Circulating cell-free DNA (cfDNA), which is released by normal cells and cancer cells, is defined as extracellular DNA in the blood. The cfDNA levels in breast cancer patients are higher than those in healthy control donors. cfDNA also carries the features of tumor tissue, such as mutations, methylations, copy number changes, and loss of heterozygosis. cfDNA is a potential bio-marker in the diagnosis, management, and prognosis of breast cancer. In this review, the authors briefly describe the biological features of cfDNA, and discuss its clinical utility as a blood biomarker in quantitative and qualitative research.

Objective To investigate the effects of down-regulation of Netrin-1 expression on cell proliferation,apoptosis and migration of a cutaneous squamous cell carcinoma(CSCC)cell line SCL-1.Methods Unspecific(control)small interference RNA(siRNA)and Netrin-1-targeting siRNA were designed and constructed.SCL-1 cells were divided into 3 groups to remain untreated(control group 1),be transfected with control siRNA(control group 2)and Netrin-l-targeting siRNA(experiment group)by using Lipofectamine 2000 respectively.After 48 hours of additional culture,flow cytometry was performed to detect the apoptosis of cells,Western blot to determine the protein expression of caspase-3 and-9,Netrin-1 and matrix metalloproteinases (MMP)in these cells,and Boyden chamber was utilized to evaluate the migration of cells.Enzyme linked immunosorbent assay(ELISA)was performed with cholecystokinin-8(CCK-8)kit to estimate the proliferation of cells at 24,48,72 and 96 hours after the transfection.Results The expression of Netrin-1 protein was significantly downregulated in SCL-1 cells in the experiment group compared with the control group 1 and 2 (0.207 ± 0.044 vs.0.741 ± 0.111 and 0.716 ± 0.109,F =31.43,P ＜ 0.05).The Netrin-1-targeting siRNA significantly downregulated the proliferation of SCL-1 cells,but induced the apoptosis in SCL-1 cells with significant differences in the percentage of early apoptotic cells and late apoptotic cells between the control group 1,2 and experiment group(11.74 ± 0.33 vs.11.55 ± 1.22 vs.24.74 ± 2.43,F=68.63,P＜ 0.01;3.82 ± 0.32 vs.4.69 ± 0.68 vs.5.33 ± 0.18,F =8.58,P ＜ 0.05).The experiment group showed a significant increase in the expression of caspase-3 and-9(1.458 ± 0.111 vs.0.398 ± 0.057 and 0.412 ± 0.049,F =183.99,P ＜ 0.01;0.967 ± 0.099 vs.0.234 ± 0.036 and 0.205 ± 0.033,F =138.11,P ＜ 0.01),but a statistical decrease in the expression of MMP-2(0.235 ± 0.038 vs.0.862 ± 0.040 and 0.892 ± 0.035,F =291.07,P ＜ 0.01)compared with the control group 1 and 2.Conclusions The down-regulation of Netrin-1 expression can suppress the proliferation and migration of SCL-1 cells,but promote the apoptosis in SCL-1 cells.

Objectlve:To assess the influential factors of decision-to-delivery inteval (DDI) in caesarean section, and its influence on neonatal outcomes.Methods:472 caesarean sections were divided into two groups according to Lucas's classification :the emergency caesarean sections as group 1 (291) ; and the e-lective caesarean sections as group 2 (181).It was divided into DDI ≤30 min group and DDI > 30 mir group in group 1.A retrospective study was performed in DDI, influential factors of DDI, neonatal Apgar score and umbilical arterial blood gas.Results: ①The mean DDI was 35.5±11.6 min in group 1,in wgich DDI≤30 min was 210 cases (72.2%) and 49.3 ±22.8 min in group 2, in which DDI≤30 min was 86 cases (47.5%).②IN group 1,umbilical artery pH and Apgar core at 1 min after birth could be improved sigbificantly in the cases of DDI ≤ 30 min (P<0.05) , but no correlation was found between the DDI and Apgar scrore at 5 min ,as compared with DDI ＞30MIN CASES(p＞0.05).③It was mainly influenced by time taken to get the patient into operation room in DDI >30 min (56 cases, 69.1 %).Concluslons :The recommended DDI ≤30 min is not routinely achieved even in emergency caesarean sections.Shortening DDI as far as pos-sible might improve the neonatal outcome.

