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

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.

Objective To investigate the effects of Shenfu injection ( SF,a Chinese herbal medicine preparation made of Codonopsis pilosula and Aconitum carmichaeli) on the cell apoptosis of focal cerebral ischemic-reperfusion injured rats and the expression of heme oxygenase-1 (HO-1). Methods Forty-two male Sprague-Dawley rats used for producing unilateral brain ischemia reperfusion model were randomly divided into three groups:sham operation group ( Sham group),ischemia reperfusion group ( IR group),and SF Injection group (SF group).The model of focal cerebral ischemia-reperfusion injury was induced by transient occlusion of middle cerebral artery (ischemia for 2 h,and reperfusion for 3,6 h respectively).In SF group,SF ( 10 mg/kg) was intraperitoneally injected duri(n)g reperfusion.Cell apoptosis rate in brain tissue was detected by the technique of Annexin-V-PI double staining and was counted in flow cytometer.Expression of HO-1 in brain was measured by RT-PCR,while the pathological and ultra structure changes of cerebral tissue were also observed.Results Cell apoptosis rate of brain tissue were significantly higher in IR group than that in Sham group (P ＜0.01 ),while SF group had less significant changes in cell apoptosis rate, HO-1 level of brain tissue than IR group (P ＜ O.01 ).The ultra structure change of brain tissue was less in SF group than that in IR group.Conclusions During early stage of brain IR injury,SF inhibits cellular apoptosis and in turn protects the brain from injury which is attributed to the increase in HO-1 expression induced by SF.

Objective To investigate the risk factors of bronchopulmonary dysplasia(BPD)in very low birth weight infant.Methods The clinical data of 49 very low birth weight infants in our NICU from Sep 2006 to Sep 2009 were reviewed,and divided into BPD group(n =15)and without BPD group(n =34).The risk factors of BPD were analysed.Results Compared with the infants without BPD,there were significant differences in gestational age[(29.30 ± 1.48)week vs(30.54 ± 1.60)week],hospital-acquired infection(9 cases vs 10 cases),intrauterine infection(9 cases vs 8 cases),the time for continuous positive airway pressure(CPAP)[(12.47 ± 5.83)d vs(4.24 ± 4.19)d],the time for hyperoxia[(1.47 ± 1.41)d vs (0.18 ±0.63)d],patent ductus arteriosus(5 cases vs 1 cases)(P＜0.05).Logistic regression revealed that intrauterine infection and the time for CPAP were independent risk factors of BPD(P ＜0.05).Conclusion Prophylaxis of intrauterine infection may decrease the mortality and severity of BPD.The prolonged time for CPAP may predict the risk of BPD.

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.

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