Objective To explore the optimal conditions for stable transfer of reporter gene containing enhanced green fluorescent protein(EGFP)into human breast cancer cells MDA-MB-231 cells under different electrotransfection conditions.Methods A PiggyBac(PB)transposon system containing EGFP reporter gene was constructed,and transfection conditions such as waveform,voltage,shock time,shock times,plasmid concentration,cell density,transposon to transposase ratio were controlled during electrotransfection of MDA-MB-231 cells.The transfection efficiency was detected by flow cytometry fluorescein isothiocyanate(FITC)channel.The expression of fluorescent protein was observed under laser scanning confocal microscope,the electrotransfection efficiency and expression of EGFP reporter gene were verified,and the electrotransfer efficiency of MDA-MB-231 cells was analyzed under different electrotransfection conditions.Results The optimal transfection conditions of MDA-MB-231 cells were as follows:voltage 280V,exponential wave,electric shock once,concentration of transposon plasmid about 1 000 ng/μl,mass ratio of transposon to transpotase 1:1,number of cells 2×106,electrotransfection rate up to 60.23%±5.63%.The state of MDA-MB-231 cells was good.Conclusion The electrotransfection conditions of MDA-MB-231 cells were successfully optimized to achieve stable and efficient transfection,which provided efficient electrotransfer experimental parameters for the relevant basic research of MDA-MB-231 cell line.

The importance of gait assessment in the rehabilitation of cancer patients is gradually being recognized. However, quantitative analysis of balance ability in cancer patients is still limited. A total of 102 cancer patients meeting the inclusion criteria were recruited from Hefei Cancer Hospital, Chinese Academy of Sciences. Their balance ability was evaluated using the Berg Balance Scale (BBS). Gait data were collected by an electronic walkway and an IMU sensor system, including spatial-temporal and kinematic gait features such as step length, cadence, support time, and range of motion. Recursive feature elimination was used for feature selection. Ridge, Elastic Net, SVR, RF, and AdaBoost models were used to predict balance ability scores. Five-fold cross-validation was used to evaluate the performance of these models. Results show that the SVR model achieves the best performance with fifteen features (RMSE=3.22, R ²=0.91), followed by Ridge (RMSE=3.63, R ²=0.89). A method for evaluating balance ability based on gait features is proposed, providing a quantitative tool for personalized rehabilitation interventions in cancer patients.
Humans ; Postural Balance ; Neoplasms/rehabilitation* ; Gait ; Gait Analysis ; Biomechanical Phenomena ; Female

Energy metabolism reprogramming is one of the pathological mechanisms involved in the occurrence and development of tumors.This article focused on the theory of spleen deficiency in TCM and energy metabolism reprogramming to elaborate the pathological mechanism of tumors.TCM believes that spleen deficiency is an important organ pathogenesis that causes cancer due to deficiency,and spleen deficiency runs through the occurrence and development of tumors.The spleen is the source of qi and blood transformation and dominates the energy metabolism of the whole body.Mitochondria are the regulatory center of energy metabolism in the body,and abnormal mitochondrial energy metabolism is the manifestation of spleen deficiency at the cellular micro level,which is mainly manifested as the limitation of mitochondrial oxidative phosphorylation and the conversion to aerobic glycolysis to provide energy for the rapid growth of tumors.Therefore,based on the theory of spleen deficiency,the treatment based on spleen strengthening is used to prevent and treat tumors by improving energy metabolism,which not only enriches the modern medical connotation of spleen deficiency in TCM,but also provides a new perspective for TCM to treat malignant tumors from the perspective of spleen strengthening.

Objective To construct CD19-targeted chimeric antigen receptor natural killer(CAR-NK)cells armored with interleukin15(IL15)or IL15 receptor-linker(RLI)and preliminarily validate their proliferative capacity and anti-tumor activity in vitro.Methods Natural killer cells(NK92 and NK92MI)from patients with human malignant non-Hodgkin lymphoma were cultured,and CD19-targeted CAR-NK cells armored with IL15 or RLI were prepared using retroviral vector particles.IL15 secretion was measured by ELISA,and proliferative capacity was assessed via CFSE dilution assays.Human B-lymphocytic leukemia cells(Nalm6-GFP-Luc)and human colon cancer cells overexpressing CD19(hCD19-SW620-GFP-Luc)were cultured,with surface CD19 expression confirmed(>99%positivity for both).Anti-tumor activity was evaluated by measuring cytotoxicity at effector-to-target(E:T)ratios using luciferase-based assays(4/12 h),detecting surface CD107a expression via flow cytometry,and quantifying cytokine release using CBA assays.Results NK92/NK92MI-CD19 CAR cells armored with IL15 or RLI were successfully generated.IL15 secretion was significantly higher in armored groups versus non-armored controls(P<0.01).Without IL-2 stimulation,IL15/RLI enhanced CAR-NK proliferation(P<0.05).Both armored designs significantly increased tumor-killing efficiency(P<0.05)and CD107a degranulation.IL15/RLI-armored NK92-CD19 CAR cells exhibited elevated release of IL2,IL10,IL6,TNF-α,sFas,IFN-γ,and Granulysin(P<0.05),while armored NK92MI-CD19 CAR cells showed additional increases in Granzyme A,Granzyme B,and Perforin(P<0.05).Conclusion IL15/RLI-armored NK92/NK92MI-CD19 CAR cells demonstrate potent anti-tumor activity,supporting their combined clinical therapeutic potential for tumors.

