Lysosomes represent a promising target for cancer therapy and reducing drug resistance. However, the short treatment time and low efficiency of lysosomal targeting have limited the application in lysosome-targeting anticancer drugs. In this study, we proposed an adhesive-bandage approach and synthesized a new lysosomal targeting drug, namely long-term lysosome-targeting anticancer drug (LLAD). It contains a SLC38A9-targeting covalently bound moiety and an alkaline component both to prolong the inhibition of SLC38A9 in lysosomes and alkalinize lysosomes. Upon short term and low-dose treatment of HeLa cells, at passage 0, with LLAD, it rapidly alkalinized lysosomes and also can be detected in lysosomes even at passage 15. LLAD induced apoptosis in HeLa cells through long-term lysosomal damage, and showed better long-term anticancer effect than cisplatin in vivo. Overall, our study paves the way for developing long-term lysosomal targeting drugs to treat cancer and overcome the drug resistance of cancer cells, and also provides a candidate drug, LLAD, for treating cancer.

Objective To evaluate the stability,safety and immune enhancement and anti-tumor effects of Wilms'tumor gene 1(WT1)peptide combined with AddaVaxTM emulsion vaccine for acute myeloid leukemia.Methods The stability of WT1 peptide in the adjuvant vaccine was evaluated using MALDI-TOF-MS time-of-flight mass spectrometry.Female C57BL/6 mice were randomly divided into PBS group,WT1 peptide group,and WT1 peptide+AddaVaxTMemulsion adjuvant vaccine group.The immunization was performed at a dose of 50 μg/mouse for antigen and 50 μg/mouse for adjuvant,with intramuscular injection on days 0,14,and 28.HE staining was used to assess the toxicity of intramuscular vaccination on mouse organ tissues.Cytokine levels were detected by ELISA,and the number of IFN-γ-secreting splenocytes was measured by ELISpot.Flow cytometry was employed to detect the maturation of bone marrow-derived dendritic cells(BMDCs)promoted by the vaccine in vitro and the promotion for lymphocyte activation,and H-2Db WT1 tetramer was utilized to detect the proportion of specific CD8+T cells.After establishing a mouse leukemia tumor model using the C1498-mWT1 stable cell line,the anti-tumor effects of the vaccine for prevention and treatment were evaluated.Results The WT1 peptide stably existed in the vaccine without causing significant organ tissue changes in mice after intramuscular injection.Compared to the mice immunized with WT1 aqueous solution,the mice after intramuscular injection of the WT1 peptide emulsion adjuvant vaccine showed stronger immune responses of Th1 cells,including IFN-γ and TNF-α,as well as Th17 cells of IL-17A(P＜0.05),and the mice had not only promoted number of IFN-γ secreting splenocytes(P＜0.01)but also enhanced maturation of BMDCs,as indicated by an increase in the proportions of CD40+/CD11c+and CD86+CD80+/CD11c+ cells(P＜0.05).Additionally,there were increases in both the proportion of CD4+/CD3+T and CD69+/CD8+T cells(P＜0.05)and the proportion of specific CD8+T cells(P＜0.05).In the anti-tumor effect study using the C1498-mWT1 mouse model,the median survival time of the WT1+AddaVaxTM group was extended by 6 d compared to the WT1 aqueous solution group.At day 50,the survival rate of mice in the WT1+AddaVaxTM group was still 28.5％,while all mice in the other groups had died(P＜0.05).Conclusion The vaccine with the WT1 peptide and AddaVaxTM emulsion adjuvant exhibits good immunological and anti-tumor effects.

