1.Role and mechanisms ofω-3 polyunsaturated fatty acids in inducing cell apoptosis and reversing drug resistance in multiple myeloma
Wei SHI ; Defang FANG ; Yonggang ZHAO
Chinese Journal of Clinical Oncology 2016;43(24):1088-1093
Objective:To explore the role and mechanisms ofω-3 polyunsaturated fatty acids (ω-3PUFAs) alone or in combination with dexamethasone (DEX) in inducing cell apoptosis and reversing drug resistance in multiple myeloma (MM). Methods:DEX-resistant MM cell line MM1R was treated with different concentrations of eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) alone or in combination with DEX for 24 or 48 h. Cel proliferation was detected by MTT assay. Cel cycle and apoptosis were measured by flow cytometry. Expression levels of apoptosis-related proteins were analyzed by Western blot. Two-tailed, unpaired Student's t-test was used to compare the two treatment groups. A value of P<0.05 was considered statistically significant. Results:MM1R proliferation was inhibited by different concentrations (10, 20, 50, and 100μM) of EPA or DHA alone or in combination with 10μM DEX in a dose-and time-dependent manner. The inhibition effect was significantly higher in combinative groups than in single EPA or DHA treatment group (P=0.014, P=0.032). The percentage of G0/G1 phase and cell apoptosis rate in MM1R treated with different concentrations of EPA or DHA alone increased in a dose-dependent manner. This percentage was also significantly higher in the combinative groups than in the single EPA or DHA treatment group (P=0.015, P=0.004). The expression levels of cleaved caspase-3 and Bax were upregulated, whereas those of pro-caspase-3 and BCL-2 were downregulated in a dose-dependent manner. Drug resistance gradually decreased in MM1R cells at different concentrations of EPA or DHA with the increase of drug concentration. The reversal fold also increased gradual y, whereas the cel s decreased in the two drug-combination groups compared with the single-drug group. Moreover, the drug-resistance reversal index increased significantly. Conclusion:ω-3PUFAs can inhibit DEX-resistant MM cell proliferation, arrest cell cycle, and induce cell apoptosis.ω-3PUFAs also exhibit a synergistic anti-resistanteffect in combination with DEX. Furthermore,ω-3PUFAs can serve as novel effective drugs for MM treatment.
2.Expression of sodium iodide symporter mRNA and its clinical significance in differentiated thyroid carcinoma
Jianlin XU ; Xiaoyan LIANG ; Yonggang NAN ; Hua ZHAO ; Changbei SHI
Journal of Xi'an Jiaotong University(Medical Sciences) 2016;(2):260-263
Objective To investigate the expression of sodium iodide symporter (NIS ) mRNA in differentiated thyroid Carcinoma (DTC)and further explore its value in clinical diagnosis and therapy of DTC. Methods The expression of NIS mRNA was detected and analyzed in 21 nodular goiter and 45 cases of DTC (including 35 cases of papillary thyroid carcinoma and 10 cases of follicular thyroid carcinoma)by using real-time fluorescent quantitative reverse transcription polymerase chain reaction (real-time RT-PCR).Results Compared with that in nodular goiter,the mRNA expression of NIS in DTC tissue was significantly decreased (P < 0.05 ). Moreover,the mRNA expression of NIS was significantly correlated with lymph node metastasis and AJCC stage, respectively.The expression of NIS mRNA in DTC with lymph node metastasis was significantly decreased compared with that in DTC without lymph node metastasis (P <0.05).In addition,the expression of NIS mRNA in Ⅱ-Ⅳstage DTC was significantly decreased compared with that in Ⅰ - Ⅱ stage DTC (P < 0.05 ).Conclusion Differential expression of NIS can provide evidence for individual 1 3 1 I therapy for DTC.