The culture of umbilical cord-derived mesenchymal stem cells is extremely important for studies on umbilical cord mesenchymal stem cells. Optimization of cellculture technology is crucial for clinical application of mesenchymal stem cells and even celltherapy. Meanwhile, the labeling and tracer technique of umbilical cord-derived mesenchymal stem cells is a hotspot in stem celltransplantation. OBJECTIVE:To review the research and development of the cellmarkers and tracer methods of umbilical cord-derived mesenchymal stem cells. METHODS:A computer-based search of VIP, CNKI, Medline, Highwire and Foreign Journals Integration System databases was performed for articles concerning culture and labeling of umbilical cord-derived mesenchymal stem cells published from January 2001 to October 2013. The keywords were“stem cells, mesenchymal stem cells, umbilical cord-derived mesenchymalstem cells, cellculture, labeling methods”in Chinese and English, respectively. Final y, 35 articles were included in result analysis. RESULTS AND CONCLUSION:Umbilical cord-derived mesenchymal stem cells have not yet been widely used, mainly because of the immature isolation, culture and staining techniques of umbilical cord-derived mesenchymal stem cells. These techniques are worthy of further optimization studies. Although in recent years, cellmarkers and tracer technology of umbilical cord-derived mesenchymal stem cell s have made great progress, there are stil many problems need to be solved.

Objective To research the applications of resting-state functional MRI in conjunction with voxel-based morphometry on cerebral functions and structures in patients with schizophrenia. Methods Thirty-two patients with schizophrenia and 32 healthy people underwent resting state functional magnetic resonance imaging (Rs-fMRI)and sagittal scanning T1WI with 3D FFE Pulse Sequence. Rest1.8 software was applied to calculate the differences of amplitude of standardized low-frequency fluctuation(ALFF)between the two groups,then the voxel-based morphometry(VBM)data of abnormal brain regions were evaluated to compare the differences of grey matter intensity. Finally,combining with ALEF and VBM,we explored the changes of cerebral functions and structures. Results As compared to the control group ,the ALFF-elevated regions were the right putamen ,the left orbital frontal cortex extending to the left putamen and left dorsal-lateral superior frontal gyrus;the ALFF-decreased regions were the right inferior tempotalgyrus,the left posterior cingulated gyrus and the left angular gyrus(P<0.05). No elevated regions of grey matter intensity were found in the patient group. Conclusions Extensive abnormal active regions of the brain under resting state could be found in patients with schizophrenia ,the grey matter intensity of abnormal regions also decreased generally. They are in accordance with the glutamate-hypothesis of schizophrenia ,which involves extensive impairments of neurons ,disorders of neurotransmitter′s circulations and balances.

Objective: To observe the impact of mind-regulating acupuncture plus donepezil on the cognitive ability, mean cerebral blood flow velocity, event-related potential P300, and activities of daily living (ADL) in the aged patients with Alzheimer disease (AD).Methods: Sixty senile AD patients were divided into a treatment group and a control group following the envelope method for random allocation, with 30 cases in each group. Based on the conventional treatment of the internal medicine, the control group received oral donepezil, and the treatment group received oral donepezil plus mind-regulating acupuncture. After 4-week treatment, the two groups were evaluated by the mini-mental state examination (MMSE), Alzheimer disease assessment scale-cognitive part (ADAS-Cog), and ADL; changes in P300 and the mean cerebral blood flow velocity were also observed.Results: Before treatment, there were no significant differences in the scores of MMSE, ADAS-Cog, or ADL between the two groups (P>0.05). The MMSE score increased after treatment in both groups and was notably higher in the treatment group than in the control group, showing intra-group and inter-group statistical significance (P<0.05). After treatment, the ADAS-Cog and ADL scores dropped in both groups and were markedly lower in the treatment group than in the control group, also showing intra-group and inter-group statistical significance (P<0.05). Compared with the same group before treatment, the latency of P300 was shortened and the amplitude was extended in both groups, all with statistical significance (P<0.05); the latency was shorter and the amplitude was larger in the treatment group than in the control group after treatment, presenting significant between-group differences (P<0.05). The mean blood flow velocity accelerated after the intervention in both groups, and the differences were statistically significant (P<0.05); the improvement in the treatment group was more notable than that in the control group (P<0.05).Conclusion: Mind-regulating acupuncture plus donepezil can regulate the latency and amplitude of P300, increase cerebral blood flow, and improve the learning and memory abilities of AD patients.

