Objective:This paper aims to analyze the monitoring model of Canadian drug price monitoring mod-el based on the international reference price, and make reference for China. Methods: Through policy analysis and literature research, the system combs the specific Canadian practices of using the international reference prices for the drug price monitoring. Results:For the introduction of new drug prices, Canada uses the international median price comparison test, the highest international price comparison test, the treatment category comparison test and the inter-national treatment category comparison test. For the listed drugs, Canada calculates the international price ratio, and then carries out bilateral comparisons and multilateral comparisons to monitor the prices of drugs already on the mar-ket. At present, Canada has achieved effective monitoring of drug prices, that is to say, the introduction of new drug prices complies with the《Excessive Price Guide》;the patented drugs prices are lower than the designed international price;the price difference between the generic drug and the international level has gradually reduced. Conclusions:The Canadian experience is worth learning, and China should add legislative about drug price monitoring, learn from this experience to identify and monitor the varieties and establish the price monitoring and early warning mechanism with the international reference price as the warning indicator.

Objective To construct a eukaryotic expression plasmid carrying the PKCI-1/HINT1 gene,to investigate its expression in A375 melanoma cells after transfection,and to evaluate its effects on apoptosis and autophagy of A375 cells.Methods The PKCI-1/HINT1 gene sequence was amplified by reverse transcription PCR (RT-PCR) with total RNA extracted from A375 cells as the template,then inserted into the eukaryotic expression plasmid PCDNA3.1 (+) to construct a recombinant plasmid,PCDNA3.1 (+)-PKCI-1/HINT1.Some A375 cells were classified into two groups to be transiently transfected with the recombinant plasmid (PCDNA3.1 (+)-PKCI-1/HINT1 group) or the empty plasmid PCDNA3.1 (+) (control group).After additional 48-hour culture,RT-PCR and Western blot analysis were performed to quantify the mRNA and protein expressions of PKCI-1/HINT1 respectively,Hoechst 33342 staining was conducted to detect apoptosis of A375 cells,Western blot analysis to detect the expressions of intracellular caspase-3 and autophagy-associated protein beclin1,and cell autophagy was observed by using the green fluorescent protein (GFP)-microtubule-associated protein 1 light chain 3 (LC3) labelling method combined with confocal laser scanning microscopy.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of A375 cells at 24,48,72 and 96 hours after transfection.Results Enzyme digestion and sequencing analysis confirmed that the eukaryotic expression plasmid PCDNA3.1 (+)-PKCI-1/HINT1 was successfully constructed and effectively expressed in the transfected A375 cells.MTT assay showed that PKCI-1/HINT1 could obviously inhibit the proliferation of A375 cells,and the number of live cells was decreased by 17.0％,25.6％ and 29.4％ in the PCDNA3.1 (+)-PKCI-1/HINT1 group at 48,72 and 96 hours,respectively,compared with the control group (all P ＜ 0.05).Hoechest 33258 staining revealed that PKCI-1/HINT1 could promote the formation of apoptotic bodies in A375 cells.Confocal laser scanning microscopy demonstrated that the overexpression of PKCI-1/HINT1 increased GFP-LC3 puncta formation in A375 cells.In addition,Western blot analysis indicated that PKCI-1/HINT1 up-regulated the protein expressions of caspase-3 and beelin1 in A375 cells.Conclusions The eukaryotic expression plasmid PCDNA3.1 (+)-PKCI-1/HINT1 was successfully constructed,and PKCI-1/HINT1 could be effectively expressed in A375 cells.High-level expression of PKCI-1/HINT1 could suppress cellular proliferation,promote apoptosis,and induce autophagy,of A375 cells.

Objective To explore the cognitive function of the theory of mind(TOM) in early-onset schizophrenia from faux pas recognition and belief understanding dimensions.Methods 41 early-onset schizophrenias (EOS) and 40 normal adolescents were interviewed by the Chinese version of faux pas recognition task and theory of mind picture-sequencing task(ToM-PST),then early-onset schizophrenias cognitive features were analyzed.Results ①In faux pas recognition task,compared with healthy subjects,EOS showed significantly less total scores (16.11±6.34 vs 20.86±7.79,P＜0.05)and subscores of understanding faux pas questions (9.88±4.11 vs 13.27± 6.07,P＜0.05).In faux pas recognition scores(6.72±2.60vs 7.58±2.20) and control questions(9.83±0.44vs 9.97± 0.16) had no statistically significant difference (P＞0.05).②In ToM-PST task,early-onset schizophrenic patients also had significantly less total score(17.29±3.38 vs 21.48± 1.73,P＜0.01),subscores of understanding first order belief,first order error belief,second order belief,second order error belief,third order error belief,sense of reality,reciprocity,fraud,detecting fraud compared to normal controls had significant difference (all P＜ 0.05 or 0.01).③ The correlation between clinical course time and PANSS score and its subscores of the theory of mind picture-sequencing and faux pas task was non-significant(P＞0.05)except the subscores of understanding the first order belief (P＜0.01).Conclusion The theory of mind is apparent damage in early onset schizophrenia and non-significant correlation is found with psychiatric symptoms.

