Objective To explore the effect of mouse proliferation-associated protein 2G4 (p38-2G4) high-expression on the proliferation and erythriod differentiation of murine erythroleukemia ( MEL ) cells.Methods To establish the recombinant lentivirus vector p 38-2G4-pLJM1, the p38-2G4-pLJM1 was cotransfected into HEK293T cells to obtain lentivirus with pCMV-VSV-G and pCMV-dR8.2.Lentivirus were infected into MEL cells to establish the stably p 38-2G4 high-expressed MEL cells.Western blotting was used to analyse the high-expression efficiency.MTT assay and benzidine staining were applied to detect the cell viability and hemoglobin synthesis of the stable cell line in presence /absence of inducers.Results Western blotting showed that the p38-2G4 high-expression stable cell strain had a higher expression of p38-2G4 as compared to the control group ( MEL) ( P <0.05).MTT result showed that there was no difference between the p38-2G4 high-expression cell strain and the control group (P>0.05), but the hemoglobin synthesis had been reduced as compared to the control group (P<0.05).Conclusion p38-2G4 high-expression does not affect the cell viability of MEL cells , but inhibits the erythriod differentiation of MEL cells in three independent experiments .

Objective To investigate the expressions of cycloxygenase-2 (COX-2 ),vascular endothelial growth factor (VEGF),matrix metalloproteinase (MMP2 )and micro-vessel density (MVD)in papillary thyroid carcinoma (PTC)and the relationship between their expressions and clinicopathological features,so as to evaluate the malignancy and prognosis of PTC.Methods The expressions of COX-2,MMP2 and VEGF and MVD count in 32 cases of PTC and 18 cases of normal thyroid tissues were detected by immunohistochemical staining.Their relationship with clinicopathological characteristics was analyzed.Results ① The expression of COX-2,MMP2, VEGF and MVD in PTC was as follows:M(Q 3 -Q 1 )=5(1),M(Q 3 -Q 1 )=5.5(2),M(Q 3 -Q 1 )=5.5(1)and M (Q 3 -Q 1 )= 28 (5.75 ),respectively.It differed significantly from that in control group (P < 0.05 ).② The expression of COX-2 in PTC was related to the sex and age of patients,clinical stage and lymph node metastasis (P <0.05).The expression of VEGF in PTC was related to the sex and age of patients,tumor size,clinical stage and lymph node metastasis (P <0.05).The expression of MMP2 and MVD count in PTC were related to the age of patients,tumor size,clinical stage and lymph node metastasis (P <0.05 ).③ The expression of COX-2,MMP2, VEGF and MVD in PTC with invasion of thyroid capsules was as follows:M(Q 3 -Q 1 =6(2),M(Q 3 -Q 1 )=6.5 (2),M(Q 3 -Q 1 )=6(1.75)and M(Q 3 -Q 1 =30(7.75).The expression of these indexes in negative group was:M (Q 3 -Q 1 )=5(1.75),M(Q 3 -Q 1 )=5(1.75),M(Q 3 -Q 1 )=5 (1.75 )and M (Q 3 -Q 1 )=26.5 (4).There was a significant difference between the two groups (P < 0.05 ).④ There was a close positive correlation between the expressions of VEGF,COX-2,MMP2 and MVD in PTC (r >0.5).Conclusion The high expressions of COX-2, VEGF and MMP2 in thyroid tissues may result in the occurrence of PTC and lymph node metastasis,which is related to the regulation of tumor new vessels.Detecting these expressions are of value in evaluating the malignancy and prognosis of PTC.

