Aim To analyze differential expression and interaction of β-catenin/ICAT proteins in HL60 cells when they were induced into monocytic differentiation, and to figure out the mechanism of NSC67657 in cellu-lar induction. Methods HL60 cells were treated by 10 μmol · L-1 NSC67657 , and cellular differentiation could be observed by cytochemical staining and flow cytometry. Then, RT-PCR and Western blot were em-ployed to determine the differential expression of β-catenin/ICAT genes and proteins. Co-immunoprecipi-tation assay was used to confirm the interaction of β-catenin/ICAT proteins, and laser co-focus light mi-croscopy technology was used to co-indentify proteins differential expression and intracellular location. Re-sults HL60 cells could be induced into monocytic dif-ferentiation after 5 days treatment using 10μM NSC67657 . The CD14 ( +)% cells could be up to o-ver 90%, and cytochemical staining reports were con-sistent with this result. The expressions of ICAT gene and protein were up-regulated significantly ( P <0. 01 ) , but the expressions ofβ-catenin gene and pro-tein, on the contrary, were down-regulated(P<0. 05) when HL60 cells were induced into monocytic differen-tiation. From co-immunoprecipitation assay findings, ICAT protein interacted with β-catenin protein, and the absorbance of protein electrophoresis bands in-creased in differentiated cells. From laser co-focus light microscopy assay findings, the fluorescence of ICAT and β-catenin protein could be both observed in cytoplasm and nucleus. In drug treated HL60 cells, the fluorescence of ICAT protein was enhanced both in cytoplasm and nucleus, however, the fluorescence ofβ-catenin protein, which looked like transferring into different organelles, decreased significantly in nucleus, but increased in cytoplasm. Conclusions HL60 cells could be induced into monocytic differentiation by NSC67657 and β-catenin/ICAT proteins differentially expressed during cellular differentiation. The enhanced interaction of β-catenin/ICAT proteins and β-catenin protein transferring from nucleus into cytoplasm indi-cates that NSC67657 probably induces HL60 cells into monocytic differentiation through down-regulating β-catenin protein and blockingβ-catenin protein from nu-cleus.

Objective To investigate the relationship between HBV genotypes and YMDD motif mutations or BCP mutations in Xiangtan of Hunan Province. Methods HBV genotypes, YMDD motif mutations and BCP mutations were analyzed in 952 HBV infected patients. Results HBV genotyping showed that 698 HBV type B patients and 115 HBV type C patients accounted for 73.32% and 12.08% respectively of all the participants. The rest 139( 14.60% )were genotype B and C mixed infection( B + C ). The analysis of YMDD motif mutations showed that 844 YMDD wild-type which accounted for 88.66% of all the subjects and the remainder were YMDD mutation types, of which 54( 5.67% ) carried YVDD, 53( 5.57% ) YIDD,and 1 YVDD and YIDD mixed infection. Basic Core Promoter mutations showed that 1762A/1764G ( wild type )accounted for 70.59% and 1762T/1764A( mutant ) accounted for 19.75%. The rest 92 patients were 1762T/1764A and 1762A/1764G mixed infection. This study showed no significant difference in the rate of YMDD mutation( 10.04% vs 10.43% ,χ2 =0.017,P＞0.05 ) ,but a significant difference in the types of YMDD mutation(χ2 = 4.836, P ＜ 0.05 )between HBV types B and C. The YVDD mutation was more commonly seen in genotype C( 9.57% ) than in genotype B( 5.88% ). The BCP mutation rate showed a significant difference( 27.36% vs 46.09%, χ2 = 16.478, P ＜ 0.01 ). Genotype C was more frequent than genotype B. The BCP mutation rate showed no significant difference between YMDD Wild-type and YMDD mutation types( 28.67% vs 35.51%, χ2 = 2.139, P ＞ 0.05 ), but most of BCP mutations happened in YVDD mutant type( 61.11% ). Conclusions ( 1 ) The predominant HBV genotypes in Xiangtan were genotype B and genotype C, the major genotype was type B, which display the characteristics of epidemiology in Southern China. ( 2 ) Determination of HBV genotypes before lamivudine therapy was probably not an important pretreatment investigation to predict antiviral responses. ( 3 ) Detection of HBV genotypes, YMDD motif mutations and BCP mutations will contribute to the correct evaluation of prognosis and timely proper management of HBV patients.