Purpose To study the expression of inhibitor of differentiation 2 (Id2) in Ewing sarcoma of bone as well as the correlation with cell cycle proteins. Methods The expression of Id2, c-Myc, Cyclin D1, p53, p16 and p27 in 45 cases of Ewing sarcoma were detected by immunohistochemical EnVision methods, the relationship between the expression of Id2 and clinicopathologic characteristics and the correlation with cell cycle proteins were analyzed. Results Total expression of Id2 was found in 88. 9% of Ewing sarcoma. Strong expression of Id2 was detected in 33. 3% cases. Higher frequencies of c-Myc and Cyclin D1 immunostaining were observed (66. 7% and 71. 1%, respectively), but lower frequencies of p53, p16 and p27 expression were present (20. 0%, 35. 6% and 28. 9%, respectively). Id2 overexpression displayed a negative relationship with p27 expression (P<0. 05). Among the follow-up of 24 cases, overexpression of Id2 was found more frequency in dead and metastases cases (41. 2%) when compared to that of progres-sion-free survival cases (14. 3%). Conclusions Id2 was extensively expressed in Ewing sarcoma of bone. Overexpression of Id2 and its correlation with p27 expression suggest that this is an important pathway involved in development of Ewing sarcoma. Moreover, Id2 overexpression may predict poor prognosis.

Objective To analyze the imaging and histopathologic features of extra-plural solitary fibrous tumor (SFT) and to improve the diagnose diagnostic ability.Methods The images and pathologic features of 32 SFT cases confirmed by pathology were retrospectively reviewed.Results Of 32 SFT cases, there were 5 cases in the head, 3 cases in the orbit, 1 case in the nasal sinuses, 1 case in the bronchus, 2 cases in the stomach,1 case in the pancreas, 3 cases in the kidney, 3 cases in the retroperitoneum, 4 cases in the pelvic cavity, 6 cases in the soft tissues, 3 cases near the vertebra.6 cases were malignant,8 cases were borderline,18 cases were benign.The tumors located in abdominal and pelvic tumors were larger,the average diameter of the tumors was 13.6 cm.The volumes of the other tumors were smaller, the average diameter was 4.2 cm.The majority of the tumors showed clear boundary, smooth edge or lobulated soft tissue mass, partial necrosis, rare cystic degeneration and calcification.MR images showed hypo to isointensity on T1WI and hyper to isointensity on T2WI.After enhanced enhancement, multiple small circuitous vessels could be found in or around the mass.The tumor showed slightly enhanced in the arterial phase and moderately to obviously heterogeneously enhancement in the venous phase.In addition, part of the tumors can could be seen positive-negative signal changes.The positive expression rate of immunohistochemical indexes were: CD34 100%, CD99 68.8%, Bcl-2 62.5%, Vimentin 46.9%, Ki-67>5% 43.8% respectively.

ObjectiveTo investigate the clinical value of transvaginal elastography in diagnosing cervical cancers.MethodsTotally,124 patients of cervical lesions with definite pathological results were enrolled.Elastograms were collected and the strain ratios of the benign and malignant lesions were record and compared.ROC curve was used to find the best cut-off point.ResultsElastography was useful in differentiating benign and malignant lesions by different color.Values of strain ratio of different pathological lesions were statistically different ( P ＜0.01).The mean value of strain ratio was 2.71 ± 1.56 for benign lesions and 8.32 ± 4.11 for malignant ones.The best cut-off point was 4.99 basing on the ROC curve.The sensitivity and specificity were 80.7％ and 89.6％,respectively.ConclusionsReal-time transvaginal elastography was a new way in differential diagnosing cervical cancers from benign lesions of the cervix.

Objective To systematically evaluate the effectiveness and safety of interleukin-2 plus cisplatin for treating ma-lignant pleural effusion(MPE)to provide a basis for clinical treatment strategy.Methods CBM,CNKI,VIP,Wanfang,Pubmed, Embase,Cochrane library clinical trial registration database were systematically retrieved.The randomized controlled trial(RCT) quality assessment criteria of Cochrane collaboration network was adopted for including the study quality.The data were extracted by meta analysis.Results (1)Thirty-four RCT involving 2 037 MPE patients were included,the quality of included RCT was ordi-nary;(2)compared with simple cisplatin,the merged RR values and their 95%CI of meta-analysis for ORR,fever,were 1.45 (1.36-1.54),2.37 (1.53 -3.66),respectively,the differences between the two groups were statistically significant(P <0.05 ). The merged RR values and their 95%CI of meta-analysis for leukopenia,myelosuppression and thoracalgia were 0.81 (0.61 -1.07),0.83(0.62-1.11)and 1.04(0.84-1.29)respectively,the differences between two groups were not statistically significant (P >0.05).Conclusion This study indicates that IL-2 plus cisplatin can significantly improve the clinical curative effect in the pa-tients with MPE,but has the adverse reactions of fever,etc.and the quality of included RCT is general.

