Objective To explore the metabolite profiles of mesenchymal stem cells(MSCs)underwent death using 9.4 T high resolution MR spectroscopy.Methods MSCs were cultured and treated for 6,12 and 24 hours in a stimulated condition which included hypoxia,serum deprivation and changes of microenvironment.Cell death and the mortality was detected by light microscopy,Hocchst staining and flow cytometry analyses.The morality of stem cells was analyzed using one-way analysis of variance (ANOVA).Cell metabolite extraction was prepared by methanol-chloroform(M/C) method and analyzed on a 9.4 T MR device.1H-MR spectroscopy was obtained and the metabolite concentration of each time point was calculated and compared using one way ANOVA,the difference between two groups was analyzed by SNK test.Results Necrosis was the major form of cell death in the built model.The morality of every time sets was 16±4(0 h),658±61 (6 h),1 571 ± 154(12 h) and 2 816± 178(24 h) respectively,and the difference between each groups were statistically significant (F=298.96,P＜0.01).After induced stem cells death for 6,12 and 24 h,the metabolite concentrations at 0.89 ppm was (1.48±0.69),(2.32±0.63)and (2.15±0.45)nmol/mg respectively,and increased compared to thc control[(1.41 ±0.25)nmol/mg]with statistical significance (F=329.57,P＜0.01).The metabolite concentrations at 1.28 ppm was (6.42±0.31),(7.26±0.32)and (7.01 ±0.61)nmol/mg,respectively,and increased compared to the control[(5.76 ±0.74)nmol/mg]with statistical significance (F=19.56,P＜0.01).The metabolite concentrations at 1.60 ppm was (2.36±0.31),(2.29±0.16)and (2.31 ± 0.24) nmol/mg respectively,and increased compared to the control[(1.96 ± 0.27)nmol/mg]with statistical significance (F=4.35,P＜0.05).After induced stem cells death for 12 hours,the metabolite concentrations at 0.89 ppm was increased compared to 6 hours with statistical significance (P＜0.05).The metabolite concentrations at 1.28 ppm was increased compared to 6 hours with statistical significance (P＜0.05).After induced stem cells death for 24 hours,the metabolite concentrations at 0.89 ppm was decreased compared to 12 hours with statistical significance (P＜0.05).Conclusions There are some specific characteristics on MRS of MSCs underwent death,and the fatty acid peak may serve as a biomarker for cell death.

Objective To investigate the imaging findings and key diagnostic points of Brunner’s gland adenoma on CT.Methods The CT imaging findings and pathological features of 9 cases of Brunner’s gland adenoma confirmed by pathology were retrospec-tively analyzed,including the lesions number,site,size,shape,margin,density and the enhancement pattern.Results Of the 9 ca-ses,1 case located in the antrum and 8 cases in the duodenaum [6 cases in the duodenal bulb (75%)and 2 in the papillary (25%)]. Of the 6 cases of duodenal lesions,3 were found at the anterior wall and 3 at the posterior wall.Except 1 case which complicated with enteritis and had an obscure margin,the other 8 cases were clear margined ,and were round or nodular in shape.The maximum size of tumors ranged 1 5-68 mm in diameters (mean 35.0 mm ± 1 6.2 mm).The density of tumors was homogeneous on CT scan without necrosis or hemorrhage.In the arterial phase after administration of contrast agent,the lesions were similar to the adjacent intestinal wall enhancement,and mucosal annular enhancement (halo sign)showed in 6 cases,and the dot-shape non-enhancement area within the lesion (black star sign)showed in 5 cases,and the thickening or tortuous enhanced blood vessel showed in 6 cases.In the venous phase,9 cases were progressive enhancement,and the “black star sign”or “the black line sign”showed more clearly in 5 cases.In the pathology,the lesions were polypoid-like,solid or cystic.Under the microscope,the hyperplasic Brunner’s glands were covered with normal duodenal mucosa and separated by bundles of smooth muscle cells with dilated duct,cyst,and adipose cells,1 case with atypical hyperplasia of the glandular epithelium and 1 case with ectopic pancreas.Conclusion There are some spe-cific CT imaging features in Brunner’s gland adenoma,which is of important clinical value in accurate preoperative diagnostic.

