Objective To study the insulin‐like growth factor binding protein (IGFBP)‐2 and IGFBP‐6 expression and clini‐cal significance in colorectal adenomas .Methods A collection of Chengde Medical College Hospital from July 2012 to March 2013 after surgical treatment of colorectal cancer confirmed by pathology (colorectal cancer ,CRC) tissue samples of 50 patients ,colorec‐tal adenomas (colorectal adenoma ,CRA) 50 cases ,20 cases of colorectal normal mucosa .Immunohistochemistry and RT‐PCR were used to detect the expression of IGFBP‐2 and IGFBP‐6 protein and mRNA ,combined with clinical and pathological data were statis‐tically analyzed .Results IGFBP‐2 protein positive expression and the amount of mRNA expression in the CRA group compared with normal colorectal mucosa had a rising trend;While compared with CRC group had a tendency to reduce ,and the differences are obvious statistical significance (P<0 .05) ,and IGFBP‐6 protein positive expression in the CRA group compared with normal color‐ectal mucosa had a lower trend;While compared with CRC group amount IGFBP‐6 mRNA expression in the CRA group compared with normal colorectal mucosa had a rising trend;While compared with CRC group had a tendency to reduce ,and the differences were obvious statistical significance (P<0 .05) .In the CRA group ,IGFBP‐2 ,IGFBP‐6 positive expression and the patient′s age , sex ,tumor and the number of parts were no significant statistical difference (P>0 .05) ,but with the degree of hyperplasia had sig‐nificant statistical difference(P<0 .05);In the CRA group ,IGFBP‐2 and IGFBP‐6 expression were negatively related to each other , and the difference was statistically significant (P<0 .01) .Conclusion Colorectal adenomas in normal colorectal mucosa of colorec‐tal cancer to the middle part of the transformation process ,and in its occurrence and development process ,insulin‐like growth factor family (IGFs) and IGFS‐R axis plays an important and irreplaceable role ,so IGFBP‐2 ,IGFBP‐6 may be used as diagnostic colorectal adenomas and early predictors of prognosis ,clinical studies on colorectal adenomas is important .

Purpose To investigate the expression of IGF-Ⅱand IGFBP-6 in co1orecta1 cancer,and to exp1ore the c1inica1 significance in co1orecta1 cancer. Methods IGF-Ⅱand IGFBP-6 were detected by immunohistochemistry and RT-PCR in 50 cases of co1orecta1 cancer(experimenta1 group)and 50 cases of the adjacent mucosa(contro1 group). Results (1)In the experimenta1 group,the ex-pression 1eve1 of IGF-Ⅱprotein and mRNA was significant1y higher than the contro1 group. The expression 1eve1s of IGF-Ⅱprotein and mRNA in co1orecta1 cancer were significant1y 1ower than the contro1 group.(2)The expression 1eve1s of IGF-Ⅱand IGFBP-6 were sig-nificant1y different between different tumor infi1tration depth,1ymph node metastasis,invasion depth and Duke’s stages( P<0. 05), but no difference between genders,age and the degree of tumor differentiation( P>0. 05). Conclusions There is a obvious corre1a-tion between of IGF-Ⅱ and IGFBP-6 in c1inica1 patho1ogica1 parameters in co1orecta1 cancer. Combined detection of the two markers may be the bio1ogica1 indicators of occurrence and prognosis of co1orecta1 cancer,and provide a new scheme for diagnosis and treatment of co1orecta1 cancer.

