Objective To investigate the expressions of TESTIN gene and Caspase-3 protein and thier relations with the clinicopathological features and prognosis of esophageal squamous cell carcinomas(ESCC). Methods The expressions of TESTIN and Caspase-3 in 50 ESCC tissues and paracancerous tissues (＞5 cm apart form the ESCC tissue) were detected by immunohistochemistry.The expression of TESTIN in 65 matched-pairs of ESCC tissues was detected by Western blotting and RT-PCR. The association of TESTIN and Caspase-3 with clinicopathological features and prognosis of ESCC were analyzed. Results The immunohistochemistry examination showed that the positive rates of TESTIN protein [30. 0% (15/50)] and Caspase-3 protein [24.0% ( 12/50)] were significantly lower in ESCC tissues than those in paracancerous tissues [(84. 0% (42/50) and 94. 0% (47/50),respectively, P＜0.01)]. The mRNA level of TESTIN was down-regulated in ESCC tissues (P＜0.05). Whereas the protein level of TESTIN was down-regulated in 45 (69.2%) of 65 ESCC tissues in comparison with paracancerous tissues. TESTIN expression was positively correlated with the differentiation of ESCC, but not associated with gender, age, tumor size, TNM stage and lymph node metastasis. There was a positive correlation between TESTIN and Caspase-3 in protein expressions (P＜0. 05). Patients with negative expression of TESTIN had lower survival rate compared to those with positive expression (P＜0. 05). Conclusions The positive relation between low-expression of TESTIN and Caspase-3 implicates that both are involved in the development of esophageal squamous cell carcinogenesis, and TESTIN might be a novel ESCC marker with prognostic significance.

Abstract BACKGROUND:Studies have found that Cu-Zn superoxide dismutase (SOD1) is associated with familial amyotrophic lateral sclerosis, opening up a new way of studying molecular genetics. So far more than 100 kinds of SOD1 gene mutations have been found. OBJECTIVE:To analyze the hot spot, research front and knowledge base of amyotrophic lateral sclerosis visualy. METHODS:Totaly 4 693 relevant articles published from 2005 to 2014 were retrieved from Web of Science in ISI with “Cu-Zn superoxide dismutase” or “SOD1” as search keywords. With the aid of CiteSpace III software, the visualization mapping of the network in co-cited articles and keywords was drawn to reveal knowledge base, hot spots and research front of SOD1. The parameters include the number of published papers and citations within 10 years, distribution of research countries and institutions, main source journals, research area of highly cited papers, keywords with high-frequency and emerging keywords with high-frequency in recent 5 years. RESULTS AND CONCLUSION:The number of published papers and citations in a year showed a trend of sustained growth. United State, China and Japan rank the top three in this area, in which Chinese Academy of Sciences has a great influence among the research institutions. The research fields of SOD1 focus on neurosciences and neurology, biochemistry and molecular biology and so on. The high impact factors of journals with a large number of articles reflect the importance and innovation of this research. Ten high-cited articles consist of the knowledge base on SOD1, directing to the finding of different sites of SOD1 mutation and the measurement of protein concentrations and activity of SOD. The hot spots of SOD1 mainly focus on oxidative stress, familial amyotrophic lateral sclerosis caused by SOD1 mutation and different types of transgenic animal models. The research fronts mainly focus on the finding of pathogenesis in amyotrophic lateral sclerosis, such as the aggregation of TDP-43, the interaction between astrocytes and motor neurons, optineurin and the inhibitor of nuclear factor-κB, hexanucleotide repeat expansion in C9ORF72 and autophagy.

