In order to explore the possible role and molecular mechanism of the combined action of leech and bear bile in liver and gallbladder diseases, this study first used network pharmacology methods to screen the components and targets of leech and bear bile, as well as the related target genes of liver and gallbladder diseases. The selected key genes were subjected to interaction network and GO/KEGG enrichment analysis. Then, using sodium oleate induced HepG2 cell lipid deposition model and DL-ethionine induced mouse fatty liver model, the activity of leech and bear bile alone and in combination in reducing liver fat was evaluated in vitro and in vivo, and the expression of key pathway related proteins suggested by network pharmacology was detected by Western blot. The results of network pharmacology analysis showed that the active ingredients of leech and bear bile have 295 intersecting targets with liver and gallbladder related diseases, involving more than 200 signaling pathways, including the PI3K/Akt signaling pathway and phospholipase D signaling pathway closely related to glucose and lipid metabolism. The results of in vitro validation experiments showed that both leech and bear bile, alone and in combination, can significantly inhibit the lipid deposition induced by sodium oleate in human liver cells, reduce the triacylglycerol level in cell culture supernatant, and inhibit the lipid content in liver cells. The observation results of Nile red staining confocal microscopy showed that the combination of leech and bear bile had better activity in reducing lipid deposition in liver cells compared to using them alone. In a mouse fatty liver model, the combination of leech and bear bile can better reduce elevated organ indices, blood lipids, and liver lipid levels, as well as lower the levels of serum liver injury biomarkers. The animals used in this experiment and related disposal meet the requirements of animal welfare. Before the experiment, it was reviewed and approved by IACUC, Institute of Materia Medica, Chinese Academy of Medical Sciences. The Western blot experiment results showed that the combination of leech and bear bile can significantly upregulate the expression levels of p-PI3K and p-Akt proteins, and increase the p-PI3K/PI3K and p-Akt/Akt ratios, which is consistent with the predicted results of network pharmacology. The combination of leech and bear bile has great potential for treating fatty liver disease, and activating the PI3K/Akt pathway may be one of the important mechanisms for reducing lipid deposition in liver cells.

OBJECTIVE To evaluate the quality of evidence in the systematic evaluation/meta-analysis of traditional Chinese medicine （TCM） for diabetes retinopathy （DR） based on the GRADE system. METHODS Chinese and English databases were searched to obtain the relevant studies of systematic evaluation/meta-analysis of traditional Chinese medicine in the treatment of DR. The search time was from the establishment of each database to January 13th， 2024. According to the inclusion and exclusion criteria， literature screening was conducted. After extracting relevant information from the included literature， the GRADE system was used to evaluate the quality level of the evidence body in the included studies， and the evidence of the outcome indicators was integrated and summarized. RESULTS A total of 51 studies were ultimately included， encompassing 135 outcome indexes. Among these， 19 indicators （14.1%） were of high quality， 87 （64.4%） were of medium quality， 26 （19.3%） were of low quality， and 3 （2.2%） were of very low quality. Overall， the evidence quality of the outcome indicators in the included studies was medium to low quality. The integrated results of evidence on the efficacy of outcome indexes showed that compared with conventional Western medicine， calcium dobesilate or placebo， TCM had significant advantages in improving overall efficacy， reducing bleeding spot area， reducing macular foveal thickness， and increasing visual improvement rate. In addition，the combination of TCM and conventional Western medicine or calcium dobesilate was significantly more effective than using conventional Western medicine or calcium dobesilate alone. CONCLUSIONS The overall quality of the evidence in the systematic evaluation/meta-analysis study on the treatment of DR with TCM is medium to low quality. Based on existing research findings， TCM demonstrates good clinical efficacy in the treatment of DR.

ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum （LDP） on epithelial-mesenchymal transition （EMT） and the expression of cancer stem cell （CSC） properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells （MDSCs）. MethodsSPF-grade female SD rats were randomly divided into three groups， which were given 0.39， 1.94， 3.89 g·kg^-1·d^-1 suspension of Liuwei Dihuangwan for 7 days， respectively， to prepare low-， medium-， and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 （CCK-8） assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry （FC）. The migration， invasion， and proliferation of 4T1 cells were detected by cell scratch assay， Transwell invasion assay， and plate colony-forming assay， respectively. Western blot （WB） was used to detect the protein expression of transforming growth factor-β （TGF-β）， nuclear factor-κB （NF-κB）， C-X-C motif chemokine ligand 2 （CXCL2）， E-cadherin， and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence （IF）. ResultsCompared with the normal group， LDP showed no significant inhibitory effect on the cell viability of 4T1 cells， but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs， as well as the expression of TGF-β （P<0.05， P<0.01）. The migration， invasion， and proliferation of 4T1 cells were increased after co-culture with MDSCs （P<0.05， P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin and the proportion of CSC （CD44⁺CD24^-） were elevated （P<0.05， P<0.01）， while the expression of E-cadherin was decreased （P<0.05）. After the intervention of MDSCs with LDP， followed by co-culture with 4T1 cells， the migration， invasion， and proliferation of 4T1 cells were obviously reduced （P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin were decreased （P<0.05， P<0.01）， and the expression of E-cadherin was increased （P<0.05， P<0.01）. There was no statistical difference in the proportion of CSC （CD44⁺CD24^-） in 4T1 cells. However， the proportion of CSC （CD44⁺CD24^-） was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention （P<0.05， P<0.01）. ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β， NF-κB， and CXCL2， reverse EMT， and reduce the characteristic expression of CSC to inhibit the migration， invasion， and proliferation of 4T1 cells.

ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum （LDP） on epithelial-mesenchymal transition （EMT） and the expression of cancer stem cell （CSC） properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells （MDSCs）. MethodsSPF-grade female SD rats were randomly divided into three groups， which were given 0.39， 1.94， 3.89 g·kg^-1·d^-1 suspension of Liuwei Dihuangwan for 7 days， respectively， to prepare low-， medium-， and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 （CCK-8） assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry （FC）. The migration， invasion， and proliferation of 4T1 cells were detected by cell scratch assay， Transwell invasion assay， and plate colony-forming assay， respectively. Western blot （WB） was used to detect the protein expression of transforming growth factor-β （TGF-β）， nuclear factor-κB （NF-κB）， C-X-C motif chemokine ligand 2 （CXCL2）， E-cadherin， and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence （IF）. ResultsCompared with the normal group， LDP showed no significant inhibitory effect on the cell viability of 4T1 cells， but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs， as well as the expression of TGF-β （P<0.05， P<0.01）. The migration， invasion， and proliferation of 4T1 cells were increased after co-culture with MDSCs （P<0.05， P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin and the proportion of CSC （CD44⁺CD24^-） were elevated （P<0.05， P<0.01）， while the expression of E-cadherin was decreased （P<0.05）. After the intervention of MDSCs with LDP， followed by co-culture with 4T1 cells， the migration， invasion， and proliferation of 4T1 cells were obviously reduced （P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin were decreased （P<0.05， P<0.01）， and the expression of E-cadherin was increased （P<0.05， P<0.01）. There was no statistical difference in the proportion of CSC （CD44⁺CD24^-） in 4T1 cells. However， the proportion of CSC （CD44⁺CD24^-） was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention （P<0.05， P<0.01）. ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β， NF-κB， and CXCL2， reverse EMT， and reduce the characteristic expression of CSC to inhibit the migration， invasion， and proliferation of 4T1 cells.

ObjectiveThis study explores the methods of lung tissue extraction and fixation required for pathological studies of fetal rats, based on the unique physiological structure of fetal rat lung tissue and existing lung tissue fixation techniques for adult rats. MethodsSix pregnant adult SD rats at 20.5 days of gestation were subjected to cesarean section to obtain fetal rats. Four healthy fetal rats with similar body weight, vital signs, and respiratory status were selected from each pregnant rat, and they were randomly divided into the following groups using a random number table: direct lung infiltration group, lung infiltration group after intratracheal infusion, whole-body infiltration group of fetal rats, and whole-body infiltration group after intratracheal infusion of fetal rats. To systematically compare and analyze the anatomical morphology under different fixation methods, lung tissues from four groups of fetal rats were harvested, perfused, and fixed, and the gross morphology of lung tissues in each group was observed. Paraffin sections were prepared and stained with Hematoxylin-Eosin (H&E). The histological morphology of the whole lung, alveoli, and bronchi was further examined under optical microscopy. ResultsIn the direct lung infiltration group, the hilar structures were unclear, lung lobation was indistinct, the shape was irregular, lung cavities were small, and alveoli and bronchi were shrunken. In the lung infiltration group after intratracheal infusion, the hilar structures were clear, lobation was pronounced, the shape was regular, lung cavities were large, and alveoli and bronchi were full. Both the whole-body infiltration group and whole-body infiltration group after intratracheal infusion of fetal rats exhibited visible lungs, hearts, skins, and other organs. The lung tissues of both groups showed obvious lobulation, irregular shape, and damage at the margins of lung lobes. In the whole-body infiltration group, the thoracic cavities of the fetus were flattened, lung cavities were small, and alveoli and bronchi were shrunken. In the whole-body infiltration group after intratracheal infusion of fetal rats, the fetal thoracic cavities were full, lung cavities were large, and alveoli and bronchi were relatively full. ConclusionThe lung infiltration after intratracheal infusion method for fetal rat lung tissue fixation outperforms direct lung infiltration, whole-body infiltration of fetal rats, and whole-body infiltration after intratracheal infusion of fetal rats in terms of preservation of the lung tissue's original morphology, paraffin sectioning, staining, and pathological observation and analysis. The embedding, sectioning, and staining processes are also simple and save consumables. Therefore, intratracheal infusion followed by lung infiltration method is recommended for fixation in histopathological observation of fetal rat lung tissue.

