AIM: To investigate the effect of CP466722, an inhibitor of ataxia telangiectasia mutated protein (ATM), on the proliferation and apoptosis of human hepatocellular carcinoma HepG2 cells.METHODS: The cell viability was detected by MTT assay.The cell growth inhibition was measured by colony formation assay.The effect of CP466722 on the cell cycle distribution of the HepG2 cells was examined by flow cytometry.The cell apoptosis was analyzed by TUNEL staining.The protein expression was examined by Western blotting.RESULTS: CP466722 inhibited the cell viability and cell proliferation in a dose-dependent manner.In CP466722-treated HepG2 cells, the cell cycle was arrested in G2/M phase, and the protein levels of phosphorylated cell division cycle protein 2 (p-Cdc2), cell division cycle protein 25C (Cdc25C) and phosphorylated Cdc25C (p-Cdc25C) were inhibited, whereas the protein expression of p27 was up-regulated.CP466722 triggered the apoptosis of HepG2 cells through cleavages of caspase-3 and poly (ADP-ribose) polymerase (PARP).In addition, CP466722 increased the phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK) and suppressed the expression of β-catenin and survivin in the HepG2 cells.CONCLUSION: CP466722 inhibits the proliferation and induces the apoptosis of HepG2 cells, which may be related to activating p38 MAPK and inhibiting the expression of β-catenin and survivin.

AIM: To study the effects of xijiao dihuang decoction, a Chinese medicine, on the expression of adhesion molecules in vascular endothelial cells (VECs) of adrenine and hypothermia-treated rats. METHODS: The expression of VCAM-1, ICAM-1, PECAM-1, iNOS and their corresponding mRNA in VECs was assayed by immunohistochemistry analysis and RT-PCR, respectively. RESULTS: Immunohistochemistry analysis showed the expression of VCAM-1, ICAM-1, PECAM-1 and iNOS are higher in adrenine and hypothermia-treated rats than that in controls, xijiao dihuang decoction decreased the expression of VCAM-1, ICAM-1, PECAM-1 and iNOS in a dose-dependent manner in adrenine and hypothermia-treated rats. CONCLUSION: Xijiao dihuang decoction can down-regulate the expression of adhesion molecules in vascular endothelial cells of adrenine and hypothermia-treated rats.

Objective To study the effect of Buyang Huanwu Decoction (BHD) on adhesion molecules in vascular endothelial cells (VECs) of blood stasis rats. Methods Expression of ICAM-1, VCAM-1, PECAM-1, iNOS in vascular endothelial cells of blood stasis rats receiving different doses of BHD was observed by immunohistochemical method and RT-PCR.Results The expression of ICAM-1, VCAM-1, PECAM-1, iNOS was increased in model group, while was decreased by BHD. There was an increase tendency on the expression of molecules and significant dose-effect relationship along with the decrease of the BHD doses. Conclusion The increased expression of adhesion molecules in vascular endothelial cells of blood stasis rats could be reduced by BHD in a dose dependent manner.

A highly sensitive and selective method was developed for both UV-vis spectrophotometric and fluo-rimetric determination of organophosphorus pesticides(OPs).This method used silver nanoparticles(AgNPs)modified with graphitic carbon nitride(g-C3N4).The AgNPs reduced the fluorescence intensity of g-C3N4.Acetylthiocholine(ATCh)could be catalytically hydrolyzed by acetylcholinesterase(AChE)to form thiocholine,which induces aggregation of the AgNPs.This aggregation led to the recovery of the blue fluorescence of g-C3N4,with excitation/emission peaks at 310/460 nm.This fluorescence intensity could be reduced again in the presence of OPs because of the inhibitory effect of OPs on the activity of AChE.The degree of reduction was found to be proportional to the concentration of OPs,and the limit of fluorometric detection was 0.0324 μg/L(S/N = 3).In addition,the absorption of the g-C3N4/AgNPs at 390 nm decreased because of the aggregation of the AgNPs,but was recovered in presence of OPs because of the inhibition of enzyme activity by OPs.This method was successfully applied to the analysis of parathion-methyl in real samples.

Objective To investigate whether chemokine CCL2(also known as monocyte chemotactic protein 1 or MCP-1)has a causal relationship with lung cancer.Methods Genetic data of chemokine CCL2 and different pathological subtypes of lung cancer were extracted from genome-wide association studies(GWAS),and inverse-variance weighted(IVW)analysis was used as main analysis,while weighted median,simple model,MR-Egger regression,and weighted model were chosen as supplementary analyses.Sensitivity analyses were performed to verify the reliability of the data.Results The result of IVW analysis on chemokine CCL2 to lung adenocarcinoma was OR = 1.065,95%CI(0.919～1.234),P = 0.401.The result of IVW analysis on chemokine CCL2 to squamous cell lung carcinoma was OR = 1.059,95%CI(0.931～1.205),P = 0.381.The result of IVW analysis on chemokine CCL2 to small cell lung carcinoma was OR = 0.959,95%CI(0.760～1.208),P = 0.720.Conclusions There is no direct causal relationship between chemokine CCL2 and lung cancer.

