1.The mechanism of extract of ginkgo biloba inducing mitochondrial autophagy in breast cancer cells MCF-7
Jianqiang SHAO ; Peng WANG ; Jie BAI ; Huixin LI ; Zunyi WANG ; Zhihong XU
Journal of International Oncology 2024;51(2):65-72
Objective:To investigate the mechanism of extract of ginkgo biloba (EGB) on mitochondrial autophagy in breast cancer cells MCF-7.Methods:Breast cancer MCF-7 cells were divided into four groups. EGB with mass concentrations of 40, 80, 120 mg/L was used to incubate breast cancer MCF-7 cells for 24 h or 48 h, as a low concentration group of EGB, a medium concentration group of EGB, and a high concentration group of EGB. Breast cancer MCF-7 cells without intervention were taken as control group. Cell proliferation was measured using MTT assay; Flow cytometry was used to detect cell apoptosis; Immunofluorescence assay was used to determine the contents of prostacyclin (P62), microtubule-associated protein light chain 3Ⅱ (LC3Ⅱ), and caspase-3; The levels of multidrug resistance-associated protein 1 (MRP1), multidrug resistance gene 1 (MDR1) and breast cancer resistance protein (BCRP) were identified by PCR; Western blotting was used to detect the expression of extracellular signal-regulated kinase (ERK), mitogen-activated protein kinase (MAPK), p-ERK, and p-MAPK proteins in cells.Results:The results of MTT assay for cell proliferation showed that cell proliferation at 24 h in control group, EGB low, medium and high concentration groups were 0.95±0.14, 0.65±0.09, 0.51±0.07, 0.37±0.04, respectively, with a statistically significant difference ( F=43.13, P<0.001), cell proliferation at 48 h were 1.32±0.19, 0.54±0.08, 0.32±0.05, 0.15±0.02, respectively, with a statistically significant difference ( F=141.30, P<0.001). Compared with 24 h, cell proliferation was decreased in EGB low, medium and high concentration groups at 48 h (all P<0.05). Pairwise comparison showed that EGB treatment significantly decreased MCF-7 cell viability and cell proliferation was decreased in turn at 24 and 48 h in control group, low, medium, high EGB groups (all P<0.05). Flow cytometry analysis revealed that the apoptosis rates of MCF-7 cells in control group, EGB low, medium and high concentration groups were 2.12%±0.23%, 9.28%±0.45%, 15.17%±1.28% and 22.21%±2.32%, respectively, with a statistically significant difference ( F=128.80, P<0.001). Pairwise comparison showed that the apoptosis rate of control group, EGB low, medium and high concentration groups were increased in turn (all P<0.05). The results of immunofluorescence assay showed that the protein relative expression levels of P62 protein in MCF-7 cells of control group, EGB low, medium and high concentration groups were 3.34±0.52, 2.85±0.47, 2.02±0.18 and 1.08±0.21, respectively, with a statistically significant difference ( F=41.55, P<0.001). LC3Ⅱ protein relative expression levels were 0.24±0.05, 1.02±0.14, 1.47±0.26, 1.95±0.21, respectively, with a statistically significant difference ( F=94.82, P<0.001). The relative expression levels of caspase-3 protein were 0.25±0.03, 0.68±0.21, 1.12±0.17 and 1.65±0.23, respectively, with a statistically significant difference ( F=68.09, P<0.001). Pairwise comparison showed that LC3Ⅱ and caspase-3 protein expression levels were increased in turn in control group, EGB low, medium and high concentration groups, while P62 protein expression levels were decreased in turn (all P<0.05). The PCR experiment results showed that the MRP1 mRNA level of MCF-7 cells in control group, EGB low, medium and high concentration groups were 1.06±0.14, 0.83±0.18, 0.71±0.11, 0.52±0.08, respectively, with a statistically significant difference ( F=17.41, P<0.001). The mRNA levels of MDR1 were 1.14±0.17, 0.75±0.13, 0.60±0.09, 0.48±0.06, respectively, with a statistically significant difference ( F=34.40, P<0.001). BCRP mRNA levels were 1.09±0.11, 0.88±0.13, 0.69±0.07, 0.57±0.05, respectively, with a statistically significant difference ( F=34.13, P<0.001). Pairwise