Objective To successfully get the full PCR alleles of the Insertion/Deletion marker rs10644346 in which a SNP-binding exists at the 3'end region of the primer.Methods Based on the AS-PCR,a common forward primer and two reverse primers with allele-specific oligonucleotides at the last second 3'end instead of the terminus were tentatively designed for typing 150 unrelated individuals and 10 father-mother--child trios from Htnan province in South-central China.Simultaneously,9 samples were typed with all the above three primers (the two primer sets which consist of the common forward primer and one of the reverse primers).Results PCR amplicons were well detected in the 150 unrelated individuals after being typed with the three primers,and the amplified fragments of parental and filial generations among 10 father-mother-child trios conformed to Mendel's principles.Allele missing was found in the two-primer group.Conclusion The primers designed by locating the specific nucleotide at the last second 3'end rather than terminal position were demonstrated also effective in getting specific alleles if perfect mismatch and PCR conditions are guaranteed,and the design strategy can provide an optional reference to rescue markers of SNP-binding primers for forensic practice.

Objective To study the early changes of the coagulation system in type 2 diabetic nephropathy.Methods Sixty-two cases of patients with type 2 diabetic nephropathy were divided into two groups:normal albuminuria group ( N-UAlb group,UACR ＜ 30 mg/g,32 cases ),microalbuminuria group ( MUAlb group,UACR:30～300 mg/g,30 cases).Thirty healthy persons constituted a control group (NC group).Fibrinogen( FIB ),antithrombin Ⅲ ( AT-Ⅲ ),protein C ( PC ),protein S ( PS ) were measured by coagulation analyzer,while yon willebrand factor (vWF) and platelet granule membrane protein 140 (GMP-140) were detected by ELISA assay,platelet count (PLT),mean platelet volume(MPV),platelet hematocrit (PCT),platelet distribution width(PDW) by hematology analyzer.Results The level of fibrinogen,GMP-140 and vWF in the M-UAlb group were (4.20 ± 1.53 ) g/L,( 30.03 ± 7.77 ) μg/L,and ( 315.53 ± 47.24 ) ％ respectively,vwhich were significantly higher than those in the N-UAlb group [ ( 3.21 ± 0.89 ) g/L,( 18.22 ± 5.08 ) μg,/L and ( 191.88 ± 57.25 ) ％ respectively ] and the NC group [ ( 2.75 ± 0.53 ) g/L,( 14.26 ± 2.29 ) μg/L and ( 138.12 ± 61.27 ) ％ respectively ] ( F =5.42,10.42,30.44,P ＜ 0.05 or 0.01 ).The fibrinogen,vWF,GMP-140 were positively correlated with UACR ( r =0.313,P ＜ 0.05 ; r =0.620,P ＜ 0.01 ; r =0.680,P ＜ 0.01 ) and PC was negatively correlated with UACR ( r =-0.255,P ＜ 0.05 ).Conclusion Hypercoagulable state in diabetic nephropathy is associated with the high fibrinogen,endothelial dysfunction and platelet activation,and these changes have already emerged in patients without albuminuria.This might mind us that we should strengthen anticoagulant therapy on patients when they are not progressing to albuminuria.

Objective To determine the content of six heavy metal elements of arsenicum(As),mercury(Hg),cuprum(Cu),plumbum(Pb),cadmium(Cd) and chromium(Cr) in Rhizoma Curcumae Longae.Methods Samples were digested with microwave digestion system,and the contents of Cu,Pb,Cd and Cr were determined by graphite furnace atomic absorption spectrometry(GFAAS);the contents of As and Hg were determined by atomic fluorescence spectrometry(AFS),and the reproducibility and recovery of the method were also examined.Results As and Hg contents in most of the products were below the limit.Only Cu,Pb and Cd contents exceed the limit.The recovery was 86.3 %～113.4 %and RSD was 10.4 %.Conclusions The method is simple and accurate,and can be used for the determination of heavy metal in Chinese herbs.

