Objective To investigate the underlying mechanisms of the effect of Fufang Changtai Decoction(FFCT)in inhibiting colorectal cancer(CRC)through the ferroptosis pathway using network pharmacology combined with experimental validation.Methods The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Swiss Target Prediction databases were employed for the systematic screening of potent active ingredients and therapeutic targets of FFCT.In addition,the identification of CRC-associated genes and ferroptosis-related genes(FRGs)was accomplished using the Gene Cards and FerrDb databases,respectively.Venn diagrams,coupled with Cytoscape software,facilitated the comprehensive analysis of key FRGs involved in FFCT's intervention in CRC by mapping the TCM compound-therapeutic target network.Transmission electron microscopy was used to examine the mitochondrial ultrastructure of SW480 and HCT116,2 Human CRC cell lines,after treatment with FFCT-containing serum.Intracellular reactive oxygen species(ROS)levels were measured using a ROS detection kit.To assess the role of ferroptosis,ferroptosis inhibitor liproxstatin-1(Lip-1)was co-administered with FFCT-containing serum.The effects on cancer cell viability and proliferation were evaluated using CCK-8 and colony formation assays.Key molecular targets involved in the regulatory effects of FFCT on the expression of FRGs were further analyzed using PCR Array and Western blot.The findings were then validated with human CRC tissue microarrays.Results A total of 103 active ingredients of FFCT,739 therapeutic targets,9 101 disease-related genes,and 564 FRGs were identified.Venn diagram analysis identified 81 FRGs associated with FFCT intervention.Network analysis revealed that NQO1,TP53,and PTGS2 served as hub nodes in the regulatory network.Findings from the in vitro experiments showed that FFCT induced ferroptosis changes,including mitochondrial condensation,membrane thickening,and cristae reduction,in SW480 and HCT116 cells.FFCT treatment significantly increased intracellular ROS levels in a dose-dependent manner(P<0.05)and reduced cancer cell viability and proliferative capacity(P<0.01).These inhibitory effects were partially reversed by Lip-1,suggesting that FFCT's antitumor activity was closely associated with the ferroptosis pathway.PCR Array and Western blot analyses further confirmed that FFCT significantly downregulated NQO1 mRNA and protein expression in cancer cells(P<0.001),which was consistent with network pharmacology predictions.Immunofluorescence analysis of clinical CRC tissue microarrays revealed that NQO1 expression was significantly higher in tumor tissues than in adjacent non-tumor tissues(P<0.001).Conclusion FFCT may induce intracellular ferroptosis by downregulating the oncogenic gene NQO1,thereby exerting anti-CRC effects.

OBJECTIVE: To explore the regulatory mechanism of human hepatocyte apoptosis induced by lysosomal membrane protein Sidt2 knockout. METHODS: The Sidt2 knockout (Sidt2^-/-) cell model was constructed in human hepatocyte HL7702 cells using Crispr-Cas9 technology.The protein levels of Sidt2 and key autophagy proteins LC3-II/I and P62 in the cell model were detected using Western blotting, and the formation of autophagosomes was observed with MDC staining.EdU incorporation assay and flow cytometry were performed to observe the effect of Sidt2 knockout on cell proliferation and apoptosis.The effect of chloroquine at the saturating concentration on autophagic flux, proliferation and apoptosis of Sidt2 knockout cells were observed. RESULTS: Sidt2^-/- HL7702 cells were successfully constructed.Sidt2 knockout significantly inhibited the proliferation and increased apoptosis of the cells, causing also increased protein expressions of LC3-II/I and P62(P < 0.05) and increased number of autophagosomes.Autophagy of the cells reached a saturated state following treatment with 50 μmol/L chloroquine, and at this concentration, chloroquine significantly increased the expressions of LC3B and P62 in Sidt2^-/- HL7702 cells. CONCLUSION Sidt2 gene knockout causes dysregulation of the autophagy pathway and induces apoptosis of HL7702 cells, and the latter effect is not mediated by inhibiting the autophagy-lysosomal pathway.
Humans ; Lysosome-Associated Membrane Glycoproteins/metabolism* ; Autophagy ; Apoptosis ; Hepatocytes ; Lysosomes/metabolism* ; Chloroquine/pharmacology* ; Nucleotide Transport Proteins/metabolism*