comparison showed that the levels of MRP1, MDR1 and BCRP mRNA were decreased in turn in control group, EGB low, medium and high concentration groups (all P<0.05). The results of Western blotting showed that the expression of ERK in MCF-7 cells in control group, EGB low, medium and high concentration groups were 2.54±0.38, 1.89±0.25, 1.55±0.21, 1.12±0.16, respectively, with a statistically significant difference ( F=31.18, P<0.001). MAPK expression were 2.47±0.34, 1.96±0.29, 1.63±0.27, 1.20±0.24, respectively, with a statistically significant difference ( F=20.90, P<0.001). p-ERK expression were 2.03±0.29, 1.74±0.21, 1.45±0.11, 1.18±0.24, respectively, with a statistically significant difference ( F=16.31, P<0.001). p-MAPK expression were 2.26±0.47, 1.90±0.41, 1.61±0.33, 1.35±0.16, respectively, with a statistically significant difference ( F=7.01, P=0.002). Pairwise comparison showed that the expressions of ERK, MAPK, p-ERK and p-MAPK in control group, EGB low, medium and high concentration groups were decreased in turn (all P<0.05) . Conclusion:EGB can inhibit the proliferation of breast cancer MCF-7 cells, promote the apoptosis of MCF-7 cells, decrease the expression of P62 protein, increase the expression of LC3Ⅱ and caspase-3 protein, induce mitochondrial autophagy.
2.Sensitivity of colorectal cancer organoids to hyperthermic intraperitoneal chemotherapy with lobaplatin
Duo LIU ; Hui WANG ; Weihao DENG ; Jianqiang LAN ; Zhiwen SONG ; Yu ZHU ; Jianling JING ; Jian CAI
Chinese Journal of Gastrointestinal Surgery 2024;27(5):486-494
Objective:To investigate the sensitivity of tumor organoids derived from samples of colorectal cancer to lobaplatin and oxaliplatin hyperthermic perfusion in vitro and to assist clinical development of hyperthermic intraperitoneal chemotherapy. Method:Tumor samples and relevant clinical data were collected from patients with pathologically confirmed colorectal cancer in the Sixth Affiliated Hospital of Sun Yat-sen University from July 2021 to December 2022. Organoids were cultured and tumor tissue were passaged. In vitro hyperthermic perfusion experiments were performed on organoids with good viability. Firstly, 10 organoids were treated with oxaliplatin and lobaplatin at the following six concentrations: 1 000, 250, 62.5, 15.6, 3.9, and 0.98 μmol/L. The organoids were exposed to oxaliplatin at 42℃ for 30 minutes and to lobaplatin at 42℃ for 60 minutes. Dose-response curves of responses to in vitro hyperthermic perfusion with these two drugs were constructed and evaluated. Clinical doses of oxaliplatin and lobaplatin were further tested on 30 organoids. This testing revealed oxaliplatin was effective at 579 μmol/L at a hyperthermic perfusion temperature of 42℃ for 30 min and lobaplatin was effective at 240 μmol/L at a hyperthermic perfusion temperature of 42℃ for 60 minutes. Result:Thirty-two tumor organoids were cultured from samples of colorectal cancer. The median concentration required for oxaliplatin to eliminate 50% of tumor cells (IC50) was 577.45 μmol/L (IQR: 1846.09 μmol/L). The median IC50 for lobaplatin was 85.04 μmol/L (IQR: 305.01 μmol/L).The difference between the two groups was not statistically significant ( Z=1.784, P=0.084). In seven of 10 organoids, lobaplatin showed a greater IC50 after in vitro hyperthermic perfusion than did oxaliplatin. Testing of 30 organoids with clinical doses of oxaliplatin and lobaplatin revealed that oxaliplatin achieved an average inhibition rate of 39.6% (95%CI: 32.1%?47.0%), whereas the average rate of inhibition for lobaplatin was 89.7% (95%CI: 87.0%?92.3%): this difference is statistically significant ( t=?15.282, P<0.001). Conclusion:The rate of inhibition achieved by hyperthermic perfusion of lobaplatin in vitro is better than that achieved by hyperthermic perfusion with oxaliplatin. Lobaplatin is more effective than oxaliplatin when administered by hyperthermic intraperitoneal perfusion and therefore has the potential to replace oxaliplatin in this setting.
