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.

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.

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 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 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 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.

In order to provide scientific basics for exploitation and sufficient application of Dendrobium officinale leaves resources, the phenol-sulfuric acid method was applied to determine the polysaccharide content. The monosaccharides were derivated by PMP and the derivatives were identified by HPLC-DAD-ESI-MS(n) and the contents of mannose and glucose were determined simultaneously. Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (2004A) was employed to generate the mean chromatogram and similarity analysis of the samples was carried out. The results demonstrated that polysaccharide content, monosaccharide compositions and composition ratio had an obvious difference between stems and leaves. The polysaccharide content of stems was higher than that of leaves. Monosaccharide composition in leaf was significantly different from that in stem. The polysaccharide from stems was composed of mannose and glucose, however the polysaccharide of leaves was acid heteropolysaccharide and was mainly composed of five monosaccharides, including mannose, galacturonic acid, glucose, galactose and arabinose. The similarity value of the 14 batches was above 0.9, indicating that similarity of fingerprints among different samples was high. The study can provide evidence for expanding the medicinal parts of D. officinale.
Chromatography, High Pressure Liquid ; Dendrobium ; chemistry ; Mass Spectrometry ; Plant Extracts ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Polysaccharides ; chemistry

ObjectiveTo observe the change of cardiac function after acute myocardial infarction (AMI) in rabbit model,and to study MRI characteristics of left ventricular remodeling (LVR).Methods Forty-five japanese white rabbits underwent chest-opening coronary artery ligation surgery to obtain rabbit myocardial infarction model The animals were scanned on a 1.5 T MR scanner ( GE Healthcare,Chalfont St.Giles,UK) at six time-points as surgery,1,2,4,6,8 weeks after surgery. Cardiac function parameters were measured,including left ventricular end diastolic volume ( EDV ),stroke volume ( SV ),ejection fraction (EF),peak ejection rate ( PER ) and peak firing rate (PFR).At each time point,5 rabbits were randomly selected and performed re-thoracotomy to obtain heart specimen.Each specimen was examined by MRI with the diffusion tensor imaging ( DTI ).The value of ADC and fractional anisotropy (FA) were collected. Cardiac function data sets of different time points were analyzed using repeated measures data of ANOVA.The FA and ADC values of infarction myocardium set of different time points or different parts of the myocardium were analyzed using one-way ANOVA. The average was compared statistically between two adjacent groups using LSD-t test. Results Left ventricular EDV increased in progress with the time.It was increased from ( 2.21 ± 0.35 ) ml preoperatively to ( 3.15 ± 0.57 ) ml 8-week postoperatively.But the difference was not statistical significant ( F =1.384,P =0.220).EF was decreased from(57.31 ± 11.11 )％ to( 34.71 ± 8.72 )％.It dropped significantly and the difference was statistically significant (F =27.134,P =0.001 ).EF showed a downward trend with the increase of EDV.By linear regression analysis,an equation was set up with y =- 5.58x + 57.7 ( F =8.855,P =0.005 ).On the other hand,PER showed a progressive decline from (27.31 ± 13.06) ml/s to ( 17.31 ± 6.41 ) ml/s ( F =2.105,P =0.037 ),and PFR decreased from ( 23.79 ± 14.15 ) ml/s to ( 12.07 ± 6.91 ) ml/s ( F =2.366,P =0.011 ).FA value decreased from 0.442 ±0.093 to 0.107 ±0.037( F =12.61,P =0.001 ),and ADC valueshowed upward trend from (5.07 ± 1.15) ×10-4 mm2/s to(6.39 ±0.78) ×10-4 mm2/s (F=4.166,P =0.022 ).FA values of infarct,adjacent and remote region were 0.201 ± 0.049,0.316 ± 0.127 and 0.323 ± 0.117 respectively( F =3.896,P =0.004 ),and the ADC values in these regions were (6.19 ± 1.78 ) ×10 -4,(5.44 ± 2.63 ) × 1 0 -4,(5.29 ± 2.02 ) × 10 -4 mm2/s respectively ( F =3.248,P =0.018 ).FA and ADC values were significantly different between the infarct region and adjacent region ( t =7.327,P =0.001 ;t =3.292,P =0.005,respectively),but there was no significantly different between adjacent region and remote region ( t =1.024,P =0.129 ; t =1.467,P =0.164,respectively ).ConclusionsMRI measurement of parameters of cardiac function can be used to monitor the process of left ventricular function remodeling after AMI.The process of micro-structural remodeling of myocardium can be reflected by DTI.MRI provides a feasible imaging modality for LVR after AMI.