Aim To investigate the effect of 2-deoxy-D-glucose(2-DG)on the sensitivity of leukemia multi-drug resistant K562/ADMcells to adriamycin by inhib-iting glycolytic pathway as well as its molecular mecha-nisms.Methods The leukemia drug-resistant K562/ADM cells and parental K562 cells were used as the target cell models.The cell proliferating activity was assessed with an MTT colorimetric assay,and the gly-colysis including glucose consumption,lactate export, and hexokinase activity was determined by glucose, lactic acid and hexokinase (HK)testing kits.The ex-pression and phosphorylation of mammalian target of rapamycin(mTOR)and glucose transporter-4 (GLUT-4)expression were analyzed by western blot.Results K562/ADM drug-resistant cells possessed higher HK activity,GLUT-4 expression level and aerobic glycolic ability than K562 sensitive cells. 2-DG treatment markedly inhibited HK activity,glucose consumption, and lactate export both in K562 cells and K562/ADM cells,and suppressed the proliferation of the two cells in a time-and concentration-dependent manner.Low concentration of 2-DG or adriamycin could increase the expression and phosphorylation of mTOR.However, the co-administration of 2-DG and adriamycin markedly counteracted adriamycin-mediated enhancement of mTOR expression and phosphorylation and down-regu-lated GLUT-4 expression in K562/ADM cells,and 2-DG dramatically improved the sensitivity of K562/ADM cells to cytotoxicity.Conclusion 2-DG inhibits the proliferation of drug-resistant K562/ADM cells and en-hances the sensitivity to adriamycin by blocking aerobic glycolysis pathway through inhibiting hexokinase activi-ty,counteracting adriamycin-stimulated increased ex-pression and phosphorylation of mTOR and downregu-lating GLUT-4 expression.

Objective To explore the level and the significance of amino-terminal propeptide of procollagen type Ⅲ(P Ⅲ NP)in the serum of hypertrophic scar patients at excessive stages.Methods The serum samples of 74 inpatients admmitted in Affiliated Fosham Hospital of Sun Yat-sen University from August 2007 to August 2009 suffering from long-persisting post-burn hypertrophic scar at various stages were collected.Serum samples of 12 were normal persons were also collected and used as control group.Hypertrophic scar group were divided into 4 subgroups according to the phase of scar and the serum concentrations of P Ⅲ NP were detected using the sensitive RIA method.Results The level of PⅢ NP increases gradually during the process of immature hypertrophic scar to mature scar before it peaking in 4 to 6 months scar subgroup.The concentration of PⅢ NP degreased gradually with the maturation of hypertrophic scar.Conclusions The high concentration of PⅢNP can reflect the high level of the synthesis of type Ⅲ collagen in the tissue of post-burn hypertrophic scar.The level of P Ⅲ NP is synonymous with the ongoing process of hypertrophic scar.PⅢ NP may be a satisfactory marker in discerning the growth and development of post-burn hypertrophic scar.

Objective To investigate the osteogenic ability of dextran sulfate/recombinant human bone morphogenetic protein-2/chitosan (DS/rhBMP2/CS) nano microspheres combined with coralline hydroxyapatite (CHA) in repair of segmental bone defects.Methods DS/rhBMP2/CS microspheres prepared by ionic crosslinking method were adsorbed into CHA by lyophilization.Seventy-two New Zealand rabbits were randomly divided into 3 equal groups after they had been made into models of bone defect at the right radius.The defects in the 3 groups were implanted respectively with CHA,rhBMP2/CHA and DS/rhBMP2/ CS/CHA.Another 18 animals served as a blank control group.Blood and bone samples were obtained at 4,8 and 12 weeks after implantation.The serum BGP was detected,and the bone grafts were scanned by micro-CT for calculation of volume ratio of the new bone.Hematoxylin and eosin (HE) staining was performed after bone decalcification.Results All the 72 animals recovered well without any infection or graft exposure.Gross observation at postoperative 12 weeks showed that the DS/rhBMP2/CS/CHA group was the best in the quality,quantity and strength of the new bone,as well as in the healing of bone defects.The serum levels of bone gamma-carboxyglutamic-acid-containing protein at all time points in the DS/rhBMP2/CS/CHA group were significantly higher than those in the CHA and rhBMP2/CHA groups (P ＜ 0.05).Micro-CT scanning demonstrated obvious progress in bone formation,cortical bone and marrow cavity at all time points in the DS/rhBMP2/CS/CHA group which showed significantly faster bone reconstruction synchronized with material degradation and significantly higher volume ratio of the new bone than the other 2 groups (P ＜ 0.05).Histological examinations showed better morphology of mature cortical bone and new marrow cavity at all time points in the DS/rhBMP2/CS/CHA group than in the other 2 groups.Conclusion Since DS/rhBMP2/CS/CHA possesses a better mechanism of sustained-releasing rhBMP2 to induce bone formation because of its reticular and hole-hole-connected structure,it may perform better in repairing segmental bone defects than simple CHA or rhBMP2/CHA.

