To construct expressive vector for human ScFv against core protein of hepatitis C virus (HCV core ScFv),and to express soluble HCV core ScFv in E.coli JM109. Using phage display technique, the recombinant phages were panned by recombinant core antigen which was coated in a microtiter plate, and after five rounds of biopanning, 86 clones were identified specific to core antigen. 750 bp fragment could be released from the plasmid of positive phage clones, and the sequence analysis indicated that we have obrained the ScFv DNA fragment. Then DNA fragment was inserted into the expressive vector pCANTAB5E, and E. coli host JM109 was transformed and induced by IPTG. The specificity of ScFv in the culture medium was evaluated by enzyme linked immunosorbent assay (ELISA).The molecular weight of expressed HCV core ScFv protein is 28 000 dalton as determined with the aid of SDS polyacrymide gel electrophoresis (PAGE). The expressed HCV core ScFv protein will be useful in the immunohistochemical study of liver tissue from patients with hepatitis C and gene therapy against HCV infection.

Objective To investigate the effects of atorvastatin combined with aspirin on carotid atherosclerotic plaque and blood biochemical parameters in patients with minor stroke.Methods 120 patients with acute minor stroke were randomly divided into A group,B group and C group,40 cases in each group in 40 cases.On the basis of conventional treatment,A B,C groups,were given 10,20,40mg atorvastatin + 100mg aspirin,continuous medication for 6 months.Before and after treatment,the lipid metabolism and oxidation indexesindices,blood rheological parameters,carotid IMT and plaque area were compared among the three groups.Results After treatment,TC,TG,LDL,HOMA-IR,IMT and plaque area were significantly decreased in all groups (t =12.687,3.653,10.590,3.675,all P ＜0.05),and TC,TG,LDL,HOMA-IR,IMT and plaque area of C group were significantly lower than those of A group and B group,the differences were statistically significant(F =63.315,5.987,4.876,4.354,3.657,3.887,all P ＜0.05).After treatment,fibrinogen,plasma viscosity,erythrocyte rigidity index and hs-CRP were significantly decreased(t =3.437,4.325,3.876,3.487,all P ＜ 0.05).And the hs-CRP of C group were was significantly lower than those that of A group and B group,the difference was statisticallysignificant(F =4.532,P ＜ 0.05).Conclusion Atorvastatin combined with aspirin can significantly improve the carotid atherosclerotic plaque and blood biochemical parametersindexes in patients with minor stroke,and high dose atorvastatin has better effect in on patients with'carotid plaque,lipid metabolism and inflammatory reaction better effect.

Objective To explore any changes in the surface electromyography (sEMG) signals measured on the spastic upper limb muscles of stroke parents during maximum isometric voluntary contraction and to analyze any abnormal synergy patterns quantitatively in order to design better rehabilitation programs for developing coordination.Methods Ten stroke survivors with hemiparesis were selected into a patient group and ten healthy counterparts were recruited into a control group.sEMG signals were recorded bilaterally from the flexor carpi ulnaris (FCU),biceps brachii (BB),triceps brachii (TB) and deltoid (D) during maximum isometric voluntary contractions involving wrist flexion and extension,elbow flexion and extension,and shoulder abduction.The two groups' co-contraction ratios (CR) and co-activation ratios were calculated and compared.Results During elbow flexion and extension the average CR of the BB on the affected side was significantly higher than that on the unaffected side and also significantly higher than the control group average.The average CR of the TB on the affected side was significantly higher than that of the healthy controls.In all cases the average CR of the BB was larger than that of the TB.The difference in CR between the TB and the BB on the affected side was significantly larger than on the unaffected side and the control group average.During elbow flexion,the co-activation ratio of the FCU,TB and D on the affected side was significantly higher than on the unaffected side and among the healthy controls,and the co-activation ratio of the FCU on the affected side was significantly higher than that of the D and TB.During elbow extension,the co-activation ratio of the FCU,BB and D on the affected side was significantly higher in the same way,and the co-activation ratio of the FCU on the affected side was again significantly higher than that of the D and BB.During wrist flexion,the average co-activation ratio of the BB and D on the affected side was significantly greater than that on the unaffected side and among the healthy controls,and the co-activation ratio of the BB on the affected side was significantly higher than that of the D and TB.During shoulder abduction,the co-activation ratio of the BB on the affected side was significantly larger than on the unaffected side and among the healthy controls.Conclusion After a stroke the upper limbs often show flexor spasticity and abnormal synergy patterns.Rehabilitation strategy should be adopted to tackle these so as to enhance overall limb coordination.