Objective To explore the mediating role of occupational well-being in the relationship between professional identity and safety behavior among nurses. Methods A total of 1 006 nurses from ten tertiary general hospitals in eight provincial administrative regions were selected as the research subjects using convenient sampling method. Their safety behavior, professional identity and occupational well-being were investigated using Nurse Safety Behavior Scale, Nurse Professional Identity Scale and Occupational Well-being Scale. Structural equation modeling was performed using AMOS 26.0 to examine the mediating effect of occupational well-being in the relationship between professional identity and safety behavior among nurses. Results The scores for safety behavior, professional identity, and occupational well-being were (53.0±6.1), (123.7±21.2) and (90.8±13.1), respectively. Safety behavior was positively correlated with both professional identity and occupational well-being (correlation coefficients were 0.50 and 0.50, respectively, both P<0.01). Professional identity was positively correlated with occupational well-being (correlation coefficient was 0.51, P<0.01). The multiple linear regression analysis results showed that the higher the professional identity and occupational well-being of nurses, the higher the level of safety behavior (both P<0.05). The result of mediating effect shows that the total effect of occupational identity on safety behavior was 0.498 [95% confidence interval (CI) was 0.405-0.576], and occupational well-being played a mediating role between professional identity and safety behavior among nurses with the mediation effect of 0.156 (95%CI was 0.112-0.205), accounting for 31.33% of the total effect. Conclusion The safety behavior of nurses is at a moderate level. Both professional identity and occupational well-being can affect the safety behavior of nurses. Professional identity can increase the safety behavior of nurses by affecting occupational well-being.

Objective: To examine the casual association between age at first sexual intercourse and gynecologic malignant tumors using Mendelian randomization (MR) approach. Methods: The single nucleotide polymorphisms (SNPs) associated with age at first sexual intercourse were obtained from a meta-analysis of genome-wide association study (GWAS), and the SNPs related to gynecologic malignant tumors (ovarian cancer, endometrial cancer, cervical cancer), and their subtypes were sourced from the IEU OpenGWAS database. Using age at first sexual intercourse as the exposure and gynecologic malignant tumors as the outcome, a MR analysis was performed with the inverse-variance weighted (IVW) method. Heterogeneity was assessed using Cochran's Q test, horizontal pleiotropy was evaluated using MR-Egger regression and MR-PRESSO test, and bias was examined using funnel plots. Results: The Mendelian randomization analysis demonstrated that younger age at first sexual intercourse was significantly associated with an increased risk of low-grade serous ovarian carcinoma (OR=0.553, 95%CI: 0.335-0.911), cervical cancer (OR=0.674, 95%CI: 0.466-0.974), endometrial cancer (OR=0.854, 95%CI: 0.730-0.999), and endometrioid carcinoma (OR=0.830, 95%CI: 0.690-0.998). No statistical association was found between the age at first sexual intercourse and ovarian cancer, high-grade serous ovarian cancer, mucinous ovarian cancer, endometrioid ovarian cancer, or non-endometrioid ovarian cancer (all P>0.05). Sensitivity analysis showed no evidence of undetected instrumental variable heterogeneity or horizontal pleiotropy (all P>0.05), and the funnel plot indicated no presence of bias. Conclusion Younger age at first sexual intercourse may be associated with an increased risk of certain gynecologic malignant tumors, highlighting the need to strengthen adolescent sex education.

Objective Radiation-induced brain injury (RIBI) is a common complication of radiotherapy for the head and neck tumors, and the current treatment methods are limited. Repetitive transcranial magnetic stimulation (rTMS), as a non-invasive neural regulation technique, has shown great potential in neuroprotection. However, the parameter selection and biological safety of rTMS in the prevention and treatment of RIBI have not been reported. Methods Using a mouse model of RIBI, this study employed three rTMS frequencies (5, 10, and 25 Hz) for intervention. Biochemical and pathological assays were conducted to identify the optimal stimulation parameter. Subsequently, this parameter was used to evaluate the biological safety in normal mice. Results Under the conditions of this experiment, rTMS interventions with all three frequencies could reduce the levels of serum brain injury markers (NSE and S100B) and inflammatory factors in mice (P < 0.001), and alleviate the morphological and structural damage of hippocampal tissue. The 10 Hz rTMS could significantly promote hippocampal neurogenesis in RIBI mice (P < 0.05). Furthermore, 10 Hz rTMS showed no significant effects on the cognitive function and mood of normal mice. The intervention did not significantly change the morphology and structure of the main organs, blood biochemical indicators, and the level of hippocampal neurogenesis in mice. Conclusion The 10 Hz rTMS is optimal for the prevention and treatment of RIBI with high biological safety.