This article was aimed to explore the effect of acupuncture on HDAC2 activity in peripheral blood of chronic obstructive pulmonary disease (COPD) rats. The COPD rat model was established by cigarette smoking. Acupuncture was given for 3 consecutive weeks. The lung ventilation of rat was detected by equipment. Peripheral blood of rat was taken after 3 weeks. The HDAC2 enzyme activity was detected by fluorescence technology. ELISA was used in the detection of IL-10, IL-8 and TNF-αexpression in peripheral blood. The results showed that after acupuncture treatment, the lung ventilation was obviously increased compared with the model group (P < 0.05). Meanwhile, the HDAC2 activity in the peripheral blood of rat was obviously increased (P < 0.05). However, the expression of inflammatory cytokines IL-10, IL-8 and TNF-α was obviously reduced (P < 0.05). It was concluded that acupuncture treatment had a good efficacy on COPD rat. It indicated that acupuncture treatment might regulate HDAC2 activity to reduce inflammation.

Objective To explore the differentially expressed proteins during erythroid differentiation .Methods The murine erythroleukemia ( MEL) cell were treated by DMSO , and the comparative proteomic was systematically analyzed and identified on different differentiating time points .ratio of cell differentiation and viability were detected by benzidine staining, MTT assay and Ter119 immunofluorescence.Using two-dimensional gel electrophoresis combined with mass spectrometry technology and bioinformatics analysis , we conducted a comparative proteomic analysis on MEL cells during the process of induced differentiation to screen and identify differential proteins .Results The MEL cells induced by 1.2%DMSO for 0 hour, 6hours, 12hours, 24hours, 36hours, 48hours, 72 hours, 96 hours, 120 hours were collected for proteomic analysis, by two-dimensional gel electrophoresis combined with mass spectrometry .A total of 87 kinds of proteins were successfully identified .MEL cells exposed to DMSO at a final concentration of 1.2% for 120 hours reached the highest differentiation rate of 67%.MTT assay showed that 1.0%, 1.2%, 1.4% DMSO had no inhibiting effect on cell vitality.Conclusion DMSO may induce MEL cells to differentiate and have no inhibiting effect on cell vitality .The 87 kinds of differentially expressed proteins from two-dimentional gel electrophoresis may be divided into twelve categories ;the most three parts are 41%enzyme protein, 15%structural protein and 13%regulatory protein.

Objective To compare gross tumor volume (GTV) of nasopharyngeal carcinoma (NPC) according to MRI and FDG PET/CT and to investigated four fixed threshold methods to delineate the GTV using FDG PET/CT.Methods Fifty patients with primary biopsy-proven NPC were prospectively were enrolled into the study.FDG PET/CT scans and MRI were carried out within one week prior to pretreatment,respectively.The GTV was named GTV-MRI (GTV were delineated according to MRI),GTV-PETvis,GTV-PET30,GTV-PET40,GTV-PET50 (GTV was delineated according to the PET-based GTVs obtained by visual interpretationor,by percentage of the SUVmax (30％,40％,50％) thresholds,respectively).The differences were compared among the GTV-MRI,GTV-PETvis,GTV-PET30,GTV-PET40 and GTV-PET50 in different by Wilcoxon test.Results Of 50 patients,the median of volume descending order were: GTV-MRI 27.8 cm3,GTV-PETvis 22.2 cm3,GTV-PET30 22.7 cm3,GTV-PET40 14.4 cm3 and GTV-PET50 9.0 cm3.However,there was no significant difference between GTV-PETvis and GTV-PET30 (Z=-0.05,P=0.958),as well as GTV-MRI and GTV-PETvis or GTV-PET30 in 25 patients who were T1-2 stage (Z =-0.93,-0.93,P=0.353,O.353),the other GTVs were all different in 50 patients' (Z=-5.74-2.09,P =0.000-0.037).Conclusions All the GTVs delineated by the different methods of using FDG PET/CT were less than the GTV delineated by MRI.The potential advantages with the GTV-PETvis or GTV-PET30 delineated by FDG PET/CT are reduction of biological metabolic tumor volume in GTV delineation and reduction of the size of the GTV in NPC patients.