Objective To evaluate the effects of different concentrations of sevoflurane anesthesia on longterm learning and memory abilities in neonatal rats.Methods Twenty-seven neonatal Sprague-Dawley rats of both sexes,aged 7 days,weighing 12-20 g,were randomly assigned into 3 groups (n =9 each):control group (C group),2％ sevoflurane group (S1 group) and 3％ sevoflurane group (S2 group).Groups C,S1 and S2 inhaled air,2 ％ sevoflurane and 3 ％ sevofluran for 4 h,respectively.The neonatal rats were reared to 35 days old and underwent open field test,to 36 days old and underwent Morris water maze test,and to 42 days old and underwent continuous multiple-trail inhibition avoidance training.Results Open field test:There was no significant difference in the movement time,movement speed and the time the animals spent in the central square among the 3 groups (P ＞ 0.05).Morris water maze test:Compared with C group,the looking for platform latency on 2nd-5th days in S2 group and on 2nd-3rd days in S1 group was significantly prolonged,and the percentage of time of staying at the platform quadrant was decreased in S1 and S2 groups (P ＜ 0.05 or 0.01).The looking for platform latency on 3rd4th days in S2 group was significantly longer than that in group S1 (P ＜ 0.05).Continuous multiple-trail inhibition avoidance training:The latency detected at 24 h after training was significantly shorter in S1 and S2 groups than in group C (P ＜ 0.05),and in group S2 than in S1 group (P ＜ 0.05).Conclusion Sevoflurane anesthesia decreases the long-term learning and memory function in neonatal rats in a concentration-dependent manner.

This article summarized the technique research and application status of near-infrared spectroscopy (NIRS), in order to explore hypoxia,qi-deficiency and their connections with typical symptoms. It also discussed NIRS acquired oxygen saturation (SaO2) and its relevant means and methods for information quantification ofqi-deficiency. The accurate obtained SaO2 value was applied in the association with the strength of chest distress in the subjective sensation due toqi-deficiency. According to current data, the normal value,qi-deficiency degree and uncomfortable degree of subjective sensation among patients of different altitudes and ages were summarized. It can be applied to assist TCM physicians in the quick quantification of qi-deficiency.

Objective To discuss the significance of region Ⅵ lymph nodes dissection in treatment of patients with clinical cervical nodes negative(cN0)papillary thyroid carcinoma(PTC).Methods Clinical data of 38 cases of cN0 PTC treated with region Ⅵ lymph nodes dissection were retrospectively analyzed.The related literature was reviewed.The cervical nodes dissection scope and the key operation points in treatment of cN0 PTC were discussed.Results No permanent recurrent laryngeal nerve injury or parathyroid damage happened.14 cases(36.84％)had occult lymph node metastasis to region Ⅵ lymph nodes.After more than 5 years of follow-up,all the cases had excellent life quality.3 cases(7.89％)were found lymph node metastasis to lateral cervical region and they were given functional neck dissection.No recurrence or metastasis occurred to the 3 cases during more than 2 years of follow-up.Conclusion Region Ⅵ lymph node dissection in treatment of cN0 PTC is safe,reliable,and with less complications.

Objective To evaluate effects of artesunate on rosacea-like inflammation in mouse models.Methods Twenty-five male BALB/c mice aged 7 weeks were injected subcutaneously with 40 μ1 antibacterial peptide LL-37 into the back once every 12 hours for 4 sessions to establish mouse models with rosacea-like inflammation.These 25 mice were randomly and equally divided into 5 groups:after each injection of LL-37,model group were gavaged with sodium chloride physiological solution,treatment groups gavaged with 25,50 and 100 mg/kg artesunate solution separately,and positive control group gavaged with 30 mg/kg doxycycline hydrochloride solution.Another 5 healthy mice injected subcutaneously with pure water into the back for 4 sessions served as blank control group.Forty-eight hours after the initial injection of LL-37,changes in skin lesions and the intensity of erythema were assessed.Skin tissues at the dorsal injection site were resected and subjected to HE staining,the tissue structure was observed and the number of inflammatory cells was counted.Enzyme-linked immunosorbent assay (ELISA) was performed to estimate the activity of myeloperoxidase (MPO) in skin lesions.Results The model group showed obvious inflammatory reactions,and significantly increased erythema score (3.20 ± 0.84),inflammatory cell count (517.27 ± 99.43) and MPO activity (0.57 ± 0.08) compared with the blank control group (all P ＜ 0.01).The positive control group showed significantly decreased erythema score (1.60 ± 0.89),inflammatory cell count (270.93 ± 124.63) and MPO activity (0.40 ± 0.05) compared with the model group (P ＜ 0.05,0.01,0.01,respectively).Moreover,the erythema score,inflammatory cell counts and MPO activity were all significantly lower in 50-(1.80 ± 0.84,286.00 ± 33.72,0.43 ± 0.05,respectively) and 100-mg/kg artesunate groups (1.40 ± 0.55,258.00 ± 36.44,0.40 ± 0.06,respectively) than in the model group (P ＜ 0.05 or 0.01).However,there were no significant differences in the erythema score,inflammatory cell count and MPO activity between 50-or 100-mg/kg artesunate group and the positive control group (P ＞ 0.05).Conclusion Artesunate can inhibit rosacea-like inflammatory reactions in mouse models,especially the middle-and high-dose artesunate.