OBJECTIVE:To observe the outcomes of umbilical cord blood mesenchymal stem call transplantation for treating neural function of multiple system atrophy (MSA) patients.METHODS:A total of 20 MSA patients were selected at the Beijing Wu Stem Cells Medical Center from January to October 2008.All patients received treatment of vessel distention,anti-free radical,trophic nerve and call membrane stabilization,as well as umbilical cord blood mesenchymal stem call transplantation via intrathecal injection.Patients at left-lateral position,and body bent at hips,knees and necks.Acupuncture was conducted at the space of lumbar vertebra 3 and 4.Following local anesthesia,No.9 needle was directly pricked into the subarachnoid cavity.2 mg dexamethasone was slowly infused,and 5 mL (5×106 stem cells) umbilical cord blood mesenchymal stern call injection was obtained and slowly infused into the subarachnoid cavity within 10 minutes,once per week,four times as a course,totally one course.We adopted Unified Multiple System Atrophy Rating Scale (UMSARS) to evaluate those MSA patients.The higher score represented a severe pathogenetic condition.RESULTS:Compared with pretransplantation,the UMSARS score was significantly decreased in 20 patients 4 weeks follwing transplantation (P ＜ 0.01).After the treatment,patient's clinical symptoms such as slow movement,balance disturbance,orthostatic hypotension,urinary and bowel disorders had full obvious improvement.Graft versus host disease was not found.CONCLUSION:It is indicated that mesenchymal stem call transplantation is effective,can partly improve MSA patients' clinical symptoms,and improve patients' life quality.

Objective To investigate the prompt value of abnormal vaginal morphology on diagnosing pelvic organ prolapse . Methods Forty eight pelvic organ prolapse female patients diagnosed by pelvic organ prolapse quantification were enrolled in the pelvic organ prolapse group and 51 normal female volunteers were enrolled in the control group in this study. Pelvic MRI T2WI were performed in all cases. The vaginal shape were evaluated according to Delancey Ⅱ level on the transverse images, which were divided into two categories:normal morphology (H-shaped) and abnormal morphology(non H-shaped). The vaginal shape distribution of different prolapse degree(0,Ⅰ,Ⅱ,Ⅲ,Ⅳstage) and types(anterior,middle, posterior pelvic prolapse) were recorded. Chi-square test was used to analyse distribution difference of vaginal shape between the two groups. The ROC curve was used to analyse the diagnostic efficiency of abnormal vaginal morphology for diagnosing pelvic organ prolapse. Results In the control group, there were 40 cases with normal vaginal morphology and 11 cases with abnormal morphology mainly including W-shaped and U-shaped abnormal morphology. In the prolapse group, there were 5 cases with normal vaginal morphology and 43 cases with abnormal morphologymainly including U-shaped (13 cases), W-shaped (26 cases) and O-shaped(4 cases) abnormal morphology. There was significant difference between the two groups(c2=46.137,P<0.01). The area under the curve (AUC) was 0.800. The sensitivity and specificity of abnormal vaginal shape for diagnosing pelvic organ prolapse were 89.6% and 78.4%respectively.The distribution of vaginal morphology in different degrees and types of prolapse were different:vaginal morphology of 0 stage prolapse showed H-typed mainly (40/51, 78.4%), Ⅰ stage prolapse showed W-shaped (16/28 57.1%), Ⅱ,Ⅲ stage prolapse all showed non H-shaped (20/20, 100%), Ⅱstage mainly showed W-shaped (9/14), Ⅲ stage mainly showed O-shaped (3/6). Anterior pelvic organ prolapse were manifested mainly with W-shaped vaginal morphology (4/9) and middle pelvic organ prolapse mainly showed O-shaped vaginal morphology (4/7). Conclusions The abnormal vaginal morphology has the prompt value on diagnosing pelvic organ prolapse.Moreover, the different shape probably indicates the different degrees and types of pelvic organ prolapse.

OBJECTIVETo search for genomic DNA sequence of the augmenter of liver regeneration (ALR) of rat.

METHODSPolymerase chain reaction (PCR) with specific primers was used to amplify the sequence from the rat genome.

RESULTSA piece of genomic DNA sequence and a piece of pseudogene of rat ALR were identified. The lengths of the gene and pseudogene are 1508 bp and 442 bp, respectively. The ALR gene of rat includes 3 exons and 2 introns. The 442 bp DNA sequence may represent a pseudogene or a ALR-related peptide. Predicted amino acid sequence analysis showed that there were 14 different amino acid residues between the gene and pseudogene. ALR-related peptide is 84 amino acid residues in length and relates closely to ALR protein.

CONCLUSIONThere might be a multigene family of ALR in rat.

Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; DNA ; genetics ; Female ; Humans ; Liver Regeneration ; genetics ; Male ; Mice ; Molecular Sequence Data ; Polymerase Chain Reaction ; Proteins ; genetics ; Pseudogenes ; genetics ; Rats

OBJECTIVE: To investigate the liver T1rho values for detecting fibrosis, and the potential impact of fatty liver on T1rho measurements. MATERIALS AND METHODS: This study included 18 healthy subjects, 18 patients with fatty liver, and 18 patients with liver fibrosis, who underwent T1rho MRI and mDIXON collections. Liver T1rho, proton density fat fraction (PDFF) and T2* values were measured and compared among the three groups. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the T1rho values for detecting liver fibrosis. Liver T1rho values were correlated with PDFF, T2* values and clinical data. RESULTS: Liver T1rho and PDFF values were significantly different (p < 0.001), whereas the T2* (p = 0.766) values were similar, among the three groups. Mean liver T1rho values in the fibrotic group (52.6 ± 6.8 ms) were significantly higher than those of healthy subjects (44.9 ± 2.8 ms, p < 0.001) and fatty liver group (45.0 ± 3.5 ms, p < 0.001). Mean liver T1rho values were similar between healthy subjects and fatty liver group (p = 0.999). PDFF values in the fatty liver group (16.07 ± 10.59%) were significantly higher than those of healthy subjects (1.43 ± 1.36%, p < 0.001) and fibrosis group (1.07 ± 1.06%, p < 0.001). PDFF values were similar in healthy subjects and fibrosis group (p = 0.984). Mean T1rho values performed well to detect fibrosis at a threshold of 49.5 ms (area under the ROC curve, 0.855), had a moderate correlation with liver stiffness (r = 0.671, p = 0.012), and no correlation with PDFF, T2* values, subject age, or body mass index (p > 0.05). CONCLUSION: T1rho MRI is useful for noninvasive detection of liver fibrosis, and may not be affected with the presence of fatty liver.
Body Mass Index ; Fatty Liver* ; Fibrosis ; Healthy Volunteers ; Humans ; Liver Cirrhosis* ; Liver* ; Magnetic Resonance Imaging ; Prospective Studies* ; Protons ; ROC Curve

Objective To improve the quality of the management of the elderly patients with cancer in Sichuan province,and to understand the attitudes and perspectives of elderly surgeons and oncologists for the management and treatment of elderly patients with cancer.Methods A face-toface questionnaire interview was conducted with oncologists (n 64) and geriatricians (n =64).128 physicians were involved in this study.Results The cancer management and therapeutics were deemed appropriate at present by 9.38％ (6/64)of the geriatricians and 25.00％ (16/64)of the oncologists.The 39.06％ (25/64) of geriatricians used to notice geriatric syndromes,while 81.25％ (52/64)of oncologists never concerned about the geriatric syndrome(P=0.011).As for the causes of the therapy-associated toxicity,oncologist versus geriatrician payed an attention to malnutrition (100.00％ vs.100.00％ in both groups),to mobility disorders(65.63％ vs.65.63％,84/128 in both groups),to cognitive impairment/mood disorder (89.06％ in geriatrician group vs.75.00％ in oncologist group,P=0.038).For the factors affecting treatment decisions,a physical ability attention (oncologists vs geriatricians:70.31％ vs.92.19％;P =0.002),and comorbidity (oncologists vs geriatricians:62.50％ vs.79.69％,P =0.032) had statistically significant difference.In addition,lack of geriatrics knowledge was also reported by more oncologists.However,one hundred percent of participants wanted very much to cooperate with each other in their clinical work.When responding to the clinical scenario,the 10.94％ (7/64)of geriatricians and 32.81％ (21/64)of oncologists chose modified treatment for 65-74 years old patient with cancer(P =0.003).When the age of the patients was 75-84 years old,only 12.50 ％ (8/64) of geriatricians prefer end-of-life care,while 31.25 ％ (20/64) of oncologists chose it (P =0.010).Conclusions Selection of treatment decisions in the elderly patients with cancer affect by ageing.Both oncologists and geriatricians are concerned with the elderly patients with cancer,ageing syndrome,total sickness and functional status.And these doctors support an establishment of a multi disciplinary team cooperation for the elderly patients with cancer.Therefore,the establishment of mutual cooperation between the two professionals is necessary and feasible.

OBJECTIVE: To investigate brain regional homogeneity (ReHo) changes of multiple sub-frequency bands in cirrhotic patients with or without hepatic encephalopathy using resting-state functional MRI. MATERIALS AND METHODS: This study recruited 46 cirrhotic patients without clinical hepatic encephalopathy (noHE), 38 cirrhotic patients with clinical hepatic encephalopathy (HE), and 37 healthy volunteers. ReHo differences were analyzed in slow-5 (0.010−0.027 Hz), slow-4 (0.027−0.073 Hz), and slow-3 (0.073−0.198 Hz) bands. Routine analysis of (0.010−0.080 Hz) band was used as a benchmark. Associations of abnormal ReHo values in each frequency band with neuropsychological scores and blood ammonia level were analyzed. Pattern classification analyses were conducted to determine whether ReHo differences in each band could differentiate the three groups of subjects (patients with or without hepatic encephalopathy and healthy controls). RESