The purpose of the present study is to establish a rapid and effective PCR method for the identification of B. multicinctus. Based on sequence alignment of B. multicinctus and its adulterants, we found that Cyt b gene is a good molecular genetic marker for the authentication of B. multicinctus. On the basis of the sequence data, a pair of highly specialized primers was designed. The templates were extracted by the DNA purification system. Key factors such as annealing temperature, concentration of Taq enzyme and cycle numbers were analyzed and optimized. The modified PCR program consisted of an initial denaturation step at 95 degrees C for 5 min, followed by 30 cycles of 95 degrees C for 30 s and 55 degrees C for 45 s and a final extension at 72 degrees C for 5 min. Thirteen samples of B. multicinctus were identified accurately from their 20 adulterants in 4 hours. The results indicated it is a highly accurate, rapid and applicable method for the authentication of B. multicinctus.

Acetyl-CoA C-acetyltransferase (AACT) is the first enzyme in the terpene synthesis pathway, catalyzed two units of acetyl-CoA to acetoacetyl-CoA. In order to study the tanshinone biosynthesis in Salvia miltiorrhiza, a novel AACT gene, SmAACT, was cloned using cDNA microarray and RACE strategy. The full length cDNA of SmAACT is 1 623 bp (accession No. EF635969), which contained a 1 200 bp open reading frame (ORF) encoding a 399 amino acid protein. Nine introns were found in the genomic sequence. SmAACT was upregulated by YE and Ag+ elicitors both with cDNA microarray and quantitative RT-PCR analyses along with the accumulation of tanshinones. Sequence homology comparison and phylogenetic analysis all suggested that SmAACT belonged to the class of acetyl-CoA C-acetyltransferase. The transcription level of SmAACT was relatively higher in root than that in stem and leaf tissues. SNP analysis revealed that SmAACT was highly variable in the region of 6 to 9 introns with 33 SNPs in the 600 bp region, there are 5 SNPs in the cDNA region while they are all synonymous cSNPs. Some special genotypes were found in Salvia miltiorrhiza from different areas. SmAACT will be an useful gene for further analyze the mechanism of gene regulation among the tanshinones biosynthesis.

Objective To summarize clinical outcomes of interventional therapy on children with common congenital heart diseases(CHD).Methods A retrospective study was conducted.One hundred and fourteen patients with CHD were selected as our subjects,who underwent catheter interventional therapy in the Second Affiliated Hospital of Zhengzhou University from Jan.2004 and Dec.2012.The size of occluder was chose according to intraoperative echocardiography or cardiac imaging measurements,and occluder was released under assisted monitoring by subtraction angiography or cardiac ultrasound.Results There are all together 112 patients got the therapy successfully,2 cases failed(occluder detachment),and the success rate of operation was 98.2％.After the success of interventional,echocardiography examination showed that 11 cases were with star point across shunt,but the function of the around valve was not affected.One months after operation,echocardiography examination showed star point across shunt of 11 cases were disappeared,and no occluder was shifted as well as no thrombosis formed.Three months after operation,chest radiograph showed pulmonary congestion decreases and heart shadow was shrink.Thirty-eight cases were with three tricuspid regurgitation before operation and 32 cases were without reflux at 3 months after operation,and 6 cases relieved significantly.The patients were followed up for 6 months or 3 years,activity endurance was significantly improved than that before operation.No occluder was shiftand hemolysis and arrhythmia occurred.Meanwhile,No thrombosis or embolism occurred.Conclusion Interventional treatment for children with congenital heart disease is proved as a safe,effective methods and it have broad prospects in clinical application.

OBJECTIVETo establish a convenient, practical and high efficient method of DNA extraction of os cervi, and lay the foundation of identification of animal bones.

METHODThe bones of sika deer, red deer, cattle, dog and pig were used to extract DNA under different decalcification time (24,48,72 h) and decalcification temperature (4,25,37,56,70 degrees C), and extract method.

RESULTIt proved by experiments that demineralization process promotes the cracking of osteocyte. In a broad of decalcification time and temperature, DNA could be extracted from all bone samples successfully while the quantity varied slightly.