Objective To investigate the optimal b value of diffusion-weighted imaging (DWI)in predicting the grade of cerebral gliomas.Methods 38 patients with pathologically-proved brain gliomas (24 high-grade gliomas and 14 low-grade ones)were studied retrospectively.All patients received conventional MRI and DWI examination with three different b values (1 000 s/mm2 ,2 000 s/mm2 and 3 000 s /mm2 ).The signal features of the tumor on DWI with three different b values were evaluated and compared.The minimum ADC values of the tumors were calculated and compared between each other.Through drawing the ROC curve of different b values, the best diagnostic threshold was found.The sensitivity and specificity in predicting the grade of brain gliomas were assessed using Chi-square test.Results On DWI with b value of 3 000 s/mm2 ,91.6% (22/24)of high-grade tumors showed hyper-intensity, while 85.7%(12/14)of low-grade tumors presented hypo-intensity signals.Regarding hyper-intensity signals as diagnostic criterion for high-grade tumors,the sensitivity and specificity were 91.6% and 100% respectively.When the b value was similar,the minimum ADC value of high-grade glioma was significantly lower than that of low-grade glioma,and statistically significant differences in differentiating high-grade glioma from low-grade glioma existed among three b values (P <0.05).When the ADC 3 000 value<0.74×10 -3 mm2/s was regarded as a standard for the identification of high-grade and low-grade glioma,the sensitivity and specificity were 100% and 87.3%respectively.Conclusion DWI with high b value is more useful than standard moderate b value in preoperative grading the gliomas.When DWI with b value of 3 000 s/mm2 is used,the minimum ADC value will provide quantitative indicators in preoperatively precisely predicting grading glioma.

Objective To study the composition of variant and point mutations of Norovirus GⅡ.4 in Guiyang regions.Methods From June to November 2010,cases information and fecal specimens were collected from guard-hospitals in Guiyang regions,who had caught the acute-gastroenteritis.Noroviruses in specimens were detected by a real-time reverse transcription polymerase chain reaction(real-time RT-PCR),and then partial genotyped norovirus-positive clinical samples (in random) were cloned and sequenced in VP1 gene code.Furthermore,the gene sequences were compared with the published variants at home and abroad of norovirus(GⅡ.4),including the phylogenetic analyses of genomes and variation of amino acids within individual sites.Results Those 267 specimens were GⅡ-norovirus-positive(62.68％) in 426 clinical samples.There were nine GⅡ.4-norovirus-positive VP1 gene-sequences available,and two subtype-norovirus variants (GⅡ.4 2008a and G Ⅱ.4 2008b variant) were epidemic in 2010,Guizhou province.The homology between and in subgroups were 95.90％-96.72％ and 99.45％-100％.Two amino acids within individual sites were apt to mutate.Conclusion Norovirus GⅡ genotype were predominant in summer and fall acute gastroenteritis in 2010 for Guiyang regions,and the variants were diversity.

OBJECTIVE: To analyze the rate of 235delC mutation in GJB2 gene in patients with idiopathic sudden hearing loss, and to explore its possible correlation with pathogenesis of idiopathic sudden hearing loss. METHOD: Two hundred and thirty-four patients with diagnosis of idiopathic sudden hearing loss in otolaryngology department were recruited as experimental group. Eighty people with normal hearing level were enrolled as control group. Their peripheral blood samples were obtained and genomic DNA was extracted. Using polymerase chain reaction, the coding region of GJB2 gene was amplified, and 235delC mutation is screened for in GJB2 gene by restriction endonuclease. At same time the clinical data of 234 patients was collected to analyze. RESULT: In 234 cases of idiopathic sudden hearing loss, 5 cases were found to have heterozygous 235delC mutation, none of them harbored homozygous 235delC mutation, the 235delC mutation rate was 2.1% (5/234). No 235delC mutation was found in control group. The rate of 235delC mutation in two group showed no statistically significant difference (P > 0.05). CONCLUSION This research shows that the rate of 235delC mutation in GJB2 is low in patients with idiopathic sudden hearing loss, and suggest that 235delC mutation possible has no correlation with idiopathic sudden hearing loss.
Adolescent ; Adult ; Aged ; Child ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; Female ; Hearing Loss, Sudden ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Young Adult

Objective To evaluate the value of dual energy computed tomography (DECT) for the diagnosis of gout.Methods Twenty-fivc consccutive patients with gout were selected as the experimental group and 20 patients with rheumatoid arthritis (RA) were selected as the control group.DECT scans were performed for every patient (all peripheral joints for the experimental group and affected joints for the control group).And every patient with gout had X-ray examination of the involved joints.x2 test and t-test were used for statistical analysis.Results All 25 patients with gout showed urate deposits on their DECT scans,whereas none of 20 controls showed urate deposits (P＜0.01).DECT scans revealed a total of 184 areas of urate deposition in 25 patients,in which 107 (58.2％) were at feet and ankle,72 (39.1％) were at knees,4 (2.2％) were at hands and wrists,1 (0.5％) was at elbows,whereas physical examination only showed 64 areas of urate deposition,in which 38(59.4％) were at feet and ankle,24(37.5％) were at knees,2(3.1％) at hands and wrists,0 was at elbows(P＜0.01 ).Only 6 patients with gout showed nonspecific manifestations on X-ray.Conclusion DECT scans may have potential value for the diagnosis of gout since it could produce evident colour displays for urate deposits and help to identify subclinical tophus deposits.

