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 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 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 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.

Objective To explore the possible relationship between alteration of cell cycle and JAK/STAT3 signal transduction pathway inhibition induced by arsenic trioxide (As_2O_3,) in myeloma cell lines U266 and RPMI8226 in vitro. Methods Multiple myeloma cell lines U266 and RPMI8226 were used as in vitro models. The influence of AS_2O_3 on myeloma cells were evaluated by MTT assay and flow cytometry. Meanwhile, methylation specific PCR and Western blotting were employed to detect the methylation status of gene SOCS-1 and protein expression level of P-STAT3 in these cells after AS_2O_3 treatment. Results AS_2O_3 significantly inhibited the growth of U266 and RPMI8226 cells in a dose-dependent manner. Furthermore, cell cycle was arrested at G0/G1 phase with inhibition of protein expression level of P-STAT3 and SOCS-1 gene demethylation after exposure to As_2O_3 for 72 h( r = 0. 85, P < 0.05). Conclusion AS_2O_3 could induce the alteration of cell cycle which might be related to JAK/STAT3 signal transduction pathway inhibition and SOCS-1 demethylation in myeloma cell lines. The study puts forward a new idea of AS_2 O_3 treatment in multiple myeloma.

ObjectiveTo study theory of mind(TOM) of youngsters in alexithymia,and to assess the impairment in TOM by analysing affective and cognitive TOM and the first and second order judgment.MethodsTOM of 31 alexithymics and 30 healthy control subjects get be examined by the Chinese version of “Yoni task” and series of tests.ResultsThere were significant differences in affective TOM accuracy scores between alexithymics and healthy controls ( ( 35.81 ± 6.30) vs ( 39.83 ± 6.02),t =-2.55,P ＜ 0.05 ).Particularly,alexithymics got significantly lower accuracy scores in the second order judgement of affective TOM ( (24.55 ±6.01 ) vs (28.80 ± 6.04),t =-2.76,P ＜ 0.01 ).However,there were no in cognitive TOM accuracy scores between alexithymics and health controls ( ( 19.00 ± 2.11 ) vs ( 19.43 ± 2.13 ),t =-0.80,P ＞ 0.05 ).ConclusionAlexithymics are impaired in affective TOM,specially the second order judgement.

Objective To investigate the effects of arsenic trioxide (AS2O3)on SOCS-1 gene methylation and expression of P-STAT3 in multiple myeloma (MM) cells.Methods MM cell lines U266 and CZ-1 were used as in vitro models.Methylation status of SOCS-1 gene was detected by the methylation specific PCR (MSP)while P-STAT3 protein expression was determined by Western blotting assay before and after AS2O3 treatment.Meanwhile growth inhibition and apoptosis of MM cells were determined by flow cytometry.Results Hypermethylation of SOCS-1 gene was observed in each MM cell line compared with wide type.After exposure to AS2O3,it was shown that SOCS-1 gene was demethylated obviously,meanwhile the expression level of P-STAT3 protein and cell proliferation was inhibited significantly in each cell line.The apoptosis rate was increased.When U266 and CZ-1 were treated with AS2O3 of 0,0.5,1.0,2.0 μmol/L respectively,the total cell apoptosisis ratio of U266 was 0.06％,0.56％,48.96％,61.07％ (X2 =9.19,P ＜ 0.05); and the total cell apoptosisis ratio of CZ-1 was 4.2％,,40.3％,,47.72％,,68.49％ (X2 =8.96,P ＜0.05 ).Conclusion AS2O3 could inhibit JAK/STAT signal transduction pathway by inducing SOCS-1 gene demethylation in MM cells which might be related to cell apoptosis.

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.