Objective Comprehensive mortality risk analyses and therapeutic assessment in real-world practice are beneficial to guide individual treatment in patients with Carbapenem-resistant Klebsiella pneumoniae bloodstream infections(CRKP-BSI).Methods Retrospective analysis of the clinical characteristics of 81 CRKP-BSI patients in our intensive care unit from July 2016 to June 2020,to indentify the risk factors of death and treatment effects of different antibiotic regimens.Results In 81 CRKP-BSI cases,the majority source were from abdominal and respiratory,accounting for 56.79%(46 cases)and 22.22%(18 cases),respectively.The 28-day mortality and hospitalization mortality of CRKP-BSI were 54.32%(44 cases)and 65.43%(53 cases).Multivariate regression analysis suggested that biliary tract disease before admission(P=0.026)and increased SOFA score at the onset of BSI(P=0.006)were independent risk factors for 28-day mortality.There was no statistically significant difference in 28-day mortality between the groups of antibiotic treatment based on tigecycline(44 cases)and polymyxin B(26 cases)[56.82%(25/44)vs.57.69%(15/26),P=0.943].Patients were evaluated based on their age(≤65 years vs.>65 years),gender,body mass index(≤25 kg/m2 vs.>25 kg/m2),and APACHEⅡ score(≤20 vs.>20),the use of renal replacement therapy and mechanical ventilation,there was no difference in the mortality among each subgroup.Conclusions Biliary tract disease before admission and SOFA score were independent risk factors for 28-day mortality.There was no significant difference outcomes between tigecycline-and polymyxin B-based therapy.

Objective:To explore the metabolic regulations of different Traditional Chinese Medicine (TCM) syndromes in the diabetic patients with high risk for foot ulceration.Methods:Based on gas chromatography-mass spectrometer and multi-dimensional data processing methods, the metabolomics analysis was used to compare the serum metabolites profile of healthy people (32 cases) and the high-risk foot patients in Cold and Blood Stagnation syndrome (44 cases), Heat-toxin hurting Yin syndrome (54 cases), and Qi-Blood deficiency syndrome (33 cases), who were hospitalized at Shanghai TCM-Integrated Hospital from Apirl to December, 2018.Results:This study suggested that compared with healthy people, the diabetic patients with high risk for foot ulceration showed significantly lower serum level of urea [(2.41 ± 1.57)×10 5vs. (3.32 ± 2.10)×10 5], L-leucine [(4.94 ± 3.15)×10 5vs. (6.39 ± 3.57)×10 5], aspartic acid [(3.94 ± 4.48)×10 5vs. (9.62 ± 6.93)×10 5], 9H-purine [(1.74 ± 1.95)×10 5vs. (3.34 ± 2.23)×10 5] ( P<0.05 or P<0.01), while higher level of d-Glucose [(3.72 ± 1.71)×10 5vs. (2.21 ± 1.32)×10 5] and d-glucopyranose [(3.32 ± 2.10)×10 5vs. (1.35 ± 1.43)×10 5] ( P<0.01). Energy metabolism, amino acid metabolism and sugar metabolism were mainly involved. the content of L-tyrosine in the group of patients with Cold and Blood Stagnation syndromewas significantly higher than that in healthy people. The urea, purine, leucine, aspartic acidcontent in patients of Heat-toxin hurting Yin syndrome were significantly lower than that in healthy people. The purine content in patients of Qi and Blood Deficiency Syndrome was significantly lower than that in healthy people. Compared with the syndrome of Heat-toxin hurting Yin syndrome, patients in Cold and Blood Stagnation syndrome showed a significantly higher content of beta-1-galactopyranoside and butanoic acid. Compared to the Qi-Blood deficiency syndrome, serum urea level in patients of Heat-toxin hurting Yin syndrome was significantly higher than those in the patients of other two TCM syndromes. Conclusions:The serum metabolomics profiling differentiate three TCM-syndrome in high-risk DF patients, which can provide objective basis for clinical TCM syndrome differentiation and treatment of high-risk diabetic foot patients.

Objective: To analyze the related factors of esophageal squamous cell carcinoma and precancerous lesions among residents aged 40-69 years old in rural areas of Shandong Province. Methods: In October 2018, 300 villages in 13 counties of the Shandong upper gastrointestinal cancerearly diagnosis and treatment projectin 2017 were selected as research areas, and 30 400 residents aged 40-69 were recruited in this study. The demographic characteristics, health status and lifestyle information were collected through the questionnaire survey, and endoscope iodine staining and indicative biopsy methods were used for cancer screening among eligible people.The multivariate logistic regression model was used to analyze the risk factors for esophageal cancer and precancerous lesions. Results: The subjects in this study were (56.42±7.24) years old, including 13 193 males (43.40%).There were 936 cases of esophageal cancer and precancerous lesions (3.08%), including 521 males and 415 females.Compared with women, 40-49 years old, high level education, drinking tap water, regular intake of meat, eggs and milk, and family average annual income more than 30 000 RMB, men (OR=1.90, 95%CI: 1.65-2.19), 60-69 years old (OR=5.28, 95%CI: 4.11-7.30), primary school education or below (OR=1.50, 95%CI: 1.20-1.89), drinking groundwater (OR=1.71, 95%CI: 1.38-2.13), never eating meat, eggs and milk (OR=1.48, 95%CI: 1.22-1.80), and family average annual income less than 30 000 RMB (OR=1.41, 95%CI: 1.16-1.70) would increase the risk of esophageal cancer and precancerous lesions. Conclusion The gender, age, educational level, annual household income, drinking water source, the frequency of eating meat, egg and milk were related to the occurrence of esophageal cancer and precancerous lesions among 40-69 years old residents in rural areas of Shandong Province.

