1.Effect of 5-aminolevulinic acid mediated photodynamic therapy on proliferation and apoptosis of gastric cancer cell line MKN-45
Xiaolong WANG ; Jun WEI ; Liwen CHENG ; Qian ZHANG ; Peng YAN ; Guiyu WANG
Cancer Research and Clinic 2016;28(4):221-225
Objective To explore the 5-aminolevulinic acid (5-ALA) mediated photodynamic therapy (PDT) for proliferation and apoptosis of gastric cancer cell line MKN-45.Methods The human gastric cancer cell line MKN-45 was cultured in vitro.The MTT method was used to detect fixed lighting processing (laser radiation dose fixed for 25.00 J/cm2,5-ALA photosensitizer concentrations were 0 mmol/L,0.25 mmol/L,0.50 mmol/L,1.00 mmol/L,2.00 mmol/L),fixed photosensitizer concentration treatment (5-ALA photosensitizer concentrations fixed to 1.00 mmol/L,laser radiation dose were 0 J/cm2,6.25 J/cm2,12.50 J/cm2,25.00 J/cm2,50.00 J/cm2,100.00 J/cm2) of the cell apoptosis rate,and then fixed 5-ALA photosensitizer 1 mmol/L and selected the control group,5-ALA group,pure illumination group and PDT group.The promoting effect on cell apoptosis was determined by MTT,electron microscope and flow cytometry.The activity of human gastric cancer cells MKN-45 apoptosis related proteins p65 and the downstream gene regulatory protein bcl-2,bax,caspase-3 and caspase-9 were detected by using Western blot method.Results When laser radiation dose was fixed,5-ALA photosensitizer concentrations in promoting gastric cancer cell apoptosis was positively related to the dominant (P < 0.01).When 5-ALA photosensitizer concentration was fixed,laser radiation dose effect had no longer dominant after 25.00 J/cm2 positive correlation (P =0.613).When 5-ALA photosensitizer concentration was fixed in 1 mmol/L,compared to control group,5-ALA group and pure illumination group,the inhibition of the MKN-45 cell proliferation in PDT group was more apparently,apoptosis rate increased obviously,signal pathways in cell apoptosis protein involved in the NF-kappa B p65 expression was reduced,the downstream gene regulation of target protein bcl-2,caspase-3 and caspase-9 expression were reduced,and bax protein expression was increased.Conclusions 5-ALA mediated photodynamic has a strong inhibitory effect on the growth of gastric cancer cell line MKN-45 in vitro.It can significantly increase the apoptosis rate of gastric cancer cells,and the effect may be carried out by activating the NF-kappa B pathway.
2.Preparation and biological evaluation of radiolabeled folate receptor-positive tumor agent 99 Tcm-(HYNIC-NHHN-FA) (EDDA)
Qian YU ; Fang XIE ; Wenjiang YANG ; Jie LU ; Zhigang LIANG ; Cheng PENG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(1):15-19
Objective To prepare a novel radiolabeled FR?positive tumor targeting agent 99 Tcm?( HYNIC?NHHN?FA) ( EDDA) and evaluate its biological properties. Methods FA derivative FA?NHHN?HYNIC was synthesized and radiolabeled with 99 Tcm using EDDA as a coligand. The radiochemical purity, octanal/water partition coefficient and in vitro stabilities of the complex were studied after purified by HPLC. In vitro cellular uptakes were performed on FR?positive KB cells ( human oral epidermoid carcinoma cells) . Biodistribution and microSPECT/CT imaging were investigated on normal Kunming mice and nude mice bearing KB tumors, respectively. Results The radiochemical purity of the complex was over 95% after pu?rified by HPLC. It displayed high stability both in saline and in serum. It also exhibited high specific FR binding in FR?positive KB cells in vitro. The binding ratio was (6.76±0.60)%1 h after incubation, and de?creased to (0.24±0.02)% after adding excessive FA. The results of biodistribution showed high kidney up?take in normal mice, and the uptake reached (21.79±9.79) %ID/g 0.5 h after injection. Flank KB tumors were clearly visualized with 99 Tcm?( HYNIC?NHHN?FA ) ( EDDA ) by microSPECT/CT imaging at 2 h postinjection, and the uptake could be inhibited by excessive FA. Conclusions 99 Tcm?( HYNIC?NHHN?FA) ( EDDA) exhibits good pharmacokinetic properties, suggesting its potential as a promising FA targeting agent for tumor imaging.