Objective To investigate whether the malignant transformation of macrophages ( Mφ) in glioma mesenchyme was induced by the fusion of glioma cells ( SU3 ) and Mφ.Methods SU3 cells transfected with red fluorescent protein genes were co-cultured in vitro with Mφexpressing enhanced green fluorescent protein.The cell lineages with RFP+/GFP+dual-color fluorescence were established by using monoclonal selection method.A series of tests for analyzing cancer-related phenotypes, tumorigenicity and specific markers for murine macrophage were performed.Results (1) A few of dual-color fluorescent cells were observed in the co-culture.Three monoclonal cell lineages (C3, C4 and C12) were obtained success-fully.(2) Three types of cells including RFP+, EGFP+and RFP+/EGFP+cells were formed during the cul-ture of monoclonal C12 cell lineage.The percentage of EGFP+cells was increased along the extended culture time and increased passages.Then, EGFP+cells gradually became the predominant cell population.Nota-bly, the percentage of RFP+/EGFP+cells were decreased and maintained at a low level, but the RFP+cells almost disappeared.(3) EGFP+cells from monoclonal C12 cell lineage showed the malignant characteristics such as loss of contact inhibition, rapid proliferation andchromosome aneuploidy, as well as high tumorigenic rate in nude mice (5/5).They also expressed macrophage specific marker CD68 and showed a large number of telocentric chromosomes.Conclusion The results of this study suggested that the malignant transforma-tion of host macrophages as previously observed in solid tumor might be induced by cell fusion occurred be-tween human glioma cells and macrophages.Along with the previous evidences showing the isolation of the malignantly transformed macrophages ( ihCTC) from solid tumor tissue of tumor-bearing mice, the results confirmed an objective existence of malignant transformation of host macrophages in tumor microenvironment. The malignant transformation of host cells induced by fusion with tumor cells revealed not only a new under-standing for the progression of tumor and cancer heterogeneity, but also new targets for cancer therapy.

Objective To investigate the ultrasonic imaging features of thyroid follicular adenoma for conducting the correct diagnosis and differentiation diagnosis.Methods The clinical and imaging data in 64 cases of pathologically proven thyroid follicular adenoma were analyzed on the maximal diameter of tumor,nodularity number,high and low echogenicity,peripheral halo,echo hom-ogeneity,calcifications,and so on.The misdiagnosis causes were investigated.Results The mass was mainly solid or cystic-solid mixed echo.The ultrasonic imaging features of thyroid follicular adenoma were non-peripheral halo or thin wall halo,hyperecho or isoecho,internal macrocalcifications and peripheral calcifications,homogeneous echo structure.Conclusion The ultrasonographic examination can provide the better diagnosis and differentiation diagnosis on thyroid follicular carcinoma.

OBJECTIVETo investigate the retrograde changes in the dorsal motor nuclei (DMV) of the vagus nerve after vagotomy in rats.

METHODSNissl staining and immunohistochemistry were used to observe the morphological and quantitative changes of the DMV and alterations of the expression of iNOS and NADPH after severing of the vagus nerve in adult male Wistar rats.

RESULTSCompared with the control group, the rats with right vagotomy showed obvious morphological changes and a significantly decreased number of neurons in the right DMV (P<0.05). Numerous iNOS- and NADPH-immunopositive cells were detected in the right DMV 5 and 10 days after right vagotomy.

CONCLUSIONVagotomy causes obvious retrograde changes in rat DMV shown by a significantly decreased number and obvious morphological changes of the neurons in the DMV.

Animals ; Male ; NADP ; metabolism ; Neurons ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Rats ; Rats, Sprague-Dawley ; Vagotomy ; Vagus Nerve ; pathology ; physiopathology ; surgery

Objective To investigate the expression and significance of bone morphogenetic protein?2 (BMP?2) and phosphatase and tensin homolog deleted on chromosome ten (PTEN) in incipient,recurrent and malignant salivary gland pleomorphic adenoma??Methods A total of 93 cases of salivary gland pleomorphic adenoma in Kailuan general hospital from 2008 to 2017 were collected,including 54 in incipient group (group A,47 cases in benign group A1,7 cases in malignant group A2),39 cases in recurrent group (group B,26 cases in benign group B1,13 cases in malignant group B2)??The expression of BMP?2 and PTEN were detected by immunohistochemical detection and western blot,the correlation of BMP?2 and PTEN was analyzed by Spearman analysis??Results The immunohistochemical and western blot analysis both showed expression of BMP?2 in recurrent group was significantly higher than that in incipient cases((129??03 ±15??52) vs??(87??88±18??11),t＝－8??094,P＝0??000),and it was lower in malignant cases than that in benign cases(( 100??24 ± 25??07) vs ( 116??66 ± 26??19), t＝2??125, P＝0??040)??There was no significant difference in PTEN expression between incipient and recurrent groups (( 89??15 ± 13??92 ) vs??( 96??19 ±28??02),t＝1??055,P＝0??279),but lower PTEN expression was found in malignant cases than benign cases ((76??06±11??16) vs??(109??28±17??05),t＝7??543,P＝0??000)??BMP?2 was positively correlated with PTEN expression (r＝0??313,P＜0??05)??Conclusion BMP?2 is associated with the recurrence of salivary gland pleomorphic adenoma, and both BMP?2 and PTEN are associated with malignant in the salivary gland pleomorphic adenoma??

