Objective To screen the key N6-methyladenosine(m6A)methylation related genes in renal clear cell carcinoma(ccRCC),and to study their expression and relationship with the prognosis,migration and invasion of renal clear cell carcinoma.Methods The RNA sequencing data and clinical data of ccRCC and ad-jacent tissues were downloaded from the Cancer Genome Atlas(TCGA)and GTEx(Genotype-Tissue Expres-sion).The expression profile and prognosis were analyzed with R 4.1.1,and the key genes were screened.Clinical specimens of 10 patients with ccRCC were collected.The mRNA and protein expressions were detec-ted by RT-qPCR and immunohistochemistry,respectively.In human ccRCC cell line RCC23,siRNA was used to knock down key genes,and CCK-8 was used to detect the survival rate of cells.Scratch test and Trans well test were used to detect the migration and invasion of cells,respectively.Results Among the 19 m6A methyl-ation related genes,only insulin-like growth factor 2 mRNA binding protein 3(IGF2BP3)was highly ex-pressed in cancer tissues,and the high expression was significantly positively correlated with poor prognosis.The high expression of IGF2BP3 was verified in clinical specimens by RT-qPCR and immunohistochemistry.After knockdown of IGF2BP3 by siRNA,the survival rate of RCC23 cells decreased significantly,and the mi-gration and invasion ability of cut cells decreased.Conclusion These results suggest that IGF2BP3 may be an effective biomarker and potential drug target for predicting the prognosis of patients with ccRCC.

Objective:To evaluate the feasibility of using a visual positioning system for both motion phantom and clinical quality control.Methods:A phantom experiment was conducted using the Dynamic Thorax Phantom from CIRS. Different ranges of motion were simulated to assess the discrepancies between camera-recorded positions and actual movements. Visual markers were also attached to the treatment bed and the collimator head, and their movements were simulated as part of the experiment. The experiment was repeated for three times. Discrepancies between system measurements and manual measurements were recorded and analyzed to assess the accuracy and reliability of the system.Results:In the motion phantom test, the deviation between the actual motion distance of the phantom and the system's recorded measurement was (0.18±0.07) mm. For linear motion analysis along the X, Y, and Z axes on the treatment table, the measurement errors were (0.14±0.08) mm, (0.15±0.09) mm, and (0.16±0.08) mm, respectively. Additionally, the measurement error in the rotational direction of the treatment couch was 0.18°±0.09°. For the rotational direction of the collimator head, the measurement error was 0.11°±0.02°. Conclusion:The system demonstrates good accuracy and stability, and has potential clinical application value.

Objective:To develop a novel α vβ 3-targeted theranostic agent 177Lu-Evans blue (EB)-Arg-Gly-Asp (RGD) and evaluate its value for SPECT imaging and targeted radionuclide therapy in the non-small cell lung cancer (NSCLC)-patient-derived xenografts (PDX). Methods:The α vβ 3-targeted molecule RGD was conjugated with the albumin binding moiety EB to obtain EB-RGD, and EB-RGD was further conjugated with the chelator 1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid (DOTA) for 177Lu radiolabeling. NSCLC-PDX mice models ( n=68) were established. 177Lu-EB-RGD SPECT imaging, biodistribution study were performed in 28 PDX mice models after being injected with 177Lu-EB-RGD or 177Lu-RGD. Targeted radionuclide therapy were subsequently performed in NSCLC-PDX mice models, saline group (group A), 18.5 MBq 177Lu-RGD group (group B), 18.5 MBq 177Lu-EB-RGD group (group C), 29.6 MBq 177Lu-EB-RGD group (group D), n=10 in each group; tumor volumes of PDX mice models in each group were observed within 50 d. Differences between 2 groups were compared using independent-sample t test. Results:177Lu-EB-RGD was radiolabeled at a specific activity of (55±14) GBq/μmol, with a radiochemical yield of more than 95% and a radiochemical purity of more than 95%. Regarding the SPECT imaging, tumors in NSCLC-PDX mice were clearly observed from 4 to 96 h post-injection and the tumor to muscle ratio (T/M) reached 7.34±0.67, 14.63±3.82, 15.69±3.58 and 15.99±5.42 at 4, 24, 72, 96 h post-injection, respectively. Biodistribution study further confirmed the findings from SPECT imaging, and the tumor uptake of 177Lu-EB-RGD were markedly increased compared to 177Lu-RGD 4 h post-injection ((10.15±1.17) vs (3.30±1.47) percent injection dose per gram (%ID/g); t=18.60, P<0.05). Regarding targeted radiotherapy, the tumor volumes were quickly increased within 50 d after treatment in group A and B, while the tumor volumes were decreased in group C and D, until the tumors in group C and D disappeared at the 28th day after initial treatment with no sign of recurrence during the observation period. Conclusions:177Lu-EB-RGD can target α vβ 3-positive NSCLC-PDX with intense tumor to background ratio and strong tumor inhibition efficacy. The preclinical data suggests that 177Lu-EB-RGD may be an effective new treatment option for advanced NSCLC patients with resistance or ineffective results for targeted therapy.