Objective To study the expression of high mobility group box 1 (HMGB1),matrix metalloproteinase-9(MMP-9) and vascular endothelial growth factor-C (VEGF-C)gene in papillary thyroid carcinoma (PTC) and their significance.Methods Expression of HMGB1,MMP-9,VEGF-C proteins and HMGB1 mRNA was detected by technology of immunohistochemistry(SP) and in situ hybridization in 58 cases of PTC tissues,20 cases of thyroid adenoma tissues,25 cases of nodular goiter tissues and 10 cases of normal thyroid tissues.Results Expression of HMGB1,MMP-9,VEGF-C proteins and HMGB1 mRNA in PTC tissues was much higher than that in the 3 non-cancerous tissues (P ＜ 0.05).The positive expression rate of HMGB1,MMP-9 proteins and HMGB1 mRNA was closely associated with the thyroid carcinoma size and lymph node metastasis (P ＜0.05).There was also closely association between the positive expression rate of VEGF-C protein and lymph node metastasis(P ＜ 0.05).The positive expression rate of HMGB1,MMP-9,VEGF-C proteins and HMGB1 mRNA had no association with patients' sex or age(P ＞ 0.05).The expression of HMGB1,MMP-9 and VEGF-C proteins had positive correlation with each other(P ＜ 0.05).The expression of HMGB1 protein and HMGB1 mRNA was also positively correlated with each other(P ＜0.05).Conclusion Expression of HMGB1,MMP-9,VEGFC proteins and HMGB1 mRNA in PTC is correlated with the progress,invasion,metastasis of PTC,by detecting them may help to predict the clinical progress and prognosis.

Objective To investigate the role of polymorphisms of scavenger receptor class B gene CD36 in affecting the progress of subclinical atherosclerosis (AS) and the associated factors affecting the expression of CD36 on the surface of peripheral blood monouclear cells (PBMC) and the association between CD36 expression and progress of subclinical AS in type 2 diabetic patients.Methods CD36 polymorphisms (CD36-rs1984112, CD36-T620C) were typed by PCR-RFLP in 470 cases of type 2 diabetes mellitus and 220 non-diabetic controls of Hans in Hunan area.The genotypes and allele frequencies were compared between cases and controls.Fluorescence intensity of CD36 on the surface of PBMC was analyzed in 102 cases of type 2 diabetes mellitus by flow cytometry and was compared between the patients without AS and the patients with subclinical AS.Multiple linear regression was applied to evaluate the relevant factors contributing to CD36 expression.Results The genotypes and allele frequencies of CD36-rs1984112 in type 2 diabetes mellitus were not significantly different between cases and controls (P＞0.05), either did CD36-T620C (P＞0.05).The mean florescence intensity (MFI) of CD36 in type 2 diabetics with subclinical AS was higher than that without AS (1 382±659 vs 1 173±340, P＜0.05).Factors affecting the CD36 expression were: age (P=0.005), gender (P=0.021), systolic blood pressure (SBP) (P=0.027), standardized coefficients Beta was 0.28, 0.31 and -0.21, respectively.Age contributed to the CD36 expression level in males (P=0.002) and diastolic blood pressure in females (P=0.001) respectively.Conclusion CD36-rs1984112 and T620C seem not to be a functional polymorphism sites in Hans of Hunan, southern China.CD36 expression level is higher in type 2 diabetics with subclinical AS in contrast with those without AS.CD36 expression on PBMC surface is higher in aged males with lower SBP.

