ObjectiveTo investigate the migration and distribution characteristics of chemical constituents in blood and hippocampal tissues before and after vinegar processing of Citri Reticulatae Pericarpium Viride（CRPV）， and to explore the potential material basis and mechanisms underlying their regulatory effects on emotional disorders by comparing the effects of raw and vinegar-processed products of CRPV. MethodsUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） was employed to characterize and identify the chemical constituents of raw and vinegar-processed products of CRPV extracts， as well as their migrating components in blood and hippocampal tissues after oral administration. Reference standards， databases， and relevant literature were utilized for compound annotation， with data processing performed using PeakView 1.2 software. Seventy male C57BL/6 mice were randomly divided into seven groups， including the blank group， model group， diazepam group（2.5 mg·kg^-1）， raw CRPV low/high dose groups（0.6， 1.2 g·kg^-1）， and vinegar-processed CRPV low/high dose groups（0.6， 1.2 g·kg^-1）， with 10 mice per group. Except for the blank group， all other groups underwent chronic restraint stress（2 h·d^-1） for 20 d. Each drug-treated group received oral administration at the predetermined dose starting 10 d after modeling， with a total treatment duration of 10 d. Following model-based drug administration， mice underwent open-field， forced swimming， and elevated plus maze tests. After anesthesia with isoflurane， whole brains were collected from each group of mice， and hippocampi were dissected. Reactive oxygen species（ROS） level in hippocampal tissues was quantified by enzyme-linked immunosorbent assay（ELISA）. Hematoxylin-eosin（HE） staining was used to observe hippocampal tissue morphology. Immunofluorescence was performed to detect neuronal nuclei（NeuN） and peroxisome proliferator-activated receptor alpha（PPARα） expressions in hippocampal tissue. Then， pharmacodynamic evaluations were conducted to assess the effects of raw and vinegar-processed CRPV on mood disorders， exploring the potential mechanisms. ResultsVinegar processing caused significant changes in the chemical composition of CRPV， with 18 components showing increased relative content and 35 components showing decreased relative content. The primary changes occurred in flavonoid compounds， including 20 flavonoids， 20 flavonoid glycosides， 3 triterpenes， 3 phenolic acids， 1 alkaloid， and 6 other compounds. Twenty-one components were detected in blood（15 methoxyflavones， 4 flavonoid glycosides， and 2 phenolic acids）， with 17 shared between raw and vinegar-processed CRPV. Seven components reached hippocampal tissues（all common to both forms）. In regulating emotional disorders， Vinegar-processed CRPV exhibited superior antidepressant-like effects compared to raw products. HE staining revealed that both treatments improved hippocampal neuronal morphology， particularly in the damaged CA1 and CA3 regions. Immunofluorescence and ELISA analyses demonstrated that both raw and vinegar-processed CRPV significantly modulated NeuN and PPARα expressions in hippocampal tissue while alleviating oxidative stress induced by excessive ROS（P<0.05）. ConclusionThe chemical composition of CRPV undergoes changes after vinegar processing， but the migrating components in blood and hippocampus are primarily methoxyflavonoids. These components may serve as the potential material basis for activating the PPARα pathway， thereby negatively regulating ROS generation in the hippocampus， reducing oxidative stress， and promoting the development of NeuN-positive neurons. These findings provide experimental evidence for enhancing quality standards， pharmacodynamic material research， and active drug development of raw and vinegar-processed CRPV.

