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2018 Vol. 48, No. 5 Published: 2018-05-25   313 1H NMR based metabonomics analysis of serum from mice with diabetic nephropathy WEI Tingting, ZHAO Liangcai, GAO Hongchang. Objective: To study the characteristic metabolites and the related pathways in serum of db/db diabetic mice by 1H nuclear magnetic resonance (1H NMR)-based metabonomics with correlative analysis. Methods: The urine and serum of eight db/db mice and nine C57BL/6J mice were collected at week 17. Then the blood glucose, urine creatinine (UCr) and urinary albumin to creatinine ratio (UACR) were measured. The characteristic alterations in serum metabolites were explored by the projection to latent structure discriminant analysis based on the 1H NMR spectra. The correlation between UACR, UCr and serum metabolites was performed by Pearson correlation analysis. Results: At 17-week, db/db mice showed apparent obesity, with the weight more than twice that of wild type mice. The blood glucose and UACR of db/db mice were also doubled compared with wild type mice, but the UCr decreased. It was demonstrated that the db/db mice showed typical symptoms of diabetic nephropathy at week 17. The metabolic profile of the two kinds of mice showed clear differences along the PC1 direction. Compared with wild type mice, pyruvate, lactate, citrate, 3-hydroxybutyrate, acetoacetate, glycine, glutamine, tyrosine, phenylalanine, creatine, choline were reduced, LDL/VLDL and isoleucine/leucine were increased in db/db mice (P<0.05). UACR showed a positive correlation with choline in the serum of db/db mice (r=0.717, P<0.05). Conclusion: In the pathogenic process of diabetic nephropathy, the tricarboxylic acid cycle, glycolysis and lipid metabolic pathways are suppressed and glycogenesis is elevated. Choline may be a potential biomarker of diabetic nephropathy. 2018 Vol. 48 (5): 313-318 [Abstract] ( 773 ) HTML (1 KB)  PDF (1502 KB)  ( 1467 ) 319 The mapping of B-cell epitopes of human PD-L2 and their three-dimensional positions CHENG Kai, CHEN Xudong, LYU Kaiji, JIN Jinji, YE Xiaoxian, ZHANG Lifang, ZHU Guanbao. Objective: To predict the B-cell epitopes of human programmed death factor 1 ligand 2 (PD-L2) and to analyze its 3D structure  in combination with PD-1. Methods: The secondary structure, the flexible region, the hydrophilicity of Hopp & Woods, the Zimmerman polarity parameter, the Jameson-Wolf antigen index and the Emini surface accessibility protocol of PD-L2 were predicted by bioinformatics analysis software and on-line analysis system on server. The results were used in the Multiplicative analysis of the dominant B-cell epitopes in PD-L2. Finally, using antigenic index analysis, B-cell epitopes of PD-L2 were predicted. Homology of human PD-L2 and mouse PD-L2 was analyzed by BLAST to match the B-epitopes of human PD-L2 with the sequence of mouse PD-L2. The sequences matching human PD-L2 epitopes were mapped on the binding model of mouse PD-1 and mouse PD-L2, and the stereostructure was simulated to further analyze the relationship between B cell epitopes and protein binding site. Results: The full-length amino acid sequence of PD-L2 contains a total of 273 amino acids and a molecular weight of 31 kDa soluble protein. Through comprehensive analysis, the dominant epitopes of B-cell may be the amino acid sequence of aa60-72, aa93-97, aa133-139, aa164-171 section. Simulated stereostructure analysis revealed that the epitope aa60-72 segment “QKVENDTSPHRER” is located in the binding region of PD-1 and PD-L2. Conclusion: Bioinformatics predicted that B-cell epitopes of PD-L2 in the predominant B-cell epitopes of PD-L2 were aa60-72, aa93-97, aa133-139,164-171, of which the epitope aa60-72(QKVENDTSPHRER) was located in the area where PD-1 and PD-L2 are combined. The above can provide theoretical basis for PD-L2 antibody design and research. 