3.Setup errors of external-beam and intracavitary radiotherapy for cervical cancer:an analysis based on cone-beam computed tomography
Qian PENG ; Pei WANG ; Xianliang WANG ; Chuandong CHENG ; Lihao LIU ; Chengjian HUANG
Chinese Journal of Radiation Oncology 2016;25(10):1113-1116
Objective To analyze the setup errors of external?beam and intracavitary radiotherapy for cervical cancer based on the data of cone?beam computed tomography ( CBCT) , and to provide a basis for clinical practice. Methods Sixty patients with stage ⅡA?ⅢB cervical cancer who were admitted to our hospital and underwent external?beam and intracavitary radiotherapy from March to June, 2015 were enrolled as subjects. Sixty patients with stage ⅡA?ⅢB cervical cancer undergoing conventional external?beam radiotherapy within the same period were also enrolled. The CBCT?based setup errors, containing setup errors in x, y, and z directions and three?dimensional vector deviation, in the initial treatment were obtained from each patient. Comparison of errors between the two treatment approaches was made by independent?samples t test. Results Fitting the data to a linear model revealed that the setup errors in x, y, and z directions increased with the increase in the distance between the position reference point and the center point of the target volume. External?beam radiation combined with intracavitary radiation had significantly reduced setup errors in x, y, and z directions than the conventional external?beam radiation ( 0.13± 0?12 vs. 0.31± 0?24, P=0?000;0.23±0?18 vs. 0.47±0?36, P=0?001;0.18±0?11 vs. 0.27±0?18, P=0?001). Conclusions In order to reduce the setup errors, CT scan needs not only a reference marker as close as possible to the center of the tumor, but also a reliable and accurate approach for postural fixation.
4.Effects of electro-acupuncture on functional performance and the expression of apoptosis-related proteins Bcl-2 and Bax in the hippocampal CA1 region of rats with cerebral infarction
Qian YU ; Xiaohong LI ; Ling HUANG ; Ming CHENG ; Yi LI ; Xinpu PENG ; Ke YU
Chinese Journal of Physical Medicine and Rehabilitation 2012;34(4):245-249
Objective To observe effects of electro-acupuncture (EA) on the expression of apoptosis related protein B-cell lymphoma/leukemia gene 2 (Bcl-2) and Bcl-associated protein X (Bax) in the hippoeampal CA1 region of rats with cerebral infarction,and on their behavior. MethodsAn animal model of cerebral ischemia was established by right side middle cerebral artery occlusion using thread in 48 male,adult Wistar rats,who were then randomly divided equally into an EA group and a control group.Both groups were sub-divided into 1 week,2 weeks and 3 weeks subgroups.The EA group began receiving EA 24h after the occlusion,applied at the Baihui (DU20) and Dazhui (BU14) points,once daily,for one,two or three weeks.The control group was reared conventionally and was not given any special treatment.The rats'learning and memory,motion and neural function were evaluated.The expression of Bcl-2 and Bax in the CA1 region of the hippocampus on the infarcted side and changes of apoptosis indexes were detected using immunohistochemical techniques.ResultsThe learning,memory,motion and neural function of the EA group rats were better on average than those of the control group at all observation time points.The expression of Bcl-2 protein increased along with reduced expression of Bax protein in the EA group significantly more than among the controls.TUNEL positive cells in the CA1 region of the hippocampus were reduced significantly in the EA group compared with the control group.ConclusionsBehavioral ability after cerebral infarction can be improved by EA at the Baihui (DU20) and Dazhui (BU14) points.EA can up-regulate the expression of Bcl-2 and down-regulate the expression of Bax and reduce TUNEL positive cells in the CA1 region of the hippocampus after cerebral infarction,which might be the mechanism of its neuroprotection.
