Objective To analyze the risk factors of central compartment lymph node (CCLN)metastasis in papillary thyroid carcinoma(PTC) without clinically suspected cervical lymph node metastasis (cN0).Methods Clinical data of 272 PTC (cN0) patients who underwent radical thyroidectomy and CCLN dissection between January 2008 and December 2012 in Huashan hospital were retrospectively analyzed.Patients were divided into CCLN positive group (115 cases)and CCLN negative group(157 cases) according to postoperative pathology.Chi-square test,wilcoxon test and multivariate logistic regression analysis were used to analyze risk factors.Results CCLN metastasis of papillary micro thyroid cancer (PMTC) and papillary thyroid cancer (PTC) was 34.9％ and 48.3％,respectively.Tumor size (x2 =10.26,P ＜ 0.01),position(x2 =13.87,P ＜ 0.01),capsular invasion(x2 =20.19,P ＜ 0.01),multifocal PTC(x2 =7.42,P ＜ 0.01) and unmicro-carcinoma (x2 =5.12,P ＜ 0.05) were significantly correlated to lymph node metastasis.Middle area or lower pole of thyroid carcinoma,capsular invasion and multifocal PTC were independent risk factors of CCLN metastasis in PTC.Conclusions The cN0 PTC has a high rate of pathological CCLN metastasis,it is imperative to conduct thyroidectomy with ipsilateral level CCLN dissection in PTC patients.

Objective To find out prevelence of thyroid goiter on school-aged children in excessive iodine region of Shandong Province.Methods By employing B-ultrasonography and palpation method,the thyroid volume were investigated and the thyroid goiter rate were calculated among 6415 children aged 8～10 years old in execssire iodine region.Results By employing B-ultrasonography method.The volumes of thyroid in goiter children aged 8,9,10 years were(5.67±1.49),(6.07±1.24),(7.30±2.01)ml,respectively.The goiter rates were 28.84%(683/2368),20.89%(448/2144),11.82%(225/1903).The volumes of thyroid in non-goiter children aged 8,9,10 years were(3.36±0.67),(3.64±0.77),(4.02±0.94)ml.When the normal value of thyroid volume in 8 and 9 year-groups lowered 0.5 ml,the goiter rate would be changed to 18.20%(430/2368)and 12.92%(277/2144) respectively.The thyroid volumes of boys and girls were(4.20±1.38),(4.18±1.73)ml,respectively.The goiter rates of boys and girls were 21.92%(732/3340)and 20.29%(624/3075)respectively.The rates of goiter detected by palpation in 8,9 and 10 year-groups were 10.05%(238/2368),10.31%(221/2144)and 14.45%(275/1903) respectively.Conclusions Along with the increasing age,the volume of thyroid in children aged 8～10 is increasing in excessive iodine region of Shandong Province.But the goiter rate is decreasing ahmg the increasing age.This phenomena may be caused by improper normal value of thyroid volume.Therefore,the normal value of thyroid volume should be revised urgently.

Lysine decarboxylase is a key enzyme involved in the upstream biosynthesis of lycopodium alkaloids (LAs) such as huperzine A, contributing to the decarboxylation of lysine to 1,5-pentanediamine (cadaverine). Three lysine decarboxylase genes (HsLDC-L1, HsLDC-L2, HsLDC-L3) were successfully cloned from Huperzia serrata using transcriptomic sequence data mining strategy combined with reverse transcription PCR. The physicochemical properties, secondary and tertiary structures, amino acid identities, and evolutionary relationship of the three LDCs were analyzed by online bioinformatics analysis platforms and DNAMAN, MEGA 7.0 software, revealing that all of these proteins had the conserved PLP binding domain and active site residues were completely conserved in LDCs. Phylogenetic analysis showed that these LDCs were located in the same branch as other known LDCs from LA-producing plants. Accordingly, the ORFs of these three HsLDCs were inserted into different expression plasmids for further expression in E. coli. However, only HsLDC-L1 was successfully expressed in E. coli BL21 (DE3) by inserting into a pCold TF vector. The recombinant protein was purified by Ni²⁺ affinity chromatography purification. HsLDC-L1 contains 469 amino acid residues, with a calculated molecular weight of 50.50 kDa. HsLDC-L1 expectedly catalyzed the decarboxylation of lysine to produce cadaverine. In addition, HsLDC-L1 can also catalyze the generation of putrescine from ornithine. However, it cannot catalyze the decarboxylation of tyrosine, phenylalanine, tryptophan and histidine. The results not only provide insight into the biosynthesis of LAs including huperzine A, but also provide a critical genetic element for the overproduction of Δ¹-piperideine and pelletierine, the essential biosynthetic precursors of LAs, using synthetic biology strategies.

