Objective To study the application of autologous natural killer(NK)cells in the treatment of advanced renal cell carcinoma and the changes of immune function and tumor markers in patients.Methods 61 patients with advanced renal cell carcinoma admitted to Gansu Wuwei Tumour Hospital from March 2023 to April 2024 were divided into targeting group(31 cases,given supportive treatment combined with sunitinib malate capsules)and cell group(30 cases,autologous NK cells combined with targeting group)according to the patient's willingness to treat combined with propensity score matching.6 weeks was a treatment cycle,and all patients were treated for 4 cycles.The clinical efficacy of the two groups after 4 cycles of treatment and the occurrence of adverse reactions during treatment were statistically analyzed.The levels of serum cytokines,tumor markers,lymphocyte function and immune function were compared between the two groups before and after 4 cycles of treatment.Results After 4 cycles of treatment,the objective remission rate and disease control rate in the cell group were higher than those in the targeting group(P<0.05).After 4 cycles of treatment,the levels of serum hypoxia-inducible factor-1α(HIF-1α),vascular endothelial growth factor(VEGF),platelet-derived growth factor(PDGF),carcinoembryonic antigen,carbohydrate antigen 125,carbohydrate antigen 199 and alpha-fetoprotein in the two groups were lower than those before treatment,and those in the cell group were lower than those in the targeting group(P<0.012 5).After 4 cycles of treatment,the levels of serum interleukin-2,interleukin-6,tumor necrosis factor-β,interferon-γ,peripheral blood CD3+,CD4+,NK cells and CD4+/CD8+in the two groups were higher than those before treatment.The levels of serum interleukin-2,tumor necrosis factor-β,interferon-γ,peripheral blood CD3+and NK cells in the cell group were higher than those in the targeting group(P<0.012 5).The level of CD8+in peripheral blood was lower than that before treatment(P<0.012 5),but there was no significant difference between the two groups(P>0.012 5).During the treatment,there was no significant difference in the incidence of diarrhea,nausea and vomiting,alopecia,liver function damage,decreased platelet level and decreased neutrophil level between the cell group and the targeting group(P>0.05).Conclusion The treatment of advanced renal cell carcinoma with autologous NK cells could improve the level of serum cytokines,reduce the level of tumor markers,regulate the function of lymphocytes and immune function,and had a good therapeutic effect.At the same time,it would not increase the incidence of adverse reactions,and the safety was good.

Objective To investigate the role and molecular mechanisms of bladder cancer-derived exosomal long non-coding RNA(lncRNA)CIAT1 in mediating collective invasion and to evaluate its clinical significance and potential therapeutic value.Methods High-throughput sequencing was used to identify lncRNAs that are highly expressed in exosomes from bladder cancer and lymph node metastatic tissues.CIAT1 was selected for further validation in clinical bladder cancer samples.By constructing CIAT1-overexpressing and knockdown bladder cancer cells,we demonstrated in vitro that CIAT1-contained exosomes target cancer-associated fibroblasts(CAFs)to induce collective invasion.The underlying mechanism of CIAT1 in bladder cancer collective invasion was explored through RNA pull-down,RNA immunoprecipitation(RIP),dual-luciferase reporter assays,chromatin isolation by RNA purification(ChIRP)and chromatin immunoprecipitation(ChIP).Results CIAT1 was significantly upregulated in exosomes derived from bladder cancer tissues compared to adjacent normal tissues by High-throughput sequencing(fold change>1.5,P<0.05)and clinical sample validation(P<0.01).In vitro experiments,exosomal CIAT1 was selectively internalized by cancer-associated fibroblasts(CAFs),significantly enhancing collective invasion of bladder cancer via regulating CAFs.In co-culture models,CIAT1 overexpression group showed increased total number and total length of collective invasion chains compared to the control group(P<0.01 for both).Mechanistically,CIAT1 was packaged into exosomes via binding to hnRNPA2B1,and internalized by CAFs,where it activated N-cadherin transcription by modulating H3K4me3 histone modification at the N-cadherin promoter.Consistently,the CIAT1 overexpression group exhibited elevated collective invasion chain numbers and lengths compared to the control group(P<0.01 for both).However,blocking N-cadherin reversed the pro-invasive effects of CIAT1,with no significant differences in chain numbers or lengths between the CIAT1 overexpression+N-cadherin blockade group and controls(P>0.05 for both).Further clinical correlation analysis confirmed that CIAT1-regulated N-cadherin is closely associated with collective invasion in bladder cancer patients(P<0.01).Conclusions Exosomal CIAT1 derived from bladder cancer cells targets CAFs to activate N-cadherin transcription,thereby promoting bladder cancer collective invasion.

