ObjectiveTo explore the role of cucurbitacin B （CuB） in inducing ferroptosis in 4T1 cells and its mechanism. MethodsThe effects of CuB（0.2， 0.4， 0.8 μmol·L^-1）on the proliferation ability of 4T1 cells in vitro were detected using the methyl thiazolyl tetrazolium （MTT） assay. The clonogenic ability of 4T1 cells was detected by the plate cloning assay， and the levels of lactate dehydrogenase （LDH） in 4T1 cells were detected by the use of a kit. The mitochondrial membrane potential and reactive oxygen species （ROS） levels in 4T1 cells were detected by flow cytometry， and the mitochondrial ultrastructure of 4T1 cells was observed by transmission electron microscopy. The western blot was used to detect the expression of ferroptosis-related protein p53 in 4T1 cells， solute carrier family 7 member 11 （SCL7A11）， glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4）， transferrin receptor protein 1 （TFR1）， and ferritin heavy chain 1 （FTH1）. ResultsCompared with that in the blank group， the survival rate of 4T1 cells in CuB groups was significantly decreased （P<0.05）， and the number of cell clones in CuB groups was significantly reduced （P<0.01）. In addition， compared with that in the blank group， the leakage of LDH in cells in CuB groups was significantly increased （P<0.01）， and the mitochondrial membrane potential of cells in CuB groups decreased significantly （P<0.01）. Cellular ROS levels were significantly elevated in CuB groups （P<0.01）. The mitochondria of cells in CuB groups were obviously wrinkled， and the mitochondrial cristae were reduced or even disappeared. Compared with that in the blank group， the protein expression of p53， ACSL4， and TFR1 were significantly up-regulated in CuB groups （P<0.05）， and that of SLC7A11， GPX4， and FTH1 were significantly down-regulated （P<0.05）. ConclusionCuB may inhibit SLC7A11 and GPX4 expression by up-regulating the expression of p53， which in turn regulates the p53/SLC7A11/GPX4 signaling pathway axis and accelerates the generation of lipid peroxidation substrate by up-regulating the expression of ACSL4. It up-regulates TFR1 expression to promote cellular uptake of Fe³⁺ and down-regulates the expression of FTH1 to reduce the ability of iron storage， resulting in an elevated free Fe²⁺ level. It catalyzes the Fenton reaction， generates excess ROS， imbalances the antioxidant system and iron metabolism， and then induces ferroptosis in 4T1 cells.

ObjectiveTo explore the role of cucurbitacin B （CuB） in inducing ferroptosis in 4T1 cells and its mechanism. MethodsThe effects of CuB（0.2， 0.4， 0.8 μmol·L^-1）on the proliferation ability of 4T1 cells in vitro were detected using the methyl thiazolyl tetrazolium （MTT） assay. The clonogenic ability of 4T1 cells was detected by the plate cloning assay， and the levels of lactate dehydrogenase （LDH） in 4T1 cells were detected by the use of a kit. The mitochondrial membrane potential and reactive oxygen species （ROS） levels in 4T1 cells were detected by flow cytometry， and the mitochondrial ultrastructure of 4T1 cells was observed by transmission electron microscopy. The western blot was used to detect the expression of ferroptosis-related protein p53 in 4T1 cells， solute carrier family 7 member 11 （SCL7A11）， glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4）， transferrin receptor protein 1 （TFR1）， and ferritin heavy chain 1 （FTH1）. ResultsCompared with that in the blank group， the survival rate of 4T1 cells in CuB groups was significantly decreased （P<0.05）， and the number of cell clones in CuB groups was significantly reduced （P<0.01）. In addition， compared with that in the blank group， the leakage of LDH in cells in CuB groups was significantly increased （P<0.01）， and the mitochondrial membrane potential of cells in CuB groups decreased significantly （P<0.01）. Cellular ROS levels were significantly elevated in CuB groups （P<0.01）. The mitochondria of cells in CuB groups were obviously wrinkled， and the mitochondrial cristae were reduced or even disappeared. Compared with that in the blank group， the protein expression of p53， ACSL4， and TFR1 were significantly up-regulated in CuB groups （P<0.05）， and that of SLC7A11， GPX4， and FTH1 were significantly down-regulated （P<0.05）. ConclusionCuB may inhibit SLC7A11 and GPX4 expression by up-regulating the expression of p53， which in turn regulates the p53/SLC7A11/GPX4 signaling pathway axis and accelerates the generation of lipid peroxidation substrate by up-regulating the expression of ACSL4. It up-regulates TFR1 expression to promote cellular uptake of Fe³⁺ and down-regulates the expression of FTH1 to reduce the ability of iron storage， resulting in an elevated free Fe²⁺ level. It catalyzes the Fenton reaction， generates excess ROS， imbalances the antioxidant system and iron metabolism， and then induces ferroptosis in 4T1 cells.

