1.Identification of the effect of cisplatin on the transcriptome of human hepatocellular carcinoma cell lines
Xin GUO ; Mengdie JI ; Qi WANG ; Xueyuan LI ; Yang CHEN
Basic & Clinical Medicine 2024;44(3):352-360
Objective To investigate the effect of cisplatin treatment on the transcriptional level of human liver cancer cells by conducting transcriptome sequencing analysis after treating human liver cancer cell lines with differ-ent concentrations of cisplatin(CDDP).Methods Liver cancer cell lines HepG2 and Huh7 were incubated with cisplatin at different final concentrations of 0,20,50,100 and 200 μmol/L.After 12 hours,cell viability,immuno-fluorescence and RNA-sequencing(RNA-seq)were performed.Differential gene expression analysis(DEG),KEGG pathway analysis,and protein-protein interaction network analysis were conducted.Results Cisplatin de-creased cell viability and increased DNA damage in HepG2,Huh7 cells.Among the genes regulated after cisplatin treatment at different concentrations,59 genes were commonly up-regulated in both HepG2 and Huh7 cells,while 81 genes were commonly down-regulated.The commonly upregulated genes were mainly enriched in cancer initiation and progression pathways.The 81 commonly down-regulated genes were mainly enriched in Rap1 signaling pathway,Ras signaling pathway,signaling pathways regulating pluripotency of stem cells,axon guidance,and cell adhesion-related pathways.Survival analysis of key nodes in the protein-protein interaction network of commonly up-regulated and downregulated genes revealed a significant correlation between high expression of Jun proto-oncogene,AP-1 transcription factor subunit(JUN)and prolonged patient survival and a significant correlation between low ex-pression of growth arrest and DNA damage inducible alpha(GADD45A)and prolonged patient survival.Conclu-sions The study revealed common transcriptional changes in liver cancer cells under cisplatin treatment.Differential expression of JUN and GADD45A is a potential core mechanism to explain drug resistance.This conclusion provides some important prognostic indicators for clinical treatment.
2.The effect of Bushen Huoxue prescription on proliferation and extracellular matrix synthesis of nucleus pulposus cells by regulating circ_0036763/miR-583 axis
Canrong LIANG ; Junjie HE ; Xueyuan ZHU ; Haojian FENG ; Weijun GUO ; Dongling CAI ; Chenghua ZHU ; Xiaofeng CHEN
Journal of Chinese Physician 2024;26(1):63-69
Objective:To explore the possible effects of Bushen Huoxue Formula (the kidney tonifying and blood activating prescription) on the proliferation and extracellular matrix synthesis of nucleus pulposus cells by regulating the circ_0036763/miR-583 axis.Methods:Real time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of circ0036763 and miR-583 in normal and intervertebral disc degeneration (IDD) nucleus pulposus cells; IDD nucleus pulposus cells were divided into pcDNA group, pcDNA circ_0036763 group, pcDNA circ_0036763+ mimic NC group, and pcDNA circ_0036763+ miR-583 mimic group. qRT-PCR was used to detect the expression levels of circ_0036763 and miR-583 in nucleus pulposus cells in each group, methyl thiazolyl tetrazolium (MTT) was used to detect cell proliferation (A value), and Western blot was used to detect the expression of proliferating cell nuclear antigen (PCNA), collagen Ⅰ, and collagen Ⅱ proteins in nucleus pulposus cells, The dual luciferase assay reported experimental validation of the targeting relationship between circ_0036763 and miR-583. 27 mice were divided into sham surgery group, IDD group, and kidney tonifying and blood activating formula group. IDD models were established in all groups except for the sham surgery group. After successful modeling, the sham surgery group and IDD group were given physiological saline by gavage, while the kidney tonifying and blood activating formula group was given 1.5 g/ml of kidney tonifying and blood activating formula by gavage for 3 consecutive weeks. QRT-PCR was used to detect the expression levels of circ0036763 and miR-583 in the nucleus pulposus cells of mice in each group, MTT was used to detect cell proliferation, and Western blot was used to detect the expression of PCNA, collagen Ⅰ, and collagen Ⅱ proteins.Results:The expression level of circ_0036763 in IDD nucleus pulposus cells decreased, while the expression level of miR-583 increased (all P<0.05); Overexpression of circ_0036763 can promote proliferation and extracellular matrix synthesis of nucleus pulposus cells (all P<0.05); Circ_0036763 targets miR-583 and upregulates miR-583 reversible overexpression. Circ_0036763 enhances the proliferation and extracellular matrix synthesis ability of IDD nucleus pulposus cells. Compared with the sham surgery group, the IDD group showed an increase in collagen Ⅰ protein expression and miR-583 expression levels (all P<0.05), while the cell A value, PCNA and collagen Ⅱ protein expression, and circ_0036763 expression levels decreased (all P<0.05); Compared with the IDD group, the Kidney Tonifying and Blood Activating Formula group showed a decrease in collagen Ⅰ protein expression and miR-583 expression levels (all P<0.05), while the cell A value, PCNA and collagen Ⅱ protein expression, and circ_0036763 expression levels increased (all P<0.05). Conclusions:The kidney tonifying and blood activating formula (Bushen Huoxue) may induce proliferation and extracellular matrix synthesis of nucleus pulposus cells by regulating the circ_0036763/miR-583 axis.