Objective To investigate the expression of exogenous gene transferred by piggyBac (PB) transposon in various gynecological malignant cell lines and reveal its potential application of gene therapy in gynecological cancer.Methods Amplified herpes simplex virus thymidine kinase (HSV-tk) gene coding region by PCR and integrated it into PB expression vector, PB[Act-RFP]DS, for reconstructing PB[Act-RFP, HSV-tk]DS (pPB/TK).By using different transfection reagents: FuGENE HD, jetPEI, lipofectamine 2000, pPB/TK together with helper plasmid Act-PBase were cotransfected into four mostly common gynecological malignant tumor cell lines HeLa, JEG-3, SKOV3 and HEC-1B.The mRFP1 report gene expressions was observed and detected by fluorescence microscope and flow cytometry to analyze transfection efficiency.The expressions of HSV-tk and mRFP1 gene were detected by reverse transcription PCR (RT-PCR).The cytotoxic effect of various concentration of pro-drug ganciclovir (GCV) for transfected cells was detected by methyl thiazole tetrazolium assay.The transfected cells were positive sorted by flow cytometry and limiting diluted to obtain the stable transfected cell line.The insertion sites of foreign gene tranferred by PB transposon in genome were analyzed by inverse PCR.Results (1) Double digests analysis and sequences test demonstrated that pPB/TK vector was reconstructed successfully.(2) Using three different transfective reagents, PB trausposon transferred HSV-tk gene and mRFP1 gene into HeLa, HEC-1 B, SKOV3 and JEG-3 cell efficiently, and the transfection efficiency of pPB/TK for the same cell was different by using different transfective reagents; in Hela cell, the transfection efficiency of FuGENE HD [(78.7 ± 9.2) %]was higher than that of lipofectamine 2000 [(54.1 ± 11.4) %]and jetPEI [(46.5 ± 7.4) %, all P < 0.05] ; using the same transfective reagent, the transfection efficiency of pPB/TK was also different on various cell lines, using FuGENE HD, the transfeetion efficiency of pPB/TK on HeLa, JEG-3 and SKOV3 cell was (78.7 ± 9.2) %, (74.4 ± 8.9) % and (83.2 ± 9.7) % respectively, which all were higher than that on HEC-1B [(39.5 ± 8.7) %, P < 0.05] .(3) RT-PCR showed that there were the mRNA expression of HSV-tk and mRFP1 in all cell lines.(4) 50% inhibitory concentration of GCV for transfected cells, HeLa, JEG-3, SKOV3 and HEC-1B, was 1.29, 3.35, 0.09 and 13.28 μg/ml respectively.Inhibitory effect of GCV (10 μg/ml) on SKOV3 transfected with pPB/TK was (86 ± 9) %, which was superior to that transfected with pORF-HSVtk alone [(52 ± 12)%, P < 0.05] .(5) The insertion sites of PB transposon in the target cells genome were located at TTAA sites, mRFP1 expression still could be detected in three months after transfected.Conclusions PB transposon could transfer exogenous gene into various gynecological malignant cells, which could integrated into genome and obtain a long-term and stable expression.It is expected that PB transposon may supply a more efficient and safer transgene technology platform for gene therapy in gynecological cancer.

