Objective To investigate the effect of resveratrol on the tumor microenvironment in osteosarcoma. Methods A C57BL/6 xenograft mouse model was established and treated with resveratrol. Single-cell sequencing was performed to analyze changes in the tumor microenvironment. Immunohistochemistry was used to assess immune cell infiltration, while Western blotting was conducted to examine alterations in cellular signaling pathways. Results Resveratrol significantly inhibited the proliferation of LM8 osteosarcoma cells in C57BL/6 mice compared to the control group. Additionally, CD8⁺ T cell recruitment was enhanced. The Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway was notably downregulated in LM8 osteosarcoma cells following resveratrol treatment. Conclusion Resveratrol promotes CD8⁺ T cell infiltration by inhibiting the JAK2/STAT3 signaling pathway, suggesting its potential as a therapeutic agent in osteosarcoma treatment.
Osteosarcoma/genetics* ; STAT3 Transcription Factor/genetics* ; Resveratrol/pharmacology* ; Animals ; Janus Kinase 2/genetics* ; Signal Transduction/drug effects* ; Tumor Microenvironment/immunology* ; Cell Line, Tumor ; Mice, Inbred C57BL ; Mice ; Humans ; Cell Proliferation/drug effects* ; Bone Neoplasms/metabolism* ; CD8-Positive T-Lymphocytes/drug effects* ; Xenograft Model Antitumor Assays

Objective:To explore the clinical value of 18F-fluoromisonidazole (FMISO) PET/CT hypoxia imaging in early response to heavy ion radiotherapy in patients with non-small cell lung cancer(NSCLC). Methods:From April 2018 to January 2021, the 18F-FMISO PET/CT images of 23 NSCLC patients (19 males, 4 females; age (64.9±10.3) years) who received heavy ion radiotherapy in Shanghai Proton and Heavy Ion Center were retrospectively analyzed. The evaluation parameters included tumor volume (TV), tumor to background ratio (TBR) before and after radiotherapy. Patients were divided into hypoxia group and non-hypoxia group with the baseline TBR value≥1.4 as hypoxia threshold. Wilcoxon signed rank test was used to compare the differences of TV and TBR before and after radiotherapy in 2 groups. Results:Of 23 NSCLC patients, 17 were hypoxia and 6 were non-hypoxia. Compared with the baseline, TV after the radiotherapy (59.44(22.86, 99.43) and 33.78(8.68, 54.44) cm 3; z=-3.05, P=0.002) and TBR after the radiotherapy (2.25(2.09, 2.82) and 1.42(1.24, 1.67); z=-3.39, P=0.001) of the hypoxia group were significantly lower, while TV (16.19(6.74, 36.52) and 8.59(4.38, 25.47) cm 3; z=-1.57, P=0.120) and TBR (1.19(1.05, 1.27) and 1.10 (0.97, 1.14); z=-1.89, P=0.060) of the non-hypoxia group decreased with no significant differences. Conclusions:Hypoxic NSCLC tumors are sensitive to heavy ion radiation. Compared with non-hypoxic tumors, hypoxic tumors respond more quickly, and a significant reduction in TV can be observed early after radiotherapy. Heavy ion radiation can significantly improve tumor hypoxia.

Objective:To test the usefulness of PET-range verification (RV) method for proton radiation accuracy verification in poly (methyl methacrylate) (PMMA) phantom using off-line PET/CT scanning.Methods:Proton irradiation dose of 2 Gy and 4 Gy were delivered in PMMA phantom. Given the difference of clinical target volume (CTV), 7 subgroups with different depth (5.0, 7.5, 10.0, 12.5, 15.0, 17.5, 20.0 cm) were set for each dose (14 radiation plans or radiation fields). PET/CT scan was performed 10 min after irradiation of 48-221 MeV proton beam. A co-registration between CT from treatment planning system and PET/CT was performed, as well as the smoothing and normalization of PET/CT data. The region of interest (ROI) and profile lines were drawn with the Raystation PET-RV software. The predictive induced radioactivity and the measured induced radioactivity profile lines were analyzed to evaluate the Δ R50, namely, the error at the position corresponding to 50% of the maximum predictive induced radioactivity at the end of both curves. Results:The size of each ROI was 5.0 cm×5.0 cm×2.5 cm. Profile lines were evenly distributed with the interval of 3 mm, and totally 289 pairs of profile lines were drew. The 2 Gy- and 4 Gy-dose groups yielded similar mean depth errors (Δ R50 between 1 mm and -1 mm with a standard deviation <1 mm). Conclusions:The off-line PET/CT scanning of PMMA phantom reveals a good agreement between predicted and measured PET data, with error of ±1 mm. The PET-RV method can be extended to clinical cases′ verification in human body treatment with further investigation.