3.Dynamic transcriptomic analysis of macrophages infected with Salmonella typhimurium
Boyuan SONG ; Xueli WU ; Xueyuan LI ; Lisha WANG ; Yang CHEN
Basic & Clinical Medicine 2024;44(6):779-785
Objective To comprehensively understand the dynamic transcriptional landscape during infection through investigating the temporal molecular changes in macrophages RAW 264.7 upon infection with Salmonella typhimurium SL1344.Methods Macrophages RAW 264.7 were infected with Salmonella typhimurium SL1344,and cell samples were collected at 0 h,8 h,and 16 h for RNA-sequencing(RNA-seq).Upstream and downstream analyses of the transcriptome data including differential gene expression,clustering,functional annotation,and mo-lecular network studies were conducted to elucidate the signaling pathways changes in macrophages.Results Infec-ted macrophages exhibited significant morphological and transcriptional changes.Differential gene analysis identified significant upregulation and downregulation patterns.Clustering revealed six gene clusters involving various signaling pathways,such as immune response,membrane transport,and lipid catabolic process.Conclusions Macrophages dynamically respond to Salmonella typhimurium infection,displaying distinct temporal gene expression patterns.The coordinated activation of immune response,membrane transport,and lipid catabolic process pathways implies a multifaceted cellular adaptation to external infections,providing essential insights into the molecular mechanisms of macrophage response to Salmonella typhimurium infection.
4.Progress in the treatment of free flap for repairing fingertip defects
Guoliang WU ; Xueyuan LI ; Hong CHEN
Chinese Journal of Microsurgery 2024;47(3):350-355
Fingertip injuries often involve the loss of skin, soft tissue, and sometimes even phalange, seriously affecting the patient's hand function and appearance. Therefore, fingertip reconstruction requires both functional recovery and aesthetics. And how to better restore fingertip defect has become an issue that hand surgeons should continue to explore. In recent years, free flaps have made remarkable progress in the field of fingertip repair, and free flaps such as radial artery superficial palmar branch free flap, free thenar flap and free toe flap provide a variety of effective methods to solve the problem. By referring to the literatures on free flap in fingertip repair, this paper summarises the application status and research progress of free flap in fingertip defect reconstruction, providing a reference for personalised treatment of patients with fingertip defect.
5.Postoperative hypoalbuminemia in patients with acute ischemic stroke with successful recanalization by endovascular therapy: risk factors and their impact on outcomes
Hao LU ; Qiang GUO ; Yongnan HAO ; Dongxu YANG ; Ya’nan CHEN ; Zhi ZHU ; Xueyuan LI
International Journal of Cerebrovascular Diseases 2023;31(10):721-727
Objective:To investigate the risk factors of hypoalbuminemia (HA) in patients with acute ischemic stroke (AIS) caused by large vessel occlusion (LVO) after endovascular mechanical thromboectomy (EMT) and impact on outcomes.Methods:Patients first diagnosed with anterior circulation LVO-AIS and underwent emergency EMT at the Department of Acute Stroke, the Affiliated Hospital of Jining Medical University from June 1, 2020 to April 31, 2023 were retrospectively included. According to the first follow-up serum albumin examination after procedure (6-8 d), the patients were divided into HA group (<35 g/L) and non-HA group (≥35 g/L). According to the modified Rankin Scale score at 90 d after EMT, the patients were divided into a good outcome group (0-2) and a poor outcome group (3-6). Univariate and multivariate logistic analysis was used to determine independent risk factors for HA after EMT and their impact on outcomes. Results:A total of 144 patients were enrolled, including 107 males (74.30%) with a median age of 64 years (interquartile range, 56-71 years). There were 50 patients (34.72%) in the HA group and 94 (65.28%) in the non-HA group; 60 (41.67%) in the good outcome group, and 84 (58.33%) in the poor outcome group. Multivariate logistic regression analysis showed that age (odds ratio [ OR] 1.061, 95% confidence interval [ CI] 1.014-1.111; P=0.011) and pulmonary infection ( OR 5.136, 95% CI 1.917-13.760; P=0.001) were independent risk factors for HA; HA ( OR 4.345, 95% CI 1.367-13.814; P=0.013), pneumonia ( OR 5.113, 95% CI 1.217-12.528; P=0.026), and onset to reperfusion time ( OR 5.473, 95% CI 1.090-16.05; P=0.038) were independent risk factors for poor outcomes. Conclusions:Age and pulmonary infection are independent risk factors for HA in LVO-ASI patients after EMT, and HA is the independent risk factor for poor outcomes of the patients.
