BACKGROUND: Bone sialoprotein (BSP) gene is expressed in human breast cancer cells, in which bone metastasis occurs easily outside the mineralized tissue. Clinical observation shows that the expression level of BSP of breast cancer cells at bone metastasis is higher that at the primary site;therefore, BSP may be closely related to tumor specific bone metastasis.The study on breast cancer bone metastasis can provide new drug target for clinical prevention and treatment.OBJECTIVE: To establish breast cancer cell strains of BSP with stable expression and observe the effect of BSP in the whole process of breast cancer bone metastasis.DESIGN: Controlled experiment.SETTING: College of Biological Sciences and Engineering, South China University of Science and Technology; Medical Experiment Center,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA.MATERIALS: This experiment was conducted in the Medical Experimental Center,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA,betweer November 2003 and March 2004..pIRES2-EGFP vector (5.3 kb) was purchased from BD Biosciences Clontech Inc.; E.Coli.Top10, pB-hBSP plasmid containing the coding region of hbsp, and human breast carcinoma cells, MDA-MB-231BR that was specifically transferred to brain and MDA -MB-231BO that was specifically transferred to bone.METHODS: hbsp gene was subcloned from pB-hBSP vector by PCR. Bg1Ⅱ and Pst Ⅰ restriction enzyme sites were inserted at 5' and 3' ends, orientation cloned to eukaryon expression vector pIRES2-EGFP, and constructed recombinant vector pIRES2-EGFP. The constructed recombinant vector was transfected into MDA-MB-231BR that was specifically transferred to brain and MDA-MB-231BO that was specifically transferred to bone.MAIN OUTCOME MEASURES: Construction of pIRES2-hBSP-EGFP recombinant expression vector; recombinant expression vector pIRES2-hBSP-EGFP transfecting breast cancer cells.Breast cancer strains specific in bone metastasis and brain metastasis were successfully transfected. The fluorescence labeling could be observed under the fluorescence microscope, and BSP had corresponding expression.CONCLUSION: The successful construction and transfection of pIRES2hBSP-EGFP of eukaryon expression vector would lay foundation for further study on the role of BSP in breast cancer metastasizing to bone in vivo or in vitro.

Objective To explore the effects of estrogen on stress-induced senescence of vascular smooth muscle cells (VSMCs) and the underlying mechanisms. Methods The VSMCs of passage 2-3 cultured from female SD rats were induced into senescence by exposing to 150μmol/L H2O2 in the presence or absence of different concentrations(10-10mol/L-10-8mol/L) of 17β-estradiol (E2). The expressions or activities of senescence associated marker DcR2, senescence-associated beta-galactosidase (SA-β-Gal), oncogene Ras and p21WAF1 were detected by flow cytometry, cytochemical staining, pull-down assay or Western blotting analysis. Results Flow cytometry analysis showed that in the physiological concentrations, E2 significantly inhibited the H2O2-promoted high-level expression of DcR2 of VSMCs in a dose-dependent manner, with a highest inhibitive rate at 14.48%±0.6%(E2=10-8 mol/L;P<0.05, n =3);this inhibitive effect could be blocked by a E2 receptors inhibitor ICI 182,780. Cytochemistry staining showed that the rate of SA-β-Gal positive VSMCs induced by H2O2 decreased in presence of 10-8mol/L E2 (20.5%±1.4% vs 9.6%±0.9%;P<0.05, n =9). Pull-down assay and Western blotting analysis revealed that administration of 10-8mol/L E2 obviously reduced the H2O2-induced activity of Ras (0.60±0.06 vs 0.26±0.04;P<0.05, n =3) and expression of p21WAF1 (0.46±0.04 vs 0.33±0.02;P<0.05, n =3). Conclusion E2 exerts, an inhibitive effects on stress-induced senescence of VSMCs by suppressing the activity of Ras and expression of p21WAF1. This finding suggests a novel mechanism for the hormone's anti-atheroschlerotic effects.

