Objective To analyze the expressions of astrocyte elevated gene-1 (AEG-1 )and Beclin-1 (BECN-1 )in papillary thyroid carcinoma (PTC)and their clinical significances.Methods Thirty patients with PTC who were performed total thyroidectomy in the General Hospital Affiliated to Tianjin Medical University from October 201 5 to May 201 6 were collected.All patients were diagnosed with unilateral PTC.The tumor tissues were taken from the patients′ipsilateral thyroids,and 30 cases of normal tissues were taken from the patients′contralateral normal thyroids.The expressions of AEG-1 and BECN-1 in PTC tissues were detected by reverse transcription-poly-merase chain reaction (RT-PCR).The relationships between the expression status and clinicopathologic features were analyzed.Results The positive expression levels [M(QR )]of AEG-1 in PTC and the normal tissues were 0.626 1 (0.741 4)and 0.049 8 (0.01 1 8)respectively,with a significant difference (Z=-4.488,P=0.000). The positive expression levels of BECN-1 in PTC and the normal tissues were 0.067 9 (0.1 98 1 )and 0.785 7 (0.361 7)respectively,with a significant difference (Z=-5.441 ,P=0.000).In PTC,the expressions of AEG-1 and BECN-1 were related to TNMstaging (Z=-3.980,P=0.000;Z=-2.265,P=0.023)and lymphatic metas-tasis (Z=-3.1 90,P=0.001;Z=-2.640,P=0.008),but they were not related to age (Z=-1 .203,P=0.229;Z=-1 .1 62,P=0.245),gender (Z=-1 .222,P=0.222;Z=-0.453,P=0.651 )and tumor size (Z=-1 .496,P=0.1 35;Z=-1 .1 66,P=0.244).The expression of AEG-1 was negatively correlated with that of BECN-1 (r=-0.343,P=0.007).Conclusion The expression of AEG-1 in PTC is higher than that in normal tissues and the expression of BECN-1 in PTC is lower than that in normal tissues,and the expressions of AEG-1 and BECN-1 are related to TNM staging and lymphatic metastasis,which are expected to become prognostic indicators.

AIM: To decide the effect that selected siRNA degrades mRNA of IL-1? specifically and suppression of its expression after connected with target site with homology complementary sequence. METHODS: Synthesized DNA expression box aimed directly at target site through PCR reaction in vivo was purified, and transfected into lymphocytes stimulated by LPS. siRNA was transcribed by cellular endogenous RNA polymerase Ⅲ and then evoke the degradation of target mRNA. After 48 hours of transfection, the cell culture supernatant was collected and the concentration of IL-1? was assayed using ELISA. RESULTS: Compared with blank-control and negative-control, selected sequence decreased the expression of IL-1?. Rate of the suppression was about 15%. CONCLUSION: RNAi technology produces specific interference effect in mouse spleen lymphocytes in original culture and inhibits the excretion of IL-1?.

Objective:To observe the therapeutic effect ofcapsaicin at different concentrations on chronic knee arthritis pain model in mice.Methods:Choosing 50 healthy adult male Kunming mice builded chronic knee arthritis pain model by injecting 0.01 mL CFA (Complete Freund's Adjuvant,CFA) into left joint cavity.The model would be succeed in building after 3 weeks.The successful model mice were divided into five groups randomly (n=10):The first experimental group (saline group),the second experimental group (capsaicin excipient group),the third experimental group (0.5 ％ of capsaicin),the fourth experimental group (3 ％ of capsaicin) and the fifth experimental group (8 ％ of capsaicin).All of the mice would be observed the time of withdrawal latencies from the thermal heated surface after administration of one,four and seven hours,and thermal withdrawal time within 60 days after the injection.Results:①The physiological saline group compared with excipient group,the thermal withdrawal time had no statistically significant difference (P＞0.05)after administration of one,four and seven hours,and thermal withdrawal time within 60 days.②The acute pain duration of the third group would disappear after capsaicin injection 7 hours,four hours for the fourth group,and one hour for the fifth group.③The duration of analgesia of the third group,lasted for 18.9± 1.1 days;The analgesia time of the fourth group lasted for 33.7± 1.0 days;The analgesia time of the fifth group lasted for 58.2± 1.2 days.Conclusions:Capsaicin has analgesic effects on chronic knee pain model in mice induced by CFA,and the days of analgesia increases with the concentration of capsaicin.