OBJECTIVE: To observe the clinical efficacy of the simple expansion of the spinal canal decompression, decompression plus hydroxyapatite/polyamide artificial lamina reconstruction, and decompression plus titanium mesh reconstruction in the treatment of spinal canal stenosi. METHODS: A total of 39 patients with lumbar spinal stenosis (with or without disc herniation, spondylolisthesis less than I degree), who received therapy of surgery from January, 2011 to January, 2012, were retrospectively analyzed. All patients were divided into 3 groups: a laminectomy surgery alone group (group A, n=15), a decompression plus hydroxyapatite/polyamide artificial lamina reconstruction group (group B, n=14), and a laminectomy decompression plus reconstruction with titanium mesh group (group C, n=10). Intraoperative situation, the postoperative excellent rate and JOA score were analyzed. RESULTS: The duration and blood loss in surgery in group A was much less than that in the group B and C (P<0.05), but there was no statistical significance between the group B and C. The postoperative excellent rate in three groups were similar in 3 months (P>0.05). Twelve months after the surgery, the group B and C showed advantage over the group A (P<0.05). JOA scores in 3 and 12 months after the surgery were all greater than that before the surgery (P<0.05). There was no difference in excellent rates in 3 groups in 3 months after the operation (P>0.05); the group B and C showed advantage over the group A in 12 months after the operation (P<0.05). No serious complications were related to the surgery in the 3 groups. Imaging changes were not significant difference. CONCLUSION The decompression plus hydroxyapatite/polyamide artificial lamina reconstruction and the decompression plus titanium mesh reconstruction show advantages in long-term effect over the simple vertebral canal decompression.
Decompression, Surgical ; Humans ; Intervertebral Disc Displacement ; surgery ; Laminectomy ; Laminoplasty ; Reconstructive Surgical Procedures ; Retrospective Studies ; Spinal Canal ; surgery ; Spinal Fusion ; Spinal Stenosis ; surgery ; Titanium ; Treatment Outcome

Objective To evaluate the application value of ultrasound in vascularization of different artificial bones . Methods A total of 15 New Zealand rabbits were utilized for model establishment of classic segmental bone defect in bilateral radius . Recombinant human bonemorphogenic protein-2 ( rhBMP-2 ) coralline hydroxyapatite(CHA) and CHA were implanted into left and right limbs . Each CHA was divided into 4 equal parts which were examined with conventional ultrasonography and contrast-enhanced ultrasonography( CEUS ) on 3 d ,7 d ,11 d ,15 d ,30 d and 45 d respectively . CEUS quantitative was performed by time-intensity curve(TIC) ,which parameters including the basic intensity(BI) ,peak intensity (PI) ,increased signal intensity ( ΔSI) and time to peak ( TTP) . Then the results were analyzed and compared to pathology . Results Within the same duration ,the vascularization degree in rhBMP-2 group was stronger than that in the ordinary group with advanced vascularization time . Positive correlation was detected between ΔSI and time of both groups ( r =0 .938 ,0 .890 ;P =0 .000) ,and negative correlation was found between BI/PI or TTP and time ( BI/PI: r = -0 .798 ,-0 .899 ; P = 0 .000 ;TTP= r -0 .874 ,-0 .868 ;P = 0 .000 ) . No statistical significance was observed among four observation points of both CHA ,which indicated no obvious difference in vascularization degree of each observation point . Conclusions The structure of bone graft can be clearly displayed by conventional ultrasound ,and CEUS is able to show the early blood perfusion in two CHA grafts and to accurately evaluate the difference of CHA microvascular growth before and after rhBMP-2 application . The combination of these two techniques is a promising approach of evaluating bone graft vascularization in clinical practice .

