Research progress of the roles of metabolic reprogramming in radiation-induced direct- and bystander-effects / 中华放射医学与防护杂志

Metabolic reprogramming refers to the phenomenon that tumor cells, in order to meet their own growth and energy needs, regulate their biological functions by changing their metabolic mode, help themselves resist external stresses, and thus enable cells to adapt to hypoxia, acid, nutrient deficiency and other microenvironments and rapidly proliferate. It was found that metabolic reprogramming could contribute to radiation resistance and it also could be induced in bystander cells which may result in radiation resistance and the cancellation. Investigation the mechanism of radiation-induced metabolic reprogramming may provide new ideas and a theoretical framework for radiation protection, radiotherapy, and radio-diagnosis. This article reviewed the research progress on the mechanism of metabolic reprogramming in the direct and bystander effects of radiation.

Simultaneous Determination of 9 Components in Huichun yuzi Granules by LC-MS/MS / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of 9 components in Huichun yuzi granules. METHODS:LC-MS/MS method was adopted. The determination was performed on Shim-pack XR-ODS C18column with column temperature of 30 ℃. The sample size was 5 μL,and ion source as electrospray ion source;MRM mode was adopted. The acetonitrile-water was used as mobile phase for ferulic acid,rutin,paeonol,icariin and schisandrin(gradient elution);positive ion mode monitoring was conducted. The methanol-0.1% fomic acid water was used as mobile phase for naringin,verbascoside, amygdalin and protocatechuic acid(gradient elution);negative ion mode monitoring was conducted. RESULTS:The linear ranges of ferulic acid,rutin,paeonol,icariin,schisandrin,naringin,verbascoside,amygdalin and protocatechuic acid were 0.499 5-500, 25-1 000,0.245 9-250,5.185 1-1 000,0.981 9-1 000,0.248 1-125,2.510 4-250,10-2 500,4.999 7-1 000 ng/mL(r≥0.997 8), respectively. The limits of detection were 0.075,0.30,0.072,0.015,0.015,0.075,0.15,0.30,0.15 ng/mL,respectively;the limits of quantitation were 0.25,1.00,0.24,0.51,0.49,0.25,0.50,0.99,0.49 ng/mL,respectively. The recoveries rate were 91.39%-103.56%(RSD=1.03%-3.67%,n=6).RSDs of stability test ranged 1.4%-3.4%(n=6)within 48 h at room temperature. CONCLUSIONS:The method is rapid,sensitive and reproducible,and can be used for content determination of 9 components in Huichun yuzi granules.It can be used for quality control of Huichun yuzi granules.

Detection of immunoglobulin and TCR gene rearrangements by PCR using BIOMED-2 multi-plex protocols in T lymphoblastie lymphoma and acute lymphoblastic leukemia / 临床与实验病理学杂志

Purpose To investigate the sensitivity of BIOMED-2 primer system in T lymphoblastic lymphoma ( T-LBL) and acute lym-phoblastic leukemia ( ALL) patients immunoglobulin ( Ig) and T-cell receptor ( TCR) gene rearrangement, and to analyze the co-rear-rangement pattern. Methods Amplification of rearranged Ig and TCR gene was performed in standard PCR in 35 T-LBL/ALL pa-tients. PCR products were analyzed by heteroduplex and polyacrylamide gel electrophoresis. Results 16 cases (45. 7%) of 35 sam-ples were detected to have TCR gene rearrangements, including 6 cases (37. 5%) of TCRβgene monoclonal rearrangements, 4 cases (25. 0%) of TCRγ gene monoclonal rearrangements, 3 cases (18. 8%) of TCRβ and TCRγ gene double rearrangements, 2 cases (12. 5%) of TCRδ gene monoclonal rearrangements and 1 case (6. 3%) of TCRγand TCRδgene double rearrangements were detec-ted. 4 cases (11. 4%) of 35 samples detected to have clonal immunoglobulin and TCR gene rearrangements. 11 cases (39. 3%) of 28 T-LBL patients were detected to have TCR gene rearrangements, 6 cases (85. 7%) of 7 T-ALL have TCR gene rearrangements. Con-clusions BIOMED-2 multiplex PCR analysis strategy is a useful technique in the T-LBL patients.

