Adult ; Female ; Heart Diseases ; etiology ; Humans ; Male ; Mitochondrial Diseases ; complications ; Young Adult

Objective The construction of suicide plasmid vector could be used to make mutation of pgm gene which attenuates the virulent of Brucella melitensis strain 16, the research may lay a foundation for the development of novel live attenuated vaccines. Methods Sucrose sensitive gene as forward screening sign and fusion sequences of kanamycin resistance gene were constructed based on plasmid pucl9; pucS1.6K suicide plasmid vector was established by modifying pgm gene with fusion sequences of kanamycin resistance gene (insertion mutation); pgm gene mutation of Brucella melitensis strain 16 was obtained by electro transformation and mutation was confirmed by PCR amplification. Results The results showed that the identified Brucella melitensis strain 16 pgm gene was inactivated after insertion of kanamycin resistance gene, and the mutant pgm gene DNA fragment length was approximately 3525 bp, in line with expectations, Brucella pgm gene mutant melitensis strain 16 was successfully constructed. Conclusions The construction of suicide plasmid vector and precise mutation of Brucella melitensis strain 16 is successful, the study is not only provided an effective technology platform for constructing mutants of Brucella but also lays a foundation for the development of novel live attenuated vaccines.

Objective To observe the expression of synaptotagmin I(syt I)protein in the prefrontal cortex of adult-onset hypothyroidism rats and the effects of replicated therapy in different doses of thyroid hormone on the syt I protein.Methods All 44 aduh male Sprague-Dawley rats were divided into 4 groups randomly according to their body mass:hypothyroidism group,routine dosage thyroxine treatment group,high dosage thyroxine treatment group and control group.The adult male Sprague-Dawley rats were replicated to the adult-onset hypothyroidism and treatment models with propyhhiouracil(PTU).The levels of serum T3,T4 were assayed by the radioimmunoassay method and the level of the syt I protein in the molecular layer,external granular layer,external pyramidal layer,internal granular layer and internal pyramidal layer in prefrontal cortex was analyzed by immunohistochemistry.Results In the hypothyroidism group,the levels of serum T3 and T4[(0.34±0.04),(43.01±2.95)nmol/L]were significantly lower than those in the control group[(0.65±0.15), (55.20±3.56)nmol/L, F value: 6.026,5.940,4.503,P<0.05 or <0.01 ], the levels of the syt I protein in the molecular layer(0.018±0.010), external granular layer (0.020±0.007), external pyramidal layer(0.013±0.008), internal granular layer(0.011±0.005), internal pyramidal layer(0.024±0.013) of prefrontal lobe were significantly lower compared to the control group[(0.028±0.010,0.031 ± 0.010,0.028 ± 0.010,0.022 ± 0.008,0.038 ± 0.013), F value: 5.697,8.965,14.668,13.597,6.807,P<0.05 or <0.01 ]. In the routine dosage of the thyroxine treatment group, the levels of serum T3,T4 [(0.63 ±0.05), (55.04 ± 3.77)nmol/L] were not significantly different compared to the control group(F value: 3.162,0.367,all P>0.05), and the level of the syt I protein in the molecular layer, external granular layer, external pyramidal layer, internal granular layer and internal pyramidal layer in prefrontal cortex showed a significant improvement of the syt I protein(0.027 ± 0.013,0.025 ± 0.009,0.022 ± 0.008,0.020 ± 0.010,0.033 ± 0.010), which were similar to that of the control group(F value: 0.094,2.208,2.467,0.350,0.693, all P>0.05). In the high dosage thyroxine thyroid hormone treatment group, the levels of serum T3 and T4[ (1.11 ± 0.10), (96.68 ± 6.42)nmoL/L] were higher than the control group(F value: 6.291,12.031, all P<0.01), the expression of the syt I protein(0.028 ± 0.008,0.031 ±0.011,0.026 ± 0.012,0.023 ± 0.011,0.038 ± 0.010) were not significantly different compare to the control group (F value: 0.001,0.019,0.111,0.061,0.001, all P>0.05). Conclusions The expression of the syt I protein in the prefrontal cortex of adult-onset hypothyroidism can be decreased, which can be reversed by routine dosage of thyroxine treatment.

