of apoptosis-related gene.

Objective　To study the clinic, neuro-electrophysiology and molecular biology of Machado-Joseph disease (MJD).Methods　Family visiting, physical examination and the blood samples were analysed on molecular biology in 44 members of a family with MJD.The cases of inpatients were examined on cerebrospinal fluid and neuro-electrophysiology.Results　10 patients of the family attacked,which were consisted with autosomal dominant inheritance type. Age of the onset was 8～38 years old. The clinical characteristic was progressive severe spinocerebellar of ataxia,faciolingual myokymia,bulging eyes.Change of denervated muscle was revealed by neuro-etectrophysiological examination. Light atrophy was observed in cerebellar,brain stem, spinal cord.The genetic defect of MJD was located the long arm of chromosome 14 between D14S280 and D14S81, their distance was 3.0 cm.All tested patients had their CAG repeated expansion from 72 to 84 in the MJD gene.Conclusion　MJD is a neuro-degenerative disorder of autosomal dominant inheritance. The disease was clinically characterized by progressive severe spinocerebellar ataxia, no obvious changes of cerebrospinal fluid,neuro-electrophysiology, CT and MRI.The genetic defect of MJD was located the long arm of chromosome 14.The number of CAG repeated expansion mutation was associated with the age of the onset.

Objective To detect hepatitis C virus (HCV) RNA in amniotic fluid of gravida and investigate mother-to-infant transmission of HCV. Methods Thirty-four HCV seropositive gravida (experimental group) were engaged. Fluorescence quantitative polymerase chain reaction (PCR) based on amplisensor assay and reverse transcription -PCR (RT-nPCR) was used. Serum HCV RNA positive sera were genotyped by RFLP analysis of PCR products from 5′NC region. Sera and amniotic fluid samples of 40 normal gravida were set as the control group. Results In the experimental group, HCV RNA was detected in amniotic fluid (5.9%, 2/34) of 2 cases. HCV RNA titers were 10 5 and 10 6 copy/ml respectively. No HCV RNA was detected in the amniotic fluid and sera of the control (n=40). Conclusions HCV RNA was rarely detected in amniotic fluid. The amniotic fluid is not the main route of HCV mother-to-infant transmission.

Objective To study the clinic, neuro electrophysiology and molecular biology of Machado Joseph disease (MJD).Methods Family visiting, physical examination and the blood samples were analysed on molecular biology in 44 members of a family with MJD.The cases of inpatients were examined on cerebrospinal fluid and neuro electrophysiology.Results 10 patients of the family attacked,which were consisted with autosomal dominant inheritance type. Age of the onset was 8~38 years old. The clinical characteristic was progressive severe spinocerebellar of ataxia,faciolingual myokymia,bulging eyes.Change of denervated muscle was revealed by neuro etectrophysiological examination. Light atrophy was observed in cerebellar,brain stem, spinal cord.The genetic defect of MJD was located the long arm of chromosome 14 between D 14 S 280 and D 14 S 81 , their distance was 3.0 cm.All tested patients had their CAG repeated expansion from 72 to 84 in the MJD gene.Conclusion MJD is a neuro degenerative disorder of autosomal dominant inheritance. The disease was clinically characterized by progressive severe spinocerebellar ataxia, no obvious changes of cerebrospinal fluid,neuro electrophysiology, CT and MRI.The genetic defect of MJD was located the long arm of chromosome 14.The number of CAG repeated expansion mutation was associated with the age of the onset.