Analysis of TCR gene rearrangements for diagnosis of T-cell lymphomas / 临床与实验病理学杂志

Purpose To discuss the TCR gene rearrangements in the diagnosis of T-cell lymphomas. Methods Formalin-fixed and paraffin-embedded samples including 30 cases of T-cell lymphomas and 30 cases of reactive lymphoid hyperplasia were chosen for ex-tracting genomic DNA and PCR amplification using 56 BIOMED-2 primers. PCR products were analyzed by heteroduplex and polyacryl-amide gel electrophoresis. Results In all 30 cases of T-cell lymphomas, 25 cases (83. 3%) showed TCRβ gene monoclonal rear-rangements, 28 cases (93. 3%) of TCRγ gene monoclonal rearrangements, 4 cases (13. 3%) of TCRδ gene monoclonal rearrange-ments. 29 cases (96. 7%) with TCRβ+TCRγ+TCRδ gene monoclonal rearrangements were detected. but no clonal TCR gene rear-rangements were found in 30 cases of reactive lymphoid hyperplasia. Conclusions The detection of TCR gene rearrangements using BIOMED-2 primers is a useful assistant method for the diagnosis of T-cell lymphomas.

Study on the relationship between Epstein-Barr virus and breast cancer / 临床与实验病理学杂志

Purpose To study the relationship between Epstein-Barr virus ( EBV) and breast cancer. Methods 246 cases of breast lesions at different development stages were selected and EBV DNA, RNA and protein was used by polymerase chain reaction ( PCR) , in situ hybridization ( ISH) , laser capture microdissection ( LCM) , immunohistochemistry ( IHC) EnVision technology. Results No expression of EBV latent membrane protein LMP1 was detected in all 246 cases of benign and malignant breast lesions. In 12 cases of breast cancer of EBV DNA, carcinoma in situ and breast lesions not EBV DNA was detected by PCR. However, using digoxigenin la-beled EBV DNA probe for the 48 cases ( including 12 cases of breast cancer specimens of positive PCR amplification) of benign and malignant breast lesions, no positive hybridization signal was detected in cancer cells, mammary epithelial cells and stromal lympho-cytes. Using laser capture microdissection and PCR amplification, cancer cells and stromal cells were captured respectively from 12 ca-ses of PCR positive and 12 cases of PCR negative of breast cancer specimens, we found EBV DNA was only amplified in mesenchymal cells. In the detection of the EBER expression with EBV RNA probe and in situ hybridization, the results of 75 cases of benign and malignant breast lesions ( including 12 cases of breast cancer by positive PCR amplification) were all negative. Conclusions The re-sults indicate that the tumorigenesis and development of breast cancer have nothing to do with EBV infection in all cases were chosen.

Effects of high-fat diets and rosiglitazone treatment on expressions of peroxisome proliferator-activated receptor-γ coactivator-1α and mitofusin-2 in skeletal muscle of aged rats / 中华老年医学杂志

Objective To investigate the effects of high-fat diets and rosiglitazone treatment on the expressions of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) and mitofusin-2 (Mfn2) in skeletal muscle of aged rats. Methods Male wistar rats aged 21-23 months were randomly divided into old control group (OC; n=20), high-fat diets group (HF; n=20) and high-fat diets plus rosiglitazone treatment group (RSG; n=20). Male wistar rats aged 4-5 months were selected as young control group (YC; n=20). Insulin sensitivity was evaluated by hyperinsulinemic-euglycemic clamp technique at the end of the 4th and 8th week. The expressions at mRNA and protein levels of PGC-1α and Mfn2 in skeletal muscle were determined by polymerase chain reaction and Western blotting technique. Results (1)After 8 weeks, the levels of free fatty acid [(0. 68±0. 18)mmol/L, (0.82±0. 23) mmol/L and (1. 53±0.40) mmol/L], triglyceride [(0.53±0.13) mmol/L,(0. 63±0. 17) mmol/L and (1.08±0.30) mmol/L]and muscle triglyceride [(1.09±0.17) mmol/L,(1.34±0. 20) mmol/L and (2.07±0. 30) mmol/L]in YC group, OC group and HF group were significantly increased and glucose infusion rate [(30.4±4. 2) mg·kg~(-1)·min~(-1), (20.9±2.2) mg·kg~(-1)·min~(-1) and (12. 0±1.9) mg·kg~(-1)·min~(-1)]was significantly decreased as compared with pre-high fat diet, respectively. The levels of fasting free fatty acid [(0.93±0.29) mmol/L], triglyceride [(0.62±0.12) mmol/L]and music triglyceride [(1.68±0.28) mmol/L]in RSG group were significantly decreased and glucose infusion rate [(16.7±1.7) mg·kg~(-1)·min~(-1)]was significantly higher than in HF group. (2)In skeletal muscle, the expressions at mRNA and protein levels of PGC-1α and Mfn2 decreased in OC group compared with YC group (all P<0.01). The expressions of PGC-1α and Mfn2 were lower significantly in HF group than in OC group, and were higher significantly in RSG group than in HF group (all P<0.01). Conclusions The aged rats fed the high-fat diets develop insulin resistance with decreased expressions of PGC-1α and Mfn2 in skeletal muscle. Insulin sensitivity is improved with rosiglitazone treatment by increasing expressions of PGC-1α and Mfn2 in skeletal muscle.