OBJECTIVE: To establish an SPE(solid phase extraction)-HPLC method for the determination of diclofenac sodium in human plasma.METHODS: The separation of diclofenac sodium was performed on ODS-2 column with a detection wavelength of 280nm.The mobile phase was composed of phosphate buffer(pH6.5)-methanol(40∶ 60) at a flow rate of 1.0mL? min-1.RESULTS: The linear range of diclofenac sodium was 16.2～ 2 024.0ng? mL-1(r=0.998 9).The methodological recovery ranged between 90% to 110% and the extraction recovery was above 90%.Both intraday RSD and inter-day RSD were less than 5%.CONCLUSION: The method is simple,accurate and reliable,and suitable for the pharmacokinetic study of diclofenac sodium.

Objective\ To evaluate the immediate effects and strength of allograft fusion cage(AFC) used for reconstructing stability of cervical spine. Methods\ Discs of C 5 and C 6 were resected on 8 fresh human cervical spine specimens, and autogenous iliac bone grafts(AIBG), and AFCs were implanted into the intervertebral spaces respectively. Compression test, pull out test and segmental motion measurement were studied.Results\ Comparing with intact and AIBG groups, the range of motions of C5-6 in AFC group were decreased in all directions except for extension; with (502?114) N compressive load,the vertebrae in AFC group were broken while the AFC were intact, but the AIBG were broken at (135?42) N load; with 300 N drawing load,no loosening was found between AFCs and vertebra, but it was found between AIBG and vertebra at 60 N load. Conclusion\ AFC could provide enough support,anti slide ability and could remain or increase the height of intervertebral spaces. It completely meets clinical and biomechanical requirements.

Objective:To establish the prediction model of postoperative Gleason score (GS) risk of prostatic cancer (PCa), and to compare the diagnostic efficacy of the model and each independent risk factor for PCa medium-high risk group.Methods:The clinical data of 362 patients who accepted transrectal prostate biopsy in the Run Run Shaw Hospital Affiliated to Zhejiang University School of Medicine from January 2018 to December 2019 were analyzed retrospectively, and a total of 343 patients with prostate cancer who met the enrollment criteria were selected. According to the GS grading system, these patients were divided into low risk group, moderate risk group and high risk group. At first, the single factor analysis and Spearman rank correlation were used to find out the effective indicators with good correlation with GS risk. Then, multiple linear regression equation was applied for multi-factor analysis to obtain the independent risk factors and the prediction model for predicting GS risk, and then the ROC curve was used to compare the diagnostic efficacy of each independent risk factor and prediction model for PCa medium-high risk group.Results:In the single factor analysis, the differences of all indicators in GS risk were statistically significant (all P<0.05). In the correlation analysis with GS risk, except for the indicators of prostate volume (all P>0.05), the other indexes had linear correlations with the different risks of GS (all P<0.05). Among them, the total prostate specific antigen and two-dimensional ultrasound (2D-US) score showed moderate positive correlations( rs=0.402, 0.579, all P<0.001), contrast enhanced ultrasound (CEUS) score showed a high positive correlation ( rs=0.709, P<0.001), and the rest indexes showed low positive correlations. Multiple linear regression was used to obtain two independent risk factors of 2D-US score ( X1) and CEUS score ( X2) for the prediction of GS risk, then, a prediction model was established: Y=0.863+ 0.066 X1+ 0.27 X2, the corresponding linear coefficient differences were statistically significant(all P<0.05). By the ROC analysis, the areas under the curves of 2D-US score, CEUS score and the prediction model were 0.838, 0.906 and 0.907, respectively. Conclusions:2D-US score and CEUS score are independent risk factors for predicting postoperative GS risk, and the diagnostic efficacy of the prediction model is higher than those of the 2D-US score and CEUS score for the medium-high risk group.