OBJECTIVE To explore the effect of clofenotane (DDT) on epithelial-mesenchymal transition (EMT) and the relevant molecular mechanism in human colorectal cancer cells. METHODS Human colorectal cancer cells DLD1 were treated with DDT 0.01, 0.1, 1.0, 10.0 and 100.0 nmol·L-1 for 48 h. Then, the morphology of DLD1 cells was observed. mRNA levels of E-cadherin, N-cadherin, vimentin and Snail1 were detected by real-time PCR. Protein expression of STAT3 signaling pathway of proteins STAT3 and p-STAT3 was detected by Western blotting. STAT3 inhibitor WP1006 (5μmol · L-1) was added to determine its impact on DDT-induced alternation of STAT3/Snail1 signaling and EMT-related molecules. Protein expression of STAT3 and p-STAT3 was detected by Western blotting and mRNA levels of E-cadherin, N-cadherin, Vimentin and Snail1 were detected by real-time PCR. RESULTS DLD1 cell morphology was changed after exposure to DDT 0.01-100.0 nmol · L- 1. Meanwhile, real-time PCR showed that the mRNA level of E-cadherin was significantly decreased compared with normal cell control (P<0.01), which was 42.4±2.8%of that in the normal control group. The mRNA levels of N-cadherin, Vimentin and Snail1 were significantly increased (P<0.01), which were 1.91±0.1, 1.5±0.2 and 1.5±0.1 times that of the normal control group. DDT 0.1, 1.0 and 10.0 nmol · L-1 exposure induced up-regulation of STAT3 and p-STAT3 protein levels (P<0.01), which were 2.1 and 1.8 times that of the normal control group. The addition of STAT3 inhibitor WP1066 (5 μmol · L-1) prevented STAT3 from phosphorylation as well as the up-regulation of Snail1(P<0.01), which was (56.3 ± 0.9)% that of the DDT 1.0 nmol · L-1 treat?ment group. Compared with DDT treatment alone, the mRNA levels of EMT-related molecules were remarkably reversed by WP1066 (5 μmol · L- 1) co-treatment, increasing E-cadherin but decreasing N-cadherin and vimentin in DLD1 cells(P<0.01), which were 50.2±2.9%and 61.6±6.1%of those in the DDT 1.0 nmol · L- 1 treatment group, respectively. CONCLUSION DDT alters the expressions of EMT-related molecules including E-cadherin, N-cadherin and vimentin via STAT3/Snail1 signaling, thus promoting the EMT process in human colorectal cancer cells. This progress may be closely related to DDT-induced colorectal cancer development.

Objective Getting the robust exogenous gene expression vector under the control of porcine insulin promoter, and to lay the foundation for pancreaticβ-cells specific transgene expressing pigs.Method Using porcine insu-lin promoter ( PIP, 1500 bp of the 5′UTR from the porcine INS gene including the first exon and the first intron) to con-struct expression vector, the HindIII restriction site which connected the sequences of PIP and EGFP was designed before ATG, named PIP-HindIII-EGFP.Considering that the different location of restriction site may affect the expression efficien-cy of the transgene, we optimized the expression vector.Firstly the HindIII restriction site was deleted to realize the seam-less connection of PIP and EGFP,the vector was named PIP-EGFP.Also we mutated the 3′intron splicing acceptor site( SA) of the first intron into HindIII restriction site, named as PIP-SA( M)-EGFP.Three different EGFP expression vectors were respectively transfected MIN-6 mouse pancreatic β-cells, pig ear fibroblasts and kidney cells.The transfected cells were cultured for 48 h and harvested for RT-PCR, flow cytometry and Western blot analysis, to analyze and compare the expres-sion efficiency of vectors.Results After transfection,green fluorescence was observed only in MIN-6 mouse pancreaticβ-cells.RT-PCR analysis and product sequencing showed that the three expression vectors did have different stability with in-tron splicing.The PIP-HindIII-EGFP construct and PIP-EGFP vector produced two kinds of mRNA with the first intron spliced and no spliced, indicating the instability of intron splicing.Mutation of the PIP splice site would cause the first in-tron not spliced, while flow cytometry and Western blot displayed that the mutation induced a most efficient expression of the downstream gene.Conclusions A robust and specific β-cells expression vector has been successfully generated by mutating the intron splicing acceptor site of the porcine insulin promoter.It provides the foundation for preparation of pigs with pancreaticβ-cells specifically expressing the transgene.