Objective: To investigte the efficacy of docetaxel combined with androgen deprivation therapy for the treatment of metastatic hormone-sensitive prostate cancer based on Chinese population. Methods: A total of 497 patients were enrolled from January 2004 to July 2018 in the Changhai Hospital. 459 patients received androgen deprivation therapy alone and 38 patients received androgen deprivation therapy combined with docetaxel. The mean age was (72.1±8.7)years. The median PSA level was 100.0 ng/ml, ranging 42.3-999.0 ng/ml. Patients of clinical T₂, T₃, T₄ stage were 213(42.9%), 160(32.2%), 124(24.9%), respectively. Patients of clinical N₀, N₁, N_x stage were 319(64.2%), 144(29.0%), 34(6.8%), respectively. Patients of clinical M₀, M_1a, M_1b, M_1c, M_x stage were 100(20.1%), 51(10.3%), 332(66.8%), 9(1.8%), 5(1.0%), respectively. Gleason scores of biopsy showed that 146(29.4%) patients was ≤7, 103(20.7%) was 8 and 248(49.9%)was ≥9. Propensity score matching was used to match the baseline between groups. Caliper value was set at 0.02. SPSS 22 software was used to achieve a 1∶1 match between the two groups. There were no statistical difference in the age(P=0.102), PSA(P=0.713), T stage(P=0.113), N stage(P=0.226), M stage(P=0.514), Gleason score(P=0.612), tumor loading(P=0.812)between the two groups. The castration resistance-free rate and cancer specific survival rate of the two groups were compared by log-rank and breslow-wilcoxon test. Furthermore, forest plots were used to display the analysis results of different subgroups such as age, PSA, clinical stage, Gleason score, tumor load, whether patients had received palliative resection, and the differences in castration resistance-free rate were compared between the subgroups with high tumor load. Results: The median follow-up time was 22.6 months in the androgen deprivation therapy group and 13.7 months in the combined therapy group. The number of patients with castration resistance in the two groups was 23 and 17, respectively. There were 3 and 6 deaths, respectively. There was no statistically significant difference in the overall progression time to castration resistance between the two groups (10.3 m vs. 16.5 m, P>0.05), and no statistically significant difference in the prostate cancer specific survival rate (21.9 m vs.14.8 m, P>0.05). When subgroup analysis was performed, it was found that patients in the high-metastasis-volume subgroup who received the combination therapy had a significantly longer castration resistance free lifetime (10.6 m vs. 7.2 m, P=0.044), but there was no significant difference in the low- metastasis-volume subgroup(10.5 m vs.12.6 m, P>0.05). Conclusion Docetaxel combined with androgen deprivation therapy can improve the castration resistance free rate in patients with high metastasis volume, but not in low metastasis volume group.