Objective To analyse long-term follow-up outcome of cryoballoon ablation (CBA) for atrial ifbrillation (AF) in a single center and to investigate the clinical relative factors which affecting the effect. Methods The inpatient, operating and outpatient data of patients, who were treated by CBA for AF in our center from January 2009 to April 2013, were retrospectively analyzed. Left atrium diameter (LAD) was measured by transthoracic echocardiography. Failure-treatment of CBA was defined by episode of AF, atrial lfutter, atrial tachycardia lasted for 30 seconds after 3 months. Results A total of 199 patients were enrolled. The rates of phrenic nerve paralysis, pericardial effusion, transient ischemic attack were 1.5%(n=3), 0.5%(n=1), 0.5%(n=1),respectively. All complications were resolved spontaneously.152 patients had completed follow-up data after ifrst-time CBA during a long-term follow-up of mean 23±14 months, 65 patients (42.8%) treated success. 75 patients with failure-treated were (86.2%) experienced the atrial arrhythmia recurrence in ifrst 12 month. The characteristics of failure-treated patients included with older age[(62±7) years vs. (52±10) years, P=0.0379]and larger LAD[(48±6)mm vs. (43±6) mm, P<0.0001]. The Logistic analysis showed that LAD[OR=0.896(0.842,0.953), P=0.005]and age[OR=1.037 (1.000,1.076), P=0.0488]could individually predict the treat-failure after ifrst CBA, and only LAD[OR=0.876 (0.822,0.935), P < 0.0001]could individually predict the total CBA. Conclusions CBA procedure for AF is safe and effective, and the result of long-term follow-up is preferable. Most atrial arrhythmia are recurred during ifrst 12 month after CBA. LAD can individually predict the failure in treatment of CBA.

Objective The expression of chemokine C-X-C motif ligand 13 (CXCL13) within liver in primary biliary cholangitis (PBC) patients is significantly increased, but its origin and mechanism is not clear yet.The study aimed to investigate the expression of CXCL13 in the liver of mice through establishing a mouse model of PBC.Methods C57BL/6 mice were randomly divided into experiment group (n=20) control group(n=10).The mice in the experimental group were intraperitoneally injected with polyriboinosinic polyribocytidylic acid (Poly I:C) while the mice in control group were injected with PBS of the same volume.The level of serum AMA was quantified by ELISA and intrahepatic inflammatory cells were assessed by HE staining.Kupffer cells, liver sinusoidal endothelial cells, and infiltrating lymphocytes in the liver of mice were collected by in situ perfusion enzyme digestion and magnetic bead separation methods.The transcriptional level of intrahepatic CXCL13 in liver tissues and cell subpopulations were detected by qPCR.Results The serum AMA titers of the mice in experiment group increased gradually with the prolonging of modeling time and the positive rates at the 4th, 8th, and 12th week after the first injection of Poly I:C were 5.9%, 52.9% and 76.5% respectively.While the serum AMA titers of the mice in control group were at a lower level through the modeling process, with only 2 mice presenting a little higher level above positive cutoff value at the 12th week.The results of HE staining in liver tissues of both groups showed that there were a great amount of intensely infiltrating inflammatory cells in the mice of experimental group while no inflammatory cell infiltration were found in the mice of control group.The separation purity of Kupffer cells and liver sinusoidal endothelial cells in the mice of experiment group tested by flow cytometry were 76%-80%, 68%-72% respectively.Compared with the CXCL13 mRNA level in Kupffer cells [2.34(0.22-8.64)], the expression levels in liver sinusoidal endothelial cells and infiltrating lymphocytes declined[0.27(0.03-1.64), 0.05(0-0.22), P<0.05].Conclusion The chemokine CXCL13 is predominantly produced by Kupffer cells in the liver of PBC mice established by Poly I:C injection.

Objective To study the relationship between social support and subjective well-being(SWB) of labor workers.Methods 1038 labor workers in Shenzhen were evaluated by subjective well-being scale for Chinese citizen (SWBS-CC), social support rating scale ( SSRS ).Results There was no significant difference in SWB between male and female( ( 63.51 ± 11.03 ) vs ( 63.22 ± 12.02 ), t = 0.389, P ＞ 0.05 ).The 30 ～ 49 age group's score was significantly higher than the 16 ～29 age group's( (62.80 ± 11.80) vs (64.54 ± 11.28), t=2.21, P ＜ 0.05 ).The married group's score was significantly higher than the unmarried group's ( (62.50 ± 12.18)vs (64.27 ± 10.80 ), t= 2.44, P ＜ 0.05 ).Social support was significant positively correlative with subjective well-being ( r=0.075 ～0.391, P＜0.05), except support availability was not correlative with both subscales.Subjective support, support availability and objective support were significantly positive predictions of SWB, meanwhile subjective support was the biggest( β =0.297).Conclusion Subjective well-being of the labor workers in Shenzhen have significant relationship with age and social support.To increase the emotional support for the labor workers could possibly help to improve their subjective well-being.