3.Standardized diagnosis and treatment of colorectal polyps
Renjie WANG ; Xiaolan ZHANG ; Jidong CAI ; Minghe WANG ; Jianqiang LIU ; Ye XU
Chinese Journal of Gastrointestinal Surgery 2024;27(6):583-590
This article explores the standardized management of colorectal polyps, including classification, treatment, follow-up, and preventive control. Corresponding treatment strategies, including endoscopic resection and surgical intervention, are employed for different types of polyps. Currently, there is debate over whether to choose endoscopic resection or surgical intervention for malignant polyps at pT1 stage. Drawing on the latest literature and guidelines, the article elaborates on polyp classification, treatment modalities, follow-up, and preventive measures. Standardized management of colorectal polyps is important for reducing the incidence of colorectal cancer and improving the cure rate of early-stage colorectal cancer.
4.Related factors of hypothyroidism induced by programmed death(PD)-1 treatment in elderly patients with cancer
Pan DU ; Ying YANG ; Fang WANG ; Jianqiang ZHU ; Xue ZHOU ; Cheng CHEN ; Lin KANG
Chinese Journal of Geriatrics 2024;43(5):592-596
Objective:To investigate the factors related to hypothyroidism induced by programmed death(PD)-1 treatment in elderly patients with cancer.Methods:A total of 193 older patients(≥60 years old)with advanced solid tumors who received PD-1 treatment between January 2018 and January 2021 at the Department of Oncology of Xiangyang Central Hospital were included in this study.The patients were divided into two groups based on whether they were diagnosed with hypothyroidism after PD-1 treatment: the hypothyroidism group(36 cases)and the non-hypothyroidism group(157 cases).The clinical data of both groups, including age, gender, Eastern Cooperative Oncology Group performance status(ECOG PS), PD-1 inhibitors, thyroid function, and thyroid antibody, were compared to analyze the risk factors associated with hypothyroidism.Results:Among the 193 patients, 36(18.7%)were diagnosed with hypothyroidism.The study found no significant differences between the two groups in terms of age, gender, ECOG PS, tumor type, and PD-1 type(all P>0.05).However, significant differences were observed in the baseline levels of thyroid stimulating hormone(TSH)and thyroid antibody subgroups(both P<0.05).The results of multivariate Logistic regression analysis revealed that the presence of baseline anti-thyroid peroxidase antibody(TPOAb)( OR=20.256, 95% CI: 5.709-71.868, P<0.001), the presence of both baseline thyroglobulin antibody(TGAb)and TPOAb( OR=5.853, 95% CI: 1.475-23.227, P=0.012), and an increase in baseline TSH levels( OR=3.065, 95% CI: 1.049-8.959, P=0.041)were identified as risk factors for hypothyroidism induced by PD-1 treatment.On the other hand, there was no significant association between the presence of baseline TGAb and the occurrence of hypothyroidism( OR=1.373, 95% CI: 0.353-5.341, P=0.648). Conclusions:The incidence rate of hypothyroidism induced by PD-1 inhibitors is high among elderly patients with cancer.Additionally, the risk of hypothyroidism is higher in patients with elevated baseline TSH and positive TPOAb.Therefore, it is crucial to remain vigilant for the occurrence of hypothyroidism during PD-1 treatment.Timely diagnosis and treatment of hypothyroidism are necessary to minimize the incidence of adverse events.