BACKGROUND:In recent years, tissue engineering technology as a new model for tissue regeneration has provided new ideas and methods for the repair of periodontal tissue defects. OBJECTIVE:To investigate the effect of human periodontal ligament cels-polyglycolic acid scaffold complex for repair of periodontal tissue defects. METHODS:Passage 4 human periodontal ligament cels at a density of 1.5×109/L were seeded onto the polyglycolic acid scaffold to prepare cel-scaffold complex. Then mongrel dogs were selected to make animal models of periodontal tissue defects and then randomly assigned into experimental group subjected to cel-scaffold complex implantation or control group subjected to direct coronal reset and suture of the gingival flap. Colagen content, new blood capilaries, new cementum, new alveolar bone and new periodontal ligament were detected within 4 weeks after operation; hematoxylin-eosin staining of periodontal tissue defects was done at 8 weeks after operation. RESULTS AND CONCLUSION:In the experimental group, the colagen content, number of newborn capilaries, amount of new cementum, new alveolar bone and new periodontal ligament tissues were significantly higher than those in the control group at postoperative 1, 2, 3, 4 weeks (P< 0.05). At 8 weeks after operation, in the experimental group, there were more vessels arranging on the connective tissue surface of new alveolar bone, the alveolar bone showed a sawtooth-like interlinking with the periodontal tissues in the presence of a thin layer of cementum; in the control group, only new alveolar bone and cementum formed below the incisure. These findings indicate that human periodontal ligament cels-polyglycolic acid scaffold complex can promote periodontal tissue regeneration.

In order to solve the problem of selection and in vivo delivery problem in siRNA treatment, hepatitis B virus (HBV) HBx gene which could be targeted by siRNA was studied. The siRNA expression plasmid which specific inhibits HBx expression was obtained by in vitro selection via a dual-luciferase plasmid including HBx-Fluc fusion protein expression domain. The selected siRNA expression plasmid was then encapsulated in PEG-modified cationic liposome, which was devoted into pharmacodynamic studies at both cellular and animal level. The results illustrated that the cationic liposome which encapsulated siRNA expression plasmid could effectively inhibit HBx gene expression both in vitro and in vivo.
Cations ; Gene Expression Regulation, Viral ; drug effects ; Hepatitis B virus ; genetics ; Liposomes ; chemistry ; Plasmids ; RNA, Small Interfering ; chemistry ; Trans-Activators ; genetics ; metabolism

OBJECTIVETo optimize the synthetic pathway and fermentation process of yeast cell factories for production of oleanoic acid.

METHODUsing the DNA assembler method, one copy of Glycyrrhiza glabra beta-amyrin synthase (GgbAS), Medicago truncatula oleanolic acid synthase (MtOAS) and Arabidopsis thaliana cytochrome P450 reductase 1 (AtCPR1) genes were introduced into Saccharomyces cerevisiae strain BY-OA, resulting in strain BY-20A. YPD medium with different glucose concentration were then used to cultivate strain BY-2OA.

RESULTIncreasing gene copies of GgbAS, MtOAS and AtCPR1 resulted in increased beta-amyrin and oleanolic acid production. The strain BY-2OA produced 136.5 mg x L(-1) beta-amyrin and 92.5 mg x L(-1) oleanolic acid, which were 54% and 30% higher than the parent strain BY-OA. Finally, the titer of oleanolic acid increased to 165.7 mg x L(-1) when cultivated in YPD medium with 40 mg x L(-1) glucose.