Objective To investigate the role and molecular mechanisms of bladder cancer-derived exosomal long non-coding RNA(lncRNA)CIAT1 in mediating collective invasion and to evaluate its clinical significance and potential therapeutic value.Methods High-throughput sequencing was used to identify lncRNAs that are highly expressed in exosomes from bladder cancer and lymph node metastatic tissues.CIAT1 was selected for further validation in clinical bladder cancer samples.By constructing CIAT1-overexpressing and knockdown bladder cancer cells,we demonstrated in vitro that CIAT1-contained exosomes target cancer-associated fibroblasts(CAFs)to induce collective invasion.The underlying mechanism of CIAT1 in bladder cancer collective invasion was explored through RNA pull-down,RNA immunoprecipitation(RIP),dual-luciferase reporter assays,chromatin isolation by RNA purification(ChIRP)and chromatin immunoprecipitation(ChIP).Results CIAT1 was significantly upregulated in exosomes derived from bladder cancer tissues compared to adjacent normal tissues by High-throughput sequencing(fold change>1.5,P<0.05)and clinical sample validation(P<0.01).In vitro experiments,exosomal CIAT1 was selectively internalized by cancer-associated fibroblasts(CAFs),significantly enhancing collective invasion of bladder cancer via regulating CAFs.In co-culture models,CIAT1 overexpression group showed increased total number and total length of collective invasion chains compared to the control group(P<0.01 for both).Mechanistically,CIAT1 was packaged into exosomes via binding to hnRNPA2B1,and internalized by CAFs,where it activated N-cadherin transcription by modulating H3K4me3 histone modification at the N-cadherin promoter.Consistently,the CIAT1 overexpression group exhibited elevated collective invasion chain numbers and lengths compared to the control group(P<0.01 for both).However,blocking N-cadherin reversed the pro-invasive effects of CIAT1,with no significant differences in chain numbers or lengths between the CIAT1 overexpression+N-cadherin blockade group and controls(P>0.05 for both).Further clinical correlation analysis confirmed that CIAT1-regulated N-cadherin is closely associated with collective invasion in bladder cancer patients(P<0.01).Conclusions Exosomal CIAT1 derived from bladder cancer cells targets CAFs to activate N-cadherin transcription,thereby promoting bladder cancer collective invasion.

Objective To study the application of autologous natural killer(NK)cells in the treatment of advanced renal cell carcinoma and the changes of immune function and tumor markers in patients.Methods 61 patients with advanced renal cell carcinoma admitted to Gansu Wuwei Tumour Hospital from March 2023 to April 2024 were divided into targeting group(31 cases,given supportive treatment combined with sunitinib malate capsules)and cell group(30 cases,autologous NK cells combined with targeting group)according to the patient's willingness to treat combined with propensity score matching.6 weeks was a treatment cycle,and all patients were treated for 4 cycles.The clinical efficacy of the two groups after 4 cycles of treatment and the occurrence of adverse reactions during treatment were statistically analyzed.The levels of serum cytokines,tumor markers,lymphocyte function and immune function were compared between the two groups before and after 4 cycles of treatment.Results After 4 cycles of treatment,the objective remission rate and disease control rate in the cell group were higher than those in the targeting group(P<0.05).After 4 cycles of treatment,the levels of serum hypoxia-inducible factor-1α(HIF-1α),vascular endothelial growth factor(VEGF),platelet-derived growth factor(PDGF),carcinoembryonic antigen,carbohydrate antigen 125,carbohydrate antigen 199 and alpha-fetoprotein in the two groups were lower than those before treatment,and those in the cell group were lower than those in the targeting group(P<0.012 5).After 4 cycles of treatment,the levels of serum interleukin-2,interleukin-6,tumor necrosis factor-β,interferon-γ,peripheral blood CD3+,CD4+,NK cells and CD4+/CD8+in the two groups were higher than those before treatment.The levels of serum interleukin-2,tumor necrosis factor-β,interferon-γ,peripheral blood CD3+and NK cells in the cell group were higher than those in the targeting group(P<0.012 5).The level of CD8+in peripheral blood was lower than that before treatment(P<0.012 5),but there was no significant difference between the two groups(P>0.012 5).During the treatment,there was no significant difference in the incidence of diarrhea,nausea and vomiting,alopecia,liver function damage,decreased platelet level and decreased neutrophil level between the cell group and the targeting group(P>0.05).Conclusion The treatment of advanced renal cell carcinoma with autologous NK cells could improve the level of serum cytokines,reduce the level of tumor markers,regulate the function of lymphocytes and immune function,and had a good therapeutic effect.At the same time,it would not increase the incidence of adverse reactions,and the safety was good.