3.Role of neuropeptide Y2 receptor in neuropathic pain in rats
Peisheng SHI ; Shuanke WANG ; Yonggang WANG ; Jing WANG ; Haiyu ZHOU
Chinese Journal of Anesthesiology 2013;33(5):565-568
Objective To evaluate the role of neuropeptide Y2 receptor (NPY2R) in neuropathic pain (NP) in rats.Methods Thirty-six adult male Sprague-Dawley rats,aged 8 weeks,weighing 190-210 g,were randomly divided into 3 groups (n =12 each):sham operation group (group S),group NP and NPY2R antisense ohgonucleotide group (group ODN).NP was induced by chronic constrictive injury (CCI).5 μg/μl NPY2R antisense oligonucleotide 30 μl was injected intrathecally 7 days after CCI in group ODN.While normal saline 30 μl was injected intrathecally in group S.The mechanical paw withdrawal threshold and cold allodynia were measured 3 days before CCI (T0,baseline),7 days after CCI (T1) and at 15 min,1.5,3.0,4.5 and 6.0 h after intrathecal injection (T2-6).The animals were then sacrificed after the last measurement and the lumbar segment of spinal cord was removed for determination of the expression of NPY2R and calcitonin gene-related peptide (CGRP) and co-expression of NPY2R with CGRP in spinal dorsal horn neurons (by immuno fluoresceence).Results Compared with group S,the mechanical paw withdrawal threshold was significantly decreased and cold allodynia was increased at T1-6,and the expression of NPY2R and CGRP and co-expression of NPY2R with CGRP in spinal dorsal horn neurons was up-regulated in NP and ODN groups (P < 0.05).Compared with group NP,the mechanical paw with-drawal threshold was significantly increased at T3-5,and the expression of NPY2R and co-expression of NPY2R with CGRP in spinal dorsal horn neurons was down-regulated (P < 0.05),and no significant change was found in cold allodynia and the expression of CGRP in spinal dorsal horn neurons in ODN group (P > 0.05).Conclusion NPY2R in the spinal cord dorsal horn is involved in the maintenance of mechanical hyperalgesia,but not in the maintenance of clod hyperalgesia in rats.
4.Association of galectin-3 expression with biological behaviors of squamous carcinoma of esophagus
Fei SHI ; Guiyong PENG ; Yonggang SUN ; Dapeng YAO
Journal of Third Military Medical University 2003;0(11):-
Objective To investigate the galectin-3 expression in squamous carcinioma of esophagus and analyze its relationship with clinicopathological characteristics.Methods S-P immunohistochemistry was used to detect the expression of galectin-3 in 80 paraffin-embedded specimens of esophageal carcinoma and 22 specimens of normal tissues adjacent to the benigh lesions of esophagus.RT-PCR was performed to detect galectin-3 mRNA expression in 36 pairs of fresh squamous carcinoma tissues and corresponding adjacent normal mucosa.Results Out of 80 specimens of squamous carcinoma of esophagus,62 specimens had positive expression of galectin-3,with the positive rate of 77.5%(62/80)that was significantly higher than 27.27% in normal esophageal tissues(6/22,P
5.Mechanisms of insulin resistance during ischemia reperfusion in the cardiomyocytes of rat
Yonggang LI ; Huanwen CHEN ; Eryong ZHANG ; Donghu SUI ; Yingkang SHI
Chinese Journal of Thoracic and Cardiovascular Surgery 2009;25(3):190-192
Objective Recent.studies have found a strong association of insulin resistance, which might occur during ischemia reperfusion in vitro in the experimental dogs, with disturbed function of cardiomyocytes. Obvious acute insulin resistance, along with glucose dysmetabolism in the reperfused cardiomyocytes, was furher observed in the study performed with ischemia-reperfused ventric- ular myocytes of rats. We tried to investigate preliminarily the molecular mechanisms of insulin resistance in the cardiomyocytes after ischemia reperfusion. Methods An experimental model of insulin-stimulated ischemia reperfusion (SI/R) was