2018 Vol. 48 (5): 319-324,329 [Abstract] ( 731 ) HTML (1 KB)  PDF (1534 KB)  ( 1178 ) 325 Chemotaxis of human adipose tissue derived mesenchymal stem cells to cervical cancer cells ZHENG Lihong, LIU Yi, ZHU Xueqiong. Objective: To investigate the chemotaxis of human adipose tissue derived mesenchymal stem cell (hATMSC) to human cervical cancer cells and to provide experimental basis for the development of hATMSC as a target vehicle for the treatment of cervical cancer. Methods: Primary hATMSC from subcutaneous fat of patients undergoing cesarean section was isolated. hATMSC was induced by osteogenic and adipogenic differentiation medium, and using Alkaline Phosphatase, Oil Red O staining respectively to verify the differentiative capacity. The expression of surface antigen and OCT-4 mRNA were detected by flow cytometry and qRT-PCR, respectively. The migration ability of hATMSC to cervical cancer squamous cells SiHa, MS751, C33-A, CaSki and adenocarcinoma cell Hela was detected by Transwell assay. Results: Having been incubated in osteogenic and adipogenic differentiation medium for 14 days, hATMSC was positive by Alkaline Phosphatase and Oil Red O staining. The hATMSC was positive for CD90, CD44, CD105 and HLA-ABC, but negative for CD34, CD45, HLA-DR. OCT-4 mRNA was highly expressed in hATMSC. hATMSC had high motility to migrate to cervical cancer cells, and the migration to C33-A was the strongest among them. Conclusion: hATMSC was successfully isolated and cultured, with the characteristics of stem cells having multi-directional differentiation potential. hATMSC has obvious chemotaxis to different cervical cancer cells, suggesting that hATMSC might be used as a potential carrier for targeting therapy of cervical cancer. 2018 Vol. 48 (5): 325-329 [Abstract] ( 747 ) HTML (1 KB)  PDF (1430 KB)  ( 1007 ) 330 Efficacy and mechanism of clove oil and jasmine oil on treatment of depressive mice LIN Huiyue, WAN Xinlong, DU Yongjun, XIE Hongli. Objective: To study the effects of odorants on the treatment of depression and their mechanisms. Methods: Forty-five C57BL/6J mice with similar in multiple physiological indices were selected. Eight of them were randomly picked out as norm group and the others were stimulated by the depression modeling methods: chronic unpredictable mild stress model (CUMS) and solitary rearing. After 28 days, 32 mice were screened and randomly divided into 4 groups. Three groups were treated respectively by using clove oil (clove group), jasmine oil (jasmine group) and distilled water (distilled water group) in atomization. The treatments were executed 3 times every day and one hour per time, lasting 28 days. The last group (depression group) and the normal group were not given treatment and normally reared for 28 days. Subsequently, multiple indices involve weight tail suspension test (TST) forced swim test (FST) and the serum concentration of 5-HT of mice and the expression levels of olfactory receptors (MOR11, MOR1, MOR124, MOR16, MOR378, MOR544, MOR19, MOR744, MOR63, MOR148, MOR9, MOR151, MOR15) of mice in all groups were tested. Results: The depression model of mice was successfully established after 28 days. After treatment, there were no significant difference in the indices of weight tail suspension test (TST) forced swim test (FST) and the serum concentration of 5-HT of mice between clove group and normal group, but the four indices decreased significantly, compared with the depression group and the distilled water group. However, all the indices except of the weight of jasmine group were significantly increased compared with normal group. There was no significant difference in other indices compared with the depression group and distilled water group except that the weight increased significantly, There were no significant differences in the four indices between distilled water group and depression group. The RT-qPCR results showed that the 12 of 13 olfactory gene expression levels in mice significantly decreased after depressed. After treatment, compared with the normal group, in clove group, 8 genes (MOR11, MOR124, MOR15, MOR16, MOR378, MOR63, MOR744, MOR1) expression levels increased significantly (P<0.05), 3 genes (MOR19, MOR151, MOR544) had no significant difference (P>0.05), the expression level of 2 genes (MOR148, MOR9) were significantly decreased (P<0.05); In jasmine group, only 2 genes (MOR744, MOR1) expression level was significantly increased (P<0.05), 3 genes (MOR11, MOR63, MOR151) had