Objective To investigate the effects of different energy,frequency and time of holmium laser on the ablated porcine pancreas in vitro,and to establish an ablation regression equation of holmium laser,in order to provide the experiment foundation for EUS-guided holmium laser ablation of pancreatic carcinoma.Methods According to pilot study,the range of energy,frequency and time of holmium laser was determined,and five values were chosen for the individual three parameters,therefore,a randomize table including 125 combinations (sample capacity) according to the various combinations of every parameter was constructed,then every combination of holmium laser was used for porcine pancreas ablation in vitro.EUS was applied to determine the ablation site and measure the ablation extent,and pathological evaluation was performed.Results Sonographic images showed hyperechoic cloudy area in the ablation site.It was observed that the ablation body was an approximately oval area,the middle part was carbonized area,and the out layer was grey-white necrosis area.Microscopic examination showed there was pool-like cavity in the middle of ablation site,and it was surrounded by coagulation necrosis of pancreatic tissue,and the out layer was inflammatory cells infiltration.Normal pancreatic tissue was found in the margin.The energy,frequency and time of hoimium laser were positively associated with the ablation extent,and the frequency was the main factor,followed by time and energy.The difference was statistically significant (P＜O.O1).The best combination was 25 s,25 Hz,1.8 J.An ablation regression equation was created,which was In (ablation volume) =β0 + β1 × time + β2 × frequency + β3 × energy.Conclusions The holmium laser can produce obvious tissue necrosis in porcine pancreas in vitro; the established holmium laser ablation regression equation can be the guidance for clinical practice.

Objective To investigate the effect of needle sizes and aspiration techniques on sample quantity. Methods Aspiration was performed on porcine liver in vitro for 10 times with three different sizes of needles(19 G, 22 G and 25 G) and four different aspiration techniques[non?negative pressure(NP), 10 ml NP,20 ml NP and slow?pull], 20 mm in depth. A total of six aspirations were performed with each needle by the same aspiration technique. All the obtained specimens were fixed in formalin with the cell block method. The samples were evacuated according to our grading criteria. Results The mean±standard deviation(SD) score for 19 G,22 G, 25 G were 5?71±0?69,4?63±1?24, 3?79±1?84 respectively. The mean±SD score for methods non?NP,10 ml NP,20 ml NP and slow?pull were 4?72±1?53,4?56±1?46,4?72±1?50,4?83±1?76 respectively. The multi?analysis of variance results showed that there were statistical differences between different needles size( F=12?00,P<0?001) with 19 G being the best,followed by 22 G and the least specimen obtained by 25 G needle. There were no statistical differences among aspiration techniques ( F=0?128, P=0?943).The analysis showed that the thicker the needle was,the better sample quality was 19 G yielded to the highest quantity of specimens. The most specimens could be obtained with 19 G needle and non?NP, 22 G needle and 20 ml NP and 25 G needle and slow?pull. Conclusion In clinic, aspiration technique should be selected according to different aspiration needles. 19 G is superior to others, with non?NP method. For 22 G needle, 20 ml NP is preferred and for 25 G needle,slow?pull is preferred.

Objective To detect the mutation frequency of EML4-ALK fusion gene in lung cancer patients, and to inves-tigate the distribution of mutation character for EML4-ALK fusion gene in Ⅰ stage lung cancer patients and clinical features as well as provide a reference for the individual treatment of lung cancer .Methods 256 fresh tumor tissue specimens of lung cancer patients were screened from the specimen bank of our hospital and all the patients had accepted the surgical treatment from February 2013 to December 2014.Total RNA was extracted and then be transcribed into cDNA, the amplification-refrac-tory mutation system(ARMS) was used to detect mutation of EML4-ALK fusion gene.The results according to the positive con-trol, negative control and RNA quality control for EML4-ALK fusion type were analyzed.Results During the 256 patients ofⅠ stage lung cancer, there were 17 patients(6.64%) had mutations in EML4-ALK fusion gene.In lung adenocarcinoma mu-tation rate(16/207, 7.73%) was higher than that of lung squamous cell mutation rate(1/39, 2.56%), lung adeno-squamous mutation rate(0/4, 0) and large cell carcinoma(0/5, 0) of the mutation rate;young lung cancer patients( <63 years) of the mutation rate(14/139, 10.07%) was significantly higher than the high age of lung cancer patients(≥63 years old) mutation rate(3/117, 2.56%), P =0.009.EML4-ALK fusion with tumor invasion and visceral pleura group incidence (9/80, 11. 25%) was significantly higher than that of non-invasive and visceral pleura group incidence rate(8/176, 4.55%), P =0.045.Conclusion The occurence of EML4-ALK fusion correlates with patients’ age as well as whether visceral pleura is in-vaded, type 1 EML4-ALK fusion was detected more in phase I lung cancer patients.

Objective:To observe the expressions of SENP1, SENP2and SENP6proteins in human malignant glioma tissue and cells, and to elucidate the their effects in the development of malignant glioma.Methods:The samples of normal human brain tissue and malignant glioma tissue were obtained and used as normal control group and malignant glioma group, respectively.The Cos7cells and the malignant glioma LN443and U343cells were cultured;the Cos7cells were used as normal cell control group, and the LN443and U343cells as malignant glioma cell group.Western blotting method was used to detect the expression levels of SENP1, SENP2and SENP6proteins in human malignant glioma tissue and cells.Results:In brain tissue, the expression levels of SENP1, SENP2and SENP6proteins in malignant glioma group were higher than those in normal control group (P＜0.05) .Compared with normal cell control group, the expression levels of SENP1, SENP2and SENP6proteins in the LN443and U343cells in malignant glioma cell group were significantly increased (P＜0.05) .Conclusion:SENP1, SENP2and SENP6proteins highly express in the malignant glioma tissue and cells, and they may play an important role in promoting the occurrence of malignant glioma.

Objective To explore the effectiveness of endoscopic ultrasound( EUS)-guided holmium laser ablation for primary pancreatic implantation tumor in nude mice. Methods Pancreatic cancer cell line SW1990 were implanted into 204-6-week-old male balb/c nude mice to establish primary pancreatic implantation tumor in situ models. Then the nude mice were randomly divided into two groups, the treatment group(n=10) and the control group(n=10). The treatment group underwent EUS-guided holmium laser ablation in the pancreatic tumor. And no interventions were given to the control group. The volume of tumors of the two groups were measured under EUS at time points of 7 d, 14 d and 28 d after ablation. The activities, appetites and psychosis of all nude mice were evaluated in the meantime. At 28 d after ablation, lesions of pancreas were dissected and sliced for H&E staining. Results There were no complications in the treatment group, and all nude mice could tolerate the procedure. The mental state, activities and appetites of nude mice in the experimental group were better than those in the control group. Tumors of the control group enlarged. There was significant difference in the tumor size between the two groups at 28 d after ablation. HE staining showed coagulation necrosis in the ablation area. Conclusion EUS-guided holmium laser, producing coagulative necrosis in the ablation area, is effective for primary pancreatic implantation carcinoma in nude mice for about 28 days.