Objective To explore the optimal conditions for stable transfer of reporter gene containing enhanced green fluorescent protein(EGFP)into human breast cancer cells MDA-MB-231 cells under different electrotransfection conditions.Methods A PiggyBac(PB)transposon system containing EGFP reporter gene was constructed,and transfection conditions such as waveform,voltage,shock time,shock times,plasmid concentration,cell density,transposon to transposase ratio were controlled during electrotransfection of MDA-MB-231 cells.The transfection efficiency was detected by flow cytometry fluorescein isothiocyanate(FITC)channel.The expression of fluorescent protein was observed under laser scanning confocal microscope,the electrotransfection efficiency and expression of EGFP reporter gene were verified,and the electrotransfer efficiency of MDA-MB-231 cells was analyzed under different electrotransfection conditions.Results The optimal transfection conditions of MDA-MB-231 cells were as follows:voltage 280V,exponential wave,electric shock once,concentration of transposon plasmid about 1 000 ng/μl,mass ratio of transposon to transpotase 1:1,number of cells 2×106,electrotransfection rate up to 60.23%±5.63%.The state of MDA-MB-231 cells was good.Conclusion The electrotransfection conditions of MDA-MB-231 cells were successfully optimized to achieve stable and efficient transfection,which provided efficient electrotransfer experimental parameters for the relevant basic research of MDA-MB-231 cell line.

Energy metabolism reprogramming is one of the pathological mechanisms involved in the occurrence and development of tumors.This article focused on the theory of spleen deficiency in TCM and energy metabolism reprogramming to elaborate the pathological mechanism of tumors.TCM believes that spleen deficiency is an important organ pathogenesis that causes cancer due to deficiency,and spleen deficiency runs through the occurrence and development of tumors.The spleen is the source of qi and blood transformation and dominates the energy metabolism of the whole body.Mitochondria are the regulatory center of energy metabolism in the body,and abnormal mitochondrial energy metabolism is the manifestation of spleen deficiency at the cellular micro level,which is mainly manifested as the limitation of mitochondrial oxidative phosphorylation and the conversion to aerobic glycolysis to provide energy for the rapid growth of tumors.Therefore,based on the theory of spleen deficiency,the treatment based on spleen strengthening is used to prevent and treat tumors by improving energy metabolism,which not only enriches the modern medical connotation of spleen deficiency in TCM,but also provides a new perspective for TCM to treat malignant tumors from the perspective of spleen strengthening.

Objective:To investigate the prognostic impact of different tumor invasion patterns in the surgical treatment of T3 glottic laryngeal cancer. Methods:A retrospective analysis was conducted on the clinical data of 91 patients with T3 glottic laryngeal cancer. Results:Among the 91 patients, 58 cases （63.7%） had anterior invasion and 33 cases （36.3%） had posterior invasion. The posterior invasion was significantly correlated with invasions of the dorsal plate of cricoid cartilage （P<0.001）, arytenoid cartilage （P= 0.001）, and subglottic region（P = 0.001）. There was no statistical difference in survival outcomes between the total laryngectomy group and the partial laryngectomy group. But in the partial laryngectomy group, the 5-year disease-free survival（DFS） of patients with anterior invasive tumors was better than that of patients with posterior invasion tumors （HR: 4.681, 95%CI 1.337-16.393, P=0.016）, and subglottic invasion was associated with worse loco-regional recurrence-free survival（LRRFS）（HR: 3.931, 95%CI 1.054-14.658, P=0.041）. At the same time, we found that involvement of the dorsal plate of cricoid cartilage was an independent risk factor for postoperative laryngeal stenosis in partial laryngectomy patients （HR:11.67, 95%CI 1.89-71.98,P=0.008）. Conclusion:Compared with total laryngectomy, selected partial laryngectomy can also achieve favorable oncological outcomes. Posterior invasion and subglottic extension are independent prognostic factors for recurrence of partial laryngectomy in T3 glottic laryngeal cancer, and the involvement of the dorsal plate of cricoid cartilage is associated with postoperative laryngeal stenosis. The tumor invasion pattern of laryngeal cancer should be further subdivided in order to select a more individualized treatment plan.
Humans ; Prognosis ; Laryngeal Neoplasms/pathology* ; Retrospective Studies ; Laryngostenosis/surgery* ; Carcinoma, Squamous Cell/pathology* ; Postoperative Complications/surgery* ; Laryngectomy