CONCLUSIONProduction of oleanoic acid increased significantly in the yeast strain BY-2OA, which can provide the basis for creating an alternative way for production of oleanoic acid in place of extraction from plant sources.

Biomass ; Biotechnology ; methods ; Dose-Response Relationship, Drug ; Fermentation ; Glucose ; pharmacology ; Oleanolic Acid ; biosynthesis ; Saccharomyces cerevisiae ; cytology ; drug effects ; metabolism

Objective To determine whether real-time three-dimensional transesophageal echocardiography(RT-3D TEE) is an accurate non-invasive technique for defining the morphology of atrial septal defects(ASD). Methods In 20 patients with secundum ASD, mean age 42.2 years (7 male, 13 female) ,live three dimensional surgical views of ASD were acquired,qualitative morphology of ASD such as the shape, presence of fenestrations and the defect margins were noted during the cardiac cycle. The measurements obtained from 2-dimensional transesophageal echocardiography(2D-TEE) and RT-3D TEE were compared to those obtained from stretched balloon diameter(SBD) or surgery. Measurements of the size and area change of ASD were validated during the cardiac cycle by RT-3D TEE,stepwise multiple linear regression analysis was performed to test the correlation between the maximum change ratios of area and diameter of ASD and age, atrial septal length, ASD shunting velocity, the maximum diameter of ASD and right ventricular systolic pressure. Results ① The morphology of ASD was circle-like or elliptical in systole,irregular or elliptical in diastole. ②The area and diameter of ASD measured by RT-3D TEE was minimal in isovolumetric contraction phase, maximal in isovolumetric diastole phase. The correlation coefficient of maximum diameter measured by 3D with SBD was 0.962 , greater than that by 2D and SBD (0.820). ③The change ratio of area and maximum diameter of ASD acquired by RT-3D-TEE was 11.48%-71.12% and 2.80%-43.87% respectively,and the correlation coefficient of them was 0.921. Conclusions RT-3D TEE using live 3D-Zoom mode accurately displayed the varying morphology,dimensions and spatial relations of ASD. RT-3D TEE can offer visualization morphological changing of ASD in different periods of cardiac cycle,providing more information for percutanous catheter intervention and open heart surgery.

