AIM Inhibitors of xanthine oxidase (XOD) may be potentially useful for the treatment of gout or other XOD inducing diseases, so the inhibition on XOD activity of diterpenoid tanshinones such as cyptotanshinone (CT) and methylenetanshinone (MT) from Tanshen (Salvia miltiorrhiza Bge.) was studied. METHODS The formation rate of uric acid from xanthine was determined by measuring the absorbance increment at 290 nm (ΔA290 nm) in the reactive medium of xanthine/XOD when CT or MT was added. RESULTS It was found that CT and MT inhibited XOD activity. Dixon plots showed that the inhibition mode was competitive type. The Ki values of CT and MT were 17.8 μmol·L-1 and 25.9 μmol·L-1, respectively. Their inhibitory potencies positively correlated with their concentrations and the IC50 values of CT and MT were 70 and 67 μmol·L-1, respectively. The IC50 value of allopurinol, the positive control, was 60 μmol·L-1. CONCLUSION CT and MT have inhibitory effects on XOD activity and may be potentially useful for the treatment of gout or other XOD inducing diseases.

Objective To summarize the SPECT/CT manifestation of spondylitis caused by Brucells infection and to evaluate the diagnostic value.Methods From June 2012 to October 2015,a total of 28 patients (14 males,14 females,average age 46.4 years) with Brucellosis spondylitis confirmed by laboratory test and pathology were included.The images of whole-body bone scan and SPECT/CT fusion imaging were retrospectively analyzed.According to the pathological and serologic test results,the diagnostic efficacy of imaging was calculated.x2 test was used.Results Most of the Brucellosis spondylitis happened in the lumbar(76.7％,43/56),and the most common locations were L3,L4,L5 (72.1％,31/43).Two or more involved consecutive vertebra were found in 71.4％ (20/28) of the patients.Moderate radioactive distribution was showed in 89.2％ (50/56) of lesions,high radioactive distribution was showed in 5.4％ (3/56) of lesions,and mild radioactive distribution was showed in the rest 3 lesions.Thirty-three lesions(58.9％,33/56) had diffuse increased radioactivity uptake in the affected vertebra,and 32.1％(18/56) showed diffuse increased radioactivity at the superior and inferior margin of the vertebra;only 8.9％ (5/56) of lesions were on one side of the vertebral bodies.The SPECT/CT results were as follows:(1) Bone destruction was showed in 80.4％ (45/56) of lesions,and the edge of the lesion was clear.(2) For 66.7％ (30/45) of lesions,bone hyperplasia was seen along with bone destruction and moderate radioactivity concentration on the edge of destruction area.(3) The damage of the intervertebral disc was mild,and the vertebral abscess was relatively rare (5.4％,3/56).The diagnostic accuracy of SPECT/CT was statistically higher than that of whole-body bone scan:67.8％(38/56) vs 96.2％(54/56);x2=13.1,P＜0.05.Conclusion SPECT/CT imaging has a higher diagnostic efficiency than whole-body bone scan in Brucellosis spondylitis.

Objective:To evaluate the additional value of chest thin layer CT over 99Tc m-hydrazinonicotinyl-(polyethylene glycol) 4-E[(polyethylene glycol) 4-c(RGDfK)] 2(HYNIC-PEG 4-E[PEG 4-c(RGDfK)] 2; 3PRGD 2) SPECT/CT in detecting isolated pulmonary space. Methods:This was a prospective study conducted in General Hospital of Ningxia Medical University. There were 87 patients with solitary pulmonary space occupying between July 2015 and December 2016, and 74 of those patients (49 males, 25 females, age range: 37-80 (58.4±9.6) years) who had pathological results were enrolled. 99Tc m-3PRGD 2 SPECT/CT imaging was performed routinely, and then the chest thin layer CT images were acquired. The maximum radioactive counts ratio of tumor to non-tumor (T/N)≥1.5 was the standard for positive planer 99Tc m-3PRGD 2 imaging, and that ≥2.0 was the standard for positive SPECT/CT imaging. According to the pathological results as gold standard, the diagnostic efficiencies of 99Tc m-3PRGD 2 planer and SPECT/CT imaging, chest thin layer CT and chest thin layer CT+ 99Tc m-3PRGD 2 SPECT/CT imaging for malignant pulmonary lesions were calculated. Kappa test was used to compare the consistency of the imaging methods and pathological results. Results:The post-surgery histopathology confirmed that 51 patients were with malignancy and 23 had benign lesions. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of 99Tc m-3PRGD 2 planer imaging, SPECT/CT imaging and chest thin layer CT in the diagnosis of malignant pulmonary lesions were 47.1%(24/51), 65.2%(15/23), 52.7%(39/74), 75.0%(24/32), 35.7%(15/42); 86.3%(44/51), 47.8%(11/23), 74.3%(55/74), 78.6%(44/56), 11/18 and 84.3%(43/51), 52.2%(12/23), 74.3%(55/74), 79.6%(43/54), 12/20, respectively. Those of the chest thin layer CT+ SPECT/CT were 98.0%(50/51), 73.9%(17/23), 90.5%(67/74), 89.3%(50/56) and 17/18 respectively. The Kappa values between the imaging methods ( 99Tc m-3PRGD 2 planer imaging, SPECT/CT imaging, chest thin layer CT and the chest thin layer CT+ SPECT/CT) and pathological examination were 0.100, 0.250, 0.354 and 0.765 (all P<0.001). Conclusion:Chest thin layer CT has an incremental value over 99Tc m-3PRGD 2 SPECT/CT imaging in the differential diagnosis of benign and malignant pulmonary lesions.