Objective To investigate the expression of transforming growth factor beta (TGF-β) and Smad signaling in ankylosing spondylitis (AS) and to explore their roles in the pathogenesis. Methods Fiftythree patients with AS were included in the study. In these 53 cases, 30 patients were performed computed tomography-guided needle biopsy in sacroiliac joint. Serum TGF-β_1 was determined by enzyme-linked immunosorbent assay (ELISA). Immunohistological studies were performed with the streptavidin-peroxidase conjugated methods to assess the expression of TGF-β_1, p-smad3 and Smad7 in sacroiliac joint tissue sample.One-way ANOVA, two independent samples t test and kolmogoorov-Simimov test were used to do statistical analysis. Results In 53 cases patients with AS, 20 cases were with high level Erythro-cyte sedimentation rate(ESR) and C-reactive protein (CRP), while those of the other 33 cases were normal. Serum average TGF-β_1level [ (15.9±5.6) ng/ml ], in patients with high level ESR/CRP [(5.4±5.8) ng/ml ] was significantly increased as compared to the controls and patients with normal ESR/CRP [(4.1±3.6) ng/ml] (P<0.05). There was no expression of TGF-β_1 could be detected in the pannus and bone marrow in SI joints tissue of 30 cases with AS, while decreased level of smad7 expression was detected. In addition, p-smad3 expression was found in the nuclear. Conclusion TGF-β_1 signaling may play an important role in the inflammatory erosion and cartilage fibrosis of sacrojlitis in AS.

Super-resolution image reconstruction techniques play an important role for improving image resolution of lung 4D-CT. We presents a super-resolution approach based on fast sub-pixel motion estimation to reconstruct lung 4D-CT images. A fast sub-pixel motion estimation method was used to estimate the deformation fields between "frames", and then iterative back projection (IBP) algorithm was employed to reconstruct high-resolution images. Experimental results showed that compared with traditional interpolation method and super-resolution reconstruction algorithm based on full search motion estimation, the proposed method produced clearer images with significantly enhanced image structure details and reduced time for computation.
Algorithms ; Four-Dimensional Computed Tomography ; Humans ; Image Enhancement ; Lung ; anatomy & histology ; Motion ; Tomography, X-Ray Computed

Objective To investigate the expression of connective tissue growth factor(CTGF),coll agen I and collagen Ⅲ in sacroiliac joint(SIJ)of patients with spondyloarthropathy(SpA).Methods Thirty patients with SpA,including 17 patients with grade Ⅱ saeroiliitis and 13 patients with grade Ⅰ sacroiliitis,were performed on CT guided needie biopsy of SIJ.After sacroiliitis were confirmed by staining with hematoxylin and eosin in sacroiliac joint tissue sample,immunohistochemical assay was performed to determine the expression of CTGF,collagen Ⅰ and collagen Ⅲ in sacroiliac ioint tissue.Univariate Chi-square test was used for data comparison between multiple groups and t-test was used for two group data comparison.Results Contrast to healthy controls,CTGF were found upexpressed on the cytoplasm of inflammatory cells in pannus and bone marrow of sacroiliac tissue samples of patients with SpA,while collagen I and collagen Ⅲ were found up-expressed in bone,cartilage and ligament tissue[(57.9±42.4)/HP vs(2.7±2.5)/HP P<0.05,0.298±0.080 vs 0.044±0.024 and 28.254±41.165 vs 0.105±0.054.P<0.05 respectively].Conclusion CTGF,collagen Ⅰ and collagen Ⅲ are up-expressed in SIJ of SpA patients.CTGF may play an important role in articular cartilage fibrosis and ossification of SpA.