no significant difference (P>0.05), the expression levels of other genes (MOR124, MOR15, MOR16, MOR19, MOR378, MOR544, MOR148, MOR9) decreased significantly (P<0.05); in distilled water group 4 genes (MOR11, MOR63, MOR744, MOR1) expression levels increased significantly (P<0.05), 4 genes (MOR124, MOR16, MOR378, MOR151) expression level showed no significant difference (P>0.05), and the expression level of the other genes (MOR15, MOR19, MOR544, MOR148, MOR9) decreased significantly (P<0.05). There were no significant differences in the remaining genes in each group. Conclusion: Clove oil is highly effective on depression, but jasmine oil and distilled water are rarely. Furthermore, the effect of odorants on depression is closely related to olfactory gene expression levels. 2018 Vol. 48 (5): 330-337 [Abstract] ( 822 ) HTML (1 KB)  PDF (1432 KB)  ( 1482 ) 338 The effect of ginsenoside Rb2 on the regulation of lipid metabolism in fatty liver mice and its mechanism HONG Yilian, GU Xuejiang, XU Jing, LIN Yi, SI Qiya. Objective: To investigate the role of ginsenoside Rb2 in hepatic lipid metabolism in fatty liver mice and its mechanism. Methods: Male C57BL/6J mice were randomly divided into normal control group, high-fat model group and ginsenoside Rb2 group. The normal control group was fed with 10% kcal control diet, and the rest was fed with 60% kcal high fat diet for 8 weeks. PBS or ginsenoside Rb2 (40 mg•kg-1•d-1) was injected intraperitoneally for 10 days. Realtime PCR and Western blot were used to investigate the expression and mechanism of lipid metabolism-related genes such as lipid synthesis and fatty acid oxidation in liver tissues. Results: Ginsenoside Rb2 significantly reduced liver weight, improved hepatic steatosis, down-regulated fatty acid synthase (FAS), lipoprotein lipase (LPL), sterol regulatory element binding protein (SREBP)-1c, and up-regulated carnitine palmitoyl transferase (CPT)-1 and cholesterol 7α-hydroxylase (CYP7A1) mRNA levels, increased the expression of hepatic peroxisome proliferator-activated receptor (PPAR) alpha gene and protein. Conclusion: Ginsenoside Rb2 can attenuate hyperlipidemic fatty liver, and its mechanism may be related to the increase of PPARα expression in the liver and the regulation of target genes expression such as SREBP-1c, FAS and CYP7A1 related to lipid metabolism. 2018 Vol. 48 (5): 338-341,349 [Abstract] ( 746 ) HTML (1 KB)  PDF (1336 KB)  ( 1002 ) 342 Effects of Sirt6 on chondrocyte autophagy in chondrocyte senescence: an in vitro study ZHANG Hui, SUN Jiali, CHEN Zexin, WU Yaosen, LIN Yan. Objective: To investigate the effects of Sirt6 in chondrocyte senescence and its effect on chondrocyte autophagy in chondrocyte senescence. Methods: The chondrocytes were isolated and cultured in vitro, and the chondrocyte senescence was induced by continuous passage. The chondrocyte senescence was detected by beta-galactosidase staining, and protein expression of Sirt6 was detected by Western blot. We then overexpressed or knocked down Sirt6 in chondrocytes using lentivirus, and compared the maximum number of passages and the percentages of senescent cells between different groups. The effects of Sirt6 on chondrocyte function were investigated by real-time PCR, to compare the mRNA expression of collagen-II, aggrecan, MMP-13, and ADAMTS-5 in each group. After overexpression of Sirt6 in chondrocytes, the presence of intracellular autophagosomes was analyzed by immunofluorescence, and the ratio of LC3-II/LC3-I was analyzed by Western blot. In addition, the effect of autophagy on the inhibition of senescence and functional degeneration of the chondrocytes during overexpression of Sirt6 was verified by using the autophagy inhibitor, 3-methyladenine. Results: Expression of Sirt6 decreased gradually with an increase in the number of cell generations, and expression of P16 increased significantly in chondrocytes with overexpressed Sirt6. Over expression of Sirt6 in chondrocytes significantly suppressed cellular senescence. Sirt6 promoted the autophagy of chondrocytes, and an autophagy inhibitor inhibited the protective effect of Sirt6 on chondrocytes. Conclusion: Increasing the expression level of Sirt6 can delay the senescence and degeneration of chondrocytes, and its effect is related to the promoted autophagy by Sirt6. 