Objective The purpose of this study was to investigate the relationship of p185 and p53 protein overexpression with pathological types and recurrence.Antibodies against p185 and p53 proteins were used to measure expression of these proteins in the nuclei or cell membrane of cells from sections of the giant cell tumor of bone GCT.It showed that 11 out of 52 tumors overexpressed p185 protein and 14 out of 52 tumors had abnormally high levels of p53 protein,4 cases had abnormally high levels of both p185 and p53 proteins,positive expression rates of p185 and p53 in cases with recurrence and cases without reccarrence were 46 2%,20 5% and 38 5%,15 4% respectively.However,there was no association between p185 and p53-positive cases and pathological degree.There was significant correlation between the expression of p185 and p53 protein in the giant cell tumor of bone and recurrence.(? 2=6 125,P=0 047).However,there was no statitically significance between the expression of p185 and p53 protein in the giant cell tumor of bone and pathological types.So that,we consider that the clinical significance for p185,p53 overexpression in GCT to be researched further.

Objective To investigate the diagnostic value and clinical implications of low hemoglobin density(LHD%)in the pa-tients with cervical carcinoma complicating iron deficiency anemia(IDA).Methods The iron metabolism indexes and whole blood cell parameters were detected in 148 patients with cervical carcinomas.LHD% was calculated.The results were compared with that in the healthy group and the hysteromyoma complicating IDA group.Results LHD% was(63 ±30)% in the cervical carcinoma complicating IDA group and(32±15)% in the cervical carcinoma complicating iron deficient erythropoiesis(IDE)group,which were higher than(7±6)% in the healthy group,(22 ±21)% in the cervical carcinoma non-iron deficiency group and(16 ±10)% in the cervical carcinoma non-anemia group(P <0.05 ).And which had no statistical difference between the hysteromyoma complicating IDA group and the cervical carcinoma complicating IDA group(P >0.05 ).Conclusion The new RBC parameter LHD% has the reference value in the diagnosis of cervical carcinoma complicating IDA.The LHD% increase may prompt cervical carcinoma com-plicating IDA.

To produce monoclonal antibody (mAb) specifically against human thrombomodulin (hTM), an immune-tolerizing procedure was employed to generate monoclonal antibodies specific to hTM. Female BALB/c mice were first immunized with CHO cells following at 10 min, 24 h, 48 h by intraperitoneal injection of different doses of cyclophosphamide (CP) 2 times at an interval of 2 weeks, thereby tolerizing the mice to common epitopes shared between CHO and CHO-TM5 cells. Subsequently the selected mice with the lowest titer of serum polyclonal antibody by cellular enzyme-linked immunoabsorbent assay (CELISA) were immunized with CHO-TM5 cells, which have stable high level expression of hTM, to produce antibodies specific to hTM 3 times at an interval of 2 weeks. On the third day after the third immunization, mouse with the highest titer of serum polyclonal antibody was sacrificed and spleen cells were harvested to prepare hybridoma cells with SP2/0 cells at the ratio of 10 to 1. Hybridoma cells were then cultured at 96 well plates for screening with CELISA. To improve probability to obtain specific mAb, CELISA was applied twice. The first CELISA was done with polyethylene ELISA plate with a monolayer of CHO-TM5 cells. The positive clones from the first screen were then selected by reacting with similar screening ELISA plate but having CHO cells monolayer instead. Only clones that were positive for the first screening and negative for the second screening were kept, and called as CHO-TM5 +CHO- hybridoma cells. BALB/c mice were intraperitoneally injected with the selected hybridoma cells. Ascites were collected and monoclonal antibodies were purified using FPLC, and its Ig class, subclass, and titer were then determined respectively. The specificity of the yielded mAb was identified with CELISA, flow cytometry, ABC immunohistochemistry and immunoblotting. Detection of CELISA showed that 100 mg/kg dose of CP could tolerize the mouse to common epitopes shared between CHO and CHO-TM5 cells. And CELISA also discovered that all hybridomas positive for CHO-TM5 cells were negative for CHO cells. Five lines of positive hybridoma cells had been obtained altogether and 2F7 was selected randomly for next investigation. The Ig subclass of the mAb 2F7 was IgG1 and the titer of ascitic mAb was 1?10-6. Furthermore, the content of ascitic mAb was 19.56 g/L and chromosome numbers is 98. Flow cytometry, CELISA and Western blotting assays demonstrated that mAb 2F7 could specifically recognize hTM expressed on CHO-TM5 and human umbilical vascular endothelial cells (HUVEC). Meanwhile, the tissue specificity of mAb 2F7 was also identified by immunohistochemical ABC staining. On the other hand, Western blotting assays indicated that mAb 2F7 could recognize the antigen protein with 105 ku molecular mass under reduction condition. Moreover, the dissociation constant of mAb 2F7, 1.22? 10-9 mol/L, indicated the affinity higher than some others. The results suggest that the immunotolerizing protocol provides a convenient general method for producing antibodies specific to desired protein isoforms. mAb 2F7 can specifically recognize the natural hTM expressed mainly on vascular endothelial cells, which will potentially useful for investigating the functions and clinic values of hTM.