created by isolating cardiomyocytes from adult rats. Glucose uptake of the cardiomyoctyes was evaluated with isotope-labeling technique. Glucose trans- porter 4 (GLUT4) translocation induced by insulin was investigated with Western blot analysis, and the intracellular level of free Ca2+ ([Ca2+]I) was measured quantitatively with Ca2+ indicator Fura-2. Results Insulin can stimulated glucose uptake by cardiomyo- cytes, indicating that these cells were insulin-sensitive. Cardiomyocytes were demonstrated notable acute insulin resistmce during reperfusion. Insulin-stimulated GLUT4 translocation in the cardiomyocytes 15 minutes after reperfusion was 72.2% of that in the con- trol group(P<0.05), in which the GLUT4 content in plasma membrane remained unchanged. The finding suggested that a disturbed GLUT4 translocation might happen in the cardiomyocytes during insulin-stimulated ischemia-reperfusion. Calcium overload was identi- fied in the cardiomyocytes with ischemia reperfusion. At 15 minutes of reperfusion, [Ca2+]I was significantly higher in the reperfused cardiomyocytes than that in the control cardiomyocytes[(318.66±23.06)vs(130.70±0.82) nmol/L, P<0.05], and kept at a higher level [(177.79±17.46) nmol/L] at 60 minutes of reperfusion (P<0.05, vs control). Partial correlation analysis revealed a negative correlation of[Ca2+]I with insulin-induced ghcose uptake in the cardiomyoctyes (r = -0.557,P=0.006). Conclusion Disturbed GLUT4 translocation and decreased intrinsic activity may be important molecular mechanisms for the development of insulin resistance in the cardiomyocytes of rat during insulin-simulated ischemia reperfusion,. [Ca2+]I overload may account for the de- creased intrinsic activity d GLUT4.
6.Comparison between standard and high-flexion posterior stabilized (PS) rotating-platform mobile-bearing (RP-MB) total knee system
Jinglei SHI ; Shaobo NIE ; Xu CAI ; Yonggang ZHOU ; Wei FENG ; Jingjing SHI ; Yang WANG
Chinese Journal of Orthopaedics 2011;31(4):316-320
Objective To explore the indication and the functional advantages of the high-flexion posterior stabilized (PS) rotating-platform mobile-bearing (RP-MB) total knee system. Methods A prospective randomized, controlled trial was performed. Osteoarthritis was the indicators for total knee arthroplasty.From Feb. 2009 to Apr. 2009, 75 patients (94 knees) were randomly assigned to to receive either a highflexion PS, RP-MB total knee system(PFC sigma RPF) or a standard one (PFC sigma RP). There were no statistical difference in the baselines, the preoperative scores of the Hospital for Special Surgery (HSS) and the knee range of motion (ROM) of both groups. The functional status were assessed with Hospital for Special Surgery and the ROM of the knee at the postoperative 1, 6, 12, 18 months. The satisfaction rates were assessed at the postoperative 18 months. The radiographic measurements were t assessed at the postoperative 3days and 3, 6, 12, 18 months. Results A total of 70 participants (87 knees) completed the 18-month followup. At the time of the final follow-up, the average Hospital for Special Surgery knee score was 92.4±5.0points in the standard group and 94.7±7.0 points in the high-flex ion group. The difference was not statistically significant(P >0.05). The average maximal flexion was 131.9±14 degrees in the high-flexion group and 123.0±15.3 degrees in the standard group. There was a statistical difference. But it was not enough to confirm our hypothesis that the difference should be higher than 10 degrees. Moreover, the satisfaction rate were 100% in both groups, and no statistical significant difference was found. Conclusion No significant differences were found between standard and high-flexion posterior-stabilized rotating-platform mobile-bearing total knee prostheses in terms of clinical outcomes or range of motion.