Objective:To prepare a virus-like particle (VLP),containing Hepatitis B virus core antigen (HBcAg) and N-terminal peptides of the L2 protein of human papilloma virus (HPV),and investigate the immunogenicity of the VLP in mice and the protection against different strains of HPV .Methods:A fusion gene was synthesized to insert a DNA fragment ,coding for the N-terminal epitopes of the L2 protein of HPV16,into the HBcAg coding sequence;HBc-L2 fusion protein was highly expressed in E.coli using the pET9a and BL21(DE3) expression system;the purified fusion protein was used to immunize BALB/c mice and antibody titers against the L2 epitopes in mouse sera were determined by indirect ELISA;the levels of neutralizing antibodies against both HPV 16 and 18 were also analyzed.Results:HBc-L2 fusion protein was expressed in E.coli and purified,with the purity >80%,by ammonium sulfate pre-cipitation and CL-4B gel filtration;analysis of the purified fusion protein ,using size exclusion chromatography with multi-angle laser light scattering detection ( SEC-MALS) and electron microscope ,revealed that HBc-L2 was assembled into a stable VLP structure auto-matically following its expression;immunization of BALB/c mice with the purified VLPs resulted in high antibody titers in mouse sera against the L2 epitopes;furthermore,it was demonstrated that the sera from the immunized mice had neutralization activities against both HPV16 and HPV18.Conclusion:The immunogenicity of the L2 epitopes was highly enhanced by the construction of HBc-L2 fusion protein and the formation of the VLP structure;the fusion protein was also capable of inducing protections against different serotypes of HPV,therefore,it could be a potential HPV vaccine with a broad coverage and low production cost .

Objective To explore the significance of the suppressor of cytokine signaling-1 (SOCS-1)gene methylation in the genesis, development and prognosis of diffuse large B-cell lymphoma (DLBCL).Methods The methylation state of CpG island in SOCS-1 gene were detected by methylation-specific polymerase chain reaction (PCR) and the level of SOCS-1 gene was measured by the real-time PCR. The clinical data of 30 patients with DLBCL were collected, and according to the international prognosis index (IPI), they were divided into low risk group and high risk group. Results Aberrant methylation of SOCS-1 in 17 DLBCL patients (56.7 %) were positive, however, in control group aberrant methylation was negative(P ＜0.01).The methylation level of DLBCL patients with positive SOCS-1 methylation was higher than that of patients with negative (P ＜0.05). Combined with the clinical data, the positive rate of methylation in patients with high level of serum LDH or the numbers of extra-nodal lesions＞l were significantly higher than that in patients with normal LDH level or the numbers of extra-nodal lesions ≤ 1, respectively. Hence, the positive rate of methylation in the high risk group of DLBCL was higher than that in the low risk group (P ＜0.05). Conclusion There were aberrant methylations of the SOCS-1 gene in the patients with DLBCL. The methylations of SOCS-1 can silence the gene expression, which indicates that SOCS-1 and its methylations play some role on genesis and development of DLBCL and can evaluate the prognosis of the patients with DLBCL.

Objective To investigate the effect of laparoscopic cholecystectomy (LTCBDE) in treatment of patients with secondary extrahepatic bile duct stones. Methods Eighty-seven cases of our hospital patients with secondary to extrahepatic bile duct stones were randomly divided into the laparoscopic bile duct exploration and T tube drainage surgery (LCBDE) treatment group and the laparoscopic transcystic duct exploration of common bile duct lithotomy (ltcbde) treatment group. The observation focused on the operation time, bleeding volume , postoperative transfusion , postoperative drainage time , postoperative hospitalization time , cost of hospitalization, postoperative recovery time and complications compared clinical efficacies. Results LTCBDE group of patients in operation time (2.1 ± 0.5) was longer than that of the control group (1.6 ± 0.4), (P <0.001), while the bleeding volume, postoperative fluid volume, postoperative drainage time, postoperative hospitalization time, hospitalization expenses and postoperative recovery time were (17.4 ± 5.4), (6 550.4 ± 1 076.9), (3.5. 1.6), (4.1 ± 1.7), (12 243.5 ± 2 379.6), (11.3 ± 3.5) were lower than that of the group LCBDE (22.1 ± 7.5), (8 304.2 ± 1 394.8), (32.9 ± 10.4), (6.4 ± 2.4), (14 098.1 ± 2 897.3), (16.1 ± 5.7) P, respectively (P values were defined as 0.001, 0, 0, 0.015, 0.001, 0 individually); LTCBDE group of patients with bile leakage, acute peritonitis rates were 1/46,1/46,in which those were lower than the corresponding LCBDE in 6/41, 7/41 (P values were 0.033, 0.016, separately). Conclusion According to indications, LTCBD surgery has the advantages of less injury, less cost, less complications and so on. It has important significance to improve the condition of patients with secondary extrahepatic bile duct stones.