Objective To analyze the related factors of esophageal squamous cell carcinoma and precancerous lesions among residents aged 40-69 years old in rural areas of Shandong Province. Methods In October 2018, 300 villages in 13 counties of the Shandong upper gastrointestinal cancerearly diagnosis and treatment projectin 2017 were selected as research areas, and 30 400 residents aged 40-69 were recruited in this study. The demographic characteristics, health status and lifestyle information were collected through the questionnaire survey, and endoscope iodine staining and indicative biopsy methods were used for cancer screening among eligible people.The multivariate logistic regression model was used to analyze the risk factors for esophageal cancer and precancerous lesions. Results The subjects in this study were (56.42±7.24) years old, including 13 193 males (43.40%).There were 936 cases of esophageal cancer and precancerous lesions (3.08%), including 521 males and 415 females.Compared with women, 40-49 years old, high level education, drinking tap water, regular intake of meat, eggs and milk, and family average annual income more than 30 000 RMB, men (OR=1.90, 95%CI : 1.65-2.19), 60-69 years old (OR=5.28, 95%CI : 4.11-7.30), primary school education or below ( OR=1.50, 95%CI : 1.20-1.89), drinking groundwater (OR=1.71, 95%CI: 1.38-2.13), never eating meat, eggs and milk (OR=1.48, 95%CI: 1.22-1.80), and family average annual income less than 30 000 RMB ( OR=1.41, 95%CI : 1.16-1.70) would increase the risk of esophageal cancer and precancerous lesions. Conclusion The gender, age, educational level, annual household income, drinking water source, the frequency of eating meat, egg and milk were related to the occurrence of esophageal cancer and precancerous lesions among 40-69 years old residents in rural areas of Shandong Province.

Objective To analyze the related factors of esophageal squamous cell carcinoma and precancerous lesions among residents aged 40-69 years old in rural areas of Shandong Province. Methods In October 2018, 300 villages in 13 counties of the Shandong upper gastrointestinal cancerearly diagnosis and treatment projectin 2017 were selected as research areas, and 30 400 residents aged 40-69 were recruited in this study. The demographic characteristics, health status and lifestyle information were collected through the questionnaire survey, and endoscope iodine staining and indicative biopsy methods were used for cancer screening among eligible people.The multivariate logistic regression model was used to analyze the risk factors for esophageal cancer and precancerous lesions. Results The subjects in this study were (56.42±7.24) years old, including 13 193 males (43.40%).There were 936 cases of esophageal cancer and precancerous lesions (3.08%), including 521 males and 415 females.Compared with women, 40-49 years old, high level education, drinking tap water, regular intake of meat, eggs and milk, and family average annual income more than 30 000 RMB, men (OR=1.90, 95%CI : 1.65-2.19), 60-69 years old (OR=5.28, 95%CI : 4.11-7.30), primary school education or below ( OR=1.50, 95%CI : 1.20-1.89), drinking groundwater (OR=1.71, 95%CI: 1.38-2.13), never eating meat, eggs and milk (OR=1.48, 95%CI: 1.22-1.80), and family average annual income less than 30 000 RMB ( OR=1.41, 95%CI : 1.16-1.70) would increase the risk of esophageal cancer and precancerous lesions. Conclusion The gender, age, educational level, annual household income, drinking water source, the frequency of eating meat, egg and milk were related to the occurrence of esophageal cancer and precancerous lesions among 40-69 years old residents in rural areas of Shandong Province.