Objective To assess the imaging characteristics of 18F-Alfalide II in different tumorbearing mice and pharmacokinetics in Beagle dogs.Methods BALB/c nude mice(n-24)were used for subcutaneous tumor models(A549 and U87MG),orthotopic lung cancer models(A549)and orthotopic breast cancer models(MDA-MB-231)(n=6 in each group).18F-Alfatide II and 18F-fluorodeoxyglucose(FDG)microPET/CT images were compared in the 4 types of tumor-bearing nude mice models.18F-Alfatide II blocking experiment,biodistribution experiment and imaging studies in tumors of different growth cycles were performed in A549 subcutaneous tumor-bearing nude mice models.Pharmacokinetic experiments were carried out in Beagle dogs(n = 6)and CD-1 mice(n = 9).Two-sample t test was used to analyze the data.Results Compared with 18F-FDG,18F-Alfatide II microPET/CT images showed better imaging quality and contrast in subcutaneous A549,U87MG tumors and orthotopic A549(tumor/heart:4.50±1.17 vs 0.95±0.31;t = 4.125,P<0.01),orthotopic MDA-MB-231(tumor/muscle:6.60±1.53 vs 0.92±0.43;t = 3.984,P<0.01)transplantation nude mice models.18F-Alfatide II could specifically target A549 tumors,and the tumor uptake of 18F-Alfatide II was reduced by about 75% after pre-injection with cyclo(Arg-Gly-Asp-D-Tyr-Lys)(c(RGDyk)).18F-Alfatide II was rapidly cleared from the blood of Beagle dogs(T1/2 was(57.34±11.69)min).It was cleared in the form of prototype drug and(69.24±6.82)% of cumulative dose was excreted through the urine within 4 h after administration.Conclusions 18F-Alfatide II shows a higher target/non-target ratio than,18F-FDG in the imaging of A549,MDA-MB-231 and U87MG tumor-bearing nude mice models,which is more conducive to the diagnosis of tumor.18F-Alfatide II has excellent pharmacokinetic properties.