Objective:To investigate the clinical characteristics and prognosis of cryptococcal meningitis patients with anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) autoantibodies.Methods:A total of 216 non-acquired immunodeficiency syndrome (AIDS) related cryptococcal meningitis cases with positive cultures of Cryptococcus, hospitalized at Huashan Hospital, Fudan University during January 2014 and December 2021, were retrospectively included. The serum anti-GM-CSF autoantibodies were detected by enzyme linked immunosorbent assay, and the clinical characteristics and prognosis were compared between patients with and without anti-GM-CSF autoantibodies. Statistical comparisons were mainly performed using the chi-square test or Fisher′s exact test. Cox proportional-hazards model was used to analyze the risk factors associated with prognosis. Results:Among 216 enrolled patients, 23 patients were positive of anti-GM-CSF autoantibodies, with a positive rate of 10.6%. Among 23 patients, seven cases were infected with Cryptococcus gattii, and 16 cases were infected with Cryptococcus neoformans. In the group with positive anti-GM-CSF autoantibodies, 30.4%(7/23) of the patients were infected with Cryptococcus gattii, which was higher than that of 1.6%(3/193) in the group with negative anti-GM-CSF autoantibodies, and the difference was statistically significant ( χ2=38.82, P<0.001). In the group with positive anti-GM-CSF autoantibodies, 30.0% (6/20) had mass lesions with a diameter greater than three centimeters in the lungs, and the one-year all-cause mortality rate was 50.0% (10/20), which were both higher than those of 3.4%(5/145) and 16.1% (29/180) in the negative group, respectively. The differences were both statistically significant (both Fisher′s exact test, P<0.01). Age≥60 years (hazard ratio ( HR)=4.146, P=0.002), predisposing factors ( HR=3.160, P=0.021), epilepsy ( HR=6.129, P=0.002), positive anti-GM-CSF autoantibodies ( HR=2.675, P=0.034), white blood cell count of cerebrospinal fluid (CSF)<100 ×10 6/L ( HR=2.736, P=0.039), the titers of cryptococcal capsular polysaccharide antigen of CSF≥1∶1 280 ( HR=4.361, P=0.009) were independent risk factors for one-year all-cause mortality in patients with cryptococcal meningitis. Conclusions:In non-AIDS related cryptococcal meningitis patients, the positive rate of serum anti-GM-CSF autoantibodies is as high as 10.6%. Patients with anti-GM-CSF autoantibodies could be infected with both Cryptococcus neoformans and Cryptococcus gattii, and they have higher proportion of lung mass lesions than patients with negative anti-GM-CSF autoantibodies. The one-year survival rate decreases significantly in patients with anti-GM-CSF autoantibodies, which is an independent risk factor for the prognosis of cryptococcal meningitis.

