Objective Identification of colorectal cancer specific antigens from cDNA phage expression library by recombinant expression cloning (SEREX). Methods The cDNA phage expression library derived from colorectal cancer tissue was constructed using the SMART (Switching Mechanism at 5'end of RNA Transcript) techniques. The cDNA phage expression library was screened with SEREX (serological identification of antigen by recombinant cDNA expression libraries), the positive clones encoding antigenic genes were obtained after immunoscreening, and the nucleotide sequences of cDNA inserts were determined and analyzed with BLST software in GenBank, and the antigenic gene function was analyzed by bioinformatics. Results The cDNA phage expression library derived from colorectal cancer tissues was successfully constructed. The primary library consisted of 2.39?10 6 recombinants, and the recombinant rate was more than 97.5%. The titer of the amplified cDNA phage expression library was 4.1?10 10pfu/ml, and the size of inserted cDNA varied from 0.5 to 4.0kb. Sixteen positive clones encoding antigenic genes were obtained after immunoscreening, and results showed that 16 reactive clones were derived from 12 different genes. Ten of 12 genes were highly homologous to the genes known in GenBank, such as IFITM1, CD24, Survivin, KLK6, et al. Two expressed sequence tags (ESTs) were not found in GenBank. The antigenic genes included structural gene, regulation gene and metabolizing gene. Conclusion The tumor-associated antigen genes and the two ESTs were obtained by SEREX were worthy of further study on their structures and functions.

AlM: To demonstrate the effect of comprehensive treatments for meibomian gland dysfunctional dry eye.
METHODS:Totally, 86 cases (172 eyes) in our hospital, who suffered from meibomian gland dysfunctional dry eye, were measured by BUT, CFS, the machine of infrared meibomian gland function and testing meibomian gland after extrusion. The scores of meibomian gland function missing and secretions were observed, which as a diagnosis, and assessed the effectiveness of treatment, after took comprehensive treatments in different stages ( comprehensive treatments including cleaning, hot covering, massaging, TobraDex, Pranoprofen and Lubricant, etc. ) .
RESULTS:All patients were treated and observed for 3wk, in which 46 cases (92 eyes) of cured, 29 cases (58 eyes) effective, 11 cases (22 eyes) ineffective, the total effective rate was 87. 2%. After 6wk, 68 cases (136 eyes) of cured, 13 cases (26 eyes) effective, 5 cases (10 eyes) ineffective, the total effective rate was 94. 2%, not appeared adverse ocular and systemic reactions in the process of treatment.
CONCLUSlON: Taken comprehensive treatments for meibomian gland dysfunctional dry eye could effectively shorten the course of the disease, quickly remove meibomian gland tube obstruction, restore the meibomian gland secretion and stable the lipid layer, and could reduce the discomfort of patients with dry eye positively and effectively.

Objective To evaluate whether estradiol can inhibit and cure the inflammation of experimental preeclampsia in rats. Methods Experimental preeclampsia was induced in 14-day-pregnant rats by infusion of endotoxin (1.0 ?g/kg). Rats with normal pregnancy were infused with sodium chloride solution.A group of preeclampsia rats was injected with 17?-estradiol (17?-E_2, 1 mg?kg -1 ?d -1 ). Blood pressure, albuminuria,inflammation associated adhesion molecule CD_ 49d and tumor necrosis factor-?(TNF-?) were assessed. Results On pregnant day 19, for normal pregnancy group(group C) the blood pressure was (120.4?2.0)mm Hg (1 mm Hg=0.133 kPa),urinary protein (0.47?0.06)mg/24 hours;for experimental preeclampsia group(group A) blood pressure was (134.2?2.4) mm Hg,urinary protein(0.79?0.10)mg/24 hours; for experimental preeclampsia with 17?-E_2 treatment group (group B) blood pressure was(123.3?1.7)mm Hg,urinary protein (0.51?0.08)mg/24 hours. A significant increase of blood pressure and urinary albumin was observed in group A. CD_ 49d expression and TNF-? concentration were also increased. 17?-E_2 reduced the expression of CD_ 49d , concentration of TNF-?,blood pressure and albuminuria of experimental preeclampsia. However, the weight of fetuses in 17?-E_2 treatment group were less than that in other groups. Conclusion 17?-E_2 can improve the symptoms of experimental preeclampsia,but its effects on fetus need to be further studied.