5.Protective effect of lycopene on rats with cognitive dysfunction induced by sevoflurane anesthesia through endoplasmic reticulum stress
International Journal of Biomedical Engineering 2024;47(4):349-355
Objective:To investigate the protective effect of lycopene on rats with cognitive dysfunction (CD) induced by sevoflurane anesthesia through endoplasmic reticulum stress (ERS).Methods:A rat CD model was established using sevoflurane anesthesia induction. SD rats were randomly divided into a sham operation group, model group, low-, high-dose (5 and 10 mg/kg) lycopene groups, and a lycopene + tunicamycin (10 mg/kg + 100 μg/kg) group, with 12 rats in each group. 5 and 10 mg of lycopene was dissolved in 1 ml of sodium carboxymethylcellulose to form a suspension, and 10 μg of clindamycin was dissolved in 1 and 2 ml of 0.1% dimethyl sulfoxide. Rats in the low-, high-dose lycopene groups were gavaged with 1 ml of lycopene suspension, respectively, and the rats in the lycopene + tunicamycin group were gavaged with 1 ml of lycopene suspension and received 1 ml intraperitoneal injection of clindamycin. The sham operation group and the model group received an equal amount of saline by gavage or intraperitoneal injection, respectively. The low-, high-dose lycopene groups received an equal amount of saline by intraperitoneal injection, both 1 time/d for 6 weeks. The Morris water maze test was used to determine the cognitive function of rats. HE staining was used to observe the morphological changes of rat hippocampal tissue. TUNEL staining was used to observe the apoptosis of neurons in rat hippocampal tissue. The ELISA method was used to detect the levels of brain-derived neurotrophic factor (BDNF) and S100 calcifying protein β (S100β) in hippocampal tissue. Western Blot was used to detect the levels of apoptosis-related proteins in rat hippocampal tissues, including the levels of B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) with ERS marker glucose-regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), and cysteinyl aspartate specific proteinase-12 (Caspase-12).Results:Compared with the model group, the escape latency, neuronal apoptosis rate, S100β level, GRP78, CHOP, Caspase-12, and Bax expression were decreased in the low-, high-dose lycopene groups on days 3, 4, and 5 (all P < 0.05), and the number of crossing platforms, BDNF level, and Bcl-2 expression were increased (all P < 0.05). With the increase of lycopene dose, the escape latency, neuronal apoptosis rate, S100β level, GRP78, CHOP, Caspase-12, and Bax expression were decreased in the high-dose lycopene group compared with the low-dose lycopene group on days 3, 4, and 5 (all P < 0.05), and the number of crossing platforms, BDNF level, and Bcl-2 expression were increased (all P < 0.05). Compared with the high-dose lycopene group, the escape latency, neuronal apoptosis rate, S100β level, GRP78, CHOP, Caspase-12, and Bax expression were increased in the lycopene + tunicamycin group on days 3, 4, and 5 (all P < 0.05), and the number of crossing platforms, BDNF level, and Bcl-2 expression were decreased (all P < 0.05). Hippocampal tissue neuronal loss and the inflammatory infiltration were both reduced in CD rats after lycopene intervention. The ERS activator tunicamycin attenuated the protective effect of lycopene on sevoflurane-induced CD rats. Conclusions:Lycopene may play a protective role against CD in rats anesthetized with sevoflurane by inhibiting ERS.
6.Sensitivity of colorectal cancer organoids to hyperthermic intraperitoneal chemotherapy with lobaplatin
Duo LIU ; Hui WANG ; Weihao DENG ; Jianqiang LAN ; Zhiwen SONG ; Yu ZHU ; Jianling JING ; Jian CAI
Chinese Journal of Gastrointestinal Surgery 2024;27(5):486-494
Objective:To investigate the sensitivity of tumor organoids derived from samples of colorectal cancer to lobaplatin and oxaliplatin hyperthermic perfusion in vitro and to assist clinical development of hyperthermic intraperitoneal chemotherapy. Method:Tumor samples and relevant clinical data were collected from patients with pathologically confirmed colorectal cancer in the Sixth Affiliated Hospital of Sun Yat-sen University from July 2021 to December 2022. Organoids were cultured and tumor tissue were passaged. In vitro hyperthermic perfusion experiments were performed on organoids with good viability. Firstly, 10 organoids were treated with oxaliplatin and lobaplatin at the following six concentrations: 1 000, 250, 62.5, 15.6, 3.9, and 0.98 μmol/L. The organoids were exposed to oxaliplatin at 42℃ for 30 minutes and to lobaplatin at 42℃ for 60 minutes. Dose-response curves of responses to in vitro hyperthermic perfusion with these two drugs were constructed and evaluated. Clinical doses of oxaliplatin and lobaplatin were