OBJECTIVETo screen the representing components in different germplasms of Canarium album and determine the content by HPLC for laying the foundation for the quality specification.

METHODGallic acid and scopoletin and scoparone were chosen as the main chemical components for clearing heat and relieving sore throat by reviewing literature and qualitative experiment. Gallic acid was determined according the method of Galla chinensis in Chinese pharmacopoeia. Scopoletin and scoparone were determined by HPLC on C18 column (4.6 mm x 250 mm, 5 microm) and the mobile phase was acetonitrile-water-acetic acid (16:84:0.5) with the detection wavelength at 345 nm and column temperature at 35 degrees C , the flow rate was 1.0 mL x min(-1).

RESULTThe resolution is good for gallic acid and scopoletin and scoparone. The standard curve in detection range showed good linear relation, the average recoveries were 100.22%, 101.22% and 100.42%, respectively, RSD were all less than 2.0%. The content of gallic acid in the large fruit C. album from Guangdong was higher than that in the small fruit C. album, while the result was contrary in samples from Sichuan. The content of total coumarins in the small fruit C. album was generally higher than that in the large fruit C. album, which from Sichuan was higher than those from Guangdong.

CONCLUSIONThe efficacies of clearing heat and relieving sore throat in C. album were formed by synergistic effect of gallic acid and scopoletin and scoparone. The small fruit C. album was better than the large fruit C. album as medicinal materials, the results provide reliable basis for quality control in medicinal materials and its preparation.

Burseraceae ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Coumarins ; therapeutic use ; Drug Synergism ; Drugs, Chinese Herbal ; chemistry ; Fever ; drug therapy ; Gallic Acid ; therapeutic use ; Hot Temperature ; Humans ; Pharyngitis ; drug therapy ; Scopoletin ; therapeutic use

Objective To investigate the value of CT and MRI in the differential diagnosis of various subtypes malignant epithelial tumors of the ovary,to improve the understanding of imaging manifestations of ovarian cancer.Methods Fifty four cases with malignant epithelial tumors of the ovary confirmed by clinical operation and pathology were included in this study and preoperative imaging examinations were analyzed retrospectively.Results Thirty four cases were diagnosed as ovary cystadenoma,including 26 cases of serous cystadenoma (48%),8 cases of mucous cystadenoma (14%),10 cases of endometrioid adenocarcinoma (19%)and 10 cases of clear-cell carcinoma (19%).All the tumors appeared as unilocular or multilocular cystic-solid masses,however some differences existed among tumors in calcification in masses,size of solid nodules,locular appearance,with or without associated massive ascites, and adjacent structures involvement .Calcification occurred in 5 cases of cystadenocarcinoma,and no calcification was found in endometrioid adenocarcinoma or clear-cell carcinoma.The proportion of solid component in clear-cell carcinoma was lower,usually presenting as intralumimal nodular protuberance.Endometrioid adenocarcinoma was often associated with endometrial hyperplasia or endometrial carcinoma and was the most prone to peripheral invasion and adhesion.Conclusion The different subtypes of ovary malignant epithelium-derived tumor have different characteristics of multilocular,the size of cyst wall nodule or solid nodule and the calcification.Compare and analysis of these characteristics help us to make a more accurate preoperative diagnosis.