Objective To explore whether voluntary wheel running affects liver oxidative stress by regulating the Nrf2/HO-1 pathway,thereby alleviating HFFC diet-related lipid deposition in the liver.Methods Eight-week-old C57BL/6J mice were randomly divided into a normal diet group(NC group,n=10)and high-fat,fructose,and cholesterol diet group(HFFC group,n=20)after 1 week of adaptive feeding.Ten weeks of feeding later,mice in the HFFC group were divided into a quiet group(HFFC group,n=10)and HFFC combined with exercise group(HFFC+EX group,n=10).HFFC+EX group mice were caged with voluntary running wheels for free movement,and the number of running wheels was recorded every day for 8 weeks.After the last treatment,the mice were sacrificed by fasting for 12 hours at an interval of 24 hours,and the blood and liver were collected for analysis.Results(1)Body weight,liver weight,and liver index of mice fed the HFFC diet were significantly higher than those of the NC group,which significantly decreased after exercise(P＜0.05).(2)Compared with the NC group,HDL-C and LDL-C in the HFFC group were significantly increased,and the LDL-C level was significantly decreased after 8 weeks of exercise(P＜0.05).(3)The liver fat droplet area and liver TG content in the HFFC group were significantly higher than those in the NC group,whereas those in HFFC+EX group were significantly decreased(P＜0.05).(4)Compared with the NC group,the content of oxidase MDA in the HFFC group were significantly increased,and nuclear translocation and gene expression of Nrf2 were significantly decreased.After exercise,the activities of SOD and T-AOC were significantly increased,and the nuclear translocation and gene expression of Nrf2 and expression levels of HO-1 and SOD-1 were significantly increased(P＜0.05).(5)The number of apoptotic hepatocytes and CHOP expression in the HFFC diet group were significantly higher than those in the NC group,whereas the number of apoptotic hepatocytes,and CHOP and Bax/Bcl-2 expression in the exercise group were significantly lower than those in the NC group(P＜0.05).Conclusions Voluntary wheel can alleviate HFFC diet induced liver lipid deposition by regulating the Nrf2/HO-1 pathway,thereby alleviating oxidative stress and reducing apoptosis in liver cells.

Objective To examine the current state of care services for geriatric osteoporotic fractures in Chinese hospitals and to provide a basis for the improvement of these services and the formulation of related policies.Methods In September to November 2023,a stratified convenience sampling method was used to investigate the implementation of care services for elderly patients with osteoporotic fractures in 621 hospitals across 31 provinces(autonomous regions and municipalities)in China.A self-designed questionnaire was utilized for this purpose.Results A total of 621 hospitals participated in the survey,with 432(69.57%)tertiary hospitals and 189(30.43%)secondary hospitals.Over 95%of hospitals provided health education on diet,medication,fall prevention,and early functional exercise for elderly fracture patients.Less than 80%of hospitals provide specialized training on osteoporosis treatment and secondary fracture prevention for medical staff.Only 263 hospitals(42.35%)routinely conduct bone density tests for patients over 50 years old with fractures,while 221 hospitals(35.59%)routinely conduct bone metabolic biochemical tests for such patients.Less than 50%of hospitals provide specialized services,such as geriatric osteoporotic fracture clinics,for elderly patients with osteoporotic fractures.Additionally,39.77%of hospital departments have not developed postoperative care plans for elderly patients with osteoporotic fractures.The lack of specialized care teams(91.63%),insufficient investment in care resources(88.08%),and the absence of policy support(77.45%)are identified as the primary factors impeding the provision of care services for elderly patients with osteoporotic fractures in hospitals.Although some care services in tertiary hospitals are superior to those in secondary hospitals(P<0.05),they are still far from adequate.Conclusion The development of care services for elderly patients with osteoporotic fractures in Chinese hospitals needs improvement.It is recommended to further standardize and enhance the content and methods of health education,intensify clinical assessments related to osteoporosis in elderly fracture patients,improve the professional care capabilities of medical staff,and at the same time,the state should introduce relevant policies to support and promote the construction and development of hospital care services for elderly osteoporotic fracture patients.