further tested on 30 organoids. This testing revealed oxaliplatin was effective at 579 μmol/L at a hyperthermic perfusion temperature of 42℃ for 30 min and lobaplatin was effective at 240 μmol/L at a hyperthermic perfusion temperature of 42℃ for 60 minutes. Result:Thirty-two tumor organoids were cultured from samples of colorectal cancer. The median concentration required for oxaliplatin to eliminate 50% of tumor cells (IC50) was 577.45 μmol/L (IQR: 1846.09 μmol/L). The median IC50 for lobaplatin was 85.04 μmol/L (IQR: 305.01 μmol/L).The difference between the two groups was not statistically significant ( Z=1.784, P=0.084). In seven of 10 organoids, lobaplatin showed a greater IC50 after in vitro hyperthermic perfusion than did oxaliplatin. Testing of 30 organoids with clinical doses of oxaliplatin and lobaplatin revealed that oxaliplatin achieved an average inhibition rate of 39.6% (95%CI: 32.1%?47.0%), whereas the average rate of inhibition for lobaplatin was 89.7% (95%CI: 87.0%?92.3%): this difference is statistically significant ( t=?15.282, P<0.001). Conclusion:The rate of inhibition achieved by hyperthermic perfusion of lobaplatin in vitro is better than that achieved by hyperthermic perfusion with oxaliplatin. Lobaplatin is more effective than oxaliplatin when administered by hyperthermic intraperitoneal perfusion and therefore has the potential to replace oxaliplatin in this setting.
7.Standardized diagnosis and treatment of colorectal polyps
Renjie WANG ; Xiaolan ZHANG ; Jidong CAI ; Minghe WANG ; Jianqiang LIU ; Ye XU
Chinese Journal of Gastrointestinal Surgery 2024;27(6):583-590
This article explores the standardized management of colorectal polyps, including classification, treatment, follow-up, and preventive control. Corresponding treatment strategies, including endoscopic resection and surgical intervention, are employed for different types of polyps. Currently, there is debate over whether to choose endoscopic resection or surgical intervention for malignant polyps at pT1 stage. Drawing on the latest literature and guidelines, the article elaborates on polyp classification, treatment modalities, follow-up, and preventive measures. Standardized management of colorectal polyps is important for reducing the incidence of colorectal cancer and improving the cure rate of early-stage colorectal cancer.
8.A Preliminary Study on the Efficacy of Percutaneous Coronary Intervention for Complex Left Main Stem Combined With Chronic Total Occlusion of the Right Coronary Artery
Zun HU ; Hu HU ; Yubo LONG ; Junshan LI ; Jingjing RONG ; Jin HE ; Changlu WANG ; Yu ZHANG ; Jianqiang PENG ; Hongwei PAN
Chinese Circulation Journal 2024;39(6):562-567
Objectives:to analyze the efficacy of percutaneous coronary intervention(PCI)for complex left main(LM)lesions combined with chronic total occlusion(CTO)of the right coronary artery. Methods:Ninety patients with complex left main lesions hospitalized in Hunan Provincial People's Hospital from January 2019 to December 2022 were consecutively included.According to the coronary angiographic vascular lesions,patients were divided into complex left main lesions combined with right coronary artery CTO(observation group,n=30)and complex left main lesions without right coronary artery CTO(control group,n=60).The baseline clinical data,intraoperative conditions,angiographic results,and postoperative follow-up results of the patients were analyzed and compared between the two groups. Results:Fifty-eight(64.4%)out of the 90 patients were male.There was no statistically significant difference between the two groups in terms of baseline clinical data(all P>0.05),left main lesion condition(P=1.000),left main calcification condition(P=0.249),and preoperative TIMI flow grading(P=1.000).In the comparison between observation group and the control group,intraoperative occurrence of no-reflow(3.3%vs.5.0%,P=1.000),hypotension(10.0%vs.8.3%,P=1.000),pericardial effusion(3.3%vs.0%,P=0.333),the percentage of intravascular ultrasound(IVUS)use(86.7%vs.90.0%,P=0.635),and the use of circulatory assist device(P=0.699),and the proportion of intraoperative coronary spinning(26.7%vs.21.7%,P=0.597)were all similar between the two groups.The median follow-up time was 14.50(11.83,15.85)months,and the differences in the incidence of major adverse cardiovascular events(MACE)such as recurrent angina,acute myocardial infarction,rebleeding,readmission for heart failure,and cardiac death(31.0%vs.32.1%,P=1.000)were not statistically significant between the observation group and the control group. Conclusions:PCI revascularization may be a viable approach for elderly patients with complex LM lesions with multiple underlying disease,and combined right coronary artery CTO,intolerance and reluctance to CABG.