Objective To evaluate the application of interferon-γ release assay T-SPOT.TB in diagnosis and efficacy assessment of pulmonary tuberculosis. Methods T-SPOT.TB assay was used to determine spot-forming cells (SFCs) formed by T-cells when stimulated by Mycobacterium tuberculosisspecific antigens in 55 patients with active tuberculosis,14 patients with non-tuberculosis lung diseases and 12 healthy controls. Meanwhile 20 sputum culture-positive and qualitative assay-positive pulmonary tuberculosis patients were tested with T-SPOT.TB before and at 2-month and 6-month after treatment.Kruskal-Wallis H and Mann-Whitney U test were used in group comparison.Wilcoxon test was used in comparison between pre- and post-treatment.Results The positive rate of T-SPOT.TB was significantly higher in patients with tuberculosis (85.5％,47/55 ) than that in patients with non-tuberculosis lung diseases (2/14) and the healthy controls (1/12) (x2 =40.926,P ＜0.05).The SFCs of hole A in response to ESAT-6 and hole B in response to CFP-10 in pulmonary tuberculosis group were 70.00 (27.00 -125.00) and 80.00 ( 17.00 - 180.00),respectively,which were all significantly higher than those in nontuberculosis lung diseases group and the healthy controls (x2 =35.376 and 30.485,P ＜ 0.05 ).The sensitivity,specificity,positive predictive value and negative predictive value of T-SPOT.TB in diagnosis of smear-positive tuberculosis were 88.6％,88.5％,91.2％ and 85％,while in diagnosis of sputum smearnegative tuberculosis,the sensitivity was 80％,specificity was 88.5％,positive predictive value was 84.2％ and negative predictive value was 85.2％ ( P ＞ 0.05 ).SFCs of hole A and hole B in 20 patients with sputum culture-positive and qualitative assay-positive pulmonary tuberculosis were 75.50 (41.25 -116.25 ) and 56.25 ( 105.00 -225.00) before the treatment.After 2-month antituberculosis treatment,the SFCsofhole A and hole B were 41.0 (18.0-68.75) and 72.50 (42.25- 158.75),which were significantly lower than those before treatment (Z =- 3.213 and - 3.622,P ＜ 0.05 ).Ater 6-month antituberculosis treatment,the SFCs of hole A and hole B were 25.00 (5.75 - 52.25) and 55.00 (6.25 -122.50),which were significantly lower than those before and 2-month after antituberculosis treatment (vs.before treatment:Z =- 3.921 and - 3.923,P ＜ 0.05 ; vs.2-month antituberculosis treatment:Z =- 3.926 and - 3.884,P ＜ 0.05 ).Conclusions T-SPOT.TB assay possess satisfactory sensitivity and specificity in diagnosis of tuberculosis infection,especially for sputum-negative pulmonary tuberculosis.It is also of value in monitoring antituberculosis treatment.

Aim Through interfering the expression of survivin with short hairpin RNA(shRNA) technology,to investigate the effect of downregulation of survivin on cell apoptosis and chemosensitivity to docetaxel in breast cancer MCF-7 cells.Methods(1) Using MCF-7 cells as a model system,three groups were set up transfected with lipofectamine,RNAi control plasmid and survivin RNAi plasmid,respectively.The expression of survivin in MCF-7 cells was measured at transcriptional and translational level by using RT-PCR and Western blot methods.(2) The effect on the cell cycle and apoptosis was analyzed with flow cytometry.(3) The viability of cells applied with different doses of ducetaxel was determined by using the method of 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide reduction(MTT method).Results(1) RT-PCR and Western blot demonstrated that survivin expression was significantly decreased by transfection with RNAi targeting plasmid;the expression proportion was reduced by nearly 75.4% and 79.8%(P

BACKGROUND:MicroRNAs are widely involved in the regulation of protein expression, and play a critical role in many physiological and pathological processes in the body. But microRNA expression profile in degenerative lumbar scoliosis is rarely reported and understood. OBJECTIVE:To compare the microRNA expression profile in the normal intervertebral disc and degenerative lumbar scoliosis and to identify degenerative lumbar scoliosis-specific microRNAs, folowed by functional validation. METHODS: Total RNA samples were extracted from the nucleus pulposus tissues of 57 patients with degenerative lumbar scoliosis as experimental groups and the normal nucleus pulposus tissues of 42 patients with lumbar fractures as control group. An initial screening of differentialy expressed microRNAs in the nucleus pulposus tissues by microRNA microarray was performed in 10 samples from each group. Subsequently, differentialy expressed microRNAs were validated using real-time quantitative RCR. The level of differentialy expressed microRNAs in the degenerative nucleus pulposus tissues was investigated. Then, the functional analysis of microRNAs in regulating colagen II expression was carried out. Western blot and luciferase reporter assay were also used to detect target genes. RESULTS AND CONCLUSION:We identified 22 microRNAs that were differentialy expressed (17 upregulated and 5 downregulated) in degenerative lumbar scoliosis patients compared with the controls. Folowing real-time quantitative RCR confirmation, miR-491-5p was significantly down-regulated in degenerative nucleus pulposus tissues in comparison with the controls. Moreover, its level was closely correlated with the pathological grading of disc degeneration. Overexpression of miR-491-5p promoted type II colagen expression in nucleus pulposus cels. Bioinformatics target prediction identified matrix metaloproteinase-9 as a putative target of miR-491-5p. Furthermore, luciferase reporter assays demonstrated that miR-491-5p directly targeted matrix metaloproteinase-9 and affected its protein expression in nucleus pulposus cels. These results show that the downregulation of miR-491-5p induces type II colagen loss by directly targeting matrix metaloproteinase-9, thereby resulting in degeneration of the intervertebral disc and degenerative lumbar scoliosis. This study also underscores the potential of miR-491-5p as a novel therapeutic target in degenerative lumbar scoliosis.