6.Study on activity conditions screening and mechanism of Panax ginseng-Poria cocos pair against aging
Zhulin BAI ; Daqing ZHAO ; Jingjing CHEN ; Jing ZENG ; Xueyuan BAI ; Siming WANG
China Pharmacy 2022;33(21):2584-2589
OBJECTIVE To screen the best compatibility ratio and administration conditions of Panax ginseng -Poria cocos pair against aging ,and investigate its mechanism . METHODS P. ginseng-P. cocos pair extracts with different compatibility ratios (1∶1,1∶2,2∶1,1∶4,4∶1,m/m)were prepared ;taking Saccharomyces cerevisiae as the aging model organism ,the S. cerevisiae growth curve was drawn by MTT method ,the best compatibility ratio ,administration concentration and administration time point of P. ginseng-P. cocos pair were screened out ;the activities of antioxidant related enzymes [superoxide dismutase (SOD),peroxidase (POD),catalase(CAT)],the levels of reactive oxygen species (ROS)and malondialdehyde (MDA),the content of adenosine triphosphate(ATP),and the mitochondrial membrane potential (MMP)in S. cerevisiae cells were detected ;mRNA expressions of SOD1,CTT1,GSH1,ATP1,MRS1 and CDC 19 were also detected . RESULTS The optimal ratio of P. ginseng-P. cocos pair for anti-aging activity was 1∶4(m/m),the optimal administration concentration was 220 μg/mL,and the optimal administration time point was the 28th hour . The extracts of P. ginseng-P. cocos pair(1∶4,m/m)could significantly increase the activities of SOD , POD and CAT ,ATP content ,MMP,mRNA expression of CTT 1,GSH1 and MRS 1(P<0.01),but decrease the levels of MDA and ROS ,mRNA expressions of SOD 1,ATP1 and CDC 19(P<0.05 or P<0.01). CONCLUSIONS P. ginseng-P. cocos pair(1∶4, m/m)has a good anti -aging effect on S. cerevisiae ,its mechanism may be related to the positive regulation of oxidative stress and energy metabolism of S. cerevisiae cell.
8.Free anterolateral thigh perforator flap pedicled with the oblique branch of lateral circumflex femoral artery assisted by three-dimensional CT angiography for repairing soft tissue defects of limbs
Haoliang HU ; Hong CHEN ; Miaozhong LI ; Xueyuan LI
Chinese Journal of Trauma 2021;37(9):780-785
Objective:To investigate the clinical effect of the anterolateral femoral perforator flap(ALTP)pedicled with the oblique branch of lateral circumflex femoral artery(LCFA)assisted by CT angiography(CTA)examination for repairing soft tissue defects of limbs.Methods:A retrospective case series study was made on 51 patients with soft tissue defects of limbs treated in Ningbo No.6 Hospital from March 2015 to March 2020,including 31 males and 20 females at age of 26-63 years[(42.0±8.9)years]. The defects were located at the forearm in 15 patients,at the hand in 13,at the lower leg in 15 and at the ankle in 8. The size of defects ranged from 9 cm×6 cm to 18 cm×10 cm,with the size of flaps from 10 cm×6 cm to 20 cm×12 cm. A total of 33 patients were examined with CTA scanning and Doppler ultrasound(CTA group)and 18 patients with Doppler ultrasound(Doppler group). All patients underwent debridement and negative pressure closed drainage(VSD)at stage I and were repaired by ALTP pedicled with the oblique branch of LCFA at stage II. The diameter and length of the vessel pedicle was recorded in CTA group before operationand in both groups during operation. The time of flap harvesting in both groups was recorded during operation. The survival of the flap in both groups was observed one week after operation. Zhang Hao's scoring standard was applied to evaluate the outcome at the last follow-up.Results:All patients were followed up for 6-12 months[(9.1±1.5)months]. In CTA group,the diameter of LCFA vessel pedicle measured before operation had no significant difference from that during operation( P>0.05),while the length of LCFA vessel pedicle before operation[(12.3±2.1)cm]was longer than(10.9±2.2)cm during operation( P<0.05). The two group showed no significant differences in the diameter and length of LCFA vessel pedicle during operation( P>0.05). The time of flap harvesting in CTA group was(38.5±6.2)minutes,significantly shorter than(51.4±8.4)minutes in Doppler group( P<0.05). One week after operation,all flaps survived. Two patients developed flap arterial congestion in CTA group,among whom one survived after surgical revision and one with partially necrosis was healed after dressing change. One patient was found with flap arterial congestion with partial necrosis in Doppler group,who was healed after dressing change. There was no significant difference in postoperative flap arterial congestion between the two groups( P>0.05). The patients' satisfaction score in CTA group was(8.5±1.5)points at the last follow-up,higher than(7.4±2.0)points in Doppler group( P<0.05). Conclusion:For repairing soft tissue defects of limbs,free ALTP pedicled with the oblique branch of LCFA assisted by three- dimensional CT angiography can accurately get the information of perforator,shorten the flap harvesting time,and obtain satisfactory clinical results as compared to Doppler ultrasound.