To evaluate the automated segmentation algorithm for detection of intra - retinal layers to process images obtained from ultra- high resolution optical coherence tomography ( OCT ) . Graph theory and the shortest path search based on dynamic programming were applied to automatically segment the 8 intra - retinal layers. We experimentally verified the accuracy and reliability of the algorithm. The results showed that the intra-retinal layer boundaries between automated and manual segmentations matched well. The algorithm successfully segmented the intra- retinal layers in glaucoma, high myopia, and retinitis pigmentosa patients. The proposed automatic segmentation for intra-retinal layers provides a promising tool for quantitative analysis in clinical diagnosis and treatment.

Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Fibroma ; metabolism ; pathology ; surgery ; Humans ; Keratins ; metabolism ; Middle Aged ; Vimentin ; metabolism

Objective To observe the MRI features of traumatic brainin jury to corpus callosum and provide valuable diagnostic information for clinical application. Methods Clinical and radiological data were retrospectively analyzed in 20 MRI-diagnosed patients as having traunmtic brain mjury to corpus callosum. Results Traumatic injury to corpus callosum was rather rare in brain trauma.Most traumatic injury to corpus callosum was non-hemorrhagic and its lesion mainly laid at its body or splenium,sometimes at genu,but rarely at rostrum.Traumatic injury to corpus callosum,being one of the common manifestations in diffuse axonal injury(DAI), could be demonstrated slight hyperintense signal on TI WI and T2WI in early phase.The injury site shrank and liquefaction necrosis appeared in the lesion at the late-mid-injured period with softened focus or gliai scar.MRI showed hyperintense T1 and T2 signal, similar to the signal of the cerebrospinal fluid and demonstrated ventriculomegaly in the corresponding parts.DWI showed hyperintense signal in the lesion in acute and subacutc stages:with time prolonged,isointense signal of the normal brain tissue was found and signal of the cerebrospinal fluid appeared in the present of softened focus.Conclusion Attention should be paid to the traumatic injury to corpus callosum,an indicator for severe trauma or disease.MRI,a sensitive and multi-directional imaging,is superior in detecting non-hemorrhagic injury to the corpus callosum.

Thermodynamic parameters of complexation of naphto-15-crown-5 with four alkaline earth ions in aqueous media was determined using titration microcalorimetry at 298.15 K. The stability of the complexes, thermal effect and entropy effect of the complexation is discussed on the basis of the guest ions structure and the solvent effect. The stability constants tendency to vary with ion radius was interpreted. Complex of naphtha-15-crown-5 with calcium ion is very stable due to the synergism of static electric interaction and size selectivity between the host and the guest.
Calorimetry ; methods ; Crown Ethers ; chemistry ; Ions ; chemistry ; Macromolecular Substances ; analysis ; chemistry ; Metals, Alkaline Earth ; chemistry ; Microchemistry ; methods

AIM:To investigate the inhibitory effect of sinapine thiocyanate(ST)on hyperglycemia,hyper-lipemia,atherosclerosis and hepatocellular steatosis of ApoE-/-mice with insulin resistance(IR)and the possible mecha-nisms.METHODS:ApoE-/-male mice(n=60)were assigned randomly into control group ,saline group,rosiglitazone group and ST treatment groups(at low,middle and high doses )with 10 mice in each group.The mice in control group were fed with fundamental diet ,while the mice in other groups were fed with high-fat diet for 12 weeks.The mice in ST groups were given gavage with different doses of ST(10,30 and 90 mg· kg-1· d-1)simultaneously,while the mice in rosiglitazone group received gavage with rosigltazone(1.33 mg· kg-1 · d-1 ).In the last 3 weeks,the mice in control group received daily intrape-ritoneal injection of physiological saline ,and IR was induced in other groups by daily intrape-ritoneal injection of dexamethasone(0.8 mg/kg).The blood sample was collected and fasting plasma glucose was tested weekly through tail vein.After all animals fasted for 12 h at the end of the 12th week,they were sacrificed and the levels of fasting insulin,tumor necrosis factor-α(TNF-α),triglyceride,total cholesterol and liver lipids were measured.The li-ver tissue and aortic immobilized sections were detected by HE staining.The expression of the proteins related to liver lipid metabolism and skeletal muscle MAPK signaling pathway was determined by Western blot.RESULTS:ST showed dose-dependently reduced serum lipids ,plasma glucose and TNF-α(P<0.05),delayed hepatocellular steatosis and atheroscle-rosis,and dose-dependently regulated hepatic lipid metabolism signaling molecules(HMGR and SREBP-2)and MAPK signaling molecules(ERK and p38)(P<0.05).CONCLUSION:ST has the biological potential of reducing blood li-pids and relieving IR.The mechanism may be related to the regulation of liver lipid metabolism and skeletal muscle MAPK signaling pathway.

