Alcoholism ; metabolism ; Animals ; Brain ; metabolism ; Chemokine CCL2 ; genetics ; metabolism ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, CCR2 ; genetics ; metabolism

Objective To construct plasmid vectors of calmodulin(CaM)Mg2+binding site mutants,and to express,purify and identify the mutant proteins. Methods Three kinds of cDNAs coding for the mutated CaM were cloned into pGEX?6P?3 plasmid vectors. These recombinant plasmids were transfected into Escherichia coli BL21 to express GST fusion proteins of CaM mutants. The fusion proteins were purified with Glutathione?Sep?harose 4B beads and PreScission protease. Results Both enzyme digestion analysis and DNA sequence identification proved the successful con?struction of the CaM mutant plasmids. SDS?PAGE results showed the high purity of each CaM mutant protein. The concentrations of three CaM mu?tants were around 1.0 mg/mL. Conclusion Prokayotic expression vectors of CaM Mg2+binding site mutants were successfully developed,and the eli?gible CaM mutant proteins were obtained. This study provided an important basis for further study on CaM’s biological function.

Objective To evaluate the clinical feasibility of localization CT enhanced image replacing plain CT scan image for target delineation and dose calculation.Methods Forty cases of NPC were collected and divided into two groups with different concentrations of contrast agents.The contours of planning target volume (PTV) and organs at risk (OARs) of each case were delineated in the plain scan image,and the contours of PTV and OARs were copied to the enhanced image.Two plans based on the plain scan image and the enhanced image were designed in the planning system of Eclipse.The dose distribution and OARs and MU were compared between the groups.Results No statistical differences were found in the dosimetry of PTV,OARs and MU (P＞0.05).Conclusion The image intensifier has little effect on the dose calculation of Eclipse for NPC.In the radiotherapy for NPC,the localization CT enhanced image can be used to replace the plain CT scan image for target delineation and dose calculation.

Objective To investigate a method for the purification of the N?terminal peptide fragment(NT)of the myocardial calcium channel Cav1.2,and characterize its interaction with calmodulin(CaM). Methods EscherichiacoliBL?21 cells were transformed with plasmid pGEX?6p?3/NT harboring the NT?GST fusion gene. The cells harboring pGEX?6p?3/NT were cultured and protein expression was induced with isopropyl?β?D?thiogalactoside(IPTG). Then,the GST?NT fusion protein was purified by using glutathione Sepharose 4B(GS?4B)beads. GST was cleaved off with the PreScission protease,and SDS?PAGE was performed to detect the purity and relative molecular weight of the purified peptide. Further, GST pull?down assay was performed to characterize the interaction of the NT peptide with CaM. Results SDS?PAGE analysis showed that the NT peptide was successfully purified,with high purity. Results of the GST pull?down assay showed that the NT peptide could interact with CaM. Conclusion This study establishes a method for the purification of the NT peptide and lays the foundation for further research on the interaction partners and biological functions of NT.

OBJECTIVETo explore the impact of microwave radiation on GC-2spd cells.

METHODSWe exposed cultured GC-2spd cells to microwave radiation at the average power densities of 0, 10 and 30 mW/cm2 for 15 minutes and, from I to 24 hours after the exposure, we observed the changes in cell proliferation, histology and ultrastructure, cell apoptosis, and cAMP content by MTIT, light microscopy, electron microscopy, flow cytometry and ELISA.

RESULTSCompared with the control group, the GC-2spd cells showed a significant decrease in proliferation ability at 1 -24 hours after 10 and 30 mW/cm2 microwave radiation, except at 12 hours after 30 mW/cm2 radiation (P <0.05 or P <0.01), with reduced length and number of cell enation and increased intra cytoplasm vacuoles. The rate of cell apoptosis (%) was significantly increased in the 10 and 30 mW/cm2 groups at 6 hours (4.56 +/- 2.09 vs 14.59 +/- 1.09 and 8.48 +/- 1.73, P <0.05 or P <0.01) , with agglutination and margin translocation of chromatins and obvious dilation of endo cytoplasmic reticula. The cAMP content (nmol/g) in the GC-2spd cells was remarkably reduced in the 10 and 30 mW/cm2 groups at 6 and 24 hours (2.77 +/-0.24 vs 1.65+/- 0. 17 and 1.96+/-0.10, 3.02 +/-0.47 vs 2.13 +/-0.33 and 1.69 +/-0.27, P <0.05 or P <0.01).

CONCLUSIONMicrowave radiation at 10 and 30 mW/cm2 may cause injury to GC-2spd cells, which is manifested by decreased content of intracellular cAMP, reduced activity of cell proliferation, and increased rate of cell apoptosis.

Animals ; Apoptosis ; radiation effects ; Cell Line ; radiation effects ; Cell Proliferation ; radiation effects ; Male ; Mice ; Microwaves ; adverse effects ; Spermatocytes ; radiation effects

OBJECTIVETo investigate the effect of curcumin (Cur) on radiosensitivity of nasopharyngeal carcinoma cell CNE-2 and its mechanism.

METHODThe effect of curcumin on radiosensitivity was determined by the clone formation assay. The cell survival curve was fitted by Graph prism 6. 0. The changes in cell cycle were analyzed by flow cytometry (FCM). The differential expression of long non-coding RNA was detected by gene chip technology. Part of differentially expressed genes was verified by Real-time PCR.

RESULTAfter 10 micro mol L-1 Cur had worked for 24 h, its sensitization enhancement ratio was 1. 03, indicating that low concentration of curcumin could increase the radiosensitivity of nasopharyngeal carcinoma cells; FCM displayed a significant increase of G2 phase cells and significant decrease of S phase cells in the Cur combined radiation group. In the Cur group, the GUCY2GP, H2BFXP, LINC00623 IncRNA were significantly up-regulated and ZRANB2-AS2 LOC100506835, FLJ36000 IncRNA were significantly down-regulated.