Objective To analyze the epidemiological characteristics of severe fever cases,with thrombocytopenia syndrome (SFTS) in Zhoushan,Zhejiang,2011-2013 and to provide evidence for the development of related strategies for prevention and control of the disease.Methods Data was collected from the studies and related demography of the cases with SFTS in Zhoushan,2011-2013.Clinical and epidemiological characteristics of the cases with SFTS were analyzed descriptively.Results During 2011-2013,45 SFTS cases were reported in Zhoushan city,with fatality rate as 11.11％ (5/45),including 41 cases (91.11％) reported in Daishan county.Most cases aged 50 or over,residing in hilly regions (93.33％).The epidemic peak was seen from May to July (84.44％).Incidence rates of the disease did not show significant statistical differences between males and females.Most cases were farmers (42.22％) and housewives (31.11％).Most patients complained of fever (97.78％),anorexia (91.11％),fatigue (62.22％),chill (44.44％) and other non-specific clinical manifestations but all appeared progressive thrombocytopenia and leucopenia.Sanitation condition of the patients was generally poor.68.89％ (31/45) of cases had farmed,mowed or involved in other outdoor activities in the previous two weeks,with some of them reported having exposed to tick bites.Three events were reported,epidemiologically.Conclusion SFTS in Zhoushan was a infectious disease with natural foci and caused by SFTSV.Ticks might serve as the main vectors for SFTSV,and might cause direct transmission between humans.Improving the environment for living and carrying out essential protective measures in outdoor activities seemed to be significant in decreasing the incidence rate of the disease.

Objective:To evaluate the performance of magnetic beads extraction method (MGE) for the measurement of catecholamine metabolites by liquid chromatography tandem mass spectrometry.Methods:This is a methodological evaluation study. The linearity, limit of quantitation, recovery, precision, and matrix effect of catecholamine metabolites 3-methoxyepinephrine (MN), 3-methoxynorepinephrine (NMN) and 3-methoxytyramine (3-MT) extracted by MGE method were evaluated according to CLSI C62-A. Consensus of method development and validation of liquid chromatography-tandem mass spectrometry in clinical laboratories and other guidelines, 132 clinical residual plasma samples were collected and extracted by automated MGE and traditional solid phase extraction (SPE) method to compare the harmonization of the two extraction methods.Results:The linearity of MN, NMN and 3-MT extracted by automated MGE was>0.99, and the LOQ for MN, NMN and 3-MT were 0.033 5 nmol/L, 0.054 7 nmol/L and 0.011 0 nmol/L, respectively. The repeatability of MN, NMN and 3-MT were 1.3%-5.1%, 2.2%-5.6% and 1.7%-7.1%, respectively. The total imprecision in the laboratory were 1.5%-8.2%, 2.2%-7.7%, 2.1%-11.2%. Although the absolute recovery is low, the average relative recoveries of MN, NMN and 3-MT were 91.5%-108.5%, 92.0%-108.6%, and 89.3%-104.1%, respectively, and the percentage deviation from the expected concentration was within 15%. After isotope internal standard correction, the relative matrix effect is close to 100%, which can compensate for the potential matrix effect. The results of MGE and SPE of MN, NMN and 3-MT showeda good correlation (correlation coefficient r>0.99). The average relative deviations of MN, NMN and 3-MT were 0.2%, -1.4% and 1.0%, respectively. Conclusion:The automatic MGE method hasa good performance in extracting catecholamine metabolites, and is expected to be used in high-throughput analysis of samples in clinical in the future.

Mesp family proteins comprise two members named mesodermal posterior 1 (Mesp1) and mesodermal posterior 2 (Mesp2). Both Mesp1 and Mesp2 are transcription factors and they share an almost identical basic helix-loop-helix motif. They have been shown to play critical regulating roles in mammalian heart and somite development. Mesp1 sits in the core of the complicated regulatory network for generation of cardiovascular progenitors while Mesp2 is central for somitogenesis. Here we summarize the similarities and differences in their molecular functions during mammalian early mesodermal development and discuss possible future research directions for further study of the functions of Mesp1 and Mesp2. A comprehensive knowledge of molecular functions of Mesp family proteins will eventually help us better understand mammalian heart development and somitogenesis as well as improve the production of specific cell types from pluripotent stem cells for future regenerative therapies.
Amino Acid Sequence ; Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Cell Differentiation ; genetics ; Gene Expression Regulation, Developmental ; Mesoderm ; embryology ; metabolism ; Mice, Knockout ; Molecular Sequence Data ; Pluripotent Stem Cells ; metabolism ; Sequence Homology, Amino Acid