BACKGROUND: Whether adherence to a healthy lifestyle is associated with a lower risk of developing pneumonia and a better long-term prognosis remains unclear. This study aimed to investigate associations of individual and combined lifestyle factors (LFs) with the incidence risk and long-term prognosis of pneumonia hospitalization. METHODS: Using data from the China Kadoorie Biobank study, we used the multistate models to investigate the role of five high-risk LFs, including smoking, excessive alcohol drinking, unhealthy dietary habits, physical inactivity, and unhealthy body shape, alone or in combination in the transitions from a generally healthy state at baseline to pneumonia hospitalization or cardiovascular disease (CVD, regarded as a reference outcome), and subsequently to mortality. RESULTS: Most of the five high-risk LFs were associated with increased risks of transitions from baseline to pneumonia and from pneumonia to death, but with different risk estimates. The greater the number of high-risk LFs, the higher the risk of developing pneumonia and long-term mortality risk after pneumonia, with the strength of associations comparable to that of LFs and CVD. Compared to participants with 0-1 high-risk LF, the adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) for transitions from baseline to pneumonia and from pneumonia to death in those with five high-risk LFs were 1.43 (1.28-1.60) and 1.98 (1.61-2.42), respectively. Correspondingly, the respective HRs (95% CIs) for transitions from baseline to CVD and from CVD to death were 2.00 (1.89-2.11) and 1.44 (1.30-1.59), respectively. The risk estimates changed slightly when further adjusting for the presence of major chronic diseases. CONCLUSION In this Chinese population, unhealthy LFs were associated with an increased incidence and long-term mortality risk of pneumonia.
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Cardiovascular Diseases/etiology* ; China/epidemiology* ; Life Style ; Pneumonia/etiology* ; Prognosis ; Risk Factors ; Smoking

BACKGROUND: Severe stroke has high rates of mortality and morbidity. This study aimed to investigate the clinical course, causes of worsening, and outcomes of severe ischemic stroke. METHODS: This prospective, multicenter cohort study enrolled adult patients admitted ≤30 days after ischemic stroke from nine hospitals in China between September 2017 and December 2019. Severe stroke was defined as a score of ≥15 on the National Institutes of Health Stroke Scale (NIHSS). Clinical worsening was defined as an increase of 4 in the NIHSS score from baseline. Unfavorable functional outcome was defined as a modified Rankin scale score ≥3 at 3 months and 1 year after stroke onset, respectively. We performed Logistic regression to explore baseline features and reperfusion therapies associated with clinical worsening and functional outcomes. RESULTS: Among 4201 patients enrolled, 854 patients (20.33%) had severe stroke on admission. Of 3347 patients without severe stroke on admission, 142 (4.24%) patients developed severe stroke in hospital. Of 854 patients with severe stroke on admission, 33.95% (290/854) experienced clinical worsening (median time from stroke onset: 43 h, Q1-Q3: 20-88 h), with brain edema (54.83% [159/290]) as the leading cause; 24.59% (210/854) of these patients died by 30 days, and 81.47% (677/831) and 78.44% (633/807) had unfavorable functional outcomes at 3 months and 1 year respectively. Reperfusion reduced the risk of worsening (adjusted odds ratio [OR]: 0.24, 95% confidence interval [CI]: 0.12-0.49, P  <0.01), 30-day death (adjusted OR: 0.22, 95% CI: 0.11-0.41, P  <0.01), and unfavorable functional outcomes at 3 months (adjusted OR: 0.24, 95% CI: 0.08-0.68, P <0.01) and 1 year (adjusted OR: 0.17, 95% CI: 0.06-0.50, P  <0.01). CONCLUSIONS: Approximately one-fifth of patients with ischemic stroke had severe neurological deficits on admission. Clinical worsening mainly occurred in the first 3 to 4 days after stroke onset, with brain edema as the leading cause of worsening. Reperfusion reduced the risk of clinical worsening and improved functional outcomes. REGISTRATION ClinicalTrials.gov , NCT03222024.
Humans ; Male ; Female ; Prospective Studies ; Ischemic Stroke/mortality* ; Aged ; Middle Aged ; Aged, 80 and over ; Stroke ; Brain Ischemia