The genomic diversity of Avian leukosis virus subgroup J (ALV-J) was investigated in an experimentally infected chicken. ALV-J variants in tissues from four different organs of the same bird were re-isolated in DF-1 cells, and their gp85 gene was amplified and cloned. Ten clones from each organ were sequenced and compared with the original inoculum strain, NX0101. The minimum homology of each organ ranged from 96.7 to 97.6%, and the lowest homology between organs was only 94.9%, which was much lower than the 99.1% homology of inoculum NX0101, indicating high diversity of ALV-J, even within the same bird. The gp85 mutations from the left kidney, which contained tumors, and the right kidney, which was tumor-free, had higher non-synonymous to synonymous mutation ratios than those in the tumor-bearing liver and lungs. Additionally, the mutational sites of gp85 gene in the kidney were similar, and they differed from those in the liver and lung, implying that organ- or tissue-specific selective pressure had a greater influence on the evolution of ALV-J diversity. These results suggest that more ALV-J clones from different organs and tissues should be sequenced and compared to better understand viral evolution and molecular epidemiology in the field.
Animals ; Avian Leukosis Virus* ; Avian Leukosis* ; Birds ; Chickens* ; Clone Cells ; Kidney ; Liver ; Lung ; Molecular Epidemiology ; Silent Mutation

Objective To explore the sonographic features and clinical factors associated with skip metastasis in papillary thyroid carcinoma (PTC).Methods We reviewed 276 PTC cases pathologically confirmed after operation in our hospital,analyzing the ultrasonographic features and clinical characteristics of primary site and risk factors of skip metastasis.Results The rate of skip metastasis was 16.6％,and level Ⅱ or level Ⅲ were the most common area.In univariate analysis,skip metastasis was associated with tumor location,tumor maximum size,and calcification.Multivariate analysis showed the primary tumor location in the upper portion,tumor size ≤ 10 mm,and microcalcification were independent predictive factors for skip metastasis.Conclusions In papillary thyroid carcinoma,skip metastases rate is high when primary tumor location in the upper portion,tumor size ≤ 10 mm,and there is microcalcification.

OBJECTIVETo investigate the effect and mechanism of recombinant human erythropoietin (rhEPO) on angiogenesis in chronic ischemic porcine myocardium.

METHODSA ameroid constrictor was placed around the proximal circumflex branch of the left coronary artery in 12 Bama miniatures' swine artery by thoracoscopy. Electrocardiogram and coronary angiography were used to confirm the establishment of myocardial ischemia. The animals were divided into rhEPO treatment group (n = 6) and negative control group (n = 6). Treatment group received subcutaneous injection of rhEPO at 1, 3, 7, 14, 21 days, control group received saline. The expression of vascular endothelial growth factor (VEGF) in serum was assessed by ELISA. Ultrasonography and coronary angiography were assessed 28 days after therapy. Western blot was used to detect the expression of VEGF, phosphorylated protein kinase B (p-Akt) and phosphorylated extracellular signal regulated kinases (p-Erk). The degree of angiogenesis was assessed by immunohistochemical analysis.

RESULTSSerum VEGF rose significantly in both control and treatment groups, peaking at 3 days and then returning to the near-baseline level at 28 days, but the two groups showed no significant difference at each time point (P > 0.05). Echocardiographic measurements showed that the left ventricular systolic function of animals in treatment group increase significantly after rhEPO therapy. the expression levels of VEGF, p-Akt and p-Erk had markedly increased, which resulted in a 2.5-fold increased of VEGF, 1.1-fold increased of p-Akt, 1.5-fold increased of p-Erk (t = 37.721, 10.907, 12.957, all P = 0.000). there were significant increase in capillary density and arteriole density in the two groups ((944 ± 98) %/mm² vs. (569 ± 102) %/mm², (73 ± 13) %/mm² vs. (45 ± 10) %/mm², t = 4.214, 2.869, P = 0.016, 0.023).

CONCLUSIONSrhEPO can promote angiogenesis and arteriogenesis and improve the left ventricular systolic function in porcine model of chronic myocardial ischemia. The potential mechanism is to up-regulated the expression of p-Akt and p-Erk.