Purpose　The aim is to study the effects of solubl e chit osan on antitumor factors and NK cell activity in rats.Methods　The level of NO、TNF produced by peritoneal macrophages,IFN-γ production and NK ce ll activity of spleen was detected,after different concentrations of chitosan we re injected into the sarcoma 180 rats peritoneally.Results　Th e cont ents of NO、TNF、IFN-γ and NK cell activity in sacroma 180 rats were significa ntly higher in the chitosan groups than those in the control group (P＜0.01) .Conclusion　Chitosan could impove the immune function in rats .

Objective To improve the gene targeting efficiency with C57BL/6 embryonic stem ( ES) cells.Meth-ods Three different genetically modified C57BL/6 ES cell lines, named TLX3, Ai3K and SL, were microinjected into ICR, B6( Cg)-Tyrc-2J and BALB/c mouse blastocysts, respectively.The efficiency was statistically evaluated according to three aspects:blastocyst collection, chimera production and germline transmission.Results None of the three ES cell lines was germline transmitted with B6(Cg)-Tyrc-2J mice as blastocyst donors, while it was achieved with both BALB/c and ICR mouse blastocysts.Compared in the aspect of blastocysts collection, ICR mouse was much better than BALB/c mouse (P<0.05), and the chimera production efficiency of ICR mouse was comparable to that of BALB/c mouse (P =0.115). As to the germline transmission efficiency, that of BALB/c mice is significantly higher than that of the ICR mice ( P<0.01).Conclusions The germline transmission efficiency of BALB/c mouse is highest among these three mouse strains. However, it has the disadvantages of blastocyst collection, developmental delay and zona pellucida fragility, compared with ICR mouse.Therefore, ICR mouse is also a good candidate as blastocyst donor for embryonic stem cell microinjection.

Objective To assess the expression of T cell subset in patients with hepatitis B virus-associated primary liver cancer (HBV-PLC) and its influence on the clinical outcome.Methods 136 cases with PLC were selected, and divided into HBV-PLC group (78 cases) and non-HBV-PLC group (58 cases) according to HBV infection.The percentage of CD4+T cells, CD8+T cells and Treg cells in serum was tested by flow cytometry.After 6 months of follow-up, HBV-PLC patients were divided into death group and survival group.Binary logistic regression analysis was performed to explore the factor and degree affecting clinical outcome of HBVPLC patients.Results As compared with non-HBV-PLC group, dysregualted T cell subset was observed in the HBV-PLC group, percentage of CD4+T cells and Treg cells in HBV-PLC patients was higher (P＜0.05), while CD8+T cell percentage was decreased (P＜0.05).The percentage of CD4+T cells and Treg cells was higher in death group than survival group (P＜0.05), but CD8+T cell percentage was lower than that of survival group (P＜0.05).T-lymphocyte subset (CD4+T cells and Treg cells) was a strong risk factor for adverse clinical outcome of HBV-PLC (OR values were 3.765 and 2.238, respectively, P＜0.05), but CD8+T cell was a protective factor (OR value was-3.537, P＜0.05).Conclusion Obvious dysfunction of T cellular immune function exists in HBV-PLC patients, and T cell subset may be a predictive factor for clinical outcome of HBV-PLC patients.

Objective To establish a simple , fast and economic total DNA extraction method to serve the rapid identification of model mouse genotype in large number of mice .Methods Three methods, i.e.phenol extraction, isopropyl alcohol precipitation and mouse ear boiling methods were used to extract the total DNA from ten C 57-rasmodel mice.The purity and yield of DNA obtained by the three methods were compared , and polymerase chain reaction ( PCR) assay was used to compare the efficacy of the three extraction methods .Results Among the three methods , phenol extraction was the best and isopropyl alcohol precipitation was the poorest in DNA yield .In terms of DNA purity , the phenol extraction was the best and the mouse ear boiling method was the poorest .All the three methods could be used to extract the total DNA from mice serving as template of PCR reaction for the mouse genotype identification .The time consumption of three methods are 12.5 hr ,13 hr and 0.18 hr.Mouse ear boiling method was significantly lower than that of the other two methods ( P <0.01 ) ,.The obtained total DNA can be stored at conventional -20℃for 7 days and 30 days later still can be used as a template for PCR reaction .Conclusions Among the three methods studied , the mouse ear boiling method is simple and with the lowest cost , so it is feasible for total DNA extraction in scaled genotyping experiments .

