Objective: To explore the clinical value of cytokeratin 19 (CK19) and human mammaglobin (hMAM) in the detection of circulating tumor cells (CTCs) in peripheral blood of patients with breast cancer. Methods: Expressions of CK19 and hMAM in blood were identified by real-time fluorescent quantitative PCR, and the correlations between CK19, hMAM and the clinicopathologic features were analyzed. Results: In breast cancer patients, the rates of diagnostic positive detection of individual CK19 or hMAM and the combination of these two biomarkers were 45.5% (20/44), 38.6% (17/44) and 56.8% (25/44), respectively. The expression of combined biomarkers was correlated with lymph node metastasis (P = 0.031). The rates of diagnostic positive detection of CK19, hMAM and the combination biomarkers were significantly higher in breast cancer patients prior to treatment than those in the healthy control group (P = 0.000 3, P = 0.000 1 and P = 0.000 1, respectively). After two-cycle chemotherapy, CTCs which were positive at baseline presented as negative in 5 stage III patients and 4 stage IV patients (P = 0.025 3 and P = 0.045 5). However, the number of CTCs-positive patients at stagesI-II decreased no matter using CK19, hMAM or CK19 plus hMAM as biomarker, leaving more CTCs-positive patients in combined biomarker group than that in single biomarker group, but there was no statistical significance. Conclusion: The combined panel of both hMAM and CK19 may serve as representative biomarkers for CTCs, thus it presents potentially significant value for monitoring early metastasis, therapeutic efficacy and prognosis for the patients with breast cancer. Copyright © 2014 by TUMOR.

Objective:To investigate the arginine (Arg) sites in splicing factor 2/alternative splicing factor (SF2/ASF) methylated by protein arginine methyltransferase 1 (PRMT1). Methods:Wild-type and Arg93,Arg97,Arg109 mutant SF2/ASF plasmids were constructed,and GST-PRMT1,GST-SF2/ASF and arginine mutant GST-SF2/ASF fusion proteins were induced and purified. Methylation activity of PRMT1 on wild-type or mutant SF2/ASF protein and methylated sites of SF2/ASF were examined by methylation assay. The effect of SF2/ASF methylation on its subcellular localization was analyzed by immunofluorescence assay.Results:PRMT1 induced methylation of SF2/ASF at arginine,and PRMT1 did not methylate SF2/ASF when SF2/ASF was mutant at Arg93,Arg97 or Arg109,with Arg97 mutation showing the most profound inhibitory effect. Methylation of SF2/ASF did not affect its subcellular localization.Conclusion:SF2/ASF is a newly identified substrate of PRMT1; Arg93,Arg97 and Arg 109 are the three methylation sites in SF2/ASF,and Arg97 is the main methylation site. Methylation of SF2/ASF does not affect its subcellular localization.

OBJECTIVE: To determine the chlorpyrifos in human blood by liquid chromatography-tandem mass spectrometry and to validate its application in poisoning cases. METHODS: The samples were extracted by a simple one-step protein precipitation procedure. Chromatography was performed on a Capcell Pack C18 MGII column (250 mm x 2.0 mm, 5 μm) using an isocratic elution of solvent A (0.1% formic acid-water with 2 mmol/L ammonium acetate) and solvent B (methanol with 2 mmol/L ammonium acetate) at 5:95 V:V). RESULTS: The linear ranged from 5 to 500 ng/mL (r = 0.998 7). The limit of detection (LOD) and the lower limit of quantification (LLOQ) were 2 ng/mL and 4 ng/mL, respectively. For this method, the precision and accuracy of intra-day and inter-day were < 10% and 97.44%-101.10%, respectively. The results in stability test of long-term frozen were satisfied. The matrix effect, recovery and process efficiency were 64.97%-86.81%, 76.70%-85.52%, and 55.57%-66.58%, respectively. CONCLUSION This method can provide a rapid approach to chlorpyrifos extraction and determination in toxicological analysis of forensic and clinical treatment.
Chlorpyrifos/blood* ; Chromatography, High Pressure Liquid/methods* ; Chromatography, Liquid/methods* ; Humans ; Limit of Detection ; Poisoning ; Reproducibility of Results ; Tandem Mass Spectrometry/methods*

OBJECTIVETo study the clinical manifestation and its pathogenesis of the developmental enamel defects in primary dentition of children with low birth weight and premature birth history.

METHODSOne hundred and seventy-six children (aged 3-8 years old) were studied about the clinical manifestation of the developmental enamel defects in the primary dentition and its relationship with their medical history.

RESULTSThe prevalence of enamel defects in primary dentition in these children was 77.3%. There was no significant correlation between enamel defects and gender. Enamel opacity mostly affected the upper and lower second primary molars. Enamel hypoplasia mostly affected the maxillary and mandibular primary incisors and the maxillary first primary molars.

CONCLUSIONEnamel defects mainly result from children's general disorder at birth or within one year after birth.