Objective To investigate the biodistribution of 18F-Alfatide II in patients with breast diseases and to compare its uptake with 18F-fluorodeoxyglucose(FDG)uptake.Methods A total of 44 female patients(age:(50.7±8.0)years)with clinically suspected breast cancer from December 2015 to May 2017 were prospectively enrolled and underwent 18 F-Alfatide II and 18F-FDG PET/CT prior to treatment.By drawing regions of interest in normal organs and breast lesions,differences between 18F-Alfatide II uptake and l8F-FDG uptake were evaluated in all patients.Paired t test,two-sample t test and Wilcoxon rank sum test were used for data analysis.Results There were 53 breast lesions confirmed by histopathology in 44 patients.Among them,42 lesions were malignant and the others were benign.The uptake of 18F-Alfatide II was very low in the brain,vocal cords,lungs,blood pool and muscle.But the renal cortex and bladder had high 18F-Alfatide II accumulation.Different levels of 18F-Alfatide II uptake were found in other normal organs including normal breast tissue.There were differences(t values:2.04-41.65,all P<0.05)between 18F-Alfatide II and 18F-FDG maximum standardized uptake value(SUVmax)and mean standardized uptake value(SUVmean)in many normal organs except for the choroid plexus,salivary glands,liver,colon and normal breast tissue.The uptake of 18F-Alfatide II was significantly lower than 18F-FDG in breast cancer lesions(SUVmax:3.77±1.78 vs 7.37±4.48,SUVmean:2.25±0.98 vs 4.54±2,82;t values:4.89,4.82,both P< 0.05),but it was still higher in benign breast lesions(SUVmax:2.37±1.62,SUVmean:1.50±0.92;t val-ues:2.35,2.29,both P<0.05).Also,target/non-target(T/NT)of 18F-Alfatide II in breast cancer lesions was higher than that in benign breast lesions(5.32±3.08 vs 2.60±2.37;t = 2.72,P<0.05).Condusion The biodistribution of 18F-Alfatide II in patients is favorable and 18F-Alfatide II can be clinically used for breast cancer imaging.

Objective:To investigate the content and mode of pharmaceutical care for the patients with severe burn and promote the rational use of drugs. Methods:Taking the treatment for one patient with severe burn as example,pharmacists provided pharmaceu-tical care in respects of anti-infection therapy,organ preservation,nutritional support, drug interactions, drug precautions, drug incom-patibility, adverse drug reactions and the effect evaluation . Results:The program of rational drug use was provided for clinics through the implementation of pharmaceutical care. The vital signs of the patient were stable, and then the patient left on pass and continued to be treated with rehabilitation therapy. Conclusion:It is necessary for clinical pharmacists to perform pharmaceutical care for the pa-tients with severe burn,reduce the abuse of drugs and improve the medication safety and effectiveness. Meanwhile, the collaborative service of doctors,pharmacists,nurses and inspectors to patients reflects the value of clinical pharmacists.

Objective To synthesize 18F-AlF-NOTA-G-TMTP1 and evaluate its potential for PET imaging on nude mice bearing high-metastatic potential hepatoma cells.Methods NOTA-G-TMTP1 was synthesized by the standard Fmoc-solid phase synthetic protocols and radiolabeled with 18F using NOTA-AlF chelation method.The nude mice models bearing low-metastatic potential HCC97L and high-metastatic potential HCCLM3 xenografts were established separately.The tumor-targeting characteristics of 18F-AlF-NOTA-G-TMTP1 were assessed by microPET/CT and biodistribution assay.Results NOTA-G-TMTP1 was labeled with 18F in one step with (25±6)％ labeling yield (n=5).The radiochemical purity of 18F-AlF-NOTA-G-TMTP1 was more than 95％ with a specific activity more than 11.1 GBq/μmol.The octanol/water partition coefficient (logP) for 18F-AlF-NOTA-G-TMTP1 was-3.166±0.022.The tumor to muscle ratios were 1.8± 0.4 and 4.7±0.2 at 35 min post injection for HCC97L and HCCLM3,respectively.The uptake of 18F-AlF-NOTA-G-TMTP1 in HCCLM3 tumor was inhibited (61.4％) by unlabeled G-TMTP1.Conclusion 18F-AlF-NOTA-G-TMTP1 has been successfully synthesized.It shows specific uptake by tumor induced by the high-metastatic potential hepatoma cells.