A polydimethylsiloxane ( PDMS )/glass microfluidic chip consisting of a three-layer sandwich structure and three-parallel micro-electrode system was fabricated for the separation of polystyrene microspheres according to the particle size in high conductive solution by electric field produced by alternating current. The principle of electrokinetics of microspheres directional movement was investigated. The results showed that, when the applied voltage was 14 V at 100 kHz, the separation efficiency of 10 and 25 μm polystyrene microspheres was the best. Similarly, with a voltage of 10 V at 2 MHz, the separation efficiency of 5 and 25 μm polystyrene microspheres could achieve the highest value. Meanwhile, the voltage of 11 V at 1 MHz was suitable for the separation of 5 , 10 and 25 μm polystyrene microspheres with the separation efficiency of over 90%. At the same time, the formation of the laminar region in the middle of the electrode gap was the key role of microsphere separation.

Objective To investigate the usefulness of Utstein template to guide the assessment and study of cardiopulmonary resuscitation (CPR) in our medical practice because it has been popularized in many countries since 1991.Methods A prospective observational study using Utstein CPR registry form to evaluate the epidemiological features and outcomes of 511 patients resuscitated in the emergency department.Results Of 511 CPR patients registered,higher cardiac arrest rates were observed in the group of patients aged 40- 70 years. In 511 CPR patients registered,preexisting chronic diseases were common including cardiovascular diseases ( 190,37.2％ ) cerebrovascular diseases (48,9.4％ ) and respiratory diseases (39,7.6％ ).Of them,173 cardiac arrest patients (33.9％) had underlying cardiac causes,such as acute myocardial infarction (AMI) found in 109 (21.3％) patients,and ventricular fibrillation witnessed during first cardiac monitoring in eighty ( 15.7％ ) patients.The restoration of spontaneous circulation (ROSC) rate and survival rate at discharge of in - hospital cardiac arrest (IHCA) patients were 47.0％ and 13.5％respectively,but 16.7％ and 4.7％ respectively in the out - hospital cardiac arrest (OHCA) patients (P ＜0.01,both ). Conclusions This study indicated that the cardiovascular diseases, cerebrovascular diseases,and respiratory diseases were the most common preexisting chronic diseases found in cardiac arrest patients.Myocardial infarct,stroke and trauma were the most common precipitation causes of cardiac arrest in the recruited patients.The rates of ROSC and survival at discharge were significantly higher in statistics in patients with IHCA than those in ones with OHCA.

ObjectiveTo demonstrate the project scope of the afferent nerves of the lumbar intervertebral disc,on which basis to explore the mechanism of the symptoms of discgenic low back pain.MethodsThirty Wistar rats were divided randomly into three groups of 10 rats each:the L4-5,L5-6,and L6 S1 group.Each group was further divided randomly into two subgroups,the experimental group and the control group,5 rats for each group.Intervertebral disc was exposed through the posterior approach under peritoneal cavity anesthesia,after the nerve roots were pull away,2 μl of 30％ cholera toxin-horseradish peroxidase (CT-HRP) was injected into the inner layer of the intervertebral disc in the experimental group,while 2 μl of 0.9％ Nacl was used in the control group.Forty-eight hours after the surgery,all rats were perfused and bilateral dorsal root ganglions(DRGs) of T10-L3 were resected and fixxied.Each DRG was sectioned at 30 μm thickness and processed by DAB method.The sections of DRGs were coverslipped and observed by optical microscopy for the neurons or axons labelled by CT-HRP.It was judged as positive that brownish-black particles were in the neurons or axons.ResultsNot in a single dorsal root ganglions,but in a scope of dorsal root ganglions axons labled by CT-HRP could be seen in the rats in the experimental groups.No CT-HRP labled neurons or axons were seen in dorsal root ganglions in the contral groups.ConclusionAfferent nerves of the lumbar intervertebral disc project to a scope of dorsal root ganglions,which is the anatomic basis of the mechanism of the symptoms of discgenic low back pain.