Malnutrition in liver cirrhosis is associated with ascites, hepatic encephalopathy, infection, and other complications even death. To date, the hazard and disease burden of malnutrition in cirrhosis patients have been severely underestimated. This review summarized the most recent advancement in the field and discussed the techniques and methodologies in detection and evaluation of malnutrition in cirrhosis patients, nutritional support therapy, and future research directions and clinical care of the patients.

Objective To analyze the characteristics of death and disease burden of residents in Pingshan District of Shenzhen City from 2019 to 2020 and provide data support for disease prevention and control. Methods The data of death surveillance and demography were collected. The causes of death were classified and coded according to ICD-10. The crude mortality, standardized mortality and potential years of life lost were calculated. Results From 2019 to 2020, 910 people died in Pingshan district.The average age of death was 64.94(47.06 - 82.34) years, the crude mortality was 102.04/100 000, the standardized mortality was 263.97/100 000, and the average life expectancy was 86.00 years. 558 men died and 352 women died , the crude mortality were 113.72/100 000 and 87.76 /100 000 , the standardized mortality rate 313.05/100 000 and 211.97/100 000 ; the average life expectancy were 84.66 years and 87.55 years . The crude mortality of male was higher than that of female (χ²=14.594, P<0.001). The standardized mortality of men was also higher than that of women. The top three causes of death in the whole population, men and women were circulatory system diseases, malignant tumors, diseases and injury from high to low. And the top three diseases with standardized potential life lost years and standardized potential life lost rate from high to low were injury, circulatory system diseases and malignant tumors. Conclusion Circulatory system diseases, malignant tumors, injury are the main causes of death and the three kinds of diseases with the heaviest disease burden in Pingshan District of Shenzhen city. Men are the key population for prevention and control. The prevention and control of the above three kinds of diseases should be done to reduce the mortality of local population.

Objective: To observe the effect of moxibustion on the expression of N-methyl-D-aspartic acid (NMDA) receptor subtype 2B (NR2B) in the hippocampus of rheumatoid arthritis (RA) rats, and to explore the analgesic mechanisms of moxibustion in RA treatment. Methods: Sixty male Sprague-Dawley rats were randomly divided into a normal group, a model group, a moxibustion group, a moxibustion + NMDA receptor antagonist (AP-5) group, and a moxibustion + NMDA receptor agonist (NMDA) group, with 12 rats in each group. Except for the normal group, rats in the other four groups were treated with complete Freund's adjuvant in a windy, cold, and damp environment to replicate RA models. Rats in the moxibustion group received suspended moxibustion with moxa sticks at Shenshu (BL23) and Zusanli (ST36), and the two points were used alternately. After intraperitoneal injection of AP-5 or NMDA, rats in the moxibustion + AP-5 group and the moxibustion + NMDA group received the same moxibustion intervention as in the moxibustion group, once a day for 15 d. The thermal withdrawal latency (TWL) of rats in each group was detected before and after modeling and after the 15-day intervention. After the 15-day intervention, hematoxylin-eosin staining was performed to observe the pathological changes in knee joints. The real-time fluorescence quantitative polymerase chain reaction method was used to detect the mRNA expression of NR2B in the hippocampus; Western blotting assay was used to detect the protein and the phosphorylated protein expression of hippocampal NR2B. Results: The synovial tissue was proliferated, the synovial lining was significantly thickened, the pannus was formed, and the cartilage and bone tissues were significantly damaged in the model group. After intervention, the pathological morphology of the knee joints in the moxibustion group, the moxibustion + AP-5 group, and the moxibustion + NMDA group was significantly improved, and the improvement in the moxibustion + AP-5 group was more notable than that in the moxibustion + NMDA group. Compared with the normal group, the TWL was significantly decreased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the model group (P<0.01). Compared with the model group, the TWL of each intervention group was significantly increased (P<0.01 or P<0.05), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased (P<0.01). Compared with the moxibustion group, the TWL was significantly increased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased in the moxibustion + AP-5 group (P<0.01); the TWL was significantly decreased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the moxibustion + NMDA group (P<0.01). Conclusion: Moxibustion reduces hyperalgesia in RA inflammatory rats. The analgesic effect may be related to the decrease in the expression and phosphorylation levels of NR2B in the hippocampus.