OBJECTIVES: Esophageal squamous cell carcinoma (ESCC) is characterized by complex pathogenesis and poor prognosis. In recent years, epithelial-mesenchymal transition (EMT) in tumor initiation and progression has attracted increasing attention. Enhancer of zeste homolog 2 (EZH2), which is aberrantly expressed in various tumors, may be closely related to the EMT process. This study aims to examine the expression and correlation of EZH2 and EMT markers in ESCC cells and tissues, evaluate the effects of EZH2 knockdown on ESCC cell proliferation, invasion, and migration, and explore how EZH2 contributes to the malignant biological behavior of ESCC. METHODS: Bioinformatics analyses were used to assess EZH2 expression levels in ESCC. Small interfering RNA was used to knock down EZH2 in ESCC cell lines EC109 and EC9706. Cell proliferation, invasion, and migration were evaluated using cell counting kit-8 (CCK-8), wound healing, and Transwell assays. Protein and mRNA expression levels of EZH2, E-cadherin (E-cad), and vimentin (Vim) were detected by Western blotting and real time fluorogenic quantitative PCR (RT-qPCR), respectively. Immunohistochemical (IHC) staining was performed on 70 ESCC tissue samples and 40 paired adjacent normal tissues collected from the First Affiliated Hospital of Shihezi University between 2010 and 2016 to assess the expression of EZH2, E-cad, and Vim, and to analyze their associations with clinicopathological feature and patient prognosis. RESULTS: Bioinformatics analysis showed that EZH2 was highly expressed in ESCC (P<0.001), and high EZH2 expression was associated with worse prognosis (P<0.001). CCK-8, wound healing, and Transwell assays demonstrated that EZH2 knockdown significantly suppressed the proliferation, invasion, and migration of ESCC cells (P<0.001). In addition, Vim expression was significantly reduced, while E-cad expression was significantly increased at both protein and mRNA levels in EZH2-silenced cells (all P<0.05). IHC staining analysis revealed higher expression of EZH2 and Vim and lower expression of E-cad in ESCC tissues compared to adjacent normal tissues. Kaplan-Meier survival analysis showed that low expression of EZH2 and Vim and high expression of E-cad were associated with longer survival (all P<0.05). CONCLUSIONS EZH2 promotes malignant biological behavior in ESCC by mediating EMT. Elevated EZH2 expression is associated with poor prognosis in ESCC patients.
Humans ; Enhancer of Zeste Homolog 2 Protein/physiology* ; Esophageal Squamous Cell Carcinoma/pathology* ; Epithelial-Mesenchymal Transition/genetics* ; Esophageal Neoplasms/metabolism* ; Cell Proliferation ; Cell Line, Tumor ; Cell Movement ; Cadherins/genetics* ; Vimentin/genetics* ; Male ; Female ; Middle Aged ; Neoplasm Invasiveness ; Prognosis ; RNA, Small Interfering/genetics* ; Gene Expression Regulation, Neoplastic

The anterior cingulate cortex (ACC) has recently been proposed as a key player in the representation of itch stimuli. However, to date, little is known about the contribution of specific ACC interneuron populations to itch processing. Using c-Fos immunolabeling and in vivo Ca²⁺ imaging, we reported that both histamine and chloroquine stimuli-induced acute itch caused a marked enhancement of vasoactive intestinal peptide (VIP)-expressing interneuron activity in the ACC. Behavioral data indicated that optogenetic and chemogenetic activation of these neurons reduced scratching responses related to histaminergic and non-histaminergic acute itch. Similar neural activity and modulatory role of these neurons were seen in mice with chronic itch induced by contact dermatitis. Together, this study highlights the importance of ACC VIP⁺ neurons in modulating itch-related affect and behavior, which may help us to develop novel mechanism-based strategies to treat refractory chronic itch in the clinic.
Animals ; Pruritus/physiopathology* ; Vasoactive Intestinal Peptide/metabolism* ; Interneurons/metabolism* ; Gyrus Cinguli/metabolism* ; Mice ; Male ; Mice, Inbred C57BL ; Histamine ; Chloroquine ; Optogenetics ; Mice, Transgenic

Objective:To explore the competitive attitude of students of medical college and its influencing factors.Methods:A sample of 382 first to third year medical students was selected with the method of random sampling. The questionnaire consists of three parts: demographics, Competitive Attitude Scale of Chinese Version and Jefferson Scale of Physician Empathy. SPSS 25.0 was applied to perform the statistical analysis. Concrete methods that used involved statistical interpretation, Kruskal-Wallis test, Mann-Whitney U test and Spearman rank sum correlation analysis. Results:The benign competitive attitude of students was 51.00(45.00，56.00) and the excessive competitive attitude was 35.00(31.00，38.00). There was a statistically significant difference in the score of benign competitive attitude among medical students of different grades( H=22.31, P<0.001). There were statistically significant differences in the scores of benign competitive attitudes among the medical students who enjoy teamwork ( Z=-4.53, P<0.001), who had role models to follow in the medical field ( Z=-3.99, P<0.001), and who had careful career planning ( Z=-2.19, P=0.029). There were statistically significant differences in the scores of excessive competitive attitudes among the medical students who enjoy teamwork ( Z=-2.47, P=0.014), who had role models to follow in the medical field ( Z=-2.89, P=0.004), and who had careful career planning ( Z=-2.32, P=0.021). There was a weak positive correlation between empathy and benign competitive attitude ( r=0.20, P<0.001), but a negative correlation between empathy and excessive attitude ( r=-0.36, P<0.001). Conclusions:Medical students had a good competitive attitude. Medical college should pay more attention to improving medical students' sense of teamwork, career planning ability and willingness to learn from role models, guide students to develop a benign competitive attitude and focus on improving their professional quality.