Objective To investigate the antibacterial effect and its preliminary mechanism of lavender essential oil on multi-drug resistant Acinetobacter baumannii.Methods Micro-dilution method was used to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC)of lavender essential oil against multi-drug resistant Acinetobacter baumannii,and bactericidal kinetic study was employed to determine the onset and maintenance time of lavender essential oil.Meanwhile,the promoting and therapeutic effects of lavender essential oil on wound healing were observed in a mouse model of infection.Subsequently,crystal violet staining was used to determine the inhibition and clearance of multi-drug resistant Acinetobacter baumannii biofilm by lavender essential oil,and laser confocal microscopy was utilized to observe the survival of bacteria in biofilms.NanoDrop instrument was utilized to quantify the leakage of bacterial DNA nucleic acid and protein after intervention with 3 and 6 mg/mL lavender essential oil,and the leakage of bacterial potassium ion was measured by potassium ion test kit.Proteomics technology combined with bio-informatics were applied to explore the action mechanism of lavender essential oil against multi-drug resistant Acinetobacter baumannii.Results The MIC and MBC of lavender essential oil were both 6 mg/mL,which could kill almost all multi-drug resistant Acinetobacter baumannii at the time point of 120 min,and showed an obvious dose-and time-dependent manner.The overall animal model evaluation showed that both 3 and 6 mg/mL lavender essential oil could promote wound healing,and the curative effect was obvious.Further studies confirmed that 3 mg/mL lavender essential oil had a certain biofilm inhibitory effect on multi-drug resistant Acinetobacter baumannii,and 6 mg/mL also had a certain biofilm clearance effect under the same conditions.Meanwhile,when incubated at 37℃ for 1 h,the dose of 3 mg/mL could increase the leakage of DNA nucleic acid and protein,and significantly promote the efflux of potassium ions.Proteomic analysis suggested that the antibacterial effect of lavender essential oil may be related to affecting the oxidorereductase activity and cell metabolic process of multi-drug resistant Acinetobacter baumannii,and interfering with the biosynthesis of cell wall/membrane/envelope and other structures.Conclusion Lavender essential oil at 3 mg/mL can play an antibacterial effect against multi-drug resistant Acinetobacter baumannii,and its mechanism may be related to the destruction of bacterial biofilm and interference with bacterial metabolism.

Objective:To investigate the effect of different acquisition duration of brain image of 18F-florbetaben(18F-FBB)positron emission tomography(PET)on standardized uptake value(SUV).Methods:Eight subjects who underwent 18F-FBB PET examination in Chinese PLA General Hospital from May 2021 to June 2021 were selected,including 5 persons of healthy control and 3 patients with mild cognitive impairment(MCI).All subjects underwent 18F-FBB PET imaging,and the dynamic PET images of them on brains were continuously acquired for 20 min between 90 and 110 min after the 18F-FBB injection was injected as(3.7-5.5 MBq/kg).Under the situation that other reconstruction parameters did not change,the images were reconstructed at 0-1,0-3,0-5,0-10,0-15 and 0-20 min,respectively.The same of region of interest(ROI)ranges were delineated in bilateral frontal cortex,bilateral temporal cortex,bilateral parietal cortex,posterior cingulate gyrus and cerebellar cortex of each group of images.And then,the corresponding mean standardized uptake value(SUVmean)of each region was obtained.The differences of SUVmean values of different ROI values between each group of data images and the images of 0-20 min were compared and analyzed.Results:There were significant differences in SUVmean between the acquired images in 0-1,0-3,0-5 and 0-10 min and the acquired standard images of 0-20 min(t=-7.569--2.410,P<0.05),respectively.There were no significant differences in SUVmean between the acquired images of 0-15 min and the acquired standard images of 0-20 min in the bilateral frontal lobe,bilateral temporal lobe,bilateral parietal cortex and posterior cingulate gyrus(P>0.05),only there was significant difference in the cerebellar cortex area between them(t=-5.597,P<0.001).Conclusion:The results of 15 min can reach to the similar results of 20 min in acquiring images,which can shorten the time of examination,and enhance the degrees of comfort and cooperation of patients in examination.It has clinical application value.

Objective:To compare the accuracy of visual qualitative assessment and semi-quantitative analysis for 18F-florbetaben ( 18F-FBB) β-amyloid (Aβ) imaging in the diagnosis of Alzheimer′s disease (AD) and to explore their clinical application value. Methods:From January 2019 to October 2019, 17 patients (8 males, 9 females, age (74.1±8.5) years) with mild/moderate-stage clinically probable AD and 17 cognitive normal control (NC; 9 males, 8 females, age (64.5±6.3) years) were prospectively enrolled in this study. All patients underwent dynamic 18F-FBB PET/CT brain imaging in the Chinese PLA General Hospital. Visual qualitative assessment and semi-quantitative analysis methods were used to analyze PET brain imaging results. The difference of standardized uptake value ratio (SUVR) between the two methods was analyzed by using independent sample t test. The consistency of the two methods and clinical results was analyzed by Kappa test. The cut-off value of SUVR for the diagnosis of AD was determined by receiver operating characteristic (ROC) curve. Results:The sensitivity, specificity and accuracy of visual qualitative assessment to diagnose AD were 14/17, 16/17 and 88.2% (30/34). The global SUVR of NC and AD group were 1.09±0.85 and 1.75±0.25 ( t=-10.263, P<0.001), and the composite SUVR were 1.16±0.57 and 1.89±0.15 ( t=-10.789, P<0.001), respectively. The cut-off value of SUVR for the diagnosis of AD was 1.47, with the sensitivity of 15/17, the specificity of 16/17 and the accuracy of 91.2%(31/34). The visual qualitative assessment and semi-quantitative analysis had good consistency with clinical diagnosis results with Kappa value of 0.765 and 0.824 respectively (both P<0.001). Conclusion:The visual qualitative assessment and semi-quantitative analysis methods used in 18F-FBB Aβ imaging to diagnose AD patients show high accuracy and can provide effective value for clinical diagnosis, but the visual qualitative assessment method is concise and easy to grasp, which is worth further promotion and use in clinical.

