AIM: To know the variations of the cytochrome b gene in cancer tissue, paracarinoma tissue and normal tissue and to inquire into the relationship between mutations of mitochondrial genome and carcinogenesis. METHODS: Cellular total DNA was extracted.The cytochrome b genes of three tissues were amplifyed with polymerase chain reaction(PCR). PCR products were analysed by DNA auto-sequencing method. RESULTS: The cytochrome b gene of cancer tissue had the C to G mutation at nt 14931, the C to G mutation at nt 15004 and the T to C mutation at nt15435,respectively. The cytochrome b gene of paracarinoma tissue had the A to C mutation at nt 15436. The cytochrome b gene of normal tissue had not mutation. CONCLUSION: Mitochondrial DNA mutations could be the endogenous factors that induce nuclear genome mutation. It could promoto carcinogenesis. The paracarinoma tissue was abnormal in DNA molecular level.

Objective To construct the eukaryotic expression vector pEGFP-N1/IL-37b and analyze the expression of IL-37 gene in THP-1 cells. Methods Total RNA was extracted from human peripheral blood mononuclear cells ( PB-MCs) and the coding region of IL-37b gene was amplified by RT-qPCR. Then, the gene was cloned into pEGFP-N1 eu-karyotic expression vector. After transfected the recombinant plasmid into THP-1 cells, the expression of IL-37 was detec-ted by RT-qPCR and Western blot. Results Double restriction enzyme digestion and gene sequencing showed that IL-37b gene was correctly inserted into the eukaryotic expression vector pEGFP-N1. RT-qPCR and Western blot showed that the IL-37 expression level was increased significantly (P<0. 01) after transfection in THP-1 cells. Conclusions We successful-ly constructed a novel anti-inflammatory cytokine IL-37 eukaryotic expression vector pEGFP-N1/IL-37b, which lays a foun-dation for further study on IL-37 functions and its association with related diseases.

Objective To explore the role of ultrasound-guided core needle biopsy in diagnosis and treatment of thyroid nodules.Methods 778 cases undergoing ultrasound-guided core needle biopsy in our department from Jan.2012 to Dec.2014 were retrospectively analyzed,and the sensitivity,specificity and accuracy were calculated.Results All the patients were operated successfully.Except for 21 nodules undiagnosed,16.4％ (124/757) were histologically diagnosed as malignant and 83.6％ (633/757) were benign.15 samples of the 124 malignant nodules were less than 10 millimeter in diameter.The sensitivity,specificity and accuracy were 100％,94.8％ and 95.6％,respectively.Of the ultrasound features,blood flow,boundary,internal situation and calcification were statistically significant between benign and malignant nodules,but not so between the groups of different greatest dimentions.Conclusion Ultrasound-guided core needle biopsy is safe and effective for obtaining thyroid nodule tissue,and it is helpful for diagnosis of thyroid diseases.

Objective To establish a stable primary culture of rat cerebellar granule neurons in vitro for further study the toxic effects of chronic arsenic exposure on cerebellar cells.Methods Cerebellar cortices were taken from brain of Wistar rat 5-7 day old after born under stereoscopic microscope.Single cell suspension was acquired after digestion and washing with trypsin (0.25％) and DNase Ⅰ solution,respectively.Granule cells were purified from other cells by differential velocity adherence method for two times.Rat cerebellar granule neurons were seeded in culture plate pre-coated with poly-L-lysine.Neurons growth,development and synaptic connections were observed daily.The neurons were identified by neuron specific enolase (NSE) immunofluorescence technique.Results The neurons were affixed to the culture plate in 24 hours,in reticular arrangement observed under contrast microscope.Granule cells gradually turned round from oval and outlines became clearer in 2-3 days.In 4-6 days,there were a wide range of synaptic connections among the neurons and a mature nerve cell network formed.A large quantity of cerebellar granule neurons was seen by NSE identification.Few bigger cells such as purkinjes cells and glial cell outlines were also seen in the same visual field.Conclusions This is a successful primary culture method for acquirement of rat cerebellar granule neurons.The method can provide experimental basis for future studies the toxic effects of chronic arsenic exposure on cerebellar cells.

Objective A preliminary study on a new discovered proteomic fingerprint correlated with anxiety disorders, which M/Z range from 15000 to 16800. Methods 221 cases of neoplasm patients from April of 2004 to now were divided into anxious group and non-anxious group according to their scores using self-rating anxiety scale (SAS), and all the patients' sera were detected utilizing SELDI-TOF-MS. The data were analyzed with supported softwares. Results Compared the mass spectra of anxious group(n=49) and non-anxious group (n=172), 2 clusters were captured which M/Z range from 15 000 to 15 380 and 15 700 to 16 800 respectively. The coincidence with SAS was 75.51%. The M/Z range from 15 700 to 16 800 was the most frequent, then the second range from 15 000 to 15 380 and 15 700 to 16 800 which was doublet cluster, the least range from 15 000 to 15 380. The same fingerprints were captured in non-anxious group and their intensity was different. Conclusion The M/Z range from 15 000 to 16 800 was protein fingerprint correlated with anxiety disorders.