CONCLUSIONSamples (about 0.1 g) decalcify with 0. mol x L(-1) EDTA at 4 degrees C for 24 h, then water-bath for 1 h after lysis buffer added, DNA extracted via the method above is of high quality and can be used for PCR.

Animals ; Bone Demineralization Technique ; Bone and Bones ; chemistry ; metabolism ; Cattle ; DNA ; isolation & purification ; Deer ; Dogs ; Polymerase Chain Reaction ; Swine ; Temperature ; Time Factors

This paper summarized and analyzed the status quo and problems about molecular identification of animal medical material, based on the facts, we proposed some research strategies, including uniting to tackle key problems, expanding the research species, accelerating manufacture and generalization of molecular identification kit, priming the research project of DNA barcoding, and establishing standard database on animal medical material.
Animal Structures ; Animals ; DNA Barcoding, Taxonomic ; Databases, Genetic ; standards ; Materia Medica ; analysis ; Mitochondria ; genetics ; Research

OBJECTIVETo assess the association of cognitive functions with gender, age, education and polymorphism of dopamine receptor D4 (DRD4) gene in healthy adults.

METHODSFour hundred and fifty-five healthy participants have completed 3 cognitive function tests including Tower of Hanoi (TOH), Wisconsin Card Sorting Test (WCST) and Trail Making Test (TMT). Peripheral blood samples were collected from all participants, and genomic DNA was extracted according to a standard phenol-chloroform procedure. Rs3758653 in the promoter region of the DRD4 gene was genotyped using Illumina GoldenGate genotyping assay.

RESULTSMales have performed better than females in terms of TOH executive time and TOH total score, but did worse in TOH planning time. Most of the measured cognitive domains were affected by age and education. Cognitive ability has decreased along with increased age and decline of educational years. The polymorphism of rs3758653 has mainly correlated with the TOH executive time. Compared with A allele carriers, G allele carriers did worse in TOH executive time.

CONCLUSIONGender, age, education and the rs3758653 polymorphism of the DRD4 gene play an important role in cognitive functions in healthy adults.

Adolescent ; Adult ; Age Factors ; Cognition ; Education ; Female ; Humans ; Male ; Middle Aged ; Neuropsychological Tests ; Polymorphism, Single Nucleotide ; Receptors, Dopamine D4 ; genetics ; Sex Factors ; Young Adult

Objective:To screen and analyze the differentially expressed genes between lacrimal gland benign lymphoepithelial lesions (LGBLEL) and mucosa-associated lymphoid tissue (MALT) lymphoma.Methods:A cross-sectional study was performed.Ten consecutive patients were included in Beijing Tongren Hospital Affiliated to Capital Medical University from January 2015 to November 2017, including five patients with LGBLEL and five patients with MALT lymphoma.Clinical data and peripheral blood sample were collected from each patient.DNA was extracted from peripheral blood.The whole-exome sequencing (WES) was employed for gene sequencing.The BWA software was used for the screen of differentially expressed gene; GATK software was used to detect genomic variation; ANNOVAR software was used to annotate and predict the effects of the variation; Varscan software was used to analyze single nucleotide polymorphisms (SNPs) and insertion-deletions (InDels), and ExomeCNV software was used to identify copy number variations (CNVs). The mutated hub gene with the maximal clique centrality was screened out by the analysis of protein interaction network and construction of functional module network.This study was approved by an Ethics Committee of Beijing Tongren Hospital Affiliated to Capital Medical University.Written informed consent was obtained from each patient prior to any medical examination.Results:There was 16.63 Gb sequencing data per sample on average.Synonymous mutation and missense mutation were the most common SNPs mutation types in the LGBLEL group and MALT lymphoma group, and no significant difference was found in gene mumber of synonymous mutation and missense mutation between the two groups.The number of terminating codon missing mutation genes in the LGBLEL group was more than that in the MALT lymphoma group ( P<0.05). The most common InDels types were frameshift mutation, non-frameshift insertion and non-frameshift deletion, and there was no significant difference in gene number of InDels between the LGBLEL group and MALT lymphoma group.The number of exon CNVs was few in both two groups and showed no significant influence in final result.Six differentially expressed hub genes were found, including IGFN1, TCP10, SLC45A4, BTBD7, PHGR1 and PIEZ02. Conclusions:IGFN1, TCP10, SLC45A4, BTBD7, PHGR1 and PIEZ02 genes may participate in the development of LGBLEL into MALT lymphoma.