Objective:To investigate the optimal experimental conditions (including antigen retrieval time, antibody titers and antibody incubation time) for reliable detection of programmed death-ligand 1 (PD-L1) expression using PD-L1 (22C3) antibody concentrate, and to establish a laboratory developed test for PD-L1 detection.Methods:Using Dako PD-L1 IHC 22C3 pharmDX staining procedure and scoring guidelines as the standard reference (group A), the PD-L1 expression in 25 tissue specimens (including 15 lung cancer tissues, 5 tonsil tissues and 5 placenta tissues) was detected with Flex+/HRP detection kit (EnVision) under 8 different experimental conditions (groups B1 to B8). The staining results were then compared to those in group A.Results:In group B1, 3 tissue samples showed the percentages of PD-L1 positive tumor cells were similar to those in group A, while the percentages of PD-L1 positive tumor cells were lower than those in group A in the other samples. In group B7, two case showed a positive rate higher than that in group A that was also above the positive cut-off value, and the rest of the samples had a percentage of PD-L1 positive tumor cells slightly higher than that in group A, but still below the positive cut-off value. The staining results of group B8 were the closest to those of group A compared with the other groups. Although the percentages of PD-L1 positive tumor cells in the B2 to B6 groups were decreased in various degrees as compared with group A, they were still concordant with group A′s classification (positive vs. negative) and would not change the choice of clinical treatments.Conclusions:The experimental conditions are associated with detection rate of PD-L1 expression using 22C3 antibody. In the present study, the most-suitable alterative conditions in the PD-L1 detection using 22C3 antibody concentrate are those applied in the group B8 (including antigen retrieval in Dako PT Link tank at 97 ℃, pH 6.0 for 40 min and incubation with 22C3 antibodies (1∶100 dilution) at room temperature for 60 min, incubation with EnVision Flex+Linker at room temperature for 30 min, incubation with EnVision/HRP at room temperature for 30 min and DAB staining for 5 min), which could provide reliable results at minimum costs.

Objective To investigate the molecular mechanism of translocation of heterogeneous nuclear ribonucleoprotein A2B1 (hnRNPA2B1 )to the cytoplasm after glucose starvation and the effects of increased cytoplasmic translocation of hnRNPA2B1 on the survival of prostate cancer PC3 cells. Methods Human prostate cancer PC3 cells were divided into normal control group (cultured conventionally with glucose-containing medium,RPMI 1640 Medium)and glucose starvation group (cultured with glucose-free medium,RPMI 1640 Medium).The 2 types of cells were treated with deacetylase inhibitor,trichostatin A (TSA ) combined with nicotinamide (NAM),AKT inhibitor BEZ235,si-NC transfection,and si-hnRNPA2B1 transfection,respectively.Cytoplasmic and nuclear protein separation,immunoprecipitation and Western blotting were used to detect changes in hnRNPA2B1 acetylation,total AKT protein and its phosphorylation level,and expression levels of hnRNPA2B1 in the cytoplasm and nucleus.CCK-8 assay was employed to observe cell survival in each group.Results After 3～5 h of glucose starvation treatment,the acetylation of hnRNPA2B1 protein was reduced (P<0.01 ),and its cytoplasmic translocation was increased in PC3 cells (P<0.01 ),which was accompanied by enhanced AKT phosphorylation and activation of the AKT signaling pathway.TSA/NAM treatment,BEZ235 treatment,and si-hnRNPA2B1 transfection all resulted in obvious increase in acetylation of hnRNPA2B1 protein when compared with glucose starvation treated cells (P<0.01 ),which could inhibit the glucose starvation-mediated cytoplasmic translocation of hnRNPA2B1,suppress AKT phosphorylation,and consequently decrease the cell survival rate after glucose starvation (P<0.01).Conclusion Glucose starvation can maintain the survival of PC3 cells by inducing the activation of the Ac-hnRNPA2B1-AKT signaling pathway.

Objective: To explore an optimal method for granulocyte cell production from umbilical cord blood mononuclear cells. Methods: Erythrocytes were precipitated by hydroxyethyl starch. Mononuclear cells were isolated through Ficoll density gradient centrifugation. Different media, additives and cultivation model were chosen for granulocyte induction. Cell morphology was observed by microscopy, and cell phenotype was detected by flow cytometry. The CD18 expression of granulocytes was tested by immunofluorescence assay, and phagocytosis test was executed as well. Results: Compared to fetal bovine serum (FBS) treatment group, cell viability, counts and differentiation rate of granulocytes induced by X-VIVO^TM 15 combined with TPO, SCF, G-CSF but without FBS were superior. And X-VIVO^TM15 medium was better than SCGM medium at effectiveness and cost. Using two-stage mode of hematopoietic stem cell expansion followed by granulocyte induction with X-VIVO^TM15 combining TPO, SCF and G-CSF, cell proliferation was nearly 132 times at day 21. Flow cytometry showed that the differentiation was lagged in 2-stage mode than in direct induction mode, CD15 expression was (69.60± 1.06) % vs (97.73±0.39) %; Wright-Giemsa staining demonstrated mature granulocytes; immunofluorescence showed the expression of lysosomal proteins CD18. A strong phagocytic function of mature granulocytes was demonstrated by phagotrophic efficiency of (51.43±0.05) %. And granulocyte had chemotaxis ability under the role of chemotactic factor IL-8. Conclusion Optimized culture media and cultivation mode are achieved for functional granulocytes induction in vitro.