OBJECTIVEThis study was designed to investigate the significance of hTERT mRNA in breast carcinogenesis and to explore the diagnostic efficacy, and to study the effect of tumor suppressor gene p53 on the expression of hTERT mRNA.

METHODSThe expression of hTERT mRNA was examined by in situ hybridization in 12 cases of normal breast tissue nearby cancer, 7 of simple ductal hyperplasia, 20 of atypical hyperplasia, 18 of ductal carcinoma in situ and 25 with invasive ductal carcinoma. The expression of p53 protein were examined by immunohistochemistry in 43 carcinomas.

RESULTShTERT was not detected in normal breast tissue nearby cancer and simple ductal hyperplasia. The positive rate of hTERT mRNA in atypical hyperplasia, ductal carcinoma in situ and invasive ductal carcinoma were 25.0%, 83.3% and 88.0%, respectively. The prevalence and intensity of hTERT mRNA expression were much greater in carcinoma than those in simple or atypical hyperplasia and normal breast tissue nearby cancer (P < 0.05). The expression of hTERT was not correlated with tumor size and lymph node metastasis (P > 0.05). The positive correlation between hTERT mRNA and p53 was found in breast carcinoma (r = 0.5540, P < 0.01).

CONCLUSIONhTERT mRNA expression is closely related to the malignant transformation of breast tissue. Semi-quantitative detection of hTERT mRNA expression in situ is helpful in differentiated diagnosis of carcinoma in situ and atypical hyperplasia. Inactivation of p53 may play a role in the transcriptive activation of hTERT gene in breast carcinoma.

Adult ; Breast ; metabolism ; pathology ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Diagnosis, Differential ; Disease Progression ; Humans ; Hyperplasia ; Lymphatic Metastasis ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics ; Telomerase ; biosynthesis ; genetics ; Tumor Suppressor Protein p53

OBJECTIVETo study the expression of E-cadherin and beta-catenin in neuroblastomas of various degrees of differentiation, and to investigate their molecular mechanisms in correlation with clinicopathologic parameters.

METHODSImmunohistochemistry EnVision method was used to detect E-cadherin and beta-catenin expression in 90 paraffin-embedded tissue samples of neuroblastomas. The methylation status of CpG islands of E-cadherin promoter was investigated by MSP in 7 fresh tissue and 24 paraffin-embedded tissue samples. The mutation status of exon 3 of beta-catenin gene was studied by PCR in 7 fresh tissue samples. Statistical analysis of the data was performed by SPSS software.

RESULTSE-cadherin and beta-catenin were abnormally expressed in neuroblastomas in general. The expression of beta-catenin in well-differentiated neuroblastoms was markedly higher (47/70, 67.1%) than that of the poorly differentiated tumors (8/20, 40.0%). There was a markedly decreased expression of both genes in tumors with lymph node metastasis than those without. Demethylation was seen in some regions of the promoter of E-cadherin in 31 cases of nuroblatomas. PCR of the exon 3 of beta-catenin followed by DNA sequencing demonstrated rearrangements and mutations in 7 cases, including 2 cases harboring identical point mutation at gene position 27184, leading to a T-->A alteration.

CONCLUSIONSThe abnormal over-expression of E-cadherin in neuroblastomas is independent of the methylation status of their promoter sequences. The abnormal expression of beta-catenin may be related to mutational changes at exon 3 of the gene.

Cadherins ; genetics ; metabolism ; Child ; Child, Preschool ; CpG Islands ; genetics ; DNA Methylation ; DNA, Neoplasm ; genetics ; Exons ; Female ; Ganglioneuroblastoma ; genetics ; metabolism ; pathology ; Gene Rearrangement ; Humans ; Infant ; Lymphatic Metastasis ; Male ; Mediastinal Neoplasms ; genetics ; metabolism ; pathology ; Neuroblastoma ; genetics ; metabolism ; pathology ; Point Mutation ; Promoter Regions, Genetic ; genetics ; Retroperitoneal Neoplasms ; genetics ; metabolism ; pathology ; Sequence Analysis, DNA ; beta Catenin ; genetics ; metabolism