Ferroptosis， a new form of programmed cell death different from apoptosis， necrosis， and autophagy， is closely associated with a variety of physiological and pathological processes. Iron-mediated accumulation of reactive oxygen species is the main inducement of ferroptosis， the mechanism of which is related to intracellular lipid metabolism， iron metabolism， and antioxidant defense pathways. Multiple signaling axes and regulators jointly regulate the occurrence and disruption of ferroptosis. Studies have demonstrated that ferroptosis regulates the growth and proliferation of tumor cells. Inducing ferroptosis in tumor cells can control the growth， metastasis， and multi-drug resistance of tumors. Therefore， the effect and mechanism of ferroptosis on tumor cells have become a hot topic in anti-cancer research. As the research advances， a variety of ferroptosis inducers has been used in the clinical chemotherapy for cancers and demonstrate significant efficacy. Accordingly， the development of ferroptosis-inducing anticancer drugs has become a new research direction for tumor treatment. Some active ingredients such as lycorine， oleanolic acid， dihydroartemisinin， pseudolaric acid B， and ophiopogonin B of Chinese medicines can induce ferroptosis in tumor cells via lipid metabolism， iron metabolism， system X_c^-， and GPX4/GSH to regulate the development of tumors， demonstrating a promising prospect in clinical treatment. Based on the theory of the mechanism of ferroptosis， this paper reviews the research progress in ferroptosis induced by active ingredients of Chinese medicines in tumor cells and describes the metabolic regulatory network of ferroptosis from signaling pathways and regulatory factors， providing new strategies for applying active ingredients of Chinese medicines in the treatment of tumors.

Objective To investigate the intrinsic neural mechanism of aggressive behavior in CD 1 mice.Methods CD1 mice with aggressive behavior were screened out by resident intruder test.After the aggressive conditioned pair preference was further verified,the activated brain regions of the whole brain were labeled with c-Fos,and the types of neurons activated by the aggressive behavior were analyzed by double immunofluorescence labeling.Finally,the effects of activity of these neurons regulated by optogenetics on aggressive behavior were observed.Results The c-Fos screening revealed that about 82％of the CD1 mice showed aggressive behavior.After the occurrence of aggressive behavior,the main activation occured in the medial prefrontal cortex(mPFC),and the results of immunofluorescence double labeling showed that the c-Fos positive neurons in the mPFC were mainly glutamatergic neurons.Finally,glutamatergic neurons in the mPFC could be activated by optogenetics,and the activation inhibited the aggressive behavior of CD1 mice.In contrast,optogenetics could inhibit glutamatergic neurons in the mPFC and then promote the aggressive behavior of CD1 mice.Conclusion Glutamatergic neurons in the mPFC are an important component in the regulation of aggressive behavior in CD1 mice.