Although DNA 5-hydroxymethylcytosine (5hmC) is recognized as an important epige-netic mark in cancer, its precise role in lymph node metastasis remains elusive. In this study, we investigated how 5hmC associates with lymph node metastasis in breast cancer. Accompanying with high expression of TET1 and TET2 proteins, large numbers of genes in the metastasis-positive pri-mary tumors exhibit higher 5hmC levels than those in the metastasis-negative primary tumors. In contrast, the TET protein expression and DNA 5hmC decrease significantly within the metastatic lesions in the lymph nodes compared to those in their matched primary tumors. Through genome-wide analysis of 8 sets of primary tumors, we identified 100 high-confidence metastasis-associated 5hmC signatures, and it is found that increased levels of DNA 5hmC and gene expression of MAP7D1 associate with high risk of lymph node metastasis. Furthermore, we demonstrate that MAP7D1, regulated by TET1, promotes tumor growth and metastasis. In conclusion, the dynamic 5hmC profiles during lymph node metastasis suggest a link between DNA 5hmC and lymph node metastasis. Meanwhile, the role of MAP7D1 in breast cancer progression suggests that the metastasis-associated 5hmC signatures are potential biomarkers to predict the risk for lymph node metastasis, which may serve as diagnostic and therapeutic targets for metastatic breast cancer.

[Abstract] Objective: To investigate the effect of alantolactone (ALT) on proliferation, migration, invasion and apoptosis of human osteosarcoma 143B cells and the underlying mechanism. Methods: Osteosarcoma 143B cells were treated with different concentrations of ALT (0, 4, 6, 8, 10 µmol/L). Then, the cell proliferation ability was detected by crystal violet staining and MTT assay, cell migration was determined by Wound-healing test, cell invasion was analyzed by Transwell assay and cell apoptosis rate was detected by Hoechst33258 staining. The mRNA and protein expressions of E-cadherin, N-cadherin, caspase-3, cleaved caspase-3 (c-caspase-3), poly ADP-ribose polymerase (PARP) and cleaved PARP (c-PARP) in 143B cells were detected by qPCR and Western blotting (WB), respectively. TCF/LEF (T cell lymphocyte factor/lymphoid enhancer factor) transcriptional activity was examined with Luciferase reporter gene assay. The mRNA and protein expressions of β-catenin as well as MMP-7 and c-Myc were detected by qPCR and WB, respectively. Results: ALT inhibited proliferation, migration and invasion of osteosarcoma143B cells and promoted apoptosis(P<0.05or P<0.01). After the treatment with ALT at 8, 10 µmol/L, the mRNA and protein expressions of E-cadherin and PARP, as well as the protein expressions of c-caspase-3 and c-PARP were up-regulated, while the mRNA and protein expressions of N-cadherin were downregulated (P<0.05 or P<0.01);At the sametime, theTCF/LEF transcriptional activity and the mRNA and protein expressions of β-catenin, MMP-7 and c-Myc were significantly down-regulated (P<0.05 or P<0.01). Conclusion:ALT may inhibit the proliferation, migration and invasion and promote cell apoptosis possibly through suppressing Wnt/β-catenin signaling pathway in osteosarcoma 143B cells.

Immunoglobulin G4-related sialadenitis (IgG4-RS) is an immune-mediated fibro-inflammatory disease and the pathogenesis is still not fully understood. The aim of this study was to explore the role and mechanism of interleukin-13 (IL-13) in the cellular senescence during the progress of IgG4-RS. We found that the expression of IL-13 and IL-13 receptor α1 (IL-13Rα1) as well as the number of senescent cells were significantly higher in the submandibular glands (SMGs) of IgG4-RS patients. IL-13 directly induced senescence as shown by the elevated activity of senescence-associated β-galactosidase (SA-β-gal), the decreased cell proliferation, and the upregulation of senescence markers (p53 and p16) and senescence-associated secretory phenotype (SASP) factors (IL-1β and IL-6) in SMG-C6 cells. Mechanistically, IL-13 increased the level of phosphorylated signal transducer and activator of transcription 6 (p-STAT6) and mitochondrial-reactive oxygen species (mtROS), while decreased the mitochondrial membrane potential, ATP level, and the expression and activity of superoxide dismutase 2 (SOD2). Notably, the IL-13-induced cellular senescence and mitochondrial dysfunction could be inhibited by pretreatment with either STAT6 inhibitor AS1517499 or mitochondria-targeted ROS scavenger MitoTEMPO. Moreover, IL-13 increased the interaction between p-STAT6 and cAMP-response element binding protein (CREB)-binding protein (CBP) and decreased the transcriptional activity of CREB on SOD2. Taken together, our findings revealed a critical role of IL-13 in the induction of salivary gland epithelial cell senescence through the elevated mitochondrial oxidative stress in a STAT6-CREB-SOD2-dependent pathway in IgG4-RS.
Cellular Senescence/genetics* ; Humans ; Immunoglobulin G/metabolism* ; Interleukin-13/pharmacology* ; Mitochondria/metabolism* ; Sialadenitis/metabolism*