Objective To evaluate the diagnostic value of dermoscopy and reflectance confocal microscopy (RCM) alone or in combination for melanocytic nevus.Methods A total of 37 patients with clinically diagnosed melanocytic nevus were collected.Skin lesions were firstly examined by dermoscopy and RCM,then were resected to be subjected to histopathological examination for final diagnosis.The imaging features of melanocytic nevus were summarized.The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of different skin imaging techniques were calculated,and the consistency was analyzed between skin imaging techniques and histopathological examination.Results Based on the dermoscopic and RCM findings,2 kinds of nevus cells with different morphological features were observed in the dermis of intradermal nevus.One kind of nevus cells was characterized by a nonfusional,highly-refractive round structure in the papillary dermis under RCM,and by a brown or light brown homogenous pattern under dermoscopy,which was observed in 5 skin lesions.The other kind of nevus cells appeared as irregular,highly-refractive cell clumps in the papillary dermis under RCM,and by a cobblestone or globular pattern under dermoscopy,which was observed in 31 skin lesions.For the diagnosis of melanocytic nevus,the sensitivity,specificity,accuracy,positive predictive value and negative predictive value of RCM combined with dermoscopy were 91.7％,87.5％,90.9％,97.1％ and 70％ respectively,those of RCM were 86.1％,75％,84％,93.9％ and 54.5％ respectively,and those of dermoscopy were 77.8％,87.5％,75％,96.3％ and 41.2％ respectively.All the diagnostic indices of RCM combined with dermoscopy were higher than those of RCM or dermoscopy alone,except that the specificity was equal to that of dermoscopy alone.RCM showed higher sensitivity,accuracy and negative predictive value,but lower specificity and positive predictive value compared with dermoscopy.There were no significant differences in the diagnostic yield in melanocytic nevus between RCM combined with dermoscopy or RCM alone and histopathological examination (x2 =0.25,0.57,P =0.63,0.45,Kappa value =0.72,0.53,respectively).However,a significant difference in the diagnostic yield in melanocytic nevus was observed between dermoscopy and histopathological examination (x2 =5.81,P =0.012).Conclusion RCM combined with dermoscopy shows higher diagnostic accuracy for melanocytic nevus compared with RCM or dermoscopy alone.

To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.
Base Sequence ; Cloning, Molecular ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Genetic Vectors/genetics ; Hep G2 Cells ; K562 Cells ; Molecular Sequence Data ; Receptors, Transferrin/*immunology ; Recombinant Fusion Proteins/biosynthesis ; Recombinant Fusion Proteins/genetics ; Single-Chain Antibodies/*biosynthesis ; Single-Chain Antibodies/genetics

Adenocarcinoma ; metabolism ; pathology ; Chromogranin A ; metabolism ; Colonic Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; methods ; Proliferating Cell Nuclear Antigen ; metabolism ; Staining and Labeling ; methods ; Vimentin ; metabolism

This study investigated the intracellular localization of asparagine synthetase (ASNS) in the relation with chemoresistance in leukemia. pIRES-GFP-ASNS-Flag/Neo expression vector was transiently tansfected into SK-N-MC cells and 297T cells respectively. Immunofluorescence and Western blot analysis were performed for cellular localization of ASNS respectively. U937 cells were treated with L-asparaginase for 48 h and examined for endogenous ASNS expression on plasma membrane by immunofluorescence staining. Immunofluorescence staining showed that the transiently expressed ASNS was partly localized on transfected-SK-N-MC cell surface. Moreover, Western blotting exhibited that ASNS expressed both in cytosol and on plasma membrane of transfected-293T cells. Immunofluorescence staining with anti-ASNS-specific monoclonal antibody revealed that endogenous ASNS was localized on the plasma membrane of U937 cells, except for its distribution in the cytosol. In addition, ASNS exhibited a higher expression on plasma membrane after treatment with L-asparaginase as compared with the untreated cells. It was concluded that the subcellular translocation of ASNS may play an important role in L-asparaginase resistance in leukemia cells.

Objective To study the characteristics of dynamic susceptibility-contrast(DSC)MR perfusion curves,color images and perfusion values in pre-operative brain metastasis.Methods Twenty- eight brain metastases underwent DSC MR perfusion imaging by using a first-pass T_2~* echo-planar sequence. The patients' data were transferred to on-line workstation.Time-signal intensity curves,color perfusion maps and rCBV,rMTT values in both tumor parenchyma and peri-tumor edema were analyzed,and independent t- test was used and P0.05).Conclusion Different originated brain metastases have nearly same characteristics in DSC MR perfusion imaging.