Objective To get the genetic information and forensic parameters for the STR locus of D1S1656 in Hu'nan Chinese Han population . Methods All 451 unrelated individuals of Hu'nan Chinese Han population were selected to analyze genetic polymorphisms by using the Powerplex 21 system kit. The deviation of allele frequencies from Hardy-weinberg equilibrium was tested and the forensic parameters were counted by software. Results The marker of D1S1656 showed no significant deviation from Hardy-weinberg expectations, and 16 alleles were found with Power of Discrimination was 0.954 and Power of Exclusion 0.667. Conclusion The STR marker of D1S1656 present rich polymorphisms in Hu'nan Chinese Han population and could provide effective application in forensic field.

Fear memories are temporarily suppressed after repeated retrieval, a phenomenon known as memory extinction.How to reduce or even eliminate fear memory is the key to the treatment of fear related diseases such as post-traumatic stress disorder(PTSD). A single extinction training based on Pavlov's fear regulation task could only inhibit the expression of conditioned fear memory traces, but it could not eliminate the acquired conditioned fear memory. However, according to the reconsolidation theory based on memory, the retrieval-extinction paradigm has a more lasting effect on the erasure and rewriting of fear memory, and can effectively prevent the return of fear memory. Studies have shown that extraction-regression is closely related to a variety of neurotransmitter receptors such as glutamate receptor(GluR), dopamine receptor(DAR), L-type voltage-gated calcium channels(LVGCs) and cannabinoid. Moreover, its effect is closely related with factors such as retrieval-extinction memory stage. At present, most of the researches on extracted boundary conditions only stay at the level of behavior, with little understanding and exploration on the level of molecular mechanism. From the perspective of molecular neurobiology, with different stages of memory and different types of receptors and molecular mechanisms, this research reviewed the mechanisms of retrieval-extinction in recent years.It provided valuable signaling pathways, molecular targets and research directions for the treatment of fear-related diseases such as PTSD.

Nonalcoholic fatty liver disease (NAFLD) and colorectal adenomatous polyps are closely associated with the various components of metabolic syndrome. This article summarizes the recent studies on the association between NAFLD and colorectal adenomatous polyps, and the results show that NAFLD is associated with an increased risk of colorectal adenomatous polyps, while related mechanism remains unclear, which may be associated with insulin resistance, chronic inflammatory response, adipokines, and intestinal flora disturbance.

OBJECTIVETo study the effects of RhoA down-regulation by RNA interference on the invasion of tongue carcinoma Tca8113 and SCC-4.

METHODSDetermination of the human RhoA sequence as well as the design and constructionof a short specific small interfering RNAs (siRNA) were performed. The siRNA of RhoA gene was transfected into humantongue squamous cell carcinoma Tca8113 and SCC-4 cells line by Lipofectamine 2000. Quantitative real-time polymerasechain reaction was used to examine the mRNA expressionlevels of RhoA. Protein expressions of mRNA, galectin-3,and matrix metalloproteinase (MMP)-9 were evaluated byWestern blot. Transwell invasion assay was performed toassess the invasion ability of tongue carcinoma.

RESULTSRhoA expressions in Tca8113 and SCC-4 cells were reducedsignificantly after transfection of RhoA-siRNA. Protein levels f galectin-3 and MVP-9 were also down-regulated significantly. Invasion ability was inhibited as well.

CONCLUSIONRhoA-siRNA can effectively inhibit RhoA expression in Tca8113 and SCC-4 cells. The invasion ability of tongue carcinoma cells decreased with down-regulation of the protein expressions of galectin-3 and MMP-9, indicating that RhoA-siRNA can inhibit invasion of tongue carcinoma. Results show that RhoA may play an important role in the processes of invasion and metastasis of tongue carcinoma.

Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Down-Regulation ; Galectin 3 ; metabolism ; Gene Silencing ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Tongue Neoplasms ; genetics ; metabolism ; pathology ; Transfection