Objective:To investigate the epidemiological and clinical characteristics of RSV among patients aged ≥60 years in Beijing from 2015 to 2023.Methods:Based on the respiratory pathogen surveillance system, samples of upper respiratory tract infections (URTI), non-severe community-acquired pneumonia (nsCAP) and severe community-acquired pneumonia (sCAP) among people aged ≥60 years were collected from 28 sentinel hospitals in 16 districts of Beijing from January 2015 to December 2023. Swab samples were collected from URTI within one week, and lower respiratory tract samples from nsCAP and sCAP were collected. Demographic and epidemiological data were also collected. Various respiratory pathogens including RSV were detected.Results:From January 2015 to December 2023, a total of 20 349 cases of acute respiratory infections aged ≥60 years were included, with the RSV-positive rate of 1.54% (313/20 349, 95% CI: 1.39%-1.68%). Among them, the total RSV-positive rates of older people during the pre-pandemic, pandemic, and post-pandemic periods of COVID-19 were 1.59% (207/13 006, 95% CI: 1.38%-1.81%), 0.82% (38/4 650, 95% CI: 0.56%-1.08%) and 2.53% (68/2 693, 95% CI: 1.93%-3.12%), respectively. The difference in RSV-positive rate was statistically significant ( P<0.001). Based on the sampling time of cases, the RSV epidemic season for older people in Beijing was from October to March of the following year, with a peak period in December or January of the following year. In the post COVID-19 pandemic, there were very few RSV-positive cases detected in the elderly from April to June 2023, with only one positive case detected in May and one in June. The RSV-positive rate of older people increased significantly from October to December, reaching 11.75% (51/383) in December. Among 263 RSV-positive cases in the elderly, RSV-A, RSV-B and unclassified type accounted for 43.35% (114/263), 29.28% (77/263) and 27.38% (72/263), respectively. Since 2020, there has been a subtype conversion, with RSV-B being the main focus. Among 197 elderly cases that have complete clinical data, the main symptoms were cough (86.8%, 171/197), sputum (80.2%, 158/197) and fever (73.60%, 145/197). About 24.87% (49/197) of elderly cases experienced complications. The hospitalization mortality rate was 4.57% (9/197), and the hospitalization rate was 78.68% (155/197). The ICU occupancy rate was 1.99% (36/197). The mechanical ventilation usage rate was 13.32% (33/197), and the length of hospital stay [ M ( Q1, Q3)] was 12 (9, 16) days. Conclusion:In Beijing, the RSV infection rate is relatively low during the COVID-19 pandemic, and the prevalence of COVID-19 is relatively high. In 2023, there was no out-of-season outbreak of RSV infection among the elderly. Elderly RSV infection cases have multiple complications, severe diseases, and poor prognosis.