Animals ; Disease Models, Animal ; Epoetin Alfa ; Erythropoietin ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; Male ; Myocardial Ischemia ; drug therapy ; metabolism ; pathology ; Neovascularization, Physiologic ; drug effects ; Proto-Oncogene Proteins c-akt ; metabolism ; Recombinant Proteins ; pharmacology ; Swine ; Swine, Miniature ; Vascular Endothelial Growth Factor A ; metabolism

Objective:To investigate the application value of the ultrasonic elastic tissue dispersion quantitative analysis technique in differentiating thyroid nodules.Methods:A total of 164 nodules in 143 patients with thyroid nodules were examined by elastography ultrasound at Sir Run Run Shaw Hospital of Zhejiang University School of Medicine from January to November 2014. Eleven parameters were obtained by the tissue dispersion quantitative analysis software. These parameters were compared between benign and malignant groups by Mann-Whitney U test. The correlations between all the parameters and the pathologic results of thyroid nodules were analyzed by Spearman analysis. The receiver operating characteristic(ROC) curve of the parameter with the highest correlation coefficient was constructed. The cut-off value was calculated. Results:All parameters except correlation (CORR) had statistically significant differences between the groups of benign and malignant thyroid nodules(all P<0.01). Moreover, except CORR, the other parameters were correlated with the pathologic results of thyroid nodules(all P<0.05), with the highest coefficient in area ration of low-strain region (%AREA)( r s=0.818). ROC curves were constructed to estimate the clinic values of %AREA in diagnosis of thyroid cancer, the area under ROC curve was 0.991 for %AREA, the cut-off point was 74.83%, the sensitivity and specifity was 98.1% and 89.8%, respectively. Conclusions:The tissue dispersion quantitative analysis technique has high value in the differential diagnosis of benign and malignant thyroid nodules.

Objective:To establish a predictive model of lateral lymph node metastasis in patients with papillary thyroid carcinoma(PTC), and further to compare the diagnostic efficiency of this model with the suspected abnormal lymph node thyroglobulin in fine-needle aspirate fluid (FNA-Tg) for lateral lymph node metastasis.Methods:The preoperative clinical and ultrasonographic data of 110 patients (257 lymph nodes) who underwent PTC cervical lymph node dissection were retrospectively analyzed. According to the postoperative pathological results, they were divided into lateral lymph node metastasis and non-metastasis group. Regression analysis was used to screen out independent risk factors affecting lateral lymph node metastasis and establish a predictive model. The ROC curve was used to evaluate the diagnostic efficacy and the best diagnostic cut-off point.Results:Prediction model: Logit( P)=-2.987+ 2.189(S/L ratio of lymph nodes)+ 1.748(hilum absent)+ 2.030(hyperechoic)+ 1.849(vascular abnormalities). The sensitivity, specificity, accuracy and AUC of the prediction model in the diagnosis of lateral lymph node metastasis were 92.1%, 83.9%, 87.9% and 0.929, respectively. The Homser-Lemeshow goodness of fit test showed that the Logistic model has a good fitting effect. The sensitivity, specificity, accuracy, and AUC of FNA-Tg in the diagnosis of lateral lymph node metastasis were 87.4%, 95.4%, 90.3% and 0.968, respectively. The sensitivity, specificity, accuracy, and AUC of the combined diagnosis of the predictive model and FNA-Tg were 92.9%, 96.9%, 94.2% and 0.989, respectively. Conclusions:The model has a good predictive value for PTC cervical lymph node metastasis. Combined with FNA-Tg, it can improve its diagnostic efficiency and provide more valuable information for the decision-making of clinical surgical procedures.

OBJECTIVETo investigate the value of thyroglobulin measurement in ultrasound guided fine-needle aspiration(FNA-Tg)for detecting neck node metastasis in patients with papillary thyroid carcinoma(PTC)after thyroidectomy.