Objective:To analyze the types and functions of CD34 + cells in full-thickness skin defect wounds of normal mice and diabetic mice by single-cell RNA sequencing. Methods:This study was an experimental study. The CD34 + cell lineage tracing mouse was produced, and the visualization of CD34 + cells under the fluorescent condition was realized. Six male CD34 + cell lineage tracing mice aged 7-8 weeks (designated as diabetic group) were intraperitoneally injected with streptozotocin to establish a diabetic model, and full-thickness skin defect wounds were prepared on their backs when they reached 13 weeks old. Another 6 male CD34 + cell lineage tracing mice aged 13 weeks (designated as control group) were also subjected to full-thickness skin defect wounds on their backs. On post-injury day (PID) 4, wound tissue was collected from 3 mice in control group and 2 mice in diabetic group, and digested to prepare single-cell suspensions. CD34 + cells were screened using fluorescence-activated cell sorting, followed by single-cell RNA sequencing. The Seurat 4.0.2 program in the R programming language was utilized for dimensionality reduction, visualization, and cell clustering analysis of CD34 + cell types, and to screen and annotate the marker genes for each CD34 + cell subpopulation. Kyoto encyclopedia of genes and genomes (KEGG) and gene ontology (GO) enrichment analysis was performed to analyze the differentially expressed genes (DEGs) of CD34 + fibroblasts (Fbs), smooth muscle cells (SMCs), keratinocytes (KCs), and chondrocyte-like cells (CLCs) in the wound tissue of two groups of mice for exploring cellular functions. Results:On PID 4, CD34 + cells in the wound tissue of both groups of mice were consisted of 7 cell types, specifically endothelial cells, Fbs, KCs, macrophages, T cells, SMCs, and CLCs. Among these, Fbs were further classified into 5 subpopulations. Compared with those in control group, the proportions of CD34 + endothelial cells, Fbs subpopulation 1, Fbs subpopulation 4, KCs, and CLCs in the wound tissue of mice were increased in diabetic group, while the proportions of CD34 + Fbs subpopulation 2, Fbs subpopulation 3, and SMCs were decreased. The marker genes for annotating CD34 + CLCs, endothelial cells, Fbs subpopulation 1, Fbs subpopulation 2, Fbs subpopulation 3, Fbs subpopulation 4, Fbs subpopulation 5, KCs, macrophages, SMCs, and T cells were respectively metastasis-associated lung adenocarcinoma transcript 1, fatty acid binding protein 4, Gremlin 1, complement component 4B, H19 imprinted maternally expressed transcript, Dickkopf Wnt signaling pathway inhibitor 2, fibromodulin, keratin 5, CD74 molecule, regulator of G protein signaling 5, and inducible T-cell co-stimulator molecule. KEGG and GO enrichment analysis revealed that, compared with those in control group, DEGs with significant differential expression (SDE) in CD34 + Fbs from the wound tissue of mice in diabetic group on PID 4 were significantly enriched in terms related to inflammatory response, extracellular matrix (ECM) organization, regulation of cell proliferation, and aging (with Pvalues all <0.05), DEGs with SDE in CD34 + SMCs were significantly enriched in terms related to cell migration, apoptotic process, positive regulation of transcription, and phagosome (with P values all <0.05), DEGs with SDE in CD34 + KCs were significantly enriched in terms related to mitochondrial function, transcription, and neurodegenerative diseases (with P values all <0.05), and DEGs with SDE in CD34 + CLCs were significantly enriched in terms related to rhythm regulation, ECM, and viral infection (with P values all <0.05). Conclusions:CD34 + cells display high heterogeneity in the healing process of full-thickness skin defect wounds in both normal mice and diabetic mice. The significantly enriched functions of DEGs with SDE in CD34 + cell subpopulations in the wound tissue of the two mouse groups are closely related to the wound healing process.

AIM:To investigate the effects of epal-restat(Epa)on the mitochondrial oxidative stress damage of radiation pneumonitis(RP)mice and to explore its possible mechanism.METHODS:C57BL/6 mice were randomly divided into control(CON),Irradiation(IR),IR combined with Epa(10 mg/kg)and IR combined with Epa(20 mg/kg)group,16 mice in each group.Mouse models of RP were es-tablished by whole thorax irradiation at a dose of 15Gy using a 6-MV linear accelerator.Continuous intragastric administration after IR for 6 or 8 weeks.Lung histopathology was analyzed by HE staining.The expression of aldose reductase(AR)was deter-mined by immunohistochemistry.Mitochondrial morphology of lung tissues was observed by trans-mission electron microscopy.The levels of inflam-matory cytokines(IL-6,TNF-α and