Objective: To analyze the clinical characteristics and current treating status from atrial ifbrillation (AF) patients with different gender in 20 emergency departments.
Methods: A total of 2015 consecutive AF patients from 20 emergency departments nationwide from 2008-11 to 2011-10 were retrospectively investigated. The patients were divided into 2 groups: Female group,n=1104 and Male group,n=911. The baseline clinical characteristics and current treating status were compared between groups, the risk of stroke in non-valvular atrial ifbrillation (NVAF) patients was evaluated by CHADS2 score and the factors affecting walfarin application were studied by Logistic regression analysis.
Results: Compared with Male group, Female group had the elder age (69.11 ± 12.96) years vs (67.67 ± 13.63) years,P=0.015, lower body mass index (BMI) (23.24 ± 3.73) kg/m2 vs (23.89 ± 3.47) kg/m2,P=0.000, more patients combining with heart failure (39.7% vs 34.6%,P=0.019), more patients with valvular heart disease (26.6% vs 12.4%,P=0.000). In contrast, Male group had more smokers (41.4% vs 5.1%,P=0.000), more patients combining with coronary artery disease (45.1% vs 39.1%,P=0.007) and more patients with previous history of myocardial infarction (9.5% vs 5.5%,P=0.001). The average CHADS2 score was higher in Female group than Male group (2.0 ± 1.4) vs (1.8 ± 1.4),P=0.008 and the proportion of patients with CHADS2 score ≥2 was higher in Female group than Male group (58.0% vs 51%,P=0.005). There were 407 patients of valvular heart disease with AF and 167 (41%) of them received walfarin treatment including 119 female and 48 male,P=0.713; 1608 NVAF patients with CHADS2 score≥ 2 and 105 of them received anticoagulation therapy including 54 female and 51 male,P=0.636.
Conclusion: The clinical characteristics and current treating status are different by gender from AF patients in 20 emergency departments in China.

Objective To investigate the expression of CD40-CD154 co-stimulatory pathway in peripheral circulation and intestinal mucosa in patients with inflammatory bowel disease (IBD),the difference between the expression of CD40-CD154 in patients with IBD and that in healthy controls,and the correlation between CD40-CD154 levels and disease activity. Methods A total of 62 patients with Crohn's disease (CD),64 patients with ulcerative colitis (UC) and 56 healthy controls were enrolled. Enzyme-Linked Immuno Sorbent Assay (ELISA),SYBR-green real time PCR and immunohistochemical assay were respectively applied to evaluate expression of CD40-CD154 in plasma,mononuclear cells of peripheral blood and intestinal mucosa. Results Levels of CD40 (P=0. 000) and CD154 (P=0. 001) in plasma,mononuclear cells of peripheral blood and intestinal mucosa were significantly higher in patients with CD and UC than in healthy controls. However,no correlation between disease activity and CD40-CD154 expression in peripheral circulation or intestinal mucosa was detected (P ＞ 0. 05 ). Conclusion CD40-CD154 pathway activation is found in plasma,peripheral blood mononuclear cells and intestinal mucosa in patients with IBD,but is not correlated with disease activity.