Background Arresting the overexpression of vascular endothelial growth factor (VEGF) will be a new approach to the inhibition of neovascularization.RNA interference (RNAi) can inhibit the expression of specific gene,and its application in eye has little interference to other gene expression.Objective This study was to investigate the effect of small interference RNA (siRNA) targeting VEGF on the expression of VEGF and retinal neovascularization in oxygen-induced retinopathy (OIR) model.Methods psi-HITM/enhanced green fluorescent protein (EGFP)/VEGF siRNA was designed and prepared in vitro.Mouse endothelioma (EOMA) were cultured in DMEM without antibiotic and divided into 5 groups.The cells were incubated in DMEM only in the blank control group;while 1 μl of LipofectamineTM 2000 + psi-HITM/EGFP,1 μl LipofectamineTM 2000 + 40,50 or 60 nmol/L of psi-HITM/EGFP/VEGF siRNA was added into DMEM in the negative control group and siRNA groups,respectively.The expression of VEGF mRNA and protein was detected by real time PCR (RT-PCR) and Western blot.The optimal effective concentration of VEGF siRNA was assessed.OIR models were established in 48 7-day-old C57BL/6J mice by raising them at an oxygen concentration of (75±2) ％ for 5 days and then to normal air.The mice were randomized into the model group,null vector group and VEGF siRNA group.1 μl of a mixture of psi-HITM/EGFP or VEGF siRNA (60 nmol/L) and LipofectamineTM 2000 was intravitreally injected,respectively,in the null vector group and VEGF siRNA group.The normal mice were used as the normal control group.Expression of VEGF mRNA and protein in the mouse retinas was detected by RT-PCR and Western blot,respectively,and FITC-dextran stretched retinal preparation was examined to evaluate the neovascularization,and retinal sections were examined to quantify the number of vascular endothelial cell nuclei extending beyond the internal limiting membrane (ILM).Results The in vitro transfection test showed that the expression of VEGF mRNA and protein in the EOMA cells was significantly different among the blank control group,negative control group and 40,50,60 nmol/L VEGF siRNA groups (F =148.890,P ＜ 0.001; F =306.960,P ＜ 0.001),and the expression of VEGF mRNA was lower in different concentrations of VEGF siRNA groups than that in the blank control group (t=73.950,119.890,156.480,all at P＜0.001).Also,the expression of VEGF protein was less in different concentrations of VEGF siRNA groups than that in the blank control group (t =15.452,23.344,42.119,all at P＜0.001).The optimal inhibitory concentration of VEGF siRNA was 60 nmol/L.In vivo study determined that compared to the model group and null vector group,the non-perfusion zones and neovascular net in the retina were decreased,and the number of vascular endothelial cell nuclei extending beyond the ILM was less in the VEGF siRNA group.The relative expression level of VEGF mRNA in the retinas was 1.23±0.18,4.02±0.16,3.98±0.19 and 1.98±0.12 in the normal control group,model group,null vector group and VEGF siRNA group,respectively,with a significant difference among them (F=369.780,P＜0.001),and the relative expression levels of VEGF mRNA in the model group and null vector group were higher than that in the normal control group (t=37.880,37.336,both P＜0.001),and the expression of VEGF mRNA in the VEGF siRNA group declined by 50.8％ (t=10.183,P＜0.001).The difference in the expression levels of VEGF protein also was assayed among the various groups (F=408.980,P＜0.001),and VEGF level in the retina was lowered by 68.0％ in the VEGF siRNA group compared to the model group (t =11.473,P＜0.001).However,VEGF level in the VEGF siRNA group remained at a high level in comparison with the normal control group (t =2.422,P＜0.001).Conclusions Intravitreal injection of VEGF siRNA can attenuate retinal neovascularization by effectively downregulate the expression VEGF mRNA and protein in the retina.

Objective:To obtain oligonucleotide aptamers which can specifically bind to Bacillus anthracis spores by in vitro selection protocol-SELEX (system evolution of ligands by exponential enrichment).Methods:An in vitro synthesized 78 mer random DNA library (≤1014-15types of different DNAs ) was subjected to 15 rounds of selection using SELEX method against spores of B.anthracis vaccine strain A.16R. Binding of the aptamers to spores was visualized by biotin-streptavidin-horseradish peroxidase system.Results:PCR amplification band pattern of the first round selection was different from that of the ninth round. The binding assay demonstrated that D absorbance at 450 nm of the fifteenth round pool increased 9 times as compared with that of the first round , and the D absorbance increased with the increment of aptamers′ quantity binding to spores. Conclusions: A set of aptamers with considerable binding affinity to B.anthracis spores were successfully selected from the initial random DNA pool.

Objective To explore the CT features of infantile hepatic hemangioendothelioma (IHH).Methods CT imaging features of 16 IHH patients confirmed by surgical pathology and clinical diagnosis were retrospectively reviewed.Results Of 16 patients,there were single lesion in 10 cases and multiple lesions in 6 cases.All the lesions were rich in blood supply,and the arterial phase showed obviously rings enhanced.Some linear enhancement towards to the center below the annular enhancement of solitary lesions were found.There were patchy enhancement in annular enhancement center area,and the area of enhancement obvious increasing in venous phase and delayed phase in 4 cases with multiple lesions,enhancement mode was similar to single lesion in 1 case with multiple lesions.The lesions of IHH Ⅱ were small and multiple,and showed intermittent line shaped edge enhancement;some lesions below the linear enhancement showed small nodular enhancement and the area of nodular enhancement enlarged in venous phase,the range of linear enhancement was not obvious.Conclusion CT manifestations of IHH have some characteristics,and the enhancement of CT is more important in the diagnosis of this disease.