Objective: To preliminarily investigate the difference of position fixation accuracy between polyurethane styrofoam and vacuum negative pressure pad in intensity-modulated radiation therapy (IMRT) after radical mastectomy for breast cancer. Methods: Forty breast cancer patients, who received breast-conserving surgery followed by hyper-fractionated IMRT of the whole breast (42.56 Gy for 16 times) in our hospital between 2017 and 2018 were recruited and randomly divided into the polyurethane styrofoam group and vacuum negative pressure pad group. Before IMRT treatment, the anterior and lateral films of patients were taken with kilovoltage digital radiographs (KVDRs) by Varian Trilogy machine-borne OBI KV image verification system. The KVDRs images were matched with the DRR images reconstructed by the planned system to obtain the setup errors in the left and right, head and foot, and ventral and back directions between two groups. Each patient was verified for 10 times to obtain 400 sets of data. The independent sample t-test was adopted to analyze the setup errors between two groups. The external expansion value of graded setup errors from clinical target volume (CTV) and planned target volume (PTV) was calculated. Results: The setup errors in the left and right, head and foot, ventral and back directions between the styrofoam fixation and vacuum pad groups were (1.63±1.29) mm and (1.83 ±1.61) mm (P=0.18), (1.46±1.51) mm and (2.26±2.03) mm (P=0.00), and (1.30±1.35) mm and (1.91±1.67) mm (P=0.00), respectively. The external expansion values of setup errors from CTV to PTV were 2.19 mm, 2.51 mm, 1.57 mm and 2.40 mm, 3.97 mm and 2.63 mm, respectively. Conclusion Both two fixation methods meet the clinical requirements. However, the setup accuracy and reproducibility in the polyurethane styrofoam group are better than those in the vacuum negative pressure pad group.

Objective To investigate the difference of styrofoam and breast carrier in postposition fixation of intensity-modulated radiotherapy after breast conservative surgery for breast cancer patients.Methods From February 2018 to August 2018,tweenty-four patients with breast cancer in Sun Yet-Sen Memorial Hospital of Sun Yet-sen University were selected for this study,who underwent hypofactionationed radiotherapy after breast conservative surgery with total dose 42.56 Gy/16Fractions.They were randomized into styrofoam test group and breastcarrier control group.Cone beam CT as used to record the positioning error under the directions of left and right (x),head and foot (y),abdomen and back (y) within two groups at the first,third,fifth,seventh,eleventh time before irradiation.Furthermore,the PTV extension margin was calculated and the positioning time of two groups was recorded.Two sets of pendulum errors were analyzed by independent sample T-test,and the outspread value of inter-fractional set up error of the PTV was calculated.Results The errors of the test group and the control group in the direction of x,y,z were as follows:(2.36±1.89) and (2.56±2.05) mm (P=0.49),(1.76± 1.78) and (3.28±2.79) mm (P＜0.05),(1.47± 1.49) and (1.73± 1.81) mm (P=0.28).The extension values of inter-fractional set up error of CTV to PTV were 2.97,2.92,2.21 mm and 3.41,4.09,2.59 mm respectively.The time of single positioning was (3.4± 1.1) and (5.5 ± 3.1) min respectively (P=0.01).Conclusion Styrofoam has better positioning accuracy and efficiency compared with breast carrier.

Breast cancer is the most common cancer for Chinese women. Early screening is the best way to improve the rates of early diagnosis and early treatment of breast cancer. The peak ages of breast cancer in Chinese women are obviously different from those in the European and American countries. It is imperative to develop a guideline for breast cancer screening that is suitable for Chinese women. Based on the analysis and summary of breast cancer screening data in China, and the latest guidelines and consensus on breast cancer screening in Europe, the United States and East Asia, China Anti-Cancer Association and National Clinical Research Center for Cancer (Tianjin Medical University Cancer Institute and Hospital) has developed a population-based guideline for breast cancer screening in Chinese women. This guideline has provided detailed recommendations on the screening starting age, screening modalities, and screening interval in Chinese women with average risk and high risk of breast cancer, respectively. This article aims to interpret the above guideline, providing references for professionals in breast cancer screening.