Objective:To investigate the epidemiological and clinical characteristics of RSV among patients aged ≥60 years in Beijing from 2015 to 2023.Methods:Based on the respiratory pathogen surveillance system, samples of upper respiratory tract infections (URTI), non-severe community-acquired pneumonia (nsCAP) and severe community-acquired pneumonia (sCAP) among people aged ≥60 years were collected from 28 sentinel hospitals in 16 districts of Beijing from January 2015 to December 2023. Swab samples were collected from URTI within one week, and lower respiratory tract samples from nsCAP and sCAP were collected. Demographic and epidemiological data were also collected. Various respiratory pathogens including RSV were detected.Results:From January 2015 to December 2023, a total of 20 349 cases of acute respiratory infections aged ≥60 years were included, with the RSV-positive rate of 1.54% (313/20 349, 95% CI: 1.39%-1.68%). Among them, the total RSV-positive rates of older people during the pre-pandemic, pandemic, and post-pandemic periods of COVID-19 were 1.59% (207/13 006, 95% CI: 1.38%-1.81%), 0.82% (38/4 650, 95% CI: 0.56%-1.08%) and 2.53% (68/2 693, 95% CI: 1.93%-3.12%), respectively. The difference in RSV-positive rate was statistically significant ( P<0.001). Based on the sampling time of cases, the RSV epidemic season for older people in Beijing was from October to March of the following year, with a peak period in December or January of the following year. In the post COVID-19 pandemic, there were very few RSV-positive cases detected in the elderly from April to June 2023, with only one positive case detected in May and one in June. The RSV-positive rate of older people increased significantly from October to December, reaching 11.75% (51/383) in December. Among 263 RSV-positive cases in the elderly, RSV-A, RSV-B and unclassified type accounted for 43.35% (114/263), 29.28% (77/263) and 27.38% (72/263), respectively. Since 2020, there has been a subtype conversion, with RSV-B being the main focus. Among 197 elderly cases that have complete clinical data, the main symptoms were cough (86.8%, 171/197), sputum (80.2%, 158/197) and fever (73.60%, 145/197). About 24.87% (49/197) of elderly cases experienced complications. The hospitalization mortality rate was 4.57% (9/197), and the hospitalization rate was 78.68% (155/197). The ICU occupancy rate was 1.99% (36/197). The mechanical ventilation usage rate was 13.32% (33/197), and the length of hospital stay [ M ( Q1, Q3)] was 12 (9, 16) days. Conclusion:In Beijing, the RSV infection rate is relatively low during the COVID-19 pandemic, and the prevalence of COVID-19 is relatively high. In 2023, there was no out-of-season outbreak of RSV infection among the elderly. Elderly RSV infection cases have multiple complications, severe diseases, and poor prognosis.

Objective:To evaluate the clinical efficacy and safety of endoscopic ultrasound (EUS)-guided coil implantation combined with glue injection for preventing rebleeding of tumorous gastric fundal varices.Methods:A retrospective analysis was performed on the clinical data of patients who underwent EUS-guided coil implantation combined with glue injection due to tumorous gastric fundal variceal bleeding from September 2017 to December 2020 in the First Affiliated Hospital, Zhejiang University School of Medicine. The basic characteristics and endoscopic manifestations of patients treated with this technique were analyzed, and the success rate of this technique, the usage of tissue glue and coil, the variceal obliteration rate and the incidence of complications such as pulmonary embolism were analyzed. Patients were followed up over a long period of time to obtain data on rebleeding. At the same time, patients were divided into complete obliteration group and incomplete obliteration group according to the endoscopic performance after the treatment, and the rebleeding rate in the two groups was compared.Results:Among the 41 patients included in this study, 18 cases (43.90%) had single isolated gastric varices type 1, and 23 cases (56.10%) had gastroesophageal varices combined. Fourteen cases had signs of recent bleeding such as thrombi and ulcers under endoscopy, and 13 cases had combined portal hypertensive gastropathy. All patients successfully completed the operation of EUS-guided coil implantation combined with glue injection. The technical success rate for the first attempt was 92.68% (38/41). The one-time pre-installation success rate of coils was 95.12% (39/41). The success rate of target vessel aspiration under EUS guidance was 100.00% (41/41). The release success rate of coil was 97.56% (40/41). The number of coils used was 1.12±0.33, and the amount of tissue glue used after inserting the coils was 1.54±0.46 mL. The target vessel obliteration rate after operation was 97.56% (40/41), and the complete gastric fundal variceal obliteration rate was 51.22% (21/41). There were 2 cases (4.88%) of intraoperative bleeding, 1 case stopped spontaneously, and 1 case successfully stopped bleeding after additional injection of tissue glue under direct endoscopic vision. No patient had symptoms of pulmonary embolism such as decreased oxygen saturation during the operation. Postoperative computed tomography angiography of pulmonary artery or lung computed tomography was performed in 24 cases, of which 1 case (4.17%) showed distal pulmonary artery embolism, but there were no relevant clinical manifestations. The patients were followed up for 21.64±16.86 months (1-50 months) after the surgery. There were 13 cases (31.71%) of recurrent gastrointestinal bleeding after the surgery, no rebleeding within 5 days, 2 cases of rebleeding within 3 months, 5 cases from 3 to 12 months, and 6 cases more than 1 year. Among the 13 cases of recurrent bleeding after the surgery, the recurrence rate of bleeding after operation in the complete obliteration group was 23.81% (5/21), which was lower than that in the incomplete obliteration group with 40.00% (8/20), but there was no statistically significant difference ( χ2=1.240, P=0.265). Conclusion:EUS-guided coil implantation combined with glue injection has good secondary prevention value and safety for tumorous gastric fundal variceal bleeding, and is worthy of clinical promotion and application.

