drawn to analyze the predictive value of risk. Results Among 480 elderly patients with thrombolysis for acute occlusive stroke of large intracranial vessels, 92 cases (19.17%) had early neurological deterioration; the blood glucose level, white blood cell count, incidence of dysphagia, and incidence of coma in the deterioration group were significantly higher than those in the non-deterioration group (P < 0.05); the Logistic regression analysis showed that high level of blood glucose (95%CI, 1.203 to 1.478), high level of white blood cell count (95%CI, 1.159 to 1.408), dysphagia (95%CI, 7.049 to 31.475), and coma (95%CI, 8.375 to 43.195) were risk factors for early neurological deterioration in elderly patients with thrombolysis for acute occlusive stroke of large intracranial vessels (OR>1, P < 0.05). ROC curve results showed that blood glucose, white blood cell count, dysphagia, and coma had certain predictive value for early neurological deterioration in elderly patients with thrombolysis for acute occlusive stroke of large intracranial vessels[the area under the curve (AUC) was 0.733, 0.708, 0.707 and 0.701]; the ROC curve showed AUC was 0.934, indicating that the model was reliable and stable. Conclusion Early neurological deterioration in elderly patients with thrombolysis for acute occlusive stroke of large intracranial vessels may be related to factors such as blood glucose, white blood cell count, dysphagia, and coma. Early risk assessment and relevant intervention measures can be taken clinically to improve prognosis.

Objective:To systematically evaluate the correlation between chronic non-occupational arsenic exposure and hypertension.Methods:A literature search was conducted through Web of Science, Pubmed, Embase, Cochrane Library, WanFang Data, China National Knowledge Infrastructure (CNKI), VIP Chinese Journal Service Platform (VIP) Database and China Biomedical Literature Database to comprehensively collect epidemiological literature related to chronic non-occupational arsenic exposure and hypertension published domestically and internationally for inclusion in the study, with a time limit from database establishment to January 1, 2023. Meta-analysis of dichotomous variables was conducted using Stata MP15 software, with odds ratio ( OR) value [95%confidence interval( CI)] as the effect evaluation indicator. A random-effects model or a fixed-effects model was selected for comprehensive quantitative analysis according to the heterogeneity results; the sources of heterogeneity were identified through subgroup analysis; a funnel plot was used for qualitative analysis of publication bias and the results were further assessed by Egger test. Stata 15.0 software was then used to analyze the dose-response relationship between chronic non-occupational arsenic exposure and hypertension using restricted cubic spline function and generalized least squares estimation (GLST) method. Results:Twenty-nine articles ( n = 127 258) were finally included, including 24 English articles and 5 Chinese articles. Through Meta-analysis, the combined OR value (95% CI) for hypertension was 1.07 (1.04 - 1.09), with a statistically significant difference ( P < 0.05). The combined OR values (95% CI) for urinary arsenic, drinking water arsenic, and hair arsenic in subgroup analysis were 1.10 (1.04 - 1.17), 1.13 (1.07 - 1.20), and 2.55 (1.55 - 4.20), respectively. The combined OR values (95% CI) for cross-sectional studies, case-control studies and cohort studies were 1.11 (1.06 - 1.16), 1.13 (1.04 - 1.23) and 1.04 (1.00 - 1.07), respectively. For every unit (μg/L) increase in arsenic exposure in drinking water, the risk of hypertension increased by 0.13% [ OR value (95% CI): 1.001 269 (1.000 104 - 1.002 434), P < 0.05]. Conclusions:There is a correlation between chronic non-occupational arsenic exposure and adult hypertension, with urinary arsenic, drinking water arsenic and hair arsenic as possible exposure markers. There is a non-linear dose-response relationship between chronic non-occupational arsenic exposure and adult hypertension.