Objective To investigate the influencing factors and their weights of professional choice among medical school graduates in Shanghai.Methods Base on literature review and interviews with medicine students in a university of Shanghai,a questionnaire of"employment intention for general practice in medical graduates"was developed.A questionnaire survey on influencing factors of general practice among five year medical students was conducted in the medical college of a university in Shanghai during May to June 2016.The weights of each factor were calculated with analytic hierarchy process(AHP). Results The valid questionnaires were retrieved from 14 male and 26 female students.The results showed that 53.8%(21/40)preferred Beijing, Shanghai, Guangzhou and other economically developed cities as future working place,and 38.5%(15/40)chose to go home town; for general practice 22.5%(9/40) chose"very like",70%(28/40)chose"like"and 7.5%(3/40)chose"not like"; only 10%(4/40) were willing to be a general practitioner, 55.0%(22/40)were depended on the working conditions of community health center,and 35%(14/40)were not ready to be a GP.AHP analysis showed that the weights of influencing factors were: the salary(0.380 2), the doctor-patient relationship(0.262 3),professional interest and occupation development prospects(0.220 4),social status and family contribution (0.137 1).Weight analysis for the affecting factors on professional interest and occupation development showed that the guidance of mentors in theoretical study and clinical practice were the major factors (comprehensive weight was 0.067 3 and 0.062 1,respectively),and the attitude GPs to general practice in community learning was more important(comprehensive weight was 0.042 5 and 0.019 6).Conclusion The willingness to be a general practitioner is not strong in the medical school graduates in Shanghai.It is important to improve the status and working conditions of general practitioners, and to strengthen education and guidance of the general practice in theoretical study and clinical practice for medical students.

Objective: To analyze the trend of lung cancer incidence rate among rural residents in Feicheng city between the years 2000 and 2012,and predict the incidence rate between the years 2013 and 2018,and subsequently provide baseline data for lung can-cer control and prevention.Methods:With the cancer registration data in Feicheng rural areas,the time trend of lung cancer inci-dence rate and the annual percentage change(APC)were calculated by the Joinpoint model,while the incidence of lung cancer from 2013 to 2018 were predicted by the ARIMA(p,d,q)model.Results:A total of 3,908 new cases of lung cancer were diagnosed be-tween 2000 to 2012.The incidence rate was 40.77/105,the age-specific cancer incidence rate in the Chinese population(ASRC)and world population(ASRW)were 32.95/105 and 32.97/105,respectively.The incidence was 2.14 times higher among males than females (P<0.001).The incidence of lung cancer which apparently rose from 25.13/105to 64.92/105 with an APC value of 9.74%(P<0.001),was increasing every year.The change in the trend of lung cancer with respect to age could be divided into three segments,the incidence rate in the 0 to 59 years group showed a rapid upward trend(APC=113.38,P<0.001),which was lower in the 60 to 79 years group (APC=20.39,P<0.05)and began to decline in the 80 years or older group(APC=-21.20,P>0.05).The incidence of lung cancer was also observed to be increasing yearly from 2013 to 2018,and with an average annual growth rate of 4.92%,was predicted to reach 87.92/105 in 2018.Conclusions:The occurrence of lung cancer was closely related to population aging,unhealthy habits,and environmental risk factors.Due to the increasing aging population,the incidence of lung cancer will continue to increase.In order to formulate specif-ic strategies,the control and prevention of lung cancer must be based on its incidence features.

PURPOSE: c-Met and its ligand, hepatocyte growth factor (HGF), play a critical role in oncogenesis and metastatic progression. The aim of this study was to identify inhibited enzymogram and to test the antitumor activity of SIM-89 (a c-Met receptor tyrosine kinase inhibitor) in non-small cell lung cancer. MATERIALS AND METHODS: Z′-LYTE kinase assay was employed to screen the kinase enzymogram, and mechanism of action (MOA) analysis was used to identify the inhibited kinases. Cell proliferation was then analyzed by CCK8 assay, and cell migration was determined by transwell assay. The gene expression and the phosphorylation of c-Met were examined by realtime-PCR and western blotting, respectively. Finally, the secretion of HGF was detected by ELISA assay. RESULTS: c-Met, activated protein kinase (AMPK), and tyrosine kinase A (TRKA) were inhibited by SIM-89 with the IC₅₀ values of 297 nmol/L, 1.31 µmol/L, and 150.2 nmol/L, respectively. SIM-89 exerted adenosine triphosphate (ATP) competitive inhibition on c-Met. Moreover, the expressions of STAT1, JAK1, and c-Met in H460 cells were decreased by SIM-89 treatment, and c-Met phosphorylation was suppressed in A549, H441, H1299, and B16F10 cells by the treatment. In addition, SIM-89 treatment significantly decreased the level of HGF, which accounted for the activation of c-Met receptor tyrosine kinase. Finally, we showed cell proliferation inhibition and cell migration suppression in H460 and H1299 cells after SIM-89 treatment. CONCLUSION: In conclusion, SIM-89 inhibits tumor cell proliferation, migration and HGF autocrine, suggesting it's potential antitumor activity.
Adenosine Triphosphate ; Blotting, Western ; Carcinogenesis ; Carcinoma, Non-Small-Cell Lung* ; Cell Movement ; Cell Proliferation ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Hepatocyte Growth Factor ; Lung Neoplasms ; Phosphorylation ; Phosphotransferases ; Protein Kinases ; Protein-Tyrosine Kinases