7.miR-485-3p regulates radiosensitivity of gastric cancer cells by targeting TLR1/NF-κB signaling pathway
Mingjun LI ; Guangyin WU ; Huijie FAN ; Hao GU ; Ruitai FAN ; Yonggang SHI ; Mingzhi ZHANG
Chinese Journal of Radiation Oncology 2016;25(8):879-884
Objective To investigate whether miR?485?3p plays a role in regulation of radiosensitivity of gastric cancer cells by targeting TLR1. Methods Quantitative real?time PCR and Western blot were used to determine the expression of miR?485?3p and TLR1, respectively. The interaction between miR?485?3p and TLR1 was verified by target prediction software ( DIANA, TargetScan, and miRanda) and dual luciferase reporter assay. Gastric cancer MGC803 cells transfected with miR?485?3p mimic or TLR1 siRNA were exposed to irradiation. Apoptosis assay, colony formation assay, and MTT assay were used to evaluate the changes in radiosensitivity of gastric cancer cells. Dual luciferase reporter assay was used to determine the effects of miR?485?3p overexpression and TLR1 silencing on the activity of NF?κB. Western blot was used to study the effects of miR?485?3p overexpression and TLR1 silencing on NF?κB target genes. Results In gastric cancer cells exposed to radiation, the expression of miR?485?3p was downregulated and the expression of TLR1 was upregulated. TLR1 was predicted to be the target of miR?485?3p by target prediction software. Dual luciferase reporter assay further confirmed TLR1 as the direct target of miR?485?3p. miR?485?3p negatively regulated the expression of TLR1. The overexpression of miR?485?3p, as well as TLR1 silencing, increased the apoptosis rate of cells, reduced colony formation and cell proliferation, and enhanced the radiosensitivity of the cells. Both miR?485?3p overexpression and TLR1 silencing reduced the activity of NF?κB and downregulated the expression of multiple NF?κB target genes. Conclusions miR?485?3p enhances the radiosensitivity of gastric cancer cells probably by targeting TLR1 and regulating the NF?κB signaling pathway.
8.A comparative study of Utrecht interstitial applicator and ring interstitial applicator in three-dimensional conformal brachytherapy for cervical cancer
Hongfu ZHAO ; Dongmei HAN ; Guanghui CHENG ; Mingyuan HE ; Dan SHI ; Zhipeng ZHAO ; Yonggang ZHU
Chinese Journal of Radiation Oncology 2016;25(4):362-366
Objective To investigate the dosimetric differences between Utrecht applicator and ring applicator in three-dimensional (3D) conformal brachytherapy for locally advanced cervical cancer.Methods Twenty-five patients with locally advanced cervical cancer were treated with magnetic resonance imaging-guided 3D conformal brachytherapy.Utrecht applicator and ring applicator were used interchangeably for 96 cycles.Patients were divided into two groups according to the type of applicator.Each group received 48 cycles of treatment, in which ring applicator was first applied for 26 cycles and Utrecht applicator was first applied for 22 cycles.High-risk clinical target volume ( HR-CTV) , width, thickness, and D90 at the point A level, D2 cm3 of organs at risk (OARs), V7 Gy , W7 Gy,A, V7 Gy ,A, and W/T7 Gy were evaluated and analyzed using paired t-test.Results There were no significant differences in HR-CTV and the width, thickness, and D90 at the point A level between the Utrecht group and the ring group ( P=0.487;P=0.340;P=0.857;P=0.921);there were no significant differences in D2 cm3 values of bladder, rectum, sigmoid, and bowel between the two groups ( P=0.136;P=0.802;P=0.985;P=0.458);there were no significant differences in V7 Gy and T7 Gy,A between the two groups ( P=0.076;P=0.435) .The Utrecht group had a significantly larger W/T7 Gy,A than the ring group ( P=0.002 ) .Conclusions Utrecht applicator is appropriate for patients with relatively large width and width/thickness ratio of HR-CTV at the point A level.