Objective To investigate dosimetric advantage of fixed field intensity-modulated radiotherapy (FF-IMRT),volumetric modulated arc therapy (VMAT) and helical tomotherapy (HT) for cervix uteri cancer.Methods CT datasets of ten patients with cervix uteri cancer were enrolled in the study.FF-IMRT,VMAT and HT plans were designed on Eclipse and HT treatment planning system.Plans were optimized with the aim to assess OAR while enforcing highly conformal target coverage.Institutional dose-volume constraints used in cervix uteri cancer were kept the same for three techniques.The different of three plan was play by single factor analysis of variance and compared to two groups by LSD method.Results All FF-IMRT,VMAT and HT resulted in equivalent target coverage but HT had an improved homogeneity index (P =0.000) and conformity index (P =0.000),or PTV of 105％ prescription dose (47.12％,45.83％ and 0.05％,P=0.000) and lowest Dmax dose (54.53 Gy,53.65 Gy,52.69 Gy,P=0.000).Compared with FF-IMRT and VMAT,the bladder V40 and Dmax of HT were lowest (50.01％,46.84％,42.98％,P=0.001 and 54.49 Gy,52.96 Gy,52.78 Gy,P=0.000),with the rectum V40 lowest (54.61％,48.34％,46.78％,P =0.006),the intestine Dmax lowest (54.53 Gy,53.65 Gy,52.66 Gy,P =0.000) and marrow Dmax lowest (54.51 Gy,54.44 Gy,52.13 Gy,P=0.000).But the delivery MU per fraction were highest (1429.20 MU,617.80 MU,7002.04 MU,P =0.000).Conclusions HT technology is feasible for clinical applications in cervical uteri cancer and can be used as a new method to promote.

Objective To evaluate the dosage regimen of sulfasalzine combined with recombinant human tumor necrosis factor receptor-Fc fusion protein (rhTNFR) in the induction and maintenance of remission in active ankylosing spondylitis.Methods This study enrolled 100 ankylosing spondylitis patients with disease duration for less than five years who were treated with sulfasalzine and rhTNFR combination therapy.Patients were randomly assigned to three groups six weeks later:patients in the G1 group received sulfasalzine combined with rhTNFR at a 25 mg dosage twice each week.Patients shifted to monotherapy with sulfasalzine six weeks later:patients in G2 group received sulfasalzine combined with rhTNFR at 25 mg dosage per week.Patients were switched to monotherapy of sulfasalzine twelve weeks later:patients in G3 group received sulfasalzine combined with rhTNFR at 25 mg dosage once every ten days.Patients were changed to monotherapy of sulfasalzine seventeen weeks later.The whole treatment lasted for 24 weeks.All participants were followed up at week 0,6,12,18,24 respectively and were evaluated by BASDAI 50.The primary end-point of this study was the percentage of patients achieved BASDAI 50 remission.Data were analyzed with SPSS version 17.0.Independent t-test and x2 test were adopted to analyze data.Results 90％ of patients treated with combination therapy reached BASDAI50 at the 6th week.All patients in the G1 group achieved BASDAI 50 remission at 12th week,but the percentage dropped to 68.7％ at 18th week,which gradually decreased to 37.5％ at the 24th week.In G2 group,93.9％ patients reached BASDAI50,which declined to 81.8％ at the 18th week.The whole number accounted for 60％ at the end point of 24th week.In G3 group,85.7％ patients achieved BASDAI50 at the 12th week,accounted for 74.3％ at 18th week,and declined to 68.6％ at the 24th week.G3 group of patients presented a significantly higher rate than other groups(P＜0.05).Conclusion Sulfasalzine and rhTNFR combination therapy can gain remission in active AS patients after treated for six weeks.Doctors may extend TNF antagonist treatment in order to achieve long-term remission.