Objective To explore the miRNA regulating the potential cancer-promoting gene CCL18 in cutaneous malignant melanoma.Methods Bioinformatics analysis was conducted by using online software miRanda and TargetScan,so as to predict the miRNA targeting CCL18 gene.Three kinds of C CL18 3'UTR dual-luciferase reporter vectors,including mutant 3'UTR vector (mutant 3'UTR group),wildtype 3'UTR vector (wild-type 3'UTR group) and empty vector (blank control group),as well as miRNA vectors carring selected miRNAs were constructed according to human gene sequence analysis,and then were used to co-transfect 293T cells.After 48-hour treatment,the cells were lysed for detection of luciferase activity.Real-time fluorescence-based quantitative PCR was performed to measure the expression of CCL 18 and selected miRNA in 14 fresh malignant melanoma tissue specimens and 14 paracancerous normal skin tissue specimens (control tissues),and their correlations were analyzed.Results Online software analysis showed that some miRNAs were identified to target the 3'UTR of CCL18 gene,including miR-183,miR-128 and miR-33a.Luciferase reporter vectors and miRNA vectors were constructed successfully.As luciferase activity assay showed,when miR-183 and miR-128 were bound to the CCL18 3'UTR,the luciferase activities were significantly higher in their mutant 3'UTR groups (11.63 ± 0.42;8.80 ± 0.49) than in their wild-type 3'UTR groups (4.86 ± 0.39;5.01 ± 0.54;both P ＜ 0.05) and blank control groups (2.41 ± 0.13;2.39 ± 0.05;both P ＜ 0.01),while there were no significant differences between miR-33a-hinding mutant 3'UTR group (6.41 ± 0.47) and miR-33a-binding wild-type 3'UTR group (6.16 ± 0.22,P ＞ 0.05).Real-time fluorescence-based quantitative PCR revealed higher mRNA expression of the CCL18 gene (3.52 ± 1.68),but lower expression of miR-183 (0.49 ± 0.32),miR-128 (0.30 ± 0.20) and miR-33a (0.46 ± 0.40) in the malignant melanoma tissues compared with the control tissues.The mRNA expression of the CCL18 gene was negatively correlated with the expression of miR-128 (rs =-0548,P ＜ 0.05),but showed no significant correlations with the expression of miR-183 and miR-33a (both P ＞ 0.05).Conclusion miR-128 may play a role in regulating the potential malignant melanoma-promoting gene CCL18.

Objective To explore the clinical value of real‐time elastography in evaluating rabbit carotid vulnerable plaque . Methods Thirty male New Zealand rabbits ( weighted from 2 .0 to 3 .0 kg) were all fed with high cholesterol diet for one week before balloon injury of their right common carotid arteries . Then they were categorized randomly into two groups which were continued fed by high cholesterol diet for 6 and 12 weeks ,respectively . Ultrasound and real‐time elastography of their right common carotid arteries were applied after their anesthesia by ear marginal vein . Strains of the carotid plaques were measured and analyzed statistically by two independent experienced sonographers . All the rabbits were sacrificed by air embolism method after all the examinations . The atherosclerotic rabbit common carotid lesions were dissected ,fixed for pathologic examination , and graded according to the standard of American Heart Association ( AHA) ,which were compared with the real‐time elastography findings . Results Real‐time elastography had well repeatability between different operators . Consistency between elastography and the AHA pathology was quite good ,with the Cohen′s kappa= 0 .803 (95% CI 0 .669 to 0 .938) . Strains of different regions in a plaque had statistically significant difference ( P <0 .000 1) . By using strain higher than 0 .37% ,the sensitivity and specificity of detecting a lipid core were 88 .0% and 81 .6% ,and area under ROC curve was 0 .902 (95% CI 0 .810 -0 .959 , P < 0 .0001) . Conclusions Real‐time elastography can display the inner elastic characteristics of the rabbit carotid plaques ,and may help evaluate the grading of a plaque . The strain value may help detect a necrotic core ,thereby help evaluate the stability of a plaque .

Objective To investigate the MRI findings of perihip heterotopic ossification (HO) in the early, mid and late stages. Meth-ods The MRI of 44 inpatients with HO from February, 2011 to September, 2013 were reviewed, in which 20 cases (28 joints) were in early stage, 18 cases (24 joints) in mid stage and 6 cases (8 joints) in late stage. For the enhanced T1WI, 9 cases (11 joints) were in early stage, 6 cases (7 joints) in mid stage, and 3 cases (4 joints) in late stage. Theχ2 trend test was used to evaluate the MRI signal change with the HO maturity. Results With the maturity of hip HO, the signal intensity of T2WI reduced (χ2=16.773, P<0.001), fat signal on T1WI increased, the enhancement reduced (χ2=16.048, P=0.007). Conclusion The MRI findings of perihip HO are characteristic in MRI in all the stages. MRI is useful for the diagnosis of perihip HO, especially for the early HO.