OBJECTIVE: We aimed to understand the willingness and barriers to the acceptance of tuberculosis (TB) preventive treatment (TPT) among people with latent TB infection (LTBI) in China. METHODS: A multicenter cross-sectional study was conducted from May 18, 2023 to December 31, 2023 across 10 counties in China. According to a national technical guide, we included healthcare workers, students, teachers, and others occupations aged 15-65 years as our research participants. RESULTS: Overall, 17.0% (183/1,077) of participants accepted TPT. There were statistically significant differences in the acceptance rate of TPT among different sexes, ages, educational levels, and occupations ( P < 0.05). The main barriers to TPT acceptance were misconceptions that it had uncertain effects on prevention (57.8%, 517/894), and concerns about side effects (32.7%, 292/894). CONCLUSION An enhanced and comprehensive understanding of LTBI and TPT among people with LTBI is vital to further expand TPT in China. Moreover, targeted policies need to be developed to address barriers faced by different groups of people.
Humans ; China/epidemiology* ; Adult ; Male ; Female ; Cross-Sectional Studies ; Middle Aged ; Young Adult ; Adolescent ; Aged ; Latent Tuberculosis/prevention & control* ; Patient Acceptance of Health Care ; Tuberculosis/prevention & control* ; Antitubercular Agents/therapeutic use* ; Health Knowledge, Attitudes, Practice

Objective:To investigate the correlation between hyponatremia and the severity of coronavirus disease 2019 (COVID-19).Methods:Clinical data of 12 patients with COVID-19 admitted to Shantou Central Hospital from January 23 to February 5 in 2020 were retrospectively analyzed, including gender, age, symptoms, lab test and clinical outcomes, to analyze the change trend of blood Na + level in the patients with COVID-19. Results:Among the 12 patients with COVID-19, there were 8 males and 4 females with the mean age of (38.0±16.3) years old, most of them were admitted to the hospital with cough and/or fever. All patients had a positive nucleic acid test for 2019 novel coronavirus (2019-nCoV), and were discharged after clinical treatment with oxygen therapy, antiviral, antibacterial, anti-inflammatory, and nutritional support. All patients were of ordinary type when they were admitted to the hospital. Among them, 1 patient turned into a severe case during the course of the disease, and 1 patient showed a tendency to become severe case. It was found that 10 patients without severe conversion had an average blood Na + of (138.3±1.3) mmol/L at admission, and the lowest blood Na + during the course of disease was (135.9±3.1) mmol/L. However, 2 patients who became severe and had a tendency to become severe disease (Na + levels at admission were 140.0 mmol/L and 138.0 mmol/L, respectively) experienced hyponatremia during the course of the disease (the lowest blood Na + levels were 129.0 mmol/L and 122.0 mmol/L). Further analysis showed that the lower serum Na + level, the higher level of white blood cell count (WBC) and C-reactive protein (CRP), but serum Na + level was consistent with the change trend of lymphocytes, suggesting that hyponatremia was closely correlated with severe inflammation reaction. Conclusions:Serum Na + showed decreasing tendency during the development of COVID-19, and hyponatremia was closely related to the severity of COVID-19. It was necessary to pay great attention to the change trend of blood Na + level. However, further research was needed to obtain more reliable conclusions and explorer the pathophysiological mechanisms.

Objective: To investigation HER2 status in gastric adenocarcinoma of Chinese and contributing factors to the HER2 expression. Methods: HER2 status of 40 842 gastric adenocarcinomas and clinical data were retrospectively collected from 23 hospitals dated from 2013 to 2016. The association between HER2 positivity and clinicopathologic features was analyzed. Results: Of the 40 842 patients the median age was 62 years, the male female ratio was 2.6∶1.0. The rate of HER2 positivity was 8.8% (3 577/40 842). HER2 expression was related to the tissue type, tumor location, Lauren classification and tumor differentiation (P values: 0.009, 0.001, <0.01 and <0.01, respectively). Different HER2 expression status was observed between primary and recurrent tumors in 7.6% (48/635) cases. The rates of HER2 positivity ranged from 2% to 10% among different institutions. The rates of HER2 FISH amplification were dramatically different among the 23 hospitals (0-100%) with an average rate of 10% (810/8 156) in patients with HER2 IHC 2+ . Conclusions HER2 expression is associated with clinicopathologic characteristics. HER2 re-assessment of tumor tissue and use of in situ hybridization techniques increase HER2 positivity. The current retrospective study should reflect the HER2 status in gastric adenocarcinoma of Chinese patients.