9.Correlation between ICRU reference point dose and dose-volume parameters of organs at risk in three-dimensional conformal brachytherapy for locally advanced cervical cancer
Hongfu ZHAO ; Dongmei HAN ; Guanghui CHENG ; Dan SHI ; Yonggang ZHU ; Zhipeng ZHAO ; Yuxin GE
Chinese Journal of Radiation Oncology 2016;25(5):483-488
Objective To investigate the correlation between ICRU reference point dose and dosevolume parameters of organs at risk (OARs) under different bladder and rectal filling status in threedimensional conformal brachytherapy for locally advanced cervical cancer.Methods A total of 31 patients who received magnetic resonance imaging-guided three-dimensional conformal brachytherapy for cervical cancer in 96 fractions were enrolled.The ICRU rectal and bladder reference points were determined in the treatment planning system,and the doses at these points were recorded and compared with the dose-volume parameters of the rectum and bladder.The paired t-test was used to analyze the differences between them.Results Bladder DICRU was lower than bladder D0.1cm3 and D1 cm3 (P=0.000 and 0.000),higher than bladder D5 cm3 and D10cm3 (P=0.000 and 0.000),and similar to bladder D2 cm3 (P=0.345).Under the bladder filling status,bladder DICRU was lower than D2cm3.Rectal DICRU was lower than rectal D0.1 cm3 and D1cm3 (P =0.000 and 0.002),higher than rectal D5 cm3 and D 10 cm3 (P =0.000 and 0.000),and similar to rectal D2cm3 (P=0.058).The ICRU bladder and rectal reference point doses were positively correlated with corresponding D2 cm3.In the case of bladder volume ≥ 200 cm3,the ICRU bladder reference point dose underestimated bladder D2 cm3.In the case of rectal volume ≥ 37 cm3,the ICRU rectal reference point dose overestimated rectal D2 cm3.Conclusions In three-dimensional conformal brachytherapy,it is generally safe to use D2 cm3 as an index to evaluate OARs,but when the bladder or rectum is in an empty status,the ICRU bladder or rectal reference point doses should be considered.
10.A comparative study of CT-and MRI-based three-dimensional conformal brachytherapy for locally advanced cervical cancer
Zhipeng ZHAO ; Yonggang ZHU ; Hongfu ZHAO ; Guanghui CHENG ; Mingyuan HE ; Dan SHI
Chinese Journal of Radiation Oncology 2015;(4):408-413
Objective To compare target volume and dosimetry between computed tomography (CT)?and magnetic resonance imaging (MRI)?guided three?dimensional (3D) conformal brachytherapy for locally advanced cervical cancer, and to provide evidence for optimization of the image?guided approach and improvement of treatment regimen. Methods Thirteen patients with locally advanced cervical cancer who were treated with radical radiotherapy in our hospital in 2014 were enrolled as subjects. All patients received MRI?guided 3D conformal intracavitary/ interstitial brachytherapy. All patients received MRI and CT scans for each brachytherapy fraction, based on which the target volume delineation, intracavitary/ interstitial regimen design, and intracavitary?only treatment regimen design were performed. Comparison of data between MRI and CT scans was performed using paired t test. Results The width and volume of the high?risk clinical target volume ( HR?CTV) were significantly smaller in the MRI simulation than in the CT simulation ((38.0±9?? 4) mm vs. (45.1±8?? 7) mm, P= 0?? 000;(34.2±15?? 3) cm3 vs. (42.9±20?? 4) cm3 , P= 0?? 002), and the width, thickness, and volume of the intermediate?risk clinical target volume (IR?CTV) were also significantly smaller in the MRI simulation than in the CT simulation ((58.8±9?? 4) mm vs. (65.4±10?? 3) mm, P= 0?? 000;(34.8±6?? 3) mm vs. (37.5±6?? 3) mm, P= 0?? 001;(90.9±28?? 5) cm3 vs. (109.0±36?? 4) cm3 , P= 0?? 000). The D90 values for HR?CTV and IR?CTV were significantly higher in the MRI simulation than in the CT simulation (87?? 6 Gy vs. 85?? 8 Gy, P= 0?? 013;67?? 7 Gy vs. 66?? 3 Gy, P= 0?? 005), while the average D2 cm3 values for the bladder and rectum were significantly lower in the MRI simulation than in the CT simulation ( 73?? 1 Gy vs . 75?? 5 Gy , P= 0?? 011 ; 61?? 0 Gy vs . 65?? 7 Gy , P= 0?? 000 ) . Conclusions Compared with the MRI simulation, the CT simulation overestimates the width of target volume. MRI has substantial advantages in dosimetry for target volume and normal tissues. The intracavitary/ interstitial treatment can make up the reduced dose for the target volume resulting from the CT simulation.