Objective To analyze the electroencephalograph (EEG) features of 43 patients with clinically possible or probable Creutzfeldt-Jakob disease (CJD) and its correlation with megnetic resonance imaging (MRI) imaging and clinical manifestations.Methods All patients diagnosed with suspected CJD who were hospitalized in Xuanwu Hospital from January 2013 to December 2015 were collected.The clinical data, EEG and MRI imaging features were analyzed retrospectively.Based on the periodic sharp wave complexes (PSWC) appearance in EEG results, the patients were divided into typically changed group (TCG), atypically changed group (ACG) and unchanged group (UCG).Age, disease duration, clinical manifestations and MRI features among three groups were analyzed and the correlations between patient′s EEG features and age, disease duration, clinical manifestations or MRI features were explored using spearman method.Results Among the 43 patients with possible or probable CJD disease, 26 were male and 17 were female with an average disease duration of 4 months.The age of onset ranged from 31 to 80 with an average of (58.0±9.8) years old, and 86.0% of patients were 51 years old or above.Clinical characteristics of CJD patients according to occurrence rate were as follows: 35 cases (81.4%) with cognitive impairment, 29 cases (67.4%) with mental and behavior disorder, 28 cases (65.1%) with pyramidal tract damage, 24 cases (55.8%) with cerebellar symptoms, 23 cases (53.5%) with extrapyramidal symptoms,17 cases (39.5%) with myoclonic, 13 cases (30.2%) with dyssomnia, 13 cases (30.2%) with visual disorder and 2 cases (4.7%) with akinetic mutism.Regarding EEG features, 39.53% (17/43) of patients showed typical periodic sharp wave complexes (PSWC) (TCG group), 51.2% (22/43) had irregular rhythm and different forms of slow wave (ACG group) and only 9.3% (4/43) had no EEG change (UCG group).The occurrence rate of ribbon sign in MRI was 82.4% (14/17) in TCG group, 77.3% (17/22) in ACG group and none in UCG group.The rates were significantly higher in TCG and ACG group than that in UCG group (both P<0.05).Spearman correlation analysis revealed that EEG features was correlated with disease duration (r=0.351, P=0.021) and visual impairment (r=-0.377, P=0.013) for all CJD patients.There was no correlation between EEG and MRI or other clinical manifestations such as myoclonic, age and so on (all P>0.05).Conclusions EEG showed typical changes associated with disease duration in different stages of disease.EEG and MRI are two different means to evaluate different aspects of patients with CJD disease, and combination of two means could achieve better evaluation results.

Objective To evaluate the outcome of radiotherapy for locally advanced pancreatic cancer. Methods From January 2000 to December 2007,41 patients with inoperable locally advanced (stage Ⅲ) pancreatic cancer were treated with three-dimensional conformal radiation therapy(3DCRT) or intensity-modulated radiation therapy (IMRT). Among these patients, 30 received concurrent radio-chemo-therapy. Results The median survival time(MST) and 1-year overall survival were 9.2 months and 23%. Patients with pretreatment KPS≥80 ,no regional lymph nodes metastasis, and CR/PR after radiotherapy had better prognosis. The corresponding MSTs were 11.1 months vs 5.8 months (χ2 = 7.50, P = 0.006), 10. 8months vs 6.5 months (χ2 = 5.67, P = 0.017), and 19.5 months vs 9.1 months (χ2= 7.28, P = 0. 007), re-spectively. Concurrent radio-chemotherapy tended to improve the overall survival (χ2 = 3.25, P = 0. 072). After radiotherapy, 18 patients had clinical benefit response, mainly being abdominal pain relief. Neither grade 4 hematologic nor grade 3 non-hematologic toxicities were observed. Conclusions For patients with locally advanced pancreatic cancer, both 3DCRT and IMRT are effective in alleviation of disease-related symptoms. Patients with better porformance status before treatment, no regional lymph nodes metastasis, and better response to radiotherapy may have better prognosis. Concurrent radio-chemotherapy trend to improve overall survival when compared with radiotherapy alone.