METHODSA total of 128 suspicious metastatic lymph nodes in 112 patients were retrospectively analyzed. Postoperative pathologic results were taken as the gold standard. The values of FNA and FNA-Tg combined FNA in the diagnosis of metastatic lymph nodes were compared with different ultrasonic features.

RESULTSThe sensitivity, specificity and accuracy of single FNA for the diagnosis of neck node metastasis were 67.5%, 98.0% and 79.7% respectively, and those of FNA-Tg combined FNA were 87.0%, 100.0% and 92.2% respectively. Compared with single FNA, the sensitivity and accuracy of FNA-Tg combined FNA were increased significantly(χ(2)=8.319, P=0.004; χ(2)=8.275, P=0.004). When the ultrasonographic characteristics met any one of five indicators for neck node metastasis, the sensitivity, specificity and accuracy of single FNA for the diagnosis of neck node metastasis were 38.1%, 95.7% and 68.2% respectively, and those of FNA-Tg combined FNA were 71.0%, 100.0% and 86.4% respectively. Compared with single FNA, the sensitivity and accuracy of FNA-Tg combined FNA were increased significantly(χ(2)=4.709, P=0.030; χ(2)=4.141, P=0.042). When the ultrasonographic characteristics met any two of five indicators for neck node metastasis, the sensitivity, specificity and accuracy of single FNA for the diagnosis of neck node metastasis were 55.0%, 100.0% and 73.5% respectively, and those of FNA-Tg combined FNA were 90.0%, 100.0% and 94.1% respectively. Compared with single FNA, the sensitivity and accuracy of FNA-Tg combined FNA were increased significantly(χ(2)=6.140, P=0.013; χ(2)=5.314, P=0.021). The ultrasonographic characteristics met any three, four or five of five indicators or did not meet any of the indicators, there was no significant difference in the value of the diagnosis of metastatic lymph nodes between single FNA and FNA-Tg combined FNA(P>0.05).

CONCLUSIONSThe combination of FNA with FNA-Tg could be helpful in diagnosis of lymph node metastasis. When the suspicious lymph nodes had one or two ultrasound characteristics for neck node metastasis, FNA-Tg could raise the sensitivity and accuracy of FNA, and FNA-Tg could not significantly improve in the diagnosis of FNA when presenting with no or with more than 2 ultrasonographic features.

Biopsy, Fine-Needle ; Carcinoma ; pathology ; Carcinoma, Papillary ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; diagnosis ; Postoperative Period ; Retrospective Studies ; Sensitivity and Specificity ; Thyroglobulin ; analysis ; Thyroid Neoplasms ; pathology ; Thyroidectomy

Objective: To investigate the occurrence of occult carcinoma in contralateral lobes based on the ultrasonic features of unilateral papillary thyroid carcinoma. Methods: The study included 202 consecutives cases of unilateral papillary thyroid carcinoma with benign nodules in the contralateral lobe identified by preoperative ultrasound or fine-needle aspiration from June 2014 to December 2015. All patients received total thyroidectomies, and with postoperative pathological examination they were divided into two groups, one including 60 cases with positive occult cancer and another one consisting of 142 cases with negative occult cancer. Univariate and multivariate analyses were performed to analyze the sonographic features of unilateral papillary thyroid carcinoma relevant to the occurrence of occult carcinoma in the contralateral nodules. Results: Univariate analysis indicated occult carcinoma in the contralateral lobes was associated with Hashimoto's thyroiditis(χ²=3.955, P=0.047), unclear border (χ²=4.375, P=0.036)and multifocality in the ipsilateral(χ²=7.375, P=0.007), but not with tumors maximum size, location, A/T, shape, internal structure, internal echo, acoustic halo, calcification, capsular invasion and blood flow signal in the lobe with carcinoma on another side. Multivariate analysis showed unclear border (OR=2.727, P=0.010) and multifocality in the ipsilateral(OR=2.807, P=0.005)of carcinoma were independent predictive factor for contralateral occult PTC. Conclusions Unclear border and multifocality of PTC in the ipsilateral were closely relevant to the occurrence of occult carcinoma in the contralateral nodules.