Obstructive nephropathy ultimately leads to end-stage renal failure. Renovascular lesions are involved in various nephropathies, and most renal diseases have an ischemic component that underlies the resulting renal fibrosis. The aim of this study was to investigate whether morphological changes occur in the renal vasculature in hydronephrosis and the possible mechanisms involved. A model of complete unilateral ureteral obstruction (CUUO) was used. Experimental animals were divided into five groups: a normal control group (N) and groups of animals at 1st week (O1), 2nd week (O2), 4th week (O4) and 8th week (O8) after CUUO. Blood pressure was measured, renal arterial trees and glomeruli were assessed quantitatively, and renovascular three-dimensional reconstruction was performed on all groups. Glomerular ultrastructural changes were examined by transmission electron microscopy. The results showed that the systolic blood pressure was significantly increased in the obstructed groups (O1, O2, O4 and O8). Three-dimensional reconstruction showed sparse arterial trees in the O8 group, and a tortuous and sometimes ruptured glomerular basement membrane was found in the O4 and O8 groups. Furthermore, epithelial media thickness and media/lumen ratio were increased, lumen diameters were decreased, and the cross-sectional area of the media was unaltered in the segmental renal artery, interlobar artery and afferent arterioles, respectively. In conclusion, renal arterial trees and glomeruli were dramatically altered following CUUO and the changes may be partially ascribed to vascular remodeling. Elucidation of the molecular mechanisms of renovascular morphological alterations will enable the development of potential therapeutic approaches for hydronephrosis.

Objective To study the relationship between the MDR1 gene expressions and CD56 antigen expression in patients with de novo acute myeloid leukemia(AML) and to explore the role of this two factors in clinical drug resistance and their correlation. Methods A real-time quantitative RT-PCR method was established for detecting MDR1 expression levels and three-color flow cytometry analysis using CD34/ SSC gating was used to examined CD56 antigen expression in 79 de novo AML patients. Results CD56 an-tigen was recorded in 19 out of 79 cases (24.1%) and particularly in those with M5 cytotypes. Moreover, CD56 expression was significantly associated with unfavorable cytogenetic abnormalities (P<0.05), Patients with t(8:21)had a significantly higher incidence (57.1%, 4/7) of CD56 expression than those with favora-ble karyotype(P<0.05). CD56~+ AML patients had a higher incidence of splenohepatomegalia and lactate dehydrogenase level than CD56~- patients(P<0.05). The median expression levels of MDR1 was statistical-ly higher in CD56~+ AML patients than that in CD56 patients(P<0.001). Patients with both high levels of MDR1 and CD56~+ had a significantly lower CR(complete remission) rate than those with both low MDR1 level and CD56 (58.8% vs 89.2%, P<0.01). Conclusion There is a linear correlation between MDR1 gene expression and CD56 expression in AML. Quantification of the MDR1 gene expression together with CD56 antigen expression is more effective to the judgement of prognosis in AML.

ObjectiveTo evaluate the therapeutic efficacy and prognostic factors of intensity-modulated radiotherapy combined with chemotherapy for early-stage nasopharyngeal carcinoma patients in northwest China. MethodsFrom January 2006 to December 2009,58 patients with early-stage nasopharyngeal carcinoma were treated with IMRT in Xijing hospital,the clinical data were analyzed retrospectively.Survival rates was calculated by the Kaplan-Meier method and the differences was compared by the Logrank test.Univariate analysis method was use to identify all significant factors.ResultsThe follow-up rate was 100％.The follow-up time of 46 patients was more than 3 years.The 1-,2 and 3-year survival were 98％,94％ and 91％,respectively.The 3-year overall survival (OS),local recurrence-free survival (LRFS),distant metastasis-free surv ival (DMFS) for T1N0-1,T2N0 and T2N1 stage were 100％,100％,100％ and 74 ％,81％,87 ％,respectively ( x2 =5.74,P =0.01 ; x2 =4.95,P =0.03 ; x2 =4.24,P=0.04).The 3-year OS,LRFS,DMFS for IMRT combined with chemotherapy and IMRT alone were 100％,100％,100％ and 85％,85％,88％ respectively ( x2 =4.02,P =0.04; x2 =4.12,P =0.03 ; x2 =4.84,P =0.02).In T2N1 stage,IMRT combined with chemotherapy and IMRT alone were 100％,100％,100％ and 79％,79％,80％ respectively (x2 =5.28,P =0.03 ;x2 =4.84,P =0.04;x2 =4.72,P =0.04).In univariate analysis,N stage,clinical stage,IMRT combined with chemotherapy were significantly associated with the survival ( x2 =5.39,P =0.02 ; x2 =5.74,P =0.01 ; x2 =4.02,P =0.04).Conclusions In all early-stage nasopharyngeal carcinoma,T2N1 stage is a sub-group of high risk of distant metastasis.Combination of IMRT and chemotherapy may improve the LRFS,DMFS and OS in those patients.

To determine the effects of Von Hippel-Lindau (VHL) on the invasion and migration of glioma U251 cells. Methods:U251 GBM cells were transfected using VHL expression plasmid. Real-time polymerase chain reaction was conducted to de-tect VHL mRNA expression after transfection. Western blot assay was used to measure protein (VHL, MMP-2, and MMP-9) expres-sion. Tumor invasion and migration were examined by the Transwell and wound-healing experimental methods after VHL up-regula-tion. The intracranial model of nude mouse was developed using U251 cells transfected by VHL expression plasmid, and immunohisto-chemical staining was used to measure protein (VHL, MMP-2, and MMP-9) expression in the tissue sections. Results: In the U251 cells transfected by VHL expression plasmid, the expression of VHL mRNA and VHL proteins increased, and the expression of MMP-2 and MMP-9 protein decreased. Meanwhile, the invasion and migration of glioma U251 cells were also inhibited. Immunohistochemical staining results showed that the expression of MMP-2 and MMP-9 proteins decreased, and the VHL protein expression increased after transfection. Conclusion:VHL can inhibit the invasion and migration of glioma U251 cells. Thus, VHL gene can be used as a target for the gene therapy of gliomas.