Child ; Child, Preschool ; China ; epidemiology ; Dental Enamel ; abnormalities ; Dental Enamel Hypoplasia ; diagnosis ; epidemiology ; Female ; Humans ; Infant, Low Birth Weight ; Infant, Newborn ; Infant, Premature ; Male ; Molar ; abnormalities ; Prevalence ; Tooth, Deciduous ; abnormalities

Objective To explore the relationship between the inflammatory reaction and insulin function in children with critically ill.Me-thods Ninty-six children with critical disease in Oct.2003 to Oct.2006 were enrolled in the study.Blood sugar,plasma insulin,C-peptide,tumor necrosis factor(TNF)-?,C reactive protein(CRP)were measured in the peak period and convalescence.Results Blood sugar and plasma levels of insulin,C-peptide,TNF-?,CRP were significantly higher in the peak period than those in the convalescence(Pa

Objective To explore the method of clinical application and the final result of the com- pound flap of distally based of saphenous nerve-great saphenous vein nutritional vessels.Methods Basing on the anatomic basis that the nutrient vessels of saphenous nerve-great saphenous vein have the same deriva- tion as the branches of the muscle,bone and skin.Eight cases of the tissue defects of ankles and feet were re- paired by the flap.Results Seven cases were survived completely except the distal partly necrosis in one case.The calcaneal defects of one case were repaired by the iliac graft after two weeks of the operation.Follow up from 3 monthes to one year,the texture,appearance and function of the multiflap were good.Conclusion The compound flap of distally based of saphenous nerve-great saphenous vein nutritional vessels has the ad- vantages as follow:the reliable blood supply,avoidance of sacrificing the major arteries,reconstruction of sen- sation and simply procedure.Besides,the defects of soft tissue and bone can be repaired on one operation.

Aim The effects of midazolam on the whole-cell sodium currents in rat sympathetic neurons were studied to explore the mechanisms where by midazolam mediates hypotension. Methods Whole-cell patch-clamp recordings were performed on enzymatically isolated rat superior cervical sympathetic neurons. Results Midazolam dose-dependently blocked the whole-cell sodium currents evoked by a voltage step to 0 mV from a holding potential of -80 mV with a mean drug concentration required to produce 50% current inhibition (IC50) values of 18.35 ?mol?L-1; Clinically relevant concentration of midazolam(0.3 ?mol?L-1) reduced sodium peak currents by 19.98%(P

Objective To introduce the clinical types of perforator branches of anterosuperior malleolus flap and explore its application.Methods Anterosuperior malleolus flap coupling with dorsal pedal flap was used for repairing the soft tissue defect of hands in 18 patients,in which anterosuperior malleolus flap-dorsal pedal single flap in 12 cases,anterosuperior malleolus flap-dorsal pedal bilobate flap in 4 cases,anterosuperior malleolus flap-dorsal pedal trilobate flap in 2 cases;Anterosuperior malleolus retrograde island (bone) flap was used in recovering pedal soft tissue in 22 patients,the flap pedicled from stem of anterior tibial artery in 16 cases,dorsal pedal flap-anterosuperior malleolus flap in 2 cases,the flap from perforate vessels without injuring the anterior main tibial artery in 2 cases,the bone flap combined with the distal of tibia in 2 cases.Results In the 18 cases of hands,17 cases survived,and 1 case of flap mild necrosis at the distal site took a second-phase skin-grafting to repair.Twenty cases of anterosuperior malleolus retrograde island (bone) flap survived,and the other 2 cases needed secondary skin-grafting to repair the necrosis edge of flaps because of venous limited.After a follow-up from 3 to 6 months,30 cases showed the satisfied postoperative outlook,with the good healing of the donor sites.Typing the 40 cases according to the location of perforator branches of the anterosuperior malleolus flap,20 cases locate in the medial of anterior tibial muscle,16 cases locate between the anterior tibial muscle and extensor hallucis longus,4 cases locate between extensor hallucis longus and extensor digitorum longus.Conclusion Knowing the clinical types of the perforator branches of anterosuperior malleolus flap is not only helpful for the accurate processes of operations、preventing cutaneous branches,but also improving the success rate of surgery.

To obtain non-pathogenic rabies virus glycoprotein(RV-G),we expressed RV-G in Saccaromyces Cerevisiae(S.cerevisiae).In our study,tat-G fusion gene was cloned into the expression vector pYes2.0,which allows expression of a foreign gene in the yeast cells under the control of GAl1 promoter.Transforma-tion was performed by using lithium-treated yeast cells and several Ura+-tranformants were isolated.Ac-cording to the relative mobility in SDS-PAGE,we know probably two forms(designated as yGI and yGⅡ) of RV-G analogues produced in S.cerevisiae,their molecular weights were estimated as 66 kD and 56 kD,respectively.On the other hand,there was a specific band about 56 kD shown in western blot result.Com-bining precursors’ achievements,we will draw a conclusion that trans-membrane domain(TD) and cyto-plasmic domain have a negative regulation on RV-G antigen immunogenicity in S.cerevisiae.

AIM:To observe the effect of arsenic trioxide (ATO) on bleomycin-induced pulmonary fibrosis in rats. METHODS:Pulmonary fibrosis was induced in Sprague-Dawley (SD) rats by intratracheal instillation of bleomycin (BLM). The rats in ATO treatment group,steroid treatment group and model group were intraperitoneally injected with ATO,dexamethasone or normal saline (NS),respectively,while the control rats received NS both intratracheally and intraperitoneally. The effects of ATO were evaluated by analyzing the median survival time,hydroxyproline level in the lung,semiquantitative grading of alveolitis and pulmonary fibrosis,and quantitative analysis of the collagen in lung tissue (Masson's trichrome staining). Apoptosis index (AI) of the lung was detected by using the terminal transferase dUTP-digoxygenin nick endlabeling (TUNEL) method. The results of immunohistochemical staining for some cytokines were quantitatively analyzed. RESULTS:ATO (1) prolonged the median survival time of rats with BLM-induced pulmonary fibrosis at some extent; (2) attenuated the alveolitis and pulmonary fibrosis,reduced hydroxyproline level and collagen deposition in the lung tissue; (3) increased the AI of lung tissue at a certain phase; and decreased the levels of transforming growth factor-?1(TGF-?1) and tissue inhibitor of metalloproteinase-1 (TIMP-1),increased the content of in-terferon-? (IFN-?),but did not influence the concentration of matrix metalloproteinase-9 (MMP-9) significantly. CONCLUSION:ATO might attenuate BLM-induced pulmonary fibrosis in rats via increasing the AI in the lung tissue.