Objective To explore the molecular genetic characteristics of primary and recurrent glioblastomas (GBMs) from the same patient in vivo, primary glioma stem cells cultured in vitro, and patient-derived xenograft (PDX). Methods (1) The primary and recurrent GBM specimens from one patient during surgical resection were collected; and the expressions of glial fibrillary acidic protein (GFAP), nestin and Ki-67 were detected by immunohistochemical staining; the methylation of O6-methylguanine DNA methyltransferase (MGMT) gene, mutation of isocitrate dehydrogenase (IDH) gene and amplification of epidermal growth factor receptor (EGFR) gene were analyzed. (2) The primary and recurrent GBM stem cells were cultured in vitro and named as SU5-1 and SU5-2 cells, respectively; the expressions of nestin and CD133 were detected by immunohistochemical staining; GFAP expression was detected by immunohistochemical staining after induced differentiation, and the growth curve was detected by CCK-8 assay; Transwell invasion assay was used to detect the invasion ability; cell resistance to temozolomide (TMZ), carboplatin (CBP), cisplatin (DDP) and adriamycin (ADM) was detected by CCK-8 assay; the protein expression of programmed death receptor-ligand 1 (PD-L1) was detected by Western blotting. The rate of PD-L1 positive cells was detected by flow cytometry; genetic testing analysis was as above. (3) The primary and recurrent in situ PDX models in nude mice were established, and the expressions of nestin, GFAP and Ki-67 were detected by immunohistochemical staining. Results (1) As compared with the primary GBM, the recurrent GBM had significantly higher percentages of Ki-67 and nestin positive cells, while statistically lower percentage of GFAP positive cells (P<0.05); genetic analysis showed that there was no mutation in IDH gene in the primary GBM tissues and recurrent GBM tissues; the MGMT gene in the primary GBM tissues was methylated and EGFR gene was not amplified, while the MGMT gene in recurrent GBM tissues was demethylated and EGFR gene amplification was positive. (2) Both SU5-1 and SU5-2 cells expressed nestin and CD133, and GFAP was expressed after induced differentiation; the growth curve showed that the proliferation of SU5-2 cells started earlier than that of SU5-1 cells, the two were equal on the 3rd, 4th, and 5th d, and the proliferation of SU5-1 cells was faster than that of SU5-2 cells from the 6th d; the invasion ability of SU5-2 cells was statistically stronger than that of SU5-1 cells (P<0.05); the inhibition rates of SU5-2 cells treated with 5, 10, and 15 mmol/L CBP, 0.3125, 1.25, and 5 mmol/L DDP, 0.5 and 2 mmol/L ADM, and 125 and 500 mmol/L TMZ were significantly lower than those of SU5-1 cells treated with the same concentrations and same drugs (P<0.05); the protein expression of PD-L1 in SU5-2 cells was higher than that in SU5-1 cells; the positive rate of PD-L1 in SU5-2 cells was statistically higher than that in SU5-1 cells (P<0.05); the results of genetic analysis were consistent with those of the primary and recurrent GBM samples. (3) As compared with those in the primary PDX model, the nestin and Ki-67 expressions were significantly higher and GFAP expression was significantly lower in the recurrent PDX model (P<0.05); the results of genetic analysis were consistent with those of the primary and recurrent GBM samples. Conclusions Genetic differences are detected between primary and recurrent GBMs; recurrent GBM has stronger invasive capacity and multi-drug resistance. The primary stem cells derived from surgical specimens and corresponding PDX models could replicate the molecular genetic characteristics of original tumors, which provide a reliable experimental platform for both tumor translation researches and screening of molecular therapeutic targets.

OBJECTIVEThe aim of this study was to clarify whether the fusion of bone marrow mesenchymal stem cells (MSCs) with tumor cells can promote tumor angiogensis.

METHODSHuman glioma stem/progenitor cells (GSPCs) (SU3 cells) were transfected with red fluorescent protein (RFP) gene. Bone marrow mesenchymal stem cells (MSCs) were harvested from nude mice with whole-body green fluorescent protein (GFP) gene expression. Then the two kinds of cells were co-cultured in vitro. At the same time SU3-RFP was transplanted into the brain of GFP-expressing nude mice to establish xenograft tumors. The co-cultured cells, GFP/RFP double positive (yellow) cells and blood vessels obtained from the xenograft tumors were observed under fluorescent microscope and laser scanning confocal microscope.

RESULTSAfter five passages in vitro, MSCs maintained the proliferative activity and highly expressed CD105. CD105 was also expressed in the femurs of GFP-expressing nude mice, tumor cells, blood vessels of SU3 xenograft tumors, and clinical malignant gliomas. When MSCs were co-cultured with SU3-RFP, the ratio of yellow cells co-expressing RFP and GFP was significantly increased after extended time and continuous passages. According to the flow cytometry, yellow cells co-expressing RFP and GFP were 83.7% of the cultured cells. In tissue slices of the xenograft tumors, bundles of yellow vessel-like structure and cross-sectioned yellow vascular wall structures including vascular wall stroma cells were observed with RFP and GFP expression, and were identified as de novo formed vessels derived from fusion of MSCs with SU3-RFP cells.

CONCLUSIONCell fusion occurs between tumor cells and host MSCs and it promotes tumor angiogenesis.

Animals ; Bone Marrow Cells ; physiology ; Cell Communication ; Cell Fusion ; Cells, Cultured ; Glioma ; Green Fluorescent Proteins ; Humans ; Luminescent Proteins ; Mesenchymal Stromal Cells ; Mice ; Mice, Nude ; Microscopy, Fluorescence ; Neoplasms ; Neovascularization, Pathologic ; Stem Cells ; Transfection ; Transplantation, Heterologous