Objective To obtain the basic growth parameters of a self-established F1 hybrid CB6F1 C57-ras transgenic mouse model, and to provide basic information for commercialization of this mouse model. Methods F1 hybrid mice (CB6F1) were produced by crossing C57-ras heterozygous transgenic (c-Ha-ras+/-) male mice and wild-type BALB/cJ female mice.The average litter size, weaning rate, sex ratio, growth performance and C57-ras transgenic positive rate were recorded and analyzed.Results The average litter size was eight, weaning rate was 90%, and sex ratio was approximately in accordance with prediction.The average body weight of newborn mice was 1.73 ±0.05 g.The average body weight of 10-week old c-Ha-ras transgenic female and male mice in CB6F1 background was 24.38 ±1.74 g and 29.42 ±1.72g, respectively, which had a significant difference (P<0.01).The c-Ha-ras transgenic positive rate was 46.9%. which was in accordance with genetic rules.Conclusions The F1 hybrid mice (CB6F1) produced by crossing C57-ras heterozygous transgenic ( c-Ha-ras +/-) male mice and wild-type BALB/cJ female mice show normal growth performance and development characteristics, and it can be used for large-scale commercial supply.

Objective: To identify potential relationship between single uncoding RNA-25-3p (miR-25-3p) expression level and the sertraline efficacy in patients with panic disorder. Methods: Sixty cases of patients with panic disorder(case group) and sixty healthy-controls(control group) were collected with demographic data and peripheral venous blood before and after treatment.All the patients were evaluated using the 14-item Hamilton Anxiety Rating Scale (HAMA) and Panic Disorder Severity Scale (PDSS) at baseline, and then received sertraline treatment for 6 weeks.After six-week treatment, each patient was evaluated again with HAMA and PDSS.RT-PCR was used to detect the level of miR-25-3p expression. Results: There was no significant difference in the miR-25-3p levels between control group (1.27±0.32) and case group (1.73±1.09) before treatment(t=1.53, P=0.14), but the levels in case group were much higher than that in control group after the treatment (5.72±4.13 vs 1.73±1.09, t=-2.15, P=0.04). Besides, the changes of the miR-25-3p levels were positively related with both the changes of PDSS3 and PDSS7 items before and after the treatment (r=0.60, P=0.02 for PDSS3 and r=0.61, P=0.02 for PDSS7). Conclusions miR-25-3p is associated with the drug efficacy and the outcome of some clinical symptoms of panic disorder.These findings might provide some evidence for the individualized treatment of patients with panic disorder according to regulation of gene expression in the future.

Objective To investigate the protective effect and mechanism of environmental enrichment(EE)on radiation induced cognitive dysfunction in mice.Methods A total of 45 female Kunming mice(2-month old)were randomly divided into control group,irradiation group and irradiation plus EE group with 15 in each group.Irradiation group and irradiation plus EE group were treated with a single dose of 4 Gy whole body irradiation,irradiation plus EE group were housed in EE condition for 35 d after irradiation.The object recognition task was used to evaluate the cognitive function of mice.The expression of microglial marker IBA-1 in hippocampus was determined by immunohistochemical staining.The expressions of CD68 and synaptophysin(SYP)proteins in hippocampus were assayed by Western blot.Results Compared with control group,the irradiation group had a low discrimination ratio in object recognition task and had a remarkable low level of SYP expression in hippocampus(t＝3.66,6.84,P＜0.05).In addition,radiation activated microglia in hippocampus by increasing the number of IBA-1-positive cells and enhancing the expression of CD 68(t ＝6.83,5.79,P ＜0.05).Compared with irradiation group,irradiation plus EE group increased the discrimination ratio and the expression of SYP in hippocampus(t＝3.56,4.06,P＜0.05),while the number of IBA-1-positive cells and the expression of CD68 were significantly reduced(t＝7.69,4.59,P＜0.05).Conclusions A single dose of 4 Gy whole body irradiation leads to cognitive dysfunction in mice,while EE could effectively improve the animals′cognitive behavior possibly by inhibiting microglial activation and preventing synapse loss in hippocampus.