5.Application value of combined detection of serum lipoprotein-associated phospholipase A2,homocysteine and blood lipid in diagnosis of cerebral infarction
Qian ZHAO ; Xiaoning ZHANG ; Tao ZHANG ; Kuangfa LI ; Huiyu CHEN ; Ruiqing XING ; Xiaodong CHENG ; Daorong PENG
International Journal of Laboratory Medicine 2017;38(10):1341-1343,1346
Objective To investigate the correlation between serum lipoprotein-associated phospholipase A2 (LP-PLA2) and homocysteine (Hcy) level change with cerebral infarction,and clinical value of combined detection of serum LP-PLA2,Hcy and blood lipid level in the diagnosis and differentiation diagnosis of cerebral infarction.Methods The serum lipoprotein-associated phospholipase A2,homocysteine and blood lipid in sixty-five cases of cerebral infarction(cerebral infarction group) and contemporaneous 64 healthy persons qualified in physical examination(health control group) were selected as the research subjects.The levels of serum LP-PLA2,TG,HDL,LDL,CHO and Hcy were detected.The differences were compared among various groups.Results The TG and LDL levels had no statistically significant difference between the health control group and cerebral infarction group(P>0.05),but the LP-PLA2 and Hcy levels in the cerebral infarction group were higher than those in the health control group(P<0.01),while serum HDL and CHO levels were lower than those in the health control group(P<0.01).Serum High Hcy and LP-PLA2 levels were the independent risk factors for cerebral infarction,however,high HDL was a protective factor for cerebral infarction.In the combined detection,the combined detection of LP-PLA2 and Hcy was superior to single index detection and other combined detection mode.Conclusion Serum LP-PLA2 and Hcy levels in the patients with cerebral infarction are significantly higher than those in the health control group,indicating that it may be involved in the occurrence of cerebral infarction,and may become an early biological marker for predicting cerebral infarction occurrence.The combined detection of serum LP-PLA2 and Hcy has highly clinical value in the diagnosis of cerebral infarction.
6.Factors for the synthesis of 18F-MyoZone and myocardial tomography in normal swine
Tiantian MOU ; Xianzhong ZHANG ; Zuoquan ZHAO ; Cheng PENG ; Qian YU ; Jie LU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(5):260-264
Objective To study the factors affecting the synthesis of 18F-MyoZone,and to evaluate its potential as a myocardial perfusion imaging (MPI) agent in normal Chinese mini-swine.Methods 18F-MyoZone was prepared by substituting the leaving group toluenesulfonyloxy (OTs) from the precursor compound with 18F-fluoride (18F-F-).The conditions affecting the labeling yield were studied by varying the amount of K2CO3 and precursor compound,18F-fluorination reaction time and temperature.PET was performed at 5,30,60 and 120 min post-injection on normal Chinese mini-swine.Results The doses of K2CO3 and precursor,the reaction time and the reaction temperature could affect the labeling yield of 18F-MyoZone,especially K2CO3.The optimized synthetic condition was 1.0 mg K2CO3,2.0 mg mpp2-OTs,20 min reaction time at 90 ℃.The total radio-synthesis time in this condition was 60 min.The uncorrected radiochemical yield was (24.0±5.1) %.The radiochemical purity was >98%.PET imaging showed that 18F-MyoZone had high initial uptake (SUV=8.17± 1.83 at 5 min post-injection) and good retention (SUV =5.78±0.99 at 120 min post-injection) in the heart.The clearance of 18F-MyoZone from liver was very fast.The heart/liver ratios were 3.32,5.31,6.09 and 5.76 at 5,30,60 and 120 min post-injection,respectively.From 5 to 120 min post-injection,the outline of heart was clear and intact.There was almost no interference from the adjacent organs.The quality of PET images was highly satisfactory.Conelusions 18 F-MyoZone has the potential to be a good myocardial perfusion agent.The amount of K2CO3 used could significantly affect the labeling yield of 18F-MyoZone.
7.Changes of MAPKs expression in rat hippocampal neurons after sleep deprivation
Qian ZHANG ; Jiangtao CHENG ; Shuchun WANG ; Peng QIAO ; Yiling WANG ; Jinglan WU ; Yuruo WANG
Chinese Journal of Pathophysiology 2000;0(10):-
AIM: To explore the change and the possible role of MAPKs in rat hippocampus neuron after sleep deprivation. METHODS: The morphology of hippocampus neuron after sleep derivation was observed by TUNEL and HE staining, the activity of ERK was assayed by ?-liquid scintillation counting and the expression of JNK was detected by Western blot. RESULTS: In paradoxical sleep deprivation (PSD) group, the number of apoptotic cells in hippocampus was increased. The scores of ERK activity were (1 764.00)?941.56. Compared with control groups, the ERK activity was obviously decreased (P
8.Effects of total flavonoids of Chrysanthemum indicum on free radical in adjuvant arthritic rats.
Junyan ZHANG ; Lei ZHANG ; Jun LI ; Wenming CHENG ; Lei PENG ; Qian ZHANG
China Journal of Chinese Materia Medica 2010;35(3):344-347
OBJECTIVETo investigate the effects of total flavonoids of Chrysanthemum indicum (TFC) on metabolism of free radical and immunoregulatory effects in adjuvant arthritis (AA) rats.
METHODAA rats were induced by Freunds complete adjuvant. Secondary paw swelling of AA rats was measured with volume meter to observe the antirheumatic effect of TFC. The levels of SOD, MDA and NO in serum and supernatant of peritoneal macrophage were measured by commercial assay kits. ConA-induced splenocyte proliferation and IL-2 level produced by splenocyte were detected by MTT method.