AIM To study the effects of nalox-one on plasma endothelin-1 and nitric oxide during myocardiac ischemia-reperfusion ( I/R ) injury. METHODS Using myocardiac ischemia models and myocardiac ischemia -reperfusion injury models that was made by means of ligating sinistra corona-ria arteria,to investigate the change of plasma ET-1 and NO during I/R injury, and after the protection and treatment with naloxone,an antagonist of opoid receptor. 40 New Zealand rabbits were randomly assigned to 4 groupsCischemia group, nalox-one protection group, naloxone treatment group and ischemia-reperfusion group, 10 in each group). The blood was phlebotomized at different time in each group. The concentration of ET-1 was detected with radioimmunology method and NO with nitrate reductase method. RESULTS The levels of ET-1 had the trend of improvement after ischemia and were at its peak at the end of 4 h, but the levels of NO were significantly decreased. The ET-1 levels were significantly improved after 0. 5-1 h of injury compared with that before ischemia (P 0. 05). The levels of NO decreased after injury , whereas its levels in naloxone protection group increased significantly compared with that before ischemia ( P 0.05). CONCLUSION Naloxone may effectively reduce the level of ET-1 and enhance the level of NO after myocardiac ischemia and during I/R injury; whereby it decreases the injury to vascular and myocardium.

Objective To observe the interference of glucose-6-phosphate dehydrogenase (G6PD) deficiency on glycated hemoglobin (HbA1c) detected by three measurement systems.Methods A total of 286 cases of blood and serum samples were collected at Zhongshan Hospital of Sun Yat-Sun University from August 2012 to April 2016.The blood samples were divided into healthy control group (122 cases),diabetes group (82 cases),glucose-6-phosphate dehydrogenase deficiency group (61 cases) and diabetes with G6PD deficiency group (21 cases).The levels of HbA1 c were detected by three measurement systems,including Primus Ultra2,Variant lⅡ Turbo 2.0 and Modular P.The results of HbA1c were converted into the estimated average blood glucose concentration (eAG).The values of A eAG-FPG in different groups were calculated and statistical analysis was performed for evaluation of the differences from the three measurement systems.Results The HbA1c results measured by the three systems and AeAG-FPG values in G6PD deficiency group were all lower than healthy control group(all P ＜0.05).The measured results were similar in both diabetes group and diabetes with G6PD deficiency group.Conclusion G6PD deficiency may cause false H-bA1c results detected by three measurement systems.In the case of HbA1c for evaluating blood glucose control,the interference of G6PD deficiency should be noticed.

The protective roles of alpha-lipoic acid in the rat model of mitochondrial DNA (mtDNA) 4834bp deletion in inner ear were investigated. Forty female Wistar rats at 4 weeks of age were divided into four groups: group A (D-galactose group, n=10), group B (D-galactose+alpha-lipoic acid group, n=10), group C (alpha-lipoic acid group, n=10), and group D (control group, n=10). Auditory brainstem response (ABR) was used to detect the hearing threshold. Colorimetry was used to analyze activity of superoxide dismutase (SOD) and concentration of malondialdehyde (MDA). The percentage of mtDNA4834bp deletion in inner ear was identified by real-time PCR. There was no significant difference in ABR threshold shift among all groups. The percentage of mtDNA4834bp deletion in group A was higher than that in other groups, but there was no significant difference in percentage of mtDNA4834bp deletion among groups B, C, and D. The activity of SOD in group A was lower than that in other groups. The concentration of MDA in group A was higher than that in other groups. It was concluded that there was no significant hearing loss when the percentage of mtDNA4834bp deletion was lower than 12.5%. alpha-Lipoic acid could prevent the reactive oxygen species (ROS)-induced mtDNA4834bp deletion in inner ear of rats.

OBJECTIVE: To observe the expression of mGluR5 in the medial vestibular nucleus (MVN) following unilateral labyrinthectomy (UL). METHOD: Thirty Wistar rats were randomly divided into two groups. Twenty four animals received unilateral labyrinthectomy while the others maintained labyrinthine well. After setting left labyrinthine, the change of mGluR5 was induced by immunohistochemistry, in situ hybridization. RESULT: mGluR5 was increased in lesioned side MVN after unilateral labyrinthectomy. The 12 h post-UL was highest. Then it was decrease in 36 h post-UL, while 7 d post-UL was same as control group. The change of contralateral was same as that in ipsilateral. CONCLUSION UL can induce increase of mGluR5 in the MVN. The reduced resting discharge in the primary vestibular afferents or in the central vestibular neurons may be responsible for the change of mGluR5. However the significance of the change of mGluR1 in the vestibular compensation is still unknown.
Animals ; Male ; Postoperative Period ; Rats ; Rats, Wistar ; Receptor, Metabotropic Glutamate 5 ; Receptors, Metabotropic Glutamate ; metabolism ; Vestibular Nuclei ; metabolism ; Vestibule, Labyrinth ; metabolism ; surgery