Objective:To clone swine leukocyte antigen,class II,DO alpha (SLA-DOA) gene from Banna mini-pig inbred line (BMI) and detect its mRNA expression level in 19 important tissues.Methods:The complete eDNA sequence of SLA-DOA gene was cloned by RT-PCR method from BMI and the mRNA expression pattern in BMI important tissues was examined by semi-quantative RT-PCR method.Nucleotide and protein sequences of SIA-DOA were used to carry out bioinformatics analysis and construct the phylogenetic tree.Results:The eDNA length of BMI SLA-DOA was 1 079 bp,which encoded a protein of 250 amino acids with molecular weight (Mw) 27.81 kD,and isoelectric point (pI) 6.48.Genome structure analysis showed it localized to Sus scrofa chromosomes 7 and consisted of four exons and three introns.Semi-quantitative expression analysis showed that SLA-DOA gene expressed highly in the lymph nodes and stomach;weakly in the heart,skin and duodenum and none in other 14 tissues.Functional bioinformatics analysis indicated that SLA-DOA protein contained one signal poptide,one transmembrane region,three conserved domains,four O-GlcNAc glycosylation sites,18 potential phosphorylation sites and to be located in the cytoplasm with 94.1％ certainty.Phylogenetic analysis demonstrated that BMI (pig) had the closest relationship with cattle.Conclusion:This study have successfully cloned the SLA-DOA gene of Banna mini-pig inbred line,performed bioinformatics analysis and tissue expression profile analysis.It will provide a basis for the studies of BMI xenotransplantation.

Objective:To investigate the effect of staphylococcus aureus on expressions of inducible nitric oxide synthase (iNOS) and interleukin-1 beta (IL-1β) in THP-1 monocytes under high glucose concentrations. Methods:THP- 1 monocytes were incubated at different surroundings with 2×2 factorial design. There were 4 experimental groups in the study, which were analyzed by univariate analysis of variance. The total RNA was distilled. The expressions of iNOS and IL-1β were examined by SQ-RT-PCR. Results:The expressions of iNOS and IL-1β were affected by high glucose concentration, bacteria and both of them together in THP-1 monocytes. The expressions of iNOS and IL-1β were weakened in high glucose concentration compared with that of control(P < 0.01). The expressions of iNOS and IL-1β were weakened in the high glucose concentration and bacterial infection compared with that of the high glucose concentration only(P < 0.01). The expressions of iNOS and IL-1β increased in bacterial infection compared with that of control(P < 0.01). The expression of IL-1β increased in high glucose concentration and bacterial infection compared with that of the high glucose concentration only(P < 0.01) but the expression of iNOS was not distinct. Conclusion:When staphylococcus aureus infected monocytes, the expressions of iNOS and IL-1β were weakened in high glucose concentration, which suggested that the depressed function of immunocyte was related with high glucose concentration.

Objective To assess regional left ventricular systolic function in patients with hypertrophic cardiomyopathy (HCM) using real-time three-dimensional echocardiography (RT-3DE).Methods Twenty-five patients with HCM which was asymmetric septal hypertrophy,and twenty healthy subjects were enrolled in the study.The apical four-chamber view of left ventricular was acquired by RT-3DE.The left ventricular volume-time curves were analyzed quantitatively with Tomtec 4D LV-Analysis 3.0,and regional end-diastolic volume and end-systolic volume of left ventricular (rEDV,rESV),the time to minimum systolic volume (rESVT),regional stroke volume (rSV),regional ejection fraction (rEF),regional-global ejection fraction (rgEF) and the parameters of left ventricular dyssynchrony were measured.Results In the HCM group,the values of Tmsv16-Dif,Tmsv16-SD,Tmsv16-Dif％,Tmsv16-SD％ were significantly lower compared with the control group (P ＜ 0.01),and rEDV,rSV,rEF and rgEF in hypertrophic segments were lower than those in non-thickening and mild-thickening segments (P ＜0.05).In the control group,there were no significant difference of those parameters among all segments (P ＞0.05).The values of rEDV,rSV and rgEF in hypertrophic segments decreased in the HCM group (P ＜0.05),at the basal level,rEF in hypertrophic segments decreased,at the apical level,it increased,but the differences at the mid-ventricular level between the two groups were not significant;the values of rEF and rgEF in non-thickening and mild-thickening segments increased (P ＜0.05).Conclusions RT-3DE could sensitively detect left ventricular dyssynchrony and accurately assess regional left ventricular volume and function of different segments in patients with HCM.

Aim To study on the mitochondrial mass change in mouse thymocyte apoptosis.Method Control and Dexamethasone(DEX) groups were set;at 6h,we studied mitochondrial mass changes by NAO and Mitotracker Green(MG) staining flowcytometry and detected mitochondrial membrane potential change with DiOC_6(3) staining flowcytometry.We also used Annexin V-PE/MG double staining flocytometry to examine the mitochondrial changes in apoptosis progress.Results NAO staining results showed that 1 ?mol?L~(-1) DEX stimulation reduced the cardiolipin content of thymocyte mitochondria(P

Objective To compare the efficacy of ureteroscopic lithotripsy(URL)combined with occlusion device and the supine and lithotomy position mini-invasive percutaneous nephrolithotomy(mPCNL)in the treatment of upper incarcerated ureteral stones. No difference could be found in age,sex and size of stones between the two groups. Method From Jan. 2014 to Dec. 2016 in our hospital,all cases of upper incarcerated ureteral stones were diviede into two groups:52 in ureteroscopic lithotripsy combined with occlusion device group and 45 in mini-inva-sive percutaneous nephrolithotomy group. Result The hospitalization and operation time in URL group were(5.5 ± 1.4)days and(53.3 ± 12.4)mins,which were significantly shorter than that in mPCNL group with(9.1 ± 3.2)days and(78.2 ± 14.1)mins,(P<0.05). There were no differences between mPCNL and URL groups in the success rate of operation(97.8% vs 94.2%,P > 0.05),stone clearance rate(100% vs 91.8,P > 0.05)and complication rate (4.4%vs 3.9%,P>0.05). Conclusion URL combined with occlusion device can obtain satifactory results as well as the supine and lithotomy position mPCNL in the treatment of upper incarcerated ureteral stones.