OBJECTIVETo observe the clinical effects of acupuncture combined with auricular point sticking based on the western medication for post stroke depression (PSD).

METHODSSixty patients with PSD were randomly assigned into an acupuncture plus auricular application group (a combination group) and a medication group, 30 cases in each one. 20 mg paroxetine hydrochloride was prescribed orally in the medication group, once a day for continuous 8 weeks. Based on the above treatment, 30-minute acupuncture was used in the combination group for 8 weeks at Baihui (GV 20), Sishencong (EX-HN 1), Shenting (GV 24), Yintang (GV 29), Shenmen (HT 7), Neiguan (PC 6), Taichong (LR 3), Hegu (LI 4), Zusanli (ST 36), Sanyinjiao (SP 6) and Fenglong (ST 40), once the other day and three times a week. Auricular point sticking therapy for 8 weeks was applied at shenmen (TF), pizhixia (AT), xin (CO), and gan (CO), with pressing 3 times a day and once 3-5 days. The total score and each factor scores of Hamilton's depression scale (HAMD) were observed in the two groups before and after treatment, and Asberg's antidepressant side-effect rating scale (SERS) and clinical effect were evaluated.

RESULTSAfter treatment, the total HAMD scores of the two groups decreased compared with those before treatment (both<0.05), with better effect in the combination group (<0.05). The scores of the combination group after treatment were lower than those in the medication group, including the anxiety/somatization factor, sleep disturbance factor, hopelessness factor (all<0.05). The total effective rate of the combination group was 86.7% (26/30), which was better than 66.7% (20/30) of the medication group (<0.05). The SERS score of the combination group was lower than that of the medication group (<0.05).

CONCLUSIONSAcupuncture combined with auricular point sticking can improve the clinical symptoms and are effective and safe for PSD.

Objective To analyze high alumina deformation Micro-CT findings of bone in patients with skeletal fluorosis in Shuicheng Guizhou Province.Methods Bone deformation children,youth and middle-aged patients with fluorosis in Goumi and Zhichang Townships Shuicheng County,coal-burning pollution endemic fluorosis areas,were selected as case group,and non-bone deformation children,youth,and children from non-fluorosis endemic areas as controls.Tibia and the anterior superior iliac spine tissue were obtained through orthopedic surgery and etiology examination,and resin embedded without decalcification.Resin-embedded bone tissue was scanned using micro-CT; relevant parameters were analyzed with ABA special bone analysis software INVEON Research Workplace and three dimensional reconstruction processing software Micview.Results ①The anterior superior iliac spine cancellous bone:compared between bone deformation children and bone non-deformation children in the diseased areas,there was an increasing tendency of the following items:relative volume of trabecular bone(0.337％ vs 0.229％),absolute thickness (μm:139 vs 133),quantities within a unit length (number/mm:2.44 vs 1.72),density woven degree of trabecular bone(number/mm:2.22 vs 1.54) and bone mineral density(mg/cm3:1 033 vs 918),while relative bone area of trabecular bone(mm2/mm3:14.5 vs 15.1) and space pitch (μm:0.274 vs 0.567) declined.Compared between bone deformation youth and bone non-deformation youth in the diseased areas,relative volume of trabecular bone was lower(0.217％ vs 0.437％),relative area increased (mm2/mm3:16.9 vs 11.6),absolute thickness reduced(μm:118 vs 172),trabecular number reduced (number/mm:1.83 vs 2.54),and space pitch increased (μm:0.427 vs 0.222),but density woven degree of trabecular bone increased (number/mm:4.61 vs 1.54),bone mineral density decreased(mg/cm3:977 vs 1 108),osteopenia,osteoporosis,bone mineral decreased,and an increase in the number of trabeculae crossing number.② Tibia bone tissue:compared between bone deformation children and bone non-deformation children in the diseased areas,relative volume of tibia trabecular bone increased(0.435％ vs 0.206％),relative volume of trabecular bone (mm2/mm3:12.3vs 12.4),and thickness (μm:188 vs 161) not changed obviously,trabecular number increased (number/mm:2.43 vs 1.28),space pitch reduced(μm:0.238 vs 0.621),density woven degree of trabecular bone decreased(number/mm:2.40 vs 3.48),bone mineral density increased(mg/cm3:1 047 vs 952),in general presented trabecular thickening,increased number and increased bone mineral.Compared between middle-aged patients with fluorosis in the diseased areas and children in non-fluorosis endemic areas,relative volume of trabecular bone (0.346％ vs 0.206％) and area (mm2/mm3:13.8 vs 12.4) increased,thickness of the trabecular bone reduced (μm:144 vs 161),trabecular number increased (number/mm:1.98 vs 1.28),space pitch decreased (μm:0.318 vs 0.621),and density woven degree of trabecular bone decreased (number/mm:2.60 vs 3.48).Conclusions The results of trabecular bone microstructure and bone mineral density have showed that the combined effects of aluminum and fluorine on human bone tissue at different developmental stages are different.High aluminum and fluorine load before the sexual development of children for trabecular bone thick dense,shows an increasing in bone mass and bone mineral deposition of bone sclerosis image.Bone deformation youth shows osteopenia osteoporosis and bone mineral deposition is reduced.Bone volume is slightly increased,the number of trabecular bone is increasing,trabecular structure is fine in middle-aged patients with skeletal fluorosis.

