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 evaluate the change of left atrial function after mitral valvuloplasty in patients with mitral valve prolapse by measuring left atrial strain using two-dimensional speckle tracking echocardiography(2D-STE).Methods Study population consisted of 25 patients with mitral valve prolapse and 25 healthy subjects.High frame rate two-dimensional images were recorded from the apical four chamber view and two chamber view.Images of patients with mitral valve prolapse were respectively recorded at pre-operation,3 days and 3 months post-operation.Left atrial longitudinal strain was measured in two views using two-dimensional strain soft ware.Results ①Compared with the control group,global peak atrial longitudinal strain (PALS),peak atrial contraction strain (PACS) and atrial longitudinal strain during early diastole(ALSED) decreased (P ＜0.01) in patients with mitral valve prolapse,but global timeto-peak of peak atrial longitudinal strain(TPLS) increased(P ＜0.01).Regurgitant jet area/left atrium area (RJA/LAA) in patients with mitral valve prolapse correlated negatively with global PALS(r =-0.620,P ＜ 0.01).② Compared with preoperative group,global PALS,PACS,ALSED and TPLS decreased significantly at 3 days after surgery(P ＜0.01).Global PALS,ALSED(P ＜0.01) and PACS(P ＜0.05)increased at 3 months after surgery,but global TPLS did not differ significantly.Conclusions 2D-STE could accurately evaluate the change of left atrial function after mitral valvuloplasty.

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.

Objective To investigate the chest HRCT characteristics of pediatric perinuclear anti-neutrophil cytoplasmic antibody (p-ANCA) associated systemic vasculitis. Methods Retrospectively analysis of the clinical and HRCT data of 15 pediatric cases diagnosed as p-ANCA related pneumonia according to the classification and diagnostic criteria of Chapel Hill meeting of 1994 in our hospital were retrospectively analyzed. They were 13 girls and 2 boys. The age range was 1—17 years and the median was about 10 years old, In the 15 patients, 12 were diagnosed as primary MPA, 2 were secondary MPA, 1 was CSS. Two radiologists with 10 years of radiological experience read the CT imaging together. Results 1 of 12 primary MPA were found large opacities and ground glass shades in both sides of lung,especially the posterior parts on CT images with bilateral pleural effusion; 1 case had focal opacities in left lower lobe, around with thickened interlobular septa and pleural effusion; Another 1 case only had focal thickened interstitial in right middle lobe; The other 9 cases showed scattered or diffused ground glass opacity, with thickened pulmonary interstitial and (or) pleura. 1 of the 2 secondary MPA who had hemoptysis showed ground glass opacity on CT images with no thickened septal nodules or pleural effusion. The other one who had renal anomalies and no respiratory symptoms showed subpleural nodules with halo sign in right low lobe. The CSS showed diffused patchy ground glass opacity, consolidation of both lingular lobes, nodules of centri-and peri-lobular, thickened septa near the pleura, obviously thickened bronchial wall, slightly widened bronchial lumen and thickened pleura. Conclusion The classic imaging features of p-ANCA pneumonia on HRCT is the non-specific small pulmonary vessel hemorrhage.

Objective To explore the effect of transplantation of mesenchymal stem cells (MSCs) transfected with the human receptor activity modifying protein 1 (hRAMP1) gene on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) after carotid balloon angioplasty was performed in rabbits with carotid atherosclerosis.Methods Density gradient centrifugation and adherent culture were carried out to obtain MSCs,which were then transinfected with an adenovirus vector carrying the hRAMP1 gene or an empty adenovirus vector.A rabbit model of atherosclerotic stenosis and balloon angioplasty was successfully established.Results were randomly divided into three groups:the hRAMP1-MSCs group,theadipose-derived MSCs (Ad-MSCs) group and the control group.MSCs were transinfected with Ad-EGFP-hRAMP1,Ad-EGFP or PBS by transplantation into the injured carotid arteries.Homing and differentiation were assessed with MSCs harvested at 7 d.With MSCs collected at 28 d,Western blotting was used to measure the expression of the hRAMP1 target gene in the carotid artery; the neointima and media area in the injured carotid arteries were estimated; carotid artery morphology was examined with H&E staining; and the proliferation and apoptosis of VSMCs were determined by immunohistochemistry and TUNEL.Results The expression of CD31 and EGFP was found in proliferating neointima lesions at 7d in the hRAMP1-MSCs group and the Ad-MSCs group.At 28d of MSC transplantation,the level of RAMP1 significantly increased in the hRAMP1-MSCs group,compared with the Ad-MSCs and control groups [(63.0±4.9) vs.(28.3±2.5) and (27.2±7.2),all P＜0.05],but there was no differencein the RAMP1 level between the Ad MSCs group and the control group (P＞0.05).Positive expression of the α-smooth muscle antibody (α-SMA) was found in all three groups at 28 d of MSC transplantation.The thickness of the hyperplastic neointima significantly decreased in the hRAMP1-MSCs group,compared with the other two groups (P＜0.05),and was lower in the Ad-MSCs group than in the control group (P＜0.05).The expression of proliferating cell nuclear antigen (PCNA) was lower in the hRAMP1-MSCs group than in the Ad-MSCs and control groups at 28d of MSC transplantation (P ＜0.05),while the PCNA level was lower in the Ad-MSCs group than in the control group (P＜ 0.05).The VSMC apoptosis rate significantly increased in the hRAMP1-MSCs group,compared with the Ad MSCs and control groups (P＜0.05),and was the lowest in the control group (P＜0.05).Conclusions Gene-modified stem cell therapy can effectively inhibit vascular intimal hyperplasia,thereby reducing restenosis after angioplasty.