10.3969/j.issn.1000-3606.2013.06.005

Objectives To investigate the correlation between single nucleotide polymorphisms (SNP) in interleu-kin-15 (IL-15) and treatment response in childhood acute lymphoblastic leukemia (ALL). Methods Genomic DNA samples extracted from remission bone marrow cells of ALL patients were genotyped by MassArray. Five SNPs (rs10519612, rs10519613, rs17007695, rs17015014 and rs35964658) in IL-15 and their association to minimal residual disease (MRD) status in the end of induction therapy were studied. Results SNP rs17007695 was associated with the early response in children with ALL(P=0.049) and the incidence of positive MRD after induction therapy in CC genotype carriers was 1.8 times more than that in TT genotype carriers. Haplotype analysis of these five SNPs showed that the frequency of haplotype CACGG in MRD positive group was 2.1 times higher than that in MRD negative group (P=0.035). Conclusions IL-15 gene polymorphism was associated with the early treatment response in Han Chinese children with acute lymphoblastic leuke-mia.

Background Haze formation is a key factor of vision reduce following corneal refractive surgery.Transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) are documented to participate in haze formation.Laboratory study showed that bevacizumab can not only inhibit corneal neovascularization,but also promote the healing of corneal epithelial basement membrane.However,the impact of bevacizumab on corneal healing after Off-flap epipolis laser in situkeratomileusis (Off-flap Epi-LASIK) is unclear.Objective The present study was to investigate the inhibition effect of bevacizumab on corneal haze after off-flap Epi-LASIK and its active mechanism.Methods Off-flap Epi-LASIK was performed in 24 adult pigmented rabbits and these rabbits were randomized into three groups.Bevacizumab of 0.1 ml (2.5 mg) was subconjunctivally injected 10 minutes after surgery in 16 rabbits and the same amount of bevacizumab was repeatedly injected 4 days after the initial injection in 8 eyes of 16 eyes.In addition,equivalent amount of normal saline solution was used in the same way in the other 8 rabbits.Another 2 health rabbits were used as the blank controls.Operative eyes were examined by slit lamp biomicroscope daily after surgery and haze was scored based on SundarRayde criteria.Corneas were obtained 4 weeks after operation for hematoxylin & eosin and periodic acid Schiff staining.Expressions of TGF-β1 and α-SMA in corneal tissue were detected by immunochemistry.Results Corneal epithelium healed completely in all eyes 4-5 days after operation.The haze scores were lower in the bevacizumab single injection group and repeat injection group than those in the normal saline solution group (P＜0.05) in 1 week and 4 weeks after operation.However,no significant difference was seen in the haze scores between the bevacizumab single injection group and repeat injection group (P ＞ 0.05).The hostopathological examination showed that the fibrosis response of cornea tissue was slight in the bevacizumab single injection group and repeat injection group comparison with the normal saline solution group.At 1 week after operation,the expression levels of TGF-β1 were (49.8 ± 2.1) PU and (38.6 ±4.4) PU in the bevacizumab single injection group and repeat injection group,and those of 4 weeks were (37.7 ±4.8) PU and (28.3 ± 3.5) PU,indicating significant decrease in the TGF-β1 expression compared with (65.1 ±5.3) PU and (51.6±2.2) PU of the normal saline solution group in both 1 week and 4 weeks (P＜0.01).The expression levels of α-SMA in corneas were (67.2±10.0) PU and (32.7±3.1) PU at 1 week,and (34.2±5.7) PU and (22.8±3.0) PU at4 weeks after operation in the bevacizumab single injection group and repeat injection group,which were significantly lower than (87.8±7.7) PU and (59.4±5.6) PU in the normal saline solution group in both 1 week and 4 weeks (P＜0.05).Meanwhile,the expression levels of TGF-β1 and α-SMA were declined in the bevacizumab repeat injection group compared with single injection group (P＜0.01).Periodic acid Schiff staining exhibited that the basement membrane of cornea was intact and continued in bevacizumab injection group.But corneal basement membrane was discontinuous in the normal saline solution group.Conclusions Subconjunctival injection of bevacizumab downregulates the expressions of TGF-β1and α-SMA in cornea after Off-flap Epi-LASIK and thus prevents haze formation.