BACKGROUND:An effective freezing-thawing technique is crucial for the clinical application of human umbilical cord mesenchymal stem cells(UC-MSCs).OBJECTIVE:To investigate biological characteristics of UC-MSCs after cryopreservation.METHODS:UC-MSCs were isolated from human umbilical cord and frozen in liquid nitrogen.The survival rate and the suppressive effect of γ-interferon(IFN-γ)of cryopreserved-thawed and fresh human UC-MSCs were compared.Furthermore,the multiple potentials and phenotype of UC-MSCs were estimated after cryopreservation.RESULTS AND CONCLUSION:There was no significant difference between cryopreserved-thawed and fresh human UC-MSCs on the survival rate and the suppressive effect of IFN-γ of peripheral blood mononuclear cells(PBMCs).After cryopreservation,human UC-MSCs had the potential differentiation and the phenotype of mesenchymal stem cells.

Objective: To study the clinical efficacy of Jianpijiangniling combined with chemotherapy for advanced non-small-cell lung cancer(NSCLC). Methods: A total of 129 patients were randomly assigned to the treatment group and the control group.All patients received four cycles of chemotherapy.Patients in the treatment group received concurrent four-cycle Jianpijiangniling therapy.Clinical curative effect，overall survival，quality of life，body weight，and side effects were analyzed. Results: No significant difference was found in the total efficiency between the two groups.Patients'quality of life and body weight in the treatment group were superior to those in the control group(P<0.05，P<0.01).Nausea and vomiting were more frequently seen in the control group than in the treatment group(P<0.05).No statistically significance was found in the rates of diarrhea，constipation，and other adverse effects between the two groups.Marrow suppression was frequently seen in the control group.The rates of leucopenia and decrease in hemoglobin were higher in the control group(P<0.05).The difference in the rate of thrombocytopenia between the two groups was not significant. Conclusion: Jianpijiangniling combined with chemotherapy is effective for advanced non small-cell lung cancer,with fewer side effects.

Objective To strengthen the scientific management of automatic processor and promote QC, based on analyzing QC management chart for automatic processor by statistical method, evaluating and interpreting the data and trend of the chart. Methods Speed, contrast, minimum density of step wedge of film strip were measured everyday and recorded on the QC chart. Mean ( ), standard deviation ( s ) and range(R) were calculated. The data and the working trend were evaluated and interpreted for management decisions. Results Using relative frequency distribution curve constructed by measured data, we can judge whether it is a symmetric bell shaped curve or not. If not, it indicates a few extremes overstepping control limits possibly are pulling the curve to the left or right. If it is a normal distribution, standard deviation( s ) is observed. When ?2s lies in upper and lower control limits of relative performance indexes, it indicates the processor works in stable status in this period. Conclusion Guided by statistical method, QC work becomes more scientific and quantified. We can deepen understanding and application of the trend chart, and improve the quality management to a new step.