OBJECTIVES: To evaluate curative effects of coronavirus disease 2019 (COVID-19) patients by the transfusion of other convalescent plasma. METHODS: Retrospective analysis of the clinical data of 18 patients with severe and critical COVID-19, who were hospitalized in the ICU of Xianghu Branch of the First Affiliated Hospital of Nanchang University from February 1 to March 15, 2020. Patients were subdivided into an experimental group (=6, who had transfused the plasma) and an observation group (=12, who had no plasma transfusion). Basic clinical data and prognosis indexes of these two groups were compared. Moreover, for the experimental group, the dynamic changes of blood oxygen saturation before and after the transfusion, the changes of lymphocyte absolute value 48 hours after the transfusion, and the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid were analyzed. RESULTS: There were no significant differences in age, gender, blood type and other basic clinical data between the two groups (all >0.05).There were no significant differences in ventilator machine weaning time, extracorporeal membrane oxygenation (ECMO) weaning time, body temperature recovery to normal time, and hospitalization days between these two groups (all >0.05). For the experimental group, before, during and after the convalescent plasma transfusion, the blood oxygen saturation of all 6 patients at all time (1, 6, 8, 12, 24, 36, and 48 h) was more than 90%, and there was no significant fluctuation. There were 3 patients whose absolute value of lymphocyte was increased 48 hours after the transfusion, and the remaining was decreased. There were 5 patients whose SARS-CoV-2 nucleic acid detection turned negative 48 hours after the transfusion, accounting for 83.3%. CONCLUSIONS Transfusion of convalescent plasma will not affect outcomesof COVID-19 patients, which can neutralize SARS-CoV-2 in patients and reduce the loading capacity of SARS-CoV-2.
Betacoronavirus ; Blood Component Transfusion ; China ; Coronavirus Infections ; therapy ; Humans ; Immunization, Passive ; Pandemics ; Plasma ; Pneumonia, Viral ; therapy ; Retrospective Studies

Objective To investigate the regulation effects of Ngn2 gene transfection on retinal neuron differnetion in three-dimentional optic vesicle (OV) of mice.Methods OV was cultured in vitro using mouse induced pluripotent stem cells (iPSC) under specific conditions.During OV culture,it was transfected multiple times by lentivirus-mediated Ngn2 gene and then it was induced after maturation.The cells were specificly differentiated toward retinal nerve cells in OV.Using the green fluorescent protein (EGFP) gene as control,the differentiation of retinal nerve cells in OV was detected by immunohistochemistry.Reverse transcription PCR and Western blot were used to quantitatively detect the expressions of retinal neuron-specific proteins Pax6,Islet1 and Brn3b.Results The mouse iPS-derived OV was successfully cultured.The number of neural cells in the OV transfected with the Ngn2 gene was increased by the lentiviral-mediated lentivirus.The expressions of PAX6,Islet1 and Brn3b in the Ngn2 transfection group were significantly higher at the gene and protein levels than those in the control group,with significant differences between the two groups (P＜0.05).Conclusions The Ngn2 gene can effectively increase the number of retinal neuron differentiation in OV and make in vitro cultured OV more mature and form a more perfect retinal cell neural circuit.