2018 Vol. 48 (5): 342-349 [Abstract] ( 758 ) HTML (1 KB)  PDF (1957 KB)  ( 1113 ) 350 Long-term follow-up of cerebellar hemangioblastomas in von Hippel-Lindau disease TU Ming, HU Shaobo, ZHENG Weiming Objective: To observe the long-term outcome of patients with VHL associated cerebellar hemangioblastomas after tumors resection, compared with patients with sporadic cerebellar hemangioblastomas.Methods: Clinical characteristics and long-term outcomes of VHL related cerebellar gioblastomas (n=31) patients were compared with patients with sporadic cerebellar gioblastomas (n=31), matched for tumor size. Sequencing of VHL gene of patients with VHL disease was conducted to determine the gene mutations. Results: Patients with VHL disease were younger than sporadic patients (P<0.001). The median size of tumors showed no difference in two groups. Median postoperative follow-up was 99 (69, 137) months in VHL group and 101 (67, 135) months in sporadic group. Mean RFS was shorter in VHL related patients, compared with sporadic patients (P<0.001). Overall survival was also worse in VHL group (P=0.037). Nine patients in VHL group died (6 from cerebellar hemangioblastomas, 2 from renal cell carcinomas, 1 from pancreatic neuroendocrine tumors). Two sporadic patients died from recurrent cerebellar hemangioblastomas. Four different mutations were found in patients with VHL disease (559c→g, 716g→c, 697c→t, 746tgDel). Conclusion: The long-term prognosis of VHL associated cerebellar hemangioblastomas is still worse than sporadic cerebellar hemangioblastomas even after complete resection. Regular postoperative follow-up and examination of multiple organs is necessary for patients with VHL disease. 2018 Vol. 48 (5): 350-354 [Abstract] ( 750 ) HTML (1 KB)  PDF (1264 KB)  ( 847 ) 355 Micellar supramolecular hydrogel for delivery of hydrophobic anticancer drugs LIN Xiaoxiao, CAI Zhe, LIN Guangyong.  Objective: To develop a micellar supra-molecular hydrogel composed of α-cyclodextrin (α-CD) and monomethoxy poly (ethylene glycol)-b-poly (e-caplactone) (MPEG5 000-PCL5 000) micelles and explore its feasibility for hydrophobic anticancer drug delivery. Methods: PTX as an object of hydrophobic antitumor drugs, drug-loaded supramolecular hydrogel was characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The effect of α-CD amount on the gelation time, mechanical strength and thixotropic property was studied by a rheometer. Results: Pay-load of paclitaxel (PTX) to supramolecular hydrogel was achieved by encapsulation of PTX into MPEG5 000-PCL5 000 micelles prior mixing with α-CD aqueous solution. In vitro release study showed that the release behavior of PTX from hydrogel could be modulated by changing the α-CD amount in the hydrogel. Furthermore, such supramolecular hydrogel could sustain the releasing of encapsulated PTX compared to free PTX, as indicated by the cytotoxicity assay in vitro. Conclusion: Successful preparation of supramolecular hydrogels loaded with antitumor drug PTX, which mainly through interactions with α-CD. 2018 Vol. 48 (5): 355-360,365 [Abstract] ( 639 ) HTML (1 KB)  PDF (1545 KB)  ( 1272 ) 361 Value of multi b-value diffusion weighted imaging in differential diagnosis of primary and metastatic hepatocellular carcinoma ZHANG Zhijing, WU Aiqin, ZHANG Xiaoyu, FU Yao, DUAN Weimin, ZHENG Wenlong. Objective: To investigate the value of multi b-value DWI double exponential function model in differential diagnosis between primary and metastatic hepatocellular carcinoma. Methods: Forty-one cases of primary hepatocellular carcinoma (23 cases) and metastatic hepatocellular carcinoma (18 cases) confirmed by surgical pathology or puncture were scanned by multi b-value DWI of MRI, being 0, 100, 400, 800, 400 s/mm2. The quantitative values of single exponential function (Slow ADC-Mono, Fast ADC-Mono) and double exponential function (Slow ADC-bi, Fast ADC-bi) were measured and related data were analyzed. Results: Except Fast ADC-Mono, the quantitative values of Slow ADC-Mono, Slow ADC-bi, and Fast