TGF-β1)in plas-ma were detected by ELISA.The contents of Malo-ndialdehyde(MDA)and 4-hydroxynonenal(4-HNE)in lung tissues were determined by colorimetry.Sin-gle cell suspension of lung tissues was prepared and reactive oxygen species(ROS)levels in the cells was examined using a DCFH-DA fluorescent probe.Real-time quantitative PCR was used to determine the expression of AR,IL-6,TNF-α and TGF-β1.The protein levels of AR,IL-6,TNF-α,TGF-β1,BAX,Bcl2,Cleaved Caspase-3,8-oxoguanine DNA glycosylase 1(OGG1)and silent information regulator 3(SIRT3)were detected by Western blot analysis.RESULTS:Compared with the CON group,the alveolar hyper-plasia,alveolar septum thickening and inflammato-ry cell infiltration were observed in the IR group.Moreover,the content of inflammatory factors such as IL-6,TNF-α and TGF-β1 and the expression of BAX and Cleaved Caspase-3 were significantly in-creased,and the expression of Bcl2 was obviously decreased after irradiation.Compared with the IR group,Epa robustly alleviated RP.Meanwhile,Epa down-regulated inflammatory cells infiltration and the expression of inflammatory cytokines,such as IL-6,TNF-α and TGF-β1.In addition,Epa could down-regulate the expression of BAX and Cleaved Caspase-3,and up-regulate Bcl2 in lung tissues.Compared with the CON group,the expression of AR,the levels of ROS,MDA and 4-HNE were signifi-cantly increased,the expression of OGG1 and SIRT3 were significantly decreased,and mitochon-drial damage was aggravated in the IR group.Com-pared with IR group,the expression of AR was sig-nificantly down-regulated,the levels of ROS,MDA and 4-HNE were significantly decreased,the expres-sions of OGG1 and SIRT3 were significantly in-creased,and the mitochondrial damage was signifi-cantly alleviated in IR group after 6 to 8 weeks of Epa administration.CONCLUSION:Epa has a pro-tective effect on RP,which may be related to the in-hibition of AR expression,the reduction of mito-chondrial oxidative stress injury,and the inhibition of inflammatory response and cell apoptosis.

In order to understand the prevalence and antimicrobial resistance of Enterococcus faeca-lis(E.faecalis)and Enterococcus faecium(E.faecium)isolated from human,pig and environ-ment in a Xinjiang pig farm,and to investigate the prevalence and potential harm of antimicrobial resistance genes,858 fecal samples from pig farm workers,anal swabs from pig and environment were collected for isolation,identification,antimicrobial susceptibility test and drug resistance gene detection.The results showed that 429 strains of E.faecalis and 222 strains of E.faecium were i-solated.The distribution of Enterococcus species varied among different sources.The isolation rate of E.faecalis was higher in pig anal swabs(73.1％,309/423)and human fecal samples(68.4％,26/38).E.faecium(42.3％,168/397)was mainly isolated from environmental samples.The drug resistance of E.faecalis and E.faecium isolated from pigs was similar to that of E.faecium isola-ted from environment.The drug resistance rates of E.faecalis and E.faecium isolated from pigs were more serious than those from humans to tetracycline,doxycycline,florfenicol and erythromy-cin,and they were more sensitive to ciprofloxacin and levofloxacin.More than 30.0％of E.faecalis and E.faecium isolated from pigs and environment were intermediate to linezolid.E.faecalis from three sources and E.faecium from environmental sources were mainly resistant to five drugs,while E.faecium from pigs was mainly resistant to six drugs.The detection rates of tet(M)and tet(L)genes in E.faecalis and E.faecium isolates from human,animal and environmental sources were more than 70.0％,which was consistent with the results of drug sensitivity.In addition,the cfr,optrA and poxtA genes that mediate oxazolidinone resistance were detected to varying extent.The cfr gene was only detected in four E.faecalis isolates from swine,one E.faecalis isolate from environment and two E.faecium isolates from environment.The positive rate of optrA gene in E.faecium isolated from pigs and environment was higher than that from humans,and the posi-tive rate was more than 60.0％.The positive rate of poxtA gene in E.faecium isolated from pigs and humans was more than 45.0％.The similar drug resistance situation suggests that there is the phenomenon of mutual contamination of drug-resistant bacteria in human-animal-environment.Therefore,we should consider from the perspective of one health,formulate comprehensive disin-fection and control programs,block the transmission route of drug-resistant strains and drug re-sistance genes between human-animal-environment,standardize the use of antibiotics,and reduce the enrichment of antibiotics in human-animal-environment,so as to reduce the risk of drug-resist-ant bacteria.