Objective Toexploreasimpleandaccuratemethodfordeterminingthequantitativeevaluationandthevolumedegree divisionofpleuraleffusionbyMDCTtissuesegmentationtechnology.Methods Thevolumeofwaterinjectedintothreecontainersof differentshapeswasmeasuredby MDCTtissuesegmentationtechnique,andwerefurthercomparedwiththevolumeoftheactual waterinjection.Byusingthesamemethod,theliquidvolumeofpleuraleffusionbeforeandafterpumpingfrompatientswasmeasured andthedifferencebetweentheanteriorandtheposteriorwascalculated.Thenthedifferencebeforeandafterthepumpingwascomparedwiththe actualvolumeofclinicalfluidpumping.Atthesametime,thespecificmethodtodeterminethedegreedivisionofpleuraleffusionwas established.Results Forcontainersofdifferentshapes,theresultfromtheCTscanningliquidmeasurementwasthesameasactualwater injection,andhadahighcorrelation.Thedifferencebetweenbeforeandafterfluidextractionandclinicalactualfluidextractionwas linearcorrelated(positivecorrelation,r=0.986,P<0.001).Accordingtothemethodfordegreedivisionofadulthemothorax,MDCT tissuesegmentationtechnologydividedthevolumeofpleuraleffusionbeforethepumpingintosmall,mediumandlargeamounts.There were44caseswithsmallpleuraleffusion,11caseswithmoderatepleuraleffusionand7caseswithlargepleuraleffusion.Conclusion The volumeofpleuraleffusion measuredby MDCTtissuesegmentationtechnologycanreflecttheactualvolumeofpleuraleffusionin clinictoacertainextent,and MDCTtissuesegmentationtechnologycanbeusedtodividethevolumedegreeofpleuraleffusion.

Objective To design a mathematical calculation model for better understanding and grasping the logical problem of replacement fluid and citric acid anticoagulant infusion in continuous veno-venous hemofiltration (CVVH). Methods ① Parameter definition: A, B, and T were respectively called the main part of pre-replacement fluid, 5% sodium bicarbonate solution, and 4% sodium citrate infused before filter. And a and b were respectively called the main part of post-replacement fluid, and 5% sodium bicarbonate solution infused after filter. ② Logic conversion:The liquid in back terminal (Z) was artificially divided into two parts. One (X) was the original residual plasma after filtration. The second (Y) was the part excluding the plasma, including the left part of pre-replacement fluid with sodium citrate, and the post-replacement fluid. ③The mathematical formulas of liquid volume and electrolyte concentration at X, Y and Z in unit time were listed according to the principle of CVVH and the screening coefficient of filter for different substances. ④The calculation formulas were entered into Excel form, and a mathematical calculation model was made, and a simulation calculation with examples was carried out. Results An Excel model was established by inserting the calculation formulas of volume, electrolyte, and total calcium at X, Y and Z. And it was found that the concentration of Na+, K+, Cl-, HCO3- at Y point remained unchanged only when A, B and (or) a, b was kept in same side and proportion even with the change of blood flow and other parameters without sodium citrate as anticoagulant. Once any of the parameters (such as blood flow, replacement fluid volume, etc.) were adjusted in other infusion methods (such as different ratios, different directions of the same year, etc.), the calculation results at Y would vary, and the electrolyte concentration at Z would change accordingly. A change of dilution model or parameter would result in the change of the electrolyte concentration at Y and Z with sodium citrate as anticoagulant. The concentration of total calcium scarcely changed no matter in what model and parameters. Conclusions All kinds of infusion ways could be included in the Excel model. The infusion results of all kinds of infusion matching could be intuitively evaluated. It is helpful for the medical staff to make a logical analysis and risk prediction in CVVH.