ADC-bi were statistically different among primary hepatocellular carcinoma, metastatic hepatocellular carcinoma and normal tissue by single factor analysis of variance. Comparing the LSD results of primary hepatocellular carcinoma and metastatic hepatocellular carcinoma, the quantitative values of both Slow ADC-Mono and Fast ADC-Mono showed no statistical difference between the two groups, while the values of Slow ADC-bi and Fast ADC-bi were statistically different. The quantitative values of Slow ADC-bi were highest in normal tissues, followed by primary liver cancer and lowest in metastatic liver cancer, while the Fast ADC-bi values were highest in primary liver cancer, followed by metastatic liver cancer and highest in normal tissues. In the same disease or normal tissues, whether they were in single exponential function model or double exponential function model, the quantitative values of Fast ADC were far greater than those of Slow ADC. Conclusion: The Slow ADC and Fast ADC parameters of the double exponential function model reflect the cytological characteristics of the malignant tumor to a certain extent. The double exponential function model of multi-b value DWI is of significance in differential diagnosis between primary hepatocellular carcinoma and metastatic hepatocellular carcinoma. 2018 Vol. 48 (5): 361-365 [Abstract] ( 680 ) HTML (1 KB)  PDF (1571 KB)  ( 678 ) 366 Value of intravoxel incoherent motion diffusion MRI in detecting alteration of renal function in diabetic kidneys with hyperfiltration ZHOU Yongjin, YE Xinjian, GONG Enhui, HUANG Xiaoyan, CUI Shihan, SONG Jiawen, LIU Kun, YAN Zhihan Objective: To investigate whether the intravoxel incoherent motion diffusion weighted imaging (IVIM-DWI) was helpful in assessing renal function changes in early-stage diabetic patients with hyperfiltration. Methods: A total of 12 diabetic patients with eGFR≥120 mL•min-1•1.73 m-2 and 12 healthy controls were included in the study. IVIM-DWI of bilateral kidneys was acquired using 11 b values on a 3.0 T MR scanner. The diffusion coefficient of apparent diffusion coefficient (ADC), pure molecular diffusion (D), perfusion-related pseudo-diffusion coefficient (DP) and perfusion fraction (PF) of the renal cortex and medullawere calculated from IVIM-DWI maps of the two groups. Two radiologists independently analyzed all imaging data. Intra-class correlation coefficients (ICCs) and group comparisons by two-sided t-test were made between the two groups. Relationship between IVIM-DWI parameters and eGFR was examined by Spearman correlation test.Results: Inter-observer agreement was “good” (ICC=0.764-0.874) for all parameters. Mean renal cortex and medulla ADC, D and PF value of diabetic patients were significantly higher than healthy controls (P<0.05). There was a significant linear correlation between renal cortex D value and eGFR level (r=0.581, P<0.05). Conclusion: It is indicated that ADC, D and PF value could detect the renal changes of water molecular diffusion and microcirculation perfusion in diabetic kidneys which may emerge as glomerular hyperfiltration that occurs in early-stage diabetic kidneys. However, it is still a challenge for DP value to reflect microcirculation perfusion changes. 2018 Vol. 48 (5): 366-370 [Abstract] ( 723 ) HTML (1 KB)  PDF (1317 KB)  ( 1120 ) 371 The expression and significance of PI3Kp85α in papillary thyroid carcinoma GONG Xiaohua, ZHOU Qi, WU Wenjun, WANG Fang, CHEN Xiaojun Objective: To explore the expression and significance of phosphatidylinositol 3-kinase regulatory subunit alpha (PI3Kp85α) in papillary thyroid carcinoma (PTC). Methods: PI3Kp85α expressions were detected in PTC tissues (n=116) and thyroid papillary hyperplasia (PTH) tissues (n=90) by immunohistochemistry, Western blot, and enzyme-linked immunosorbent assay. The relationships between PI3Kp85α expressions and clinical pathological features in PTC were analyzed. Diagnostic values of PI3Kp85α in PTC were evaluated by receiver operating characteristic (ROC) curve. Results: PI3Kp85α expression levels in PTC tissues were statistically higher than those in PTH tissues (P<0.05). PI3Kp85α expression was associated with lymph node