Objective To discuss the operative methods and cnrative effect of calcaneal tuberosity fracture.Methods The data of 15 patients with calcaneal tuberosity fracture was retrospectively analyzed who received treatment between January 2008 and June 2011.There were 9 males and 6 female,with the age ranging from 31 to 68 years (average,51.4 years).All the patients had unilateral acute injury,with the left foot in 7 cases and the right foot in 8 cases.According to the Beavis classification,there were 3 cases in type Ⅰ and 12 cases in type Ⅱ.All the cases in type Ⅰ and 10 cases in type Ⅱ were treated with open reduction and screw fixation.The other 2 cases in type Ⅱ with larger fragment involving a portion of the subtalar joint were treated with plate and screw fixation.The effect of the treatment was assessed according to the ankle and hindfoot score system of American Orthopaedic Foot and Ankle Society (AOFAS) after the operation.Results Ten patients were followed up for 12 to 36 months (average,20 months).The healing time in these patients ranged from 8 to 25 weeks (average,12 weeks).The postoperative score ranged from 47 to 100 points (average,91.1 points).There were 7 cases which received excellent effect,2 cases,good,and 1 case,poor.The rate of excellent and good was 90％.Necrosis of skin and soft tissue and exposure of the plate happened to one patient,who eventually healed after 3 weeks by debridement with plate preserved and peroneal artery perforator flap transplantation.Loss of reduction happened to another patient,who was treated with the revision surgery by open reduction and screw fixation again.Conclusion To patients with obvious Fracture displacement whose soft tissues are irritated severely.Emergency open reduction and internal fixation operation should be offered to prevent the necrosis of the flaps as far as possible.To patients with small fractures,it is advisable to choose open reduction and large diameter screw fixation,while plate and screw fixation may be better for patients with larger tuberosity fragment,especially when the fracture line extends to the facet of the subtalar joint.

Objective To retrospectively assess clinical outcomes of buttress plating in fixation of posterior pilon fractures.Methods The study involved 16 cases of posterior pilon fractures that had undergone buttress plate fixation between January 2005 and December 2009.There were 11 males and 5 females,at mean age of 37.6 years (range,23-62 years).All cases received radiography,CT scan and three-dimensional reconstruction preoperatively.Posterior malleolar fragments were reduced and fixed through posterolateral approach or combined approach (posteromedial plus posterolateral approaches) based on effect area of fracture line on CT films.Clinical and radiographic examinations were performed in postoperative follow-up.Clinical outcomes were evaluated by American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and visual analogue scale (VAS).Results Posterior pilon fractures were classified into 3 types according to CT scans.Fourteen cases were followed up for average 37.6 months (range,16-52 months).Mean AOFAS score was 86.4 points (range,70-98 points) and mean VAS score was 1.4 points (range,0-3 points).All cases received favorable functional outcomes without hardware failure,except that one patient still suffered from peri-ankle swelling and long term walking discomfort two years after operation.Conclusion Buttress plating is effective in treatment of posterior pilon fractures and can facilitate early weight-bearing functional exercise.

Objective To investigate BAALC(brain and acute leukemia cytoplasmic)gene expression in patients with de novo acute myeloid leukemia(AML)and its clinical significance. Methods BAALC expression was determined by real-time quantitative polymerase chain reaction(RQ-PCR) in 63 de novo AML patients.The association between BAALC expression and therapeutic effect was analyzed.Results The correlation coefficiencies were over 0.99 for standard curves of RQ-PCR method. BAALC expression was detected in 49(78%)AML patients.The peripheral WBC counts,hemoglobin, platelet counts and the bone mahow blast cell percentage at onset in 31 AML patients with high BAALC expression were(26.3?18.1)?10~9/L,(78.3?21.8)g/L,(76.9?64.5)?10~9/L and(61.2?22.3)% and those of 32 AML patients with low BAALC expression were(30.2?21.7)?10~9/L,(81.6?30.9)g/L, (73.9?57.2)?10~9/L,(54.3?16.3)%,respectively.No statistic differences were found between these two groups.The AML patients with normal chromosome karyotypes are more likely to have a high BAALC expression(68%)compared with those with abnormal chromosome karyotypes(23%,?~2=12.093,P= 0.001).AML patients with normal cytogenetics and high BAALC expression shows significant lower CR rate (65%)compared with those with low BAALC expression(84%,?~2=6.573,P=0.013). Conclusion High BAALC expression may define an important risk factor in AML with normal cytogenetics and predicts an adverse prognosis.