Objective To investigate the effects of danhong injection on the apoptosis of SMMC-7721 heptoma cells.Methods The hepatocellular carcinoma cell line SMMC-7721 was incubated with different concentrations(0,2,4,8 μL·mL-1) of danhong injection, and cell morphology was studied under the microscope. The apoptosis index was detected by flow cytometry at 0,6,12,24 h after cultured with 2 μL·mL-1danhong injection. Cell proliferation was measured by MTT assay;Gene expressions of p53,Bax and Bcl-2 were detected by RT-PCR at different drug concentrations. Results The morphology of the hepatoma cells changed obviously,and the apoptosis index increased by danhong injection in a dose-dependant manner. The danhong injection decreased gene expressions of Bcl-2,and obviously increased p53 and Bax. Conclusion Danhong injection can dose-and time-dependently promote apoptosis of SMMC-7721 cells.

OBJECTIVE: To detect the expression of nucleolin in cardiac hypertrophy rats induced by pressure overload. METHODS: A total of 40 SD rats with body weight 180 g and 220 g were recruited and randomly divided into 2 groups: a transverse aortic constriction (TAC) group and a sham surgery group. Cardiac hypertrophy model was employed by transverse aortic constriction surgery. Then 2 weeks and 4 weeks after the experiment, the heart mass index (HMI), left ventricle mass index (LVMI) were measured. β-MHC mRNA in the heart tissue was detected with RT-PCR. Nucleolin in the heart, brain and kidney was respectively detected with Western blot. RESULTS: Compared with the sham surgery group, HMI, LVMI in the TAC group increased significantly (P<0.01) 4 weeks after the surgery; the expression of β-MHC mRNA in the heart tissue increased (P<0.05) in the TAC group 4 weeks after the surgery; and the expression of nucleolin protein in the heart tissue of the TAC group was remarkably upregulated (P<0.05) 2 weeks after the surgery, with no change in the brain and kidney tissue between the 2 groups. CONCLUSION Expression of nucleolin protein has been upregulated in response to pressure overload, which may suggest that nucleolin plays a role in cardiac hypertrophy induced by pressure overload.
Animals ; Blood Pressure ; Cardiomegaly ; metabolism ; Myocardium ; metabolism ; Phosphoproteins ; metabolism ; RNA, Messenger ; RNA-Binding Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To investigate the effects of exogenous pulmonary surfactant(PS)on the inflammatory response in rats with ventilator-induced lung injury(VILI).Methods Twenty-eight adult male Wistar rats weighing 310-388 g were randomly divided into 4 groups(n =7 each):normal control group(group C),group VILI,group PS and air control group(group A).VILI was produced by high-pressure ventilation(HPV)with peak inspiratory pressure(PIP)40 cm H2 O,respiratory rate(RR)20 hpm and without positive end-expiratory pressure(PEEP)for 20 min.The rats were sacrificed by exsanguination immediately after anesthesia and after VILI in groups C and VILI,respectively.In groups PS and A,PS 100 mg/kg(50 mg/ml)and the equal volume of air were injected into the trachea via the airway after the tracheal edema fluid was removed respectively,and the rats were mechanically ventilated(Vr 10 ml/kg,RR 45 bpm and PEEP 7.5 cmH2O)for 120 min and then sacrificed by exsanguination.The blood samples were taken from femoral artery for determination of the plasma levels of IL-6,IL-10,MIP-2 and TNF-α by ELISA.The tracheal edema fluid was collected to determine the protein concentration by Bradford method.The lungs were removed for microscopic examination and the number of neutrophils was counted under microscope.Results There was no significant difference in the plasma levels of TNF-α among the four groups(P ＞ 0.05).Compared to group C,the plasma levels of MIP-2,IL-10 and IL-6 were significantly increased and the number of neutrophils was significantly enlarged in group VILI(P ＜ 0.05).The number of neutrophils was significantly smaller in group PS than in group A(P ＜ 0.05).There was no significant difference in the plasma levels of MIP-2,IL-10 and IL-6 between groups PS and A(P ＞ 0.05).The inflammatory damage in lung tissues was observed obviously in groups VILI,A and PS.Conclusion When exogenous PS is used to treat the rats with VILI,PS reduces neutrophil recruitment,but can not inhibit the release of inflammatory cytokines.