metastasis (P<0.05). The diagnostic sensitivity and specificity of PI3Kp85α were 92.2% and 91.1%, respectively, with a cut-off value of 2.100 and an area under curve of 0.966. Conclusion: PI3Kp85α may be related to the tumorigenesis and progression of PTC. 2018 Vol. 48 (5): 371-374 [Abstract] ( 695 ) HTML (1 KB)  PDF (1305 KB)  ( 935 ) 375 Initial effects of platelet-rich fibrin and bone marrow aspirate concentrate on reconstruction of bone defect in an animal model YANG Ying, LIU Guo, LIU Chuantong, JIN Qiong, ZHU Fei. Objective: To assess the initial effects of platelet-rich fibrin (PRF) in conjunction with bone marrow aspirate concentrate (BMAC) on reconstruction of bone defect. Methods: Four bone defects of 5-mm in diameter were performed in the calvaria of 6 rabbits. These defects were randomly filled with blood clot (group A), autogenous bone (group B), PRF and BMAC (group C). Meanwhile, PRF was collected for histological observation. Animals were sacrificed 6 weeks later, and bone samples were harvested for histological observation and statistical analysis. Results: PRF was milk white fibrin glue with elasticity and toughness. The loose and porous three dimensional network structure and white blood cells at the bottom of PRF could be seen in the histological sections. The percentage of new bone formed in each group was 7.64%±1.02% (group A), 12.94%±0.81% (group B), 37.19%±1.26% (group C), respectively. Group C showed a statistically greater amount of new bone formation compared with group A and group B (P<0.05). Conclusion: PRF mixed with BMAC couldaccelerate the regeneration of new bone to repair the rabbit calvarial defect. 2018 Vol. 48 (5): 375-378 [Abstract] ( 712 ) HTML (1 KB)  PDF (1362 KB)  ( 820 ) 379 Clinical features and CT signs of ischemic colitis ZHAO Fenhua, SHAN Kangfei, HUANG Chaohui, ZHU Weihua. Objective: To investigate the clinical features and CT signs of ischemic colitis (IC). Methods: Retrospective data analysis of 39 patients’ with IC confirmed by CT and colonoscopy. All patients underwent CT plain scan and dual-phase enhanced scan and some underwent abdominal CTA examination. Results: Of 39 patients, ascending colon was found in 9 cases, transverse colon in 13 cases, splenic flexure of colon in 27 cases, descending colon in 36 cases, sigmoid colon in 11 cases and 1 case of rectum. The colonic wall had a different degree of thickening, with an average thickness of 14.3 mm. The density of all invovling intestinal walls decreased, and the enhancement of the intestinal wall weakened. Intestinal dilatation was observed in 7 cases and 10 cases showed mesenteric edema and exudation. A total of 25 cases of abdominal aortic calcification plaque were detected, 8 cases involved the origin of the superior mesenteric artery and inferior mesenteric artery. Conclusion: The typical CT manifestations of IC included colonic wall thickening and edema, weakened enhancement and surrounding mesenteric edema and exudation. The descending colon and the splenic flexure of colon involvement were the most common. After enhanced scan, the “Reticulated python sign” maybe valuable for the evaluation of intestinal wall ischemia. 2018 Vol. 48 (5): 379-382 [Abstract] ( 974 ) HTML (1 KB)  PDF (1289 KB)  ( 1354 ) 383 Clinical analysis of portal venous system thrombosis treated with drugs YU Zheke, WAng Jingjing, QIU Zhenming, ZHAO Zhengzheng, ZHENG Yonghua. Objective: To evaluate the effectiveness of drug therapy for portal vein thrombosis. Methods: A retrospective analysis was conducted on the clinical data of 19 patients with portal vein thrombosis treated with drug from April 2011 to April 2016 at Taizhou Hospital Affiliated to Wenzhou Medical University. Results: All 19 patients who were treated with low molecule Gansugai and Warfarin recovered well. Their average hospital stay was (12±5) d and average follow-up (20±9) months without recurrence. Conclusion: Conservative treatment with drugs is safe and effective for patients with portal vein thrombosis and without peritoneal irritation. 2018 Vol. 48 (5): 383-384,封三 [Abstract] ( 606 ) HTML (1 KB)  PDF (1229 KB)  ( 856 )

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