RESULTTFC could decrease the levels of MDA and NO, as well as increase the activity of SOD in serum and supernatant of peritoneal macrophage compared with AA model group. Meanwhile, the suppressed lymphocyte proliferation and IL-2 production of splenic lymphocytes in AA rats were reversed by treatment with TFC.
CONCLUSIONTFC showed significant therapeutical effect on adjuvant arthritis and its mechanism was at least in part related to the antioxidant and immunoregulatory effects.
Animals ; Antioxidants ; metabolism ; Arthritis, Experimental ; drug therapy ; immunology ; metabolism ; Cells, Cultured ; Chrysanthemum ; chemistry ; Flavonoids ; chemistry ; therapeutic use ; Free Radicals ; metabolism ; Interleukin-2 ; metabolism ; Lymphocyte Activation ; drug effects ; Male ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred C57BL ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spleen ; cytology ; Superoxide Dismutase ; metabolism
9.Increasing Expression of IL-3 7 in Peripheral Blood Mononuclear Cell of Patients with Primary Biliary Cirrhosis and Its Clinical Significance
Tengda LI ; Yan CHEN ; Shuping LONG ; Mingli GU ; Peng LIU ; Linhong WU ; Shunjiang DENG ; Weiwei ZHANG ; Cheng QIAN ; Anmei DENG
Journal of Modern Laboratory Medicine 2017;32(1):12-15
Objective To test the expression level of IL-37 in peripheral blood mononuclear cells (PBMCs)of patients with primary biliary cirrhosis (PBC)and further explore its clinical significance in the pathological process of PBC.Methods Pe-ripheral blood samples were collected from 42 patients diagnosed as PBC and 38 health individuals examined at the same time during June 2013 to August 2015 in Changhai Hospital.PBMCs were separated by sucrose density gradient centrifugation, qualified Real Time-Polymerase Chain Reaction (qRT-PCR)was used to measure IL-37 mRNA expression level in PBMCs. Enzyme-Linked Immuno Sorbent Assay (ELISA)was to measure the protein level of IL-37,IL-6,IL-17,TNF-α,TGF-β,IL-18 and IL-23 in plasma.Meanwhile,the pathological stages of PBC cases were recorded.Pearson correlation analysis was performed on IL-37 and IL-6,TNF-α,IL-17,TGF-β,IL-18 and IL-23.Spearman rank correlation analysis was on IL-37 and pathological stages of PBC.Results The mRNA and protein level of IL-37 in experimental and controlled group were 2.81 ±0.94 vs 1.09±0.56,356.14±169.36 pg/ml vs 86.68±48.23 pg/ml separately(t=9.811,9.462,P<0.000 1),with sta-tistical differences.The correlation analysis showed that IL-37 was positively related with IL-17,TNF-α,IL-6 and TGF-β(r=0.561 2,0.661 9,0.672 1,0.765 3,P<0.001),and disease stages (Ⅰ~Ⅳ)(rs=0.348 9,P<0.05).Conclusion IL-37 might involve in the pathological process of PBC,and it is significant for disease prediction and diagnosis.
10.Exploring the prediction model of chronic renal failure based on serum proteomics
Lei HE ; Yawei CHENG ; Ping LIAO ; Heng HU ; Yaming JIN ; Fufeng LI ; Wenjing WANG ; Peng QIAN ; Yiqin WANG
Basic & Clinical Medicine 2010;30(3):263-267
Objective To Screen serum protein markers related to CRF and establish a diagnosis model,exploring and discussing its significance in serodiagnosis by comparing differences of serum protein spectrum expression between patients with chronic renal failure (CRF) and control group.Methods The trial included 62 CRF patients and 28 control ones.Serum samples were tested by surface enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS).The data were analyzed to screen serum proteomic biomarkers.By bioinformatics analysis,decision classification tree models were to be established and tested.Results A total of 19 effective protein peaks were significantly different between CRF and normal control (P<0.001) at m/z range of 1 500 to 30 000,among which 18 showed low expression and 1 showed high expression in CRF.CRF and normal control were obviously different in the clustering;By bioinformatics analysis,a CRF-normal controls of the diagnostic decision tree model was developed,which was 87.8% in with prediction accuracy rate of 87.8% sensitivity of 87.1% and a specificity of 89.3%.Condusion Diagnostic decision tree model provides a more accurate prediction and solid experimental evidence for early clinical diagnosis.