The bFGF plays an important role in embryonic development of tendons and ligaments and in the healing of injuried tendons and ligaments. The eukaryotic expression plasmid of rat basic fibroblast growth factor (bFGF) gene was constructed in order to further investigate the bFGF function in molecular regulatory mechanism in the repair of tendons and ligaments and to provide the foundation for the clinical application. The cDNA fragments of bFGF were cloned from the skin of rats by RT-PCR, and recombinated to the pMD18-T vector. The cDNA encoding bFGF was cloned from the pMD18-T vector by RT-PCR, digested with restriction enzyme EcoR I Pst I and bound to eukaryotic expression plasmid pIRES2-EGFP to construct eukaryotic expression plasmid pIRES2-EGFP-bFGF. The pIRES2-EGFP-bFGF was transfected into the tenocytes by lipid-mediated ransfection technique. MTT test was used to detect the biological activity of bFGF in supernatants after the transfection. The expression of type I and III collagen genes was detected by using RT-PCR. It was verified that the pIRES2-EGFP-bFGF was successfully constructed, and its transfection into tenocytes could significantly enhance the biological activity of bFGF, and increase the expression of type I and III collagen mRNA, suggesting that pIRES2-EGFP-mediated bFGF gene therapy was beneficial to the repair of tendons and ligaments.

OBJECTIVETo analyze the clinical effect and related risk factors of Gartland type II-III supracondylar fractures of humerus in children in the emergency closed reduction and percutaneous Kirschner wire fixation.

METHODSFrom January 2008 to June 2013,112 children of Gartland type II to III supracondylar humeral fractures were treated in children in emergency closed reduction and percutaneous K-wire fixation, including 72 males and 40 females with an average age of 6.2 years old ranging from 2 to 11 years old. Among them,74 cases were in Gartland type II fractures,38 cases were in type III; The duration from injury to surgery time was 2.5 to 8 hours (averaged 4.6 hours). Elbow cast was applied after operation with the elbow extended of 100 degrees for 4 to 6 weeks, then the gypsum and Kirschner wires were removed.

RESULTSAll patients were follow-up from 6 to 60 months (averaged 12 months). All fractures reached clinical healing. The final follow-up was assessed by Flynn criteria, the result was excellent in 86 cases, good in 23 cases, general in 3 cases, excellent and good rate was 97.3%. Three patients had mild cubitus varus deformity without orthopedic treatment. No pin tract infections, iatrogenic ulnar nerve injury, compartment syndrome, and complications such as Volkmann ischemic contracture occurred.

CONCLUSIONClosed reduction and percutaneous Kirschner wire fixation had advantages of exact reduction, firm fixation, fewer complications ,less pain in children undergoing emergency surgery, and.high success rate, so it is a safe and efficient treatment for humeral supracondylar fracture in children.

Bone Wires ; Child ; Child, Preschool ; Female ; Fracture Fixation, Internal ; instrumentation ; methods ; Humans ; Humeral Fractures ; surgery ; Male ; Reconstructive Surgical Procedures

The aim of this study is to prepare hyaluronic acid (HA) modified core-shell liponanoparticles (pHA-LCS-NPs) as gene delivery system and investigate its gene transfection efficiency in human retinal pigment epithelium (ARPE-19) cells in vitro. The pHA-LCS-NPs was prepared by firstly hydrating dry lipid film with CS-NPs suspension to get LCS-NPs, then modifying the lipid bilayer with HA by amidation reaction between HA and dioleoyl phosphatidylethanolamine (DOPE). Its morphology, particle size and zeta potential were investigated. XTT assay was used to evaluate the cell safety of different vectors in vitro. The gene transfection efficiency of pHA-LCS-NPs modified with different contents of HA was investigated in ARPE-19 cells with green fluorescent protein (pEGFP) as the reporter gene. The results showed that the obtained pHA-LCS-NPs exhibited a clear core-shell structure with the average particles size of (214.9 +/- 7.2) nm and zeta potential of (-35 +/- 3.7) mV. The 24 h cumulative release of gene from pHA-LCS-NPs was less than 30%. After 48 h incubation, gene transfection efficiency of pHA-LCS-NPs/pEGFP was 1.81 times and 3.75 times higher than that of CS-NPs/pEGFP and naked pEGFP, respectively. Also no obvious cytotoxicity was observed on pHA-LCS-NPs. It suggested that the pHA-LCS-NPs might be promising non-viral gene delivery systems with high efficiency and low cytotoxicity.
Cell Survival ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Hyaluronic Acid ; chemistry ; pharmacology ; Lipids ; Nanoparticles ; Particle Size ; Phosphatidylethanolamines ; chemistry ; pharmacology ; Retinal Pigment Epithelium ; drug effects ; Transfection