Objective? To? analyze? the? ineffective? triggering? caused? by? nebulization? in? the? way? of?respiratory?mechanics.? Methods? A?test-lung?and?an?oxygen-driven?jet?nebulizer?were?connected?to?the?circuit?in?a?PB840?ventilator.?The?test-lung?was?pulled?outwards?in?manual?way?till?an?inspiration?was?effectively?triggered?separately?in?different?flow-trigger?modes?[flow-trigger?sensitivity?(VTrig)?3?L/min?and?5?L/min]?and?pressure-trigger?modes?[pressure-trigger?sensitivity?(PTrig)?2?cmH2O?and?4?cmH2O,?1?cmH2O?=?0.098?kPa]?with?the?nebulizer?being?closed?and?opened?in?turn.?The?corresponding?relationship?and?characteristics?between?the?flow?and?pressure?in?the?circuit?under?different?triggering?conditions?were?observed?by?adjusting?the?curve?amplitude?in?the?screen.?The?minimum?alveolar?pressure?(Pa)?which?could?cause?an?effective?triggering?and?the?variation?span?of?Pa?during?the?triggering?period?were?analyzed?in?respiratory?mechanics.? Results? ①?In?flow-trigger?mode:?Pa?was?pulled?down? from? positive? end-expiratory? pressure? (PEEP)? or? intrinsic? positive? end-expiratory? pressure? (PEEPi)? to?"PEEP－VTrigR"?(R?meant?airway?resistance)?without?nebulization,?and?the?span?of?Pa?was?"VTrigR"?or?"PEEPi－PEEP+VTrigR".?Pa?was?pulled?down?from?PEEP?or?PEEPi?to?"PEEP－(VTrig+N)?R"?(N?meant?nebulization?airflow)?with?nebulization,?and?the?span?of?Pa?was?"(VTrig+N)?R"?or?"PEEPi－PEEP+(VTrig+N)?R".?②?In?pressure-trigger?mode:?Pa?was?pulled?down?from?PEEP?or?PEEPi?to?"PEEP－PTrig－1R"?without?nebulization,?and?the?span?of?Pa?was?"PTrig+1R"?or?"PEEPi－PEEP+PTrig+1R".?Pa?was?pulled?down?from?PEEP?or?PEEPi?to?"PEEP－PTrig－(N+1)?R"?with?nebulization,?and?the?span?of?Pa?was?"PTrig+(N+1)?R"?or?"PEEPi－PEEP+PTrig+(N+1)?R".? ?Conclusions? Nebulization?airflow?increases?the?difficulty?of?inspiratory?triggering?in?mechanical?ventilation.?PEEPi?makes?it?more?difficult.

BACKGROUND: The tyrosine kinase inhibitors (TKI) treatment of non-small cell lung cancer (NSCLC) with epidermal growth factor receptor (EGFR) mutation may have a positive effect, but most patients may develop drug resistance, therefore, the detection of the developing time in drug resistance and the research of the mechanism of drug resistance are need to be solved. While the emerge of next generation sequencing (NGS) have make it possible. The aim of this study is to monitor the efficacy of targeted therapy by studying the variation of circulating tumor DNA (ctDNA) mutation frequency and gene mutation spectrum through the targeted therapy. METHODS: Our center enrolled 22 patients with EGFR mutation detected by tissue or peripheral blood, and collect 8 mL of peripheral blood of the patients for ctDNA sequencing in different phases, before systematic prior treatment, followed-up by 2 months and disease progression after TKI administration. RESULTS: Patients with EGFR gene mutation may acquire a longer median survival time after receiving targeted drug therapy, due to the drop of mutation abundances, while the therapy may have a minor effect in patients which their mutation abundances have slightly decreased compared to the statistics before the cession (P=0.015,3). The significantly reduced group median progression was associated with a longer survival [progression free survival (PFS)=390 d]. At the same time, we found out that when related to TP-53 gene mutation, the effect of targeted drug therapy for EGFR-sensitive mutation was unsatisfactory (the median PFS was 120 d compared with 630 d, P=0.000,2). CONCLUSIONS Patients who has lower mutation abundance with EGFR sensitive mutations after TKI treatment may have a longer survival period (P<0.05), and the mutation abundance were not significantly dropping or accompanied by other mutations may indicating TKI resistance.

Objective: To design and evaluate a new method for measuring the center of rotation of the hip joint, and to compare it with the traditional methods. Methods: Data of healthy hips of 120 total hip arthroplasty (THA) patients (120 hips) and bilateral hips of 20 healthy volumteers (40 hips) were collected. A series of X-rays and magnetic resonance imaging (MRI) were taken from each patient with the hip joint abductted at differenct angles (0°, 7.5°, 15°, and 22.5°). The motion track of rotation center was obtained by comparing these X-ray and MRI images. The preoperative and postoperative data of 67 surgical patients who underwent two different measurement methods before surgery were compared. Results: Compared with traditional measurement methods, the accuracy of our new method was significantly improved; the results of hip MRI verified the objectivity and accuracy of our new method; preoperative measurements and postoperative follow-up results showed that our new method had a better clinical effect than traditional measurements. Conclusions The team′s innovative new measurement method based on pelvic orthotopic X-ray photographs with different abduction angles of the hip shows more accurate data than traditional measurement methods and shows better clinical results.