Objective To investigate the relationship between Pin1 and CyclinD1 expression and the development of gastrointestinal stromal tu?mor(GIST). Methods The protein and mRNA expression of Pin1 and CyclinD1 in 85 samples of GIST and adjacent non?cancerous tissues were detected by immunohistochemistry and real?time quantitative polymerase chain reaction. Results The expression rate of Pin1 protein in GIST tis?sues(64.7%;55/85)was higher than that in adjacent non?cancerous tissues(26.7%;4/15). Similarly,the expression rate of CyclinD1 protein in GIST tissues(42.3%;36/85)was higher than that in adjacent non?cancerous tissues(6.7%;1/15). The expression of Pin1 and CyclinD1 mRNA in GIST tissues was 7.03 and 5.53 times that in adjacent non?cancerous tissues ,respectively. There was no obvious correlation between the expres?sion of Pin1 and clinicopathological parameters. The expression of CyclinD1 was positively correlated with the grade of NIH and tumor diameter (P<0.05). There was a significant correlation between the expression of Pin1 and CyclinD1 in GIST tissues. Conclusion The expression of both Pin1 and CyclinD1 was up?regulated in GIST tissues. The significant correlation between the expression of Pin1 and CyclinD1 in GIST tissues sug?gests that their synergistic effect promotes carcinogenesis and the development of GIST.

Objective: To observe the effects of transient exposure to high glucose on biological behaviors of human dermal microvascular endothelial cells cultured in vitro. Methods: The dividing method and treatment of cells for the detection of all indexes in this study were as follows. Human dermal microvascular endothelial cells of the 4th passage were divided into 3 groups according to the random number table, with 12 wells in each group. Cells in control group (C) were cultured with complete culture solution containing 5 mmol/L D-glucose for 7 d. Cells in transient high glucose group (THG) were cultured with complete culture solution containing 30 mmol/L D-glucose for 2 d and complete culture solution containing 5 mmol/L D-glucose for 5 d. Cells in prolonged high glucose group (PHG) were cultured with complete culture solution containing 30 mmol/L D-glucose for 7 d. (1) The cell morphology in groups C and PHG on culture day 7 and that in group THG on culture day 2 and 7 was observed by inverted optical microscope. (2) On culture day 0, 2, 4, and 7, cell proliferation rate was determined by cell viability analyzing counter. (3) After culture day 2, the scratch experiment was performed, and the cells were further cultured. At post scratch hour (PSH) 0, 24, 48, 72, 96, and 120, the scratch area was measured, and the cell migration rates of the latter 5 time points were calculated. (4) On culture day 0, 2, 4, and 7, the cell apoptosis rate was determined by cell analyzer. (5) Cells were seeded into Matrigel to culture for 24 h after culture day 7. The formation of vessel-like structure was observed by inverted optical microscope. The length and number of branch point of vessel-like structure were calculated. (6) On culture day 2, 4, and 7, mRNA expression of vascularization-related gene tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) was determined with real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were processed with analysis of variance of factorial design, analysis of variance for repeated measurement, one-way analysis of variance, and LSD test. Results: (1) Cells in group C exhibited ovary shape in cobble stone order on culture day 7. Cells in group THG exhibited long ovary shape and lost cobble stone order on culture day 2 and kept the same changes on culture day 7. Cells in group PHG exhibited long ovary shape and lost cobble stone order on culture day 7. (2) On culture day 0, there was no significant difference in cell proliferation rate among the 3 groups (F=0.23, P>0.05). On culture day 2, cell proliferation rates in groups THG and PHG were similar (P>0.05), which were significantly lower than the cell proliferation rate in group C (with P values below 0.01). On culture day 4 and 7, the cell proliferation rates in groups THG and C were similar (with P values above 0.05), which were significantly higher than those in group PHG (with P values below 0.01). (3) At PSH 24-120, the cell migration rates in groups THG and PHG were similar (with P values above 0.05), which were significantly lower than those in group C (with P values below 0.01). (4) On culture day 0, there was no statistically significant difference in cell apoptosis rate among the 3 groups (F=0.78, P>0.05). On culture day 2, cell apoptosis rates in groups THG and PHG were similar (P>0.05), which were significantly higher than the cell apoptosis rate in group C (with P values below 0.01). On culture day 4 and 7, the cell apoptosis rates in groups THG and C were similar (with P values above 0.05), which were significantly lower than those in group PHG (with P values below 0.01). (5) The length of vessel-like structure of cells in group THG was (1.84±0.10)×10⁵ μm, close to (1.82±0.11)×10⁵ μm in group PHG (P>0.05), both significantly shorter than (2.75±0.23)×10⁵ μm in group C (with P values below 0.01). The numbers of branch point of vessel-like structure of cells in groups THG and PHG were 43±5 and 46±8 respectively, which were close to each other (P>0.05) and both significantly less than 103±21 in group C (with P values below 0.01). (6) On culture day 2, 4, and 7, mRNA expressions of TIMP-3 of cells in groups THG and PHG were similar (with P values above 0.05), which were significantly lower than those in group C (with P values below 0.05). Conclusions Transient exposure to high glucose can cause metabolic memory of morphology, migration, and angiogenesis in human dermal microvascular endothelial cells cultured in vitro, resulting in sustained changes in biological behaviors. The mechanism may be related to the changes of vascularization-related genes.