ObjectiveTo investigate the intervention effects of moxibustion and moxa smoke on blood lipids,hepatic pathological changes and intrahepatocytic molecules related to cholesterol metabolism and analyze the regulating effects of moxibustion and moxa smoke on cholesterol metabolism and explore the mechanisms of actions of moxibustion and moxa smoke. MethodFifty-one 8-week-old ApoE-/-mice were randomized into model, moxa smoke and moxibustion groups, 17 mice each. Twenty C57BL/6 mice comprised a blank control group. The normal and model groups of mice were routinely grabbed and fastened. The moxa smoke group of mice was exposed to 10-15 mg/m3moxa smoke circumstances. The moxibustion group of mice was given moxibustion on point Guanyuan(CV4). All interventions were made 20 min daily, 6 times a week, for 12 consecutive weeks. Total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) were measured using an automatic biochemical analyzer. Hepatic pathologic morphology was observed by HE staining. Hepatic CD36 and ABCA1 expressions were determined by immunohistochemical method.ResultIn the model group of mice, serum TG and LDL-C contents were significantly higher than in the normal group (P=0.003;P=0.001);HDL-C content was significantly lower than in the normal group (P=0.007); TC content had no significant difference compared with the normal group (P>0.05). In the moxibustion group of mice, serum TG and LDL-C contents were significantly lower than in the model group (P=0.03;P=0.001) and HDL-C content had no significant difference compared with the model group (P=0.11). In the moxa smoke group of mice, serum TG and LDL contents were significantly lower than in the model group (P=0.01;P=0.008) and HDL content had no significant difference compared with the model group (P=0.11). There were no significant differences in various blood lipid indicators between the moxibustion and moxa smoke groups (P>0.05). There were hepatic cell cord and sinusoid derangement and obvious hepatocytic swelling in the model group of mice. In the moxa smoke and moxibustion groups, hepatocytic swelling subsided significantly, and inflammatory cell infiltration reduced compared with the model group. In the model group,CD36 expression was significantly higher than in the normal group (P=0.004) and ABCA1 expression was significantly lower than in the normal group (P=0.001). In the moxibustion group, CD36 expression had no significant difference compared with the model group (P=0.09) and ABCA1 expression was significantly higher than in the model group (P=0.03). In the moxa smoke group, CD36 expression was significantly lower than in the normal group (P=0.02) and ABCA1 expression was significantly higher than in the model group (P=0.002). There were no significant differences in CD36 and ABCA1 expressions between the moxibustion and moxa smoke groups (P>0.05).ConclusionEarly moxibustion on point Guanyuan can regulate disorders of blood lipid metabolism, delay the occurrence of hepatic lesions and reduce intrahepatic accumulation of cholesterol to a certain extent in an ApoE-/-mouse model of atherosclerosis. That may be one of the mechanisms by which moxibustion therapy prevents atherosclerosis. Moxa smoke as the product of moxibustion is an effective factor in moxibustion producing a therapeutic effect.

OBJECTIVETo investigate whether inhalable particulate matters can cause the damage of chromosome or mitotic apparatus to produce micronucleus, and to evaluate genetic toxicology of moxa smoke on chromosome.

METHODSBy MTT method, the 24 h half maximal inhibitory concentration (IC50) of moxa smoke condensation (MSC) on Chinese hamster ovary (CHO) cells was 0.087 mg/mL. CHO cells, which were cultured in vitro, were divided into a solvent control group, a positive control group (cyclophosphamide as solvent), a low concentration group, a moderate concentration group and a high concentration group. The low concentration group, moderate concentration group and high concentration group were set approximately 1/8, 1/4, 1/2 of IC50, respectively. Whether micronucleus had dose-effect response induced by the damage of chromosome or mitotic apparatus was observed after CHO cells were contaminated by MSC in the low concentration group, moderate concentration group and high concentration group.

RESULTSThe rate of micronucleus induced by MSC in the low concentration group, moderate concentration group and high concentration group was higher than that in the solvent control group (all P < 0.05), which presented dosage-effect response. The experiment was repeated 3 times, indicating it was repeatable with statistical significance.

CONCLUSIONHigh concentration of MSC shows toxicity to induce chromosome damage, which disappears at low concentration. The genetic toxicology is also dependent on concentration, and the concentration of moxa smoke is essential. In clinical treatment, it is noted to control the level of moxa smoke, while the clinical safety standard of moxa smoke concentration is in need of further study.

Air Pollutants ; adverse effects ; Animals ; CHO Cells ; Cell Nucleus ; drug effects ; genetics ; Cricetinae ; Cricetulus ; Inhalation Exposure ; adverse effects ; analysis ; Micronucleus Tests ; Moxibustion ; adverse effects ; Particulate Matter ; adverse effects ; Smoke ; adverse effects ; analysis