CONCLUSIONCur has radiosensitizing effect on human nasopharyngeal carcinoma CNE-2 cells. Its mechanism may be related to the changes in the cell cycle distribution and the expression of long non-coding IncRNA.

Cell Cycle ; drug effects ; radiation effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Curcumin ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; radiation effects ; Humans ; RNA, Long Noncoding ; genetics ; Radiation Tolerance ; drug effects

OBJECTIVETo prospectively evaluate the long-term effect of pancreaticoduodenectomy with regional lymphadenectomy.

METHODSOne hundred and twenty-one patients with ductal adenocarcinoma in the pancreatic head treated from 1996 to 2001 were studied prospectively. The enrollment of the patients was dependent on 7 criteria. The patients were divided into two groups: regional lymphadenectomy (group A, n = 50) and routine Whipple procedure (group B, n = 71). Their pre- and postoperative conditions, clinicopathological data, survival rates were studied.

RESULTSIt was comparable between the 2 groups in age, sex, preoperative risk factors, operative management, and postoperative complication. Clinicopathological results showed no difference in tumor size and plexus invasion; but the frequency of lymph node involvement and the amount of resected lymph node in group A were significantly higher than those in group B. The rate of local recurrence was significantly higher in group A than in group B. The survival rates of 1-, 3-, 5-year in group A were 70.8%, 31.4%, 20.9%, respectively, which were higher than those in group B. No direct relations were observed between nodal involvement and survival rate.

CONCLUSIONLymphadenectomy in radical pancreaticoduodenectomy could remove lymph nodes effectively and sufficiently, and reduce the rate of local recurrence so as to improve the long-term survival rate.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Lymph Node Excision ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; prevention & control ; Pancreatic Neoplasms ; pathology ; surgery ; Pancreaticoduodenectomy ; Prospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the factors regarding the recovery of postoperative blood pressure of aldosterone producing adenoma (APA) patients.

METHODSSixty-eight patients with APA were recruited and their data including retinal blood vessel by Doppler sonography, urinary trace albumin, pathological changes of renal biopsy and the adrenal tissues around the adenoma were analyzed in order to determine the correlation between these data and postoperative durative hypertension.

RESULTSPostoperative durative hypertension occurred in 14 cases (41.2%) with increased resistance of unilateral or bilateral central artery of retina, in 16 cases (66.7%) with increased level of urinary trace albumin. Fifteen cases underwent renal biopsy and all of them showed different pathological alterations, 11 cases (73.3%) of which presented with postoperative durative hypertension. The pathological changes of the adrenal tissues around the adenoma is either atrophy or non-atrophy (normal or hyperplasia), 8 cases (40%) and 10 cases (22.2%) of which showed postoperative durative hypertension, respectively.

CONCLUSIONThe renal pathological changes and increased resistance of retinal blood vessel are the main reasons leading to postoperative hypertension in patients with APA.

Adolescent ; Adrenal Cortex Neoplasms ; physiopathology ; surgery ; Adrenal Glands ; pathology ; Adrenocortical Adenoma ; physiopathology ; surgery ; Adult ; Blood Pressure ; physiology ; Female ; Humans ; Hyperaldosteronism ; etiology ; physiopathology ; surgery ; Hypertension ; etiology ; Kidney ; pathology ; Male ; Middle Aged ; Postoperative Period ; Retinal Artery ; physiopathology ; Retrospective Studies ; Vascular Resistance ; physiology

OBJECTIVETo clarify the clinicopathological significance of lymphatic vessel density (LVD) and distribution in pancreatic cancer.

METHODSWe measured LVD in 43 pancreatic cancer specimens by immunostaining with specific lymphatic endothelium marker, and examined their relationship with well-defined clinicopathological variables.

RESULTSIntratumoral LVD (9.4 +/- 10.0) was significantly lower than periturmoral (16.0 +/- 9.7) (P < 0.001) and nontumoral LVD (13.5 +/- 6.0) (P < 0.01). Increased peritumoral LVD correlated significantly with tumor staging (P < 0.05) and lymph node involvement (P < 0.05).

CONCLUSIONThe lymphatic vessels distribution in pancreatic cancer samples and peritumoral lymphangiogenesis may promote the malignant progression and lymph node metastasis of pancreatic cancer.

Adenocarcinoma ; pathology ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Immunohistochemistry ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; Male ; Middle Aged ; Pancreatic Neoplasms ; pathology

Objective To construct a CaME141G fusion protein-expressing plasmid,and to express,purify,and identify the activity of the recombinant protein. Methods The 141st site of the wild type CaM,E (GAG),was mutated to G (GGG),using site-specific mutagenesis technology. Escherichia coli BL-21 was transformed with the mutant plasmid. The GST-CaME141G fusion protein was mass-cultured and induced for expression. Subsequently,the GST-CaME141G fusion protein was purified using GS-4B beads. PreScission protease was applied to remove the GST,the Bradford method used to determine the concentration of purified protein,and SDS-PAGE used to detect its relative molecular weight and purity. The GST pull-down assay was used to study the protein's biological activity. Results The CaME141G protein was successfully purified at a high concentration and purity. The protein could interact with PreIQ protein fragments from the myocardial CaV1. 2 calcium channel C terminal,in a CaME141G concentration-dependent manner. Therefore,CaME141G has the ability to bind with the CaV1. 2 calcium channel. Conclusion This study successfully constructed a CaME141G fusion protein-expressing plasmid and purified the CaME141G protein. This lays a foundation for regulating the function of CaM mutations in the myocardial CaV1. 2 calcium channel,and for the study of its relationship with diseases of the cardiovascular system.