Objective To understand the causes and the ratio of cause constituents in children with chronic cough in Beijing. Methods Patients with chronic cough treated at respiratory clinic of the Children' s Hospital Affiliated to Capital Institute of Pediatrics from May 2009 to April 2011 were included in this study. Etiologic diagnostic procedure suggested for children by the Subspecialty Group of Respiratory Diseases,the Society of Pediatrics,Chinese Medical Association in 2008 was implemented to manage the patients. After three - month follow - up,the etiological data was analyzed. Results Totally 213 patients with chronic cough aged 1. 1 to 14. 0 years old were enrolled,inclu-ding 40 cases(18. 8% )aged≤3 years old,134 cases(62. 9% )aged ﹥ 3 to 6 years old,and 39 cases(18. 3% ) aged ﹥ 6 years old. The majority of patients with positive allergen tests were sensitized to inhaled allergens. One child had positive result in 24 - hour esophageal pH monitoring,but his cough didn't respond well to the specific treatment for gastroesophageal reflux,so he wasn't diagnosed as gastroesophageal reflux cough. The 4 leading causes of the 213 pa-tients with chronic cough were cough variant asthma(CVA)in 92 cases(43. 2% ),CVA associated with upper airway cough syndrome(UACS)in 57 cases(26. 8% ),UACS in 28 cases(13. 2% ),respiratory infection and post - infec-tion cough(PIC)in 26 cases(12. 2% ),while other causes were found in 8 cases(3. 7% ),and unknown etiology in 2 cases(0. 9% ). The incidence of CVA ranked top 1 in all the 3 age groups,followed by PIC in ≤3 years old group, while CVA associated with UACS in the other 2 age groups. Conclusions CVA,CVA associated with UACS,UACS and PIC were the 4 leading causes for children with chronic cough in Beijing. Children in different age groups had dif-ferent constituents ratio of causes of chronic cough.