9.Application of end-to-side anastomosis in personalised free ilioinguinal flap transfer
Keyue YANG ; Jian HUANG ; Chuan CHEN ; Heyang SUN ; Yi LI ; Xueyuan LI ; Hong CHEN ; Xin WANG ; Jianwu QI
Chinese Journal of Microsurgery 2021;44(4):403-407
Objective:To investigate the clinical application and effect of end-to-side anastomosis in personalised free ilioinguinal flap transfer.Methods:From March, 2015 to July, 2020, 88 patients with soft tissue (bone) defect of limbs were treated. Different ilioinguinal flaps were designed according to the wound condition of patients, which were 48 cases of free superficial circumflex iliacartery perforator flap, 7 cases of free superficial epigastric artery perforator flap, 19 cases of composite tissue flap with iliac bone, 8 cases of combined flap of superficial circumflex iliac artery and superficial abdominal wall artery, and 6 cases of superficial circumflex iliac artery and superficial abdominal wall artery lobulated flap. The area of the flap was 4.0 cm×6.0 cm-10.0 cm×30.0 cm. The artery and recipient artery were anastomosed end-to-side: 36 cases to radial artery; 12 cases to ulnar artery; 18 cases to dorsalis pedis artery; 15 cases to anterior tibial artery; 7 cases to posterior tibial artery. Venous anastomosis of skin flap: 42 cases were anastomosed with 2 veins, which were superficial vein of the same name and accompanying vein; 46 cases were anastomosed with 1 superficial vein of the same name. The accompanying vein of the flap was anastomosed end-to-side with the accompanying vein of the main artery of the recipient area, and the superficial vein of the same name was anastomosed end-to-end with the accompanying vein or subcutaneous superficial vein of the recipient artery. Follow-up includes flap blood supply, blood supply to the distal limbs, appearance of both the donor site and the recipient area, and patient satisfaction.Results:There were 83 cases of flaps survived successfully, and 5 cases of crisis. Among them, 2 cases had artery crisis at 48 h after surgery. After exploration, it was found that 1 case caused by arterial thrombosis, and 1 case compressed by the stapler that anastomoses the vein. The other 3 cases had venous crisis at 72 h after surgery: after exploration, it was found that caused by thrombosis at the venous anastomotic site. The average follow-up period was 10 (range, 3-24) months. All flaps survived after re anastomosis or vascular transposition. The donor site and recipient site of the flap healed well. The blood supply of the flap was good and the texture was soft. There was no blood supply disorder in the distal limb.Conclusion:The end-to-side anastomosis technique is suitable for all kinds of free flap transplantation in ilioinguinal region, with high vascular patency rate. It can not only solve the problem of thin vascular pedicle of donor site flap, but also retain the main artery of recipient limb without affecting the distal blood supply.
10. Research status of long non-coding RNA MALAT1 in breast cancer
Dandan SONG ; Xueyuan CHEN ; Ruiqi CHEN ; Hanqiao LI ; Tian WU ; Dandan SONG ; Xueyuan CHEN ; Ruiqi CHEN ; Hanqiao LI ; Tian WU
Chinese Journal of Clinical Pharmacology and Therapeutics 2021;26(1):49-57
Long non-coding RNA (LncRNA) is a kind of RNA longer than 200nt, whose abnormal expression plays a significant role in the development of tumors. Metastasis-associated lung adenocarcinoma transcription 1 (MALAT1), as one of LncRNAs, is closely related to the pathogenesis of breast cancer. MALAT1 can affect the tumorigenesis and progression of breast cancer and is expected to be an effective target for the diagnosis and treatment of breast cancer. Herein, we review the research status of MALAT1 in breast cancer.

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