9.The effectiveness of treatment based on intestine on acute lung injury/acute respiratory distress syndrome caused by sepsis:a Meta analysis
Yuan LIU ; Jianqiang MEI ; Yan DONG ; Fenqiao CHEN ; Bu WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2024;31(1):6-13
Objective To systematically evaluate the clinical efficacy of treatment based on intestine in the sepsis induced acute lung injury/acute respiratory distress syndrome(ALI/ARDS)under the traditional Chinese medicine theory"interior and exterior relationship between the lung and large intestine",and to provide evidence-based reference for clinical practice.Methods A computerized search of CNKI,Wanfang database,VIP database,China Biology Medicine disc(CBMdisc),Embase database of Dutch medical abstracts,PubMed database,Cochrane library database was performed to identify randomized controlled trial(RCT)that used the Tongfu method as an intervention for the treatment of ALI/ARDS induced by sepsis,published from the database establishment until October 8,2020.The control group was treated with conventional western medicine,and the test group was treated with the combination of Tongfu method and conventional western medicine.Screening of the literature and data extraction for those that met the inclusion criteria were performed by 2 investigators.The modified Jadad scale was used for quality assessment,and RevMan 5.4 and Stata 16.0 software were used for Meta analysis of the data.Results Thirteen articles were finally included,all of which were single center RCT studies,including 7 articles with≥4 points and 6 articles with<4 points.A total of 579 patients were included,290 in the test group and 289 in the control group.Meta-analysis showed that combined Tongfu treatment significantly increased the ALI/ARDS patient caused by sepsis oxygenation index[PaO2/FiO2;mean difference(MD)= 62.55,95%confidence interval(95%CI)was 55.74 to 69.37,P<0.05],reduced the levels of interleukin-6(IL-6;MD =-29.70,95%CIwas-48.34 to-11.06,P<0.05),tumor necrosis factor-α(TNF-α;MD=-2.94,95%CIwas-5.28 to-0.59,P<0.05),procalcitonin(PCT;MD =-1.34,95%CI =-2.17 to-0.51,P<0.05)and C-reactive protein(CRP;MD =-22.41,95%CIwas-36.52 to-8.29,P<0.05),reduced mechanical ventilation time(MD=-2.26,95%CIwas-2.86 to-1.66,P<0.05)and hospitalization time in intensive care unit(ICU;MD=-4.15,95%CIwas-7.47 to-0.84,P<0.05),reduced the 28-day mortality[relative risk(RR)= 0.43,95%CI was 0.24 to 0.76,P<0.05]and acute physiology and chronic health status scoreⅡ(APACHEⅡ;MD =-3.53,95%CI was-5.41 to-1.64,P<0.05).Conclusion Treatment based on intestine is valuable for reducing the condition and improving the prognosis of ALI/ARDS patients caused by sepsis,but further high-quality RCT studies are still needed to verify its efficacy and provide more accurate evidence-based evidence for clinical treatment.
10.Expectation of research and application of human milk science in China
Ye WANG ; Xinxin XING ; Jianqiang LAI
Chinese Journal of Perinatal Medicine 2024;27(7):529-531
Human milk is a product of human evolution and is crucial for the early growth and development of infants as well as maternal health. This paper discusses the research and application of human milk science in China, aiming to clarify the research pathways in this field and contribute to the promotion of breastfeeding initiatives and the realization of the Healthy China 2030 goals.

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