[Abstract] Objective: To investigate the expression and clinical significance of PD-1/PD-L1 in gastric cancer (GC) tissues. Methods: Paraffin embedded tumor tissues and clinical data of 82 GC patients who had undergone operation at the Fourth Hospital of Hebei Medical University from January 2007 to December 2007 were collected, and their survival status was followed. The protein expressions of PD-1 and PD-L1 in tumor tissues were detected by immunohistochemistry. Kaplan-Meier analysis and Log-Rank test were adopted to analyze the survival of GC patients, and the ROC curve was plotted. Results: The positive rate of PD-L1 protein expression was 42.68% while the positive rate of PD-1 expression was 13.41% in GC tissues. The positive rate of PD-1 and PD-L1 expression in GC tissues of patients without pre-operative distant metastasis was significantly lower than those patients with pre-operative metastasis (PD1： 3.28% vs 42.86%; PD-L1： 13.11% vs 90.48%; all P<0.01). The positive rate of PD-L1 expression in tumor stroma of patients without pre-operative distant metastasis was significantly lower than those with metastasis (PD-L1: 13.11% v s 47.62%, P<0.01). The resection range of stomach, PD-L1 over-expression and the presence of pre-operative distant metastasis were the adverse factors affecting the prognosis of patients with GC (P<0.05). Conclusion: PD-1 and PD-L1 expressions in GC tissues were closely related to the presence of pre-operative distant metastasis and the depth of tumor infiltration. The postoperative survival of patients who were PD-L1 positive was shorter than the negative ones.

Objective: To identify the function of 91-112 amino acids (aa) fragment, the interaction domain of head and stalk of Newcastle disease virus(NDV) HN glycoprotein, and clarify the role of the fragment in promoting cell specific membrane fusion. Methods: The specific gene sequences were identified by aligning 91-112 amino acids of NDV HN protein with amino acids of MeV H, RSV G, hPIV3 HN protein. The fragment deletion, fragment substitution and intermolecular homologous recombination method were combined to construct the deletion mutant, De(HN), and three chimeras, Ch(MeV), Ch(RSV), Ch(hPIV3). Cationic transfection reagent was used to transfect the plasmids into baby hamster kidney cells (BHK-21), in which vaccinia virus-T7 RNA polymerase expression system was expressed. Indirect immunofluorescence assay (IIFA) and flow cytometry (FCM) were executed to analyze the cell surface expression level. Cell fusion promotion activity, receptor recognition activity and neuraminidase activity of each mutant were also detected. Results: Cell surface expression efficiency of De(HN) and Ch(MeV), Ch(RSV), Ch(hPIV3) proteins were 9.04%, 82.20%, 70.16%, 75.65% of that of wild-type (wt) HN. Fusion promotion activity of De(HN), Ch(MeV), Ch(RSV), Ch(hPIV3) were 3.83%, 24.76%, 29.42%, 57.84% of that of wt HN. The fusion promotion activity of De(HN) almost disappeared and syncytium couldn’t be found under the microscope. Hemadsorption activity was 13.48%, 36.25%, 34.93%, 65.22%, respectively (P<0.05), which was consistent with the fusion promotion activity of mutant proteins. Neuraminidase activity was 10.81%, 54.42%, 50.13%, 60.35% of that of wt HN, respectively (P<0.05). Conclusions The amino acids fragment (91-112) of NDV HN protein plays an important role in promotion fusion. The loss of fusion promotion activity of De(HN) protein was related to the failure of effective cell surface expression of the mutant.