Specific tumor-targeted gene delivery remains an unsolved therapeutic issue due to aberrant vascularization in tumor microenvironment (TME). Some bacteria exhibit spontaneous chemotaxis toward the anaerobic and immune-suppressive TME, which makes them ideal natural vehicles for cancer gene therapy. Here, we conjugated ZIF-8 metal-organic frameworks encapsulating eukaryotic murine interleukin 2 (Il2) expression plasmid onto the surface of VNP20009, an attenuated Salmonella typhimurium strain with well-documented anti-cancer activity, and constructed a TME-targeted Il2 delivery system named Il2/ZIF-8@Salmonella. Both in vitro and in vivo experiments demonstrated that Il2/ZIF-8@Salmonella maintained the tumor-targeting feature of bacteria, and could be effectively phagocytosed by intratumoral macrophages, thus leading to the expression and secretion of IL2 in TME. The detailed analysis of tumor immune microenvironment (TIME) showed that one dose of combinatorial Il2/ZIF-8@Salmonella achieved synergistic actions on a potent remodeling of TIME, marked by the activation of cytotoxic T cells and M1-polarization of macrophages in TME, thus leading to significant anti-tumor effects in melanoma, orthotopic hepatocellular carcinoma, and pulmonary metastasis models. More importantly, Il2/ZIF-8@Salmonella exhibited high safety to major organs and hematopoietic systems. Taken together, we report a novel plasmid/ZIF-8@Salmonella system that simultaneously achieves effective TME-targeted delivery of therapeutic gene, as well as synergistic re-activation of TIME.

Objective:To explore the medication law and academic thought of Professor Hu Guohua in treating diminished ovarian reserve (DOR) based on data mining.Methods:Collecting the medical records of DOR patients treated by Prof. Hu from January 1, 2021 to March 31, 2022. The TCM symptoms, tongue and pulse conditions, as well as the frequency, taste, meridian tropism and efficacy categories of TCM were analyzed by using the Traditional Chinese Medicine Inheritance Computing Platform V3.0 software. Commonly used drug combination analysis, high-frequency drug network relationship analysis, association rule analysis based on Apriori algorithm, cluster analysis based on K-means clustering algorithm, and core prescriptions were summarized.Results:A total of 85 DOR patients were included, involving 76 TCM symptoms, 15 tongue images, 11 pulse images. A total of 415 prescriptions were included, involving 164 kinds of Chinese materia medica. High-frequency drugs included Ligustri Lucidi Fructus, Angelicae Sinensis Radix, Astragali Radix, Cuscutae Semen, Herba Leonuri and Rhizoma Atractylodis Macrocephalae. The medicines were mostly warm, mild and cold in property, and the tastes were mostly sweet, bitter and pungent, mainly belonging to the liver, spleen and kidney meridians. 42 high-frequency drug combinations and 6 core prescriptions were obtained.Conclusions:Professor Hu treats DOR with the principle of tonifying the kidney and spleen to regulate body constitution, conditioning Chong Vessel and Conception Vessel to regulate the menstruation, soothing the liver and tranquillizing the heart to regulate the spirit. Professor Hu selects the medicines which are simplified and condensed, and is good at using drugs. The prescriptions are persistent and flexible, concerning the balance.

Pulmonary fibrosis is the end-stage manifestation of a large class of lung diseases characterized by fibroblast proliferation and accumulation of a large amount of extracellular matrix accompanied by inflammatory injury and tissue structure destruction. Studies have shown that flavonoids have anti-pulmonary fibrosis effects through multiple paths， including dihydromyricetin， morin and fisetin can inhibit fibroblast differentiation； Trollius altaicus flavonoids， hesperidin and linarin can play an anti-inflammatory role； total flavonoids from Scutellaria baicalensis， scutellarin and chrysin can inhibit epithelial- mesenchymal transition； total flavonoids of Litchi chinensis， diosmin and silybin can play an anti-oxidative stress role； quercetin， baicalin and apigenin can regulate autophagy； total flavonoids of Oxytropis falcata， calycosin and dihydroquercetin can regulate apoptosis； naringin， scutellarin and total flavonoids of Dracocephalum moldavica can inhibit pyroptosis， thus exerting anti- pulmonary fibrosis effects.