BACKGROUND:MicroRNAs (miRNAs) play an important role in a variety of diseases. Investigation of miRNA expression profile in degenerative lumbar scoliosis is beneficial for understanding its pathogenesis, providing a novel therapeutic target. Therefore, we tested the hypothesis that miRNAs promote intervertebral disc degeneration through the interleukin-10/STAT3 signaling pathway, a potential regulator of intervertebral disc degeneration.
OBJECTIVE:To compare the differentialy expressed miRNAs in the intervertebral disc tissues from patients with degenerative lumbar scoliosis and normal controls and to identify specific miRNAs in degenerative lumbar scoliosis folowed by functional validation.
METHODS: An initial screening of miRNA expression in nucleus pulposus tissues by miRNA Solexa Sequencing was performed in samples from 10 patients with degenerative lumbar scoliosis and 10 controls, respectively. Subsequently, differentialy expressed miRNAs were validated using qRT-PCR. The level of differentialy expressed miRNAs in degenerative nucleus pulposus tissues was investigated. Then, functional analysis of the miRNAs in regulating type II colagen expression was carried out. Western blot and luciferase reporter assay were used to further confirm the target gene.
RESULTS AND CONCLUSION: We identified 30 miRNAs that were differentialy expressed (16 upregulated and 14 downregulated) in patients with degenerative lumbar scoliosis compared with controls. Folowing qRT-PCR confirmation, Has-let-7f was significantly down-regulated in degenerative nucleus pulposus tissues as compared with controls. Moreover, its level was correlated with the severity of disc degeneration. Overexpression of Has-let-7f promoted type II colagen expression in nucleus pulposus cels. Knockout of interleukin-10 induced effects on nucleus pulposus cels similar to Has-let-7f. Bioinformatics target prediction identified interleukin-10 as a putative target of Has-let-7f. Furthermore, luciferase reporter assays demonstrated that Has-let-7f altered the expression of STAT3 and matrix metaloproteinase-2. These findings indicate that the downregulation of Has-let-7f induces type II colagen loss by directly targeting inleukin-10, thereby resulting in intervertebral disc degeneration and degenerative lumbar scoliosis. Has-let-7f is likely to be a novel therapeutic target for degenerative lumbar scoliosis.

Objective To evaluate the effect of transcatheter hepatic artery embolization with lipiodol on perfused radiofrequency ablation (PRFA) on mini-porcine.Methods Ten Chinese mini-porcine were randomly divided into embolization group and control group (each n=5) .A standard PRFA with infusing cooling procedure was done in control group,while transcatheter hepatic artery embolization with lipiodol was done before PRFA in embolization group.CT and MR scan were performed after PRFA.Then a comparative analysis of the shape and volume of the lesions in the liver were performed.Resuits In both two groups,distinct spherical margin of the PRFA lesions was obtained.The minimal diameter was (31.76±3.43) mm in embolization group and (27.23±3.49) mm in control group (t=2.675,P＜0.05) .The average volume of the embolization group was (54.47±9.98) cm~3,while in the control group was (20.90±5.68) cm~3 (t=10.424,P＜0.05) .Conclusion Transcatheter hepatic artery embolization with lipiodol before PRFA can enlarge the ablation size of mini-porcine's liver.

Objective To evaluate the effect of electric vagal stimulation on postoperative cognitive dysfunction in aged rats.Methods Thirty healthy Sprague-Dawley rats of both sexes,aged 18-20 months,weighing 390-550 g,were randomly divided into 3 groups (n=10 each) using a random number table:control group (group C),surgery group (group S),and electric vagal stimulation group (group V).Incision of the right carotid sheath was performed,and the vagal nerve was exposed after anesthesia in S and V groups.The right vagus nerve was stimulated for 30 min with continuous electric rectangular pulses (1 ms,10 Hz,1-2 V) after surgery in group V.The Morris water maze place navigation test was performed at 4,3,2 and 1 days before surgery and 2 days after surgery.The cognitive function was assessed using spatial probe and open field tests conducted at 2 days after surgery.The escape latency,the number of crossing the platform,the number of crossing the grid,the number of standing on the back legs,and the time animals spent in the central square were recorded.After the end of behavioral testing,venous blood samples were collected from the jugular vein for determination of concentrations of serum tumor necrosis factor-alpha (TNF-α),interleukin (IL)-1β and IL-6 using enzyme-linked immunosorbent assay.Results Compared with group C,the number of crossing the platform,the number of crossing the grid,and the number of standing on the back legs were significantly decreased,the escape latency and the time animals spent in the central square were significantly prolonged,and the concentrations of serum TNF-α,IL-1β and IL-6 were significantly increased at 2 days after surgery in group S (P＜0.05).Compared with group S,the number of crossing the platform,the number of crossing the grid,and the number of standing on the back legs were significantly increased,the escape latency and the time animals spent in the central square were significantly shortened,and the concentrations of serum TNF-α,IL-1β and IL-6 were significantly decreased at 2 days after surgery in group V (P＜0.05).Conclusion Electric vagal stimulation can improve the postoperative cognitive dysfunction in aged rats.