Purpose To investigate the value of low dose CT combined with iterative model reconstruction (IMR) technique for evaluation of both breast lesion and pulmonary metastasis in patients with breast cancer,and compare the effects of IMR,hybrid iterative reconstruction (HIR) and filtered back projection (FBP) techniques on image quality.Materials and Methods Twenty-one patients with solitary tumor confirmed by mammography and/or ultrasonography at Navy General Hospital from January to May in 2016 were enrolled,and the related breast imaging reporting and data system reached to level 4 and above.All patients underwent dynamic contrast-enhanced CT with different tube current for breast (scanning 6 times within 6 minutes) and chest scans;the conventional tube current (61 mAs) was used at the 2 minutes phase after injection,and the low tube current (10 mAs) was used at the other 5 phases.CT images were reconstructed with FBP,HIR and IMR techniques,respectively.The images of breast lesions and chest in 2 minutes scans were evaluated objectively and subjectively.Results The mean effective radiation dose was (2.15 ± 0.39) mSv.The images of 3 reconstruction techniques at the 2 minutes phase were compared,and the IMR images showed the lowest image noise and the optimal contrast to noise ratio (P＜0.05).The images quality of 3 kinds of reconstruction techniques was acceptable,of which IMR was better than HIR and FBP (P＜0.05).Conclusion Low dose CT scan combined with IMR technique can remain low radiation with dose of 2 mSv,while maintaining good image quality,which can be used for preoperative evaluation of both breast lesion and pulmonary metastasis in patients with breast 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.

Objective To study the clinical significance of antinuclear antibody (ANA),anti-thyroid peroxidase antibody (TPO-Ab) and antithyroglobulin antibody (Tg-Ab),thyroid stimulating hormone (TSH),free triiodothyronine (FT3),free thyroxine (FT4) in patients with recurrent spontaneous abortion (RSA).Methods 46 women with RSA diagnosis in out patient department in Peking University Shenzhen Hospital from March 2015 to December 2016 were recruited as the study group,20 women with normal childbirth history were enrolled as the control group,ANA was detected by indirect immunofluorescence (IIF) assay and ELISA,TSH,FT3 and FT4 were detected by chemiluminescence immunoassay technology.Results The positive rates of ANA(IIF),TPO-Ab and Tg-Ab in patients with RSA were 24％,24％ and 15％ respectively which were significantly higher than those in healthy control (P＜0.05).Serum levels of TSH,TPO-Ab and Tg-Ab in patients with RSA were 2.70 ± 1.38 mIU/L,38.99 ± 10.18 IU/ml and 3.07 ± 1.69 IU/ml respectively,which were significantly higher than those in healthy cases (P＜0.05).Conclusion ANA and antithyroid antibodies were closely related with RSA.It is very important to screen ANA and antithyroid antibodies for etiological diagnosis and appropriate intervention in patients with RSA to reduce the incidence of abortion.

Objective To investigate the effect of oil with medium-long-chain triacylglycerols on blood lip- id and lipoproteins in hyperlipemic patients. Methods Totally, 112 patients with hypertriglyceridemia were en- rolled and randomly divided into MLCT group (consumed oil with medium-long-chain fatty acids) and LCT group (consumed oil with long-chain fatty acids) (both 25-30 g/d for 8 weeks). Patients in both two groups were also instructed to take exercises. Height and weight were measured at baseline and 8 weeks later. Blood glucose, serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterols (TC), triglyc- erides (TG), high density lipoprotein cholesterol, low density lipoprotein cholesterol, apolipoprotein Al (ApoAl), ApoB, ApoA II , ApoC2, ApoC3, and ApoE were measured and compared. Results At the end of study, 101 subjects were included. There were 50 subjects left in LCT group and 51 subjects left in MLCT group, respectively. There was no significant difference in weight, ALT, AST, TC, and TG at baseline between two groups (P>0.05). Eight weeks later, weight, serum TG, ApoC2, and ApoC3 were significantly lower and ApoAl level was significantly higher than those at baseline in MLCT group (P < 0.05). At the end of study, the decreases in body weight and blood biochemical variables including TG, ApoB, ApoA II , ApoC2, ApoC3 were significantly much greater in MLCT groups than those in LCT group (P < 0. 05). Conclusion When the diet is reasonably controlled, oil of medium-long-chain triacylglycerols may reduce the concentration of TG and improve the levels of apolipoproteins.

· Basal ceil carcinoma (BCC) is one of the most common human cancers. The giant ulcerated BCC invading the eyeball and orbit has been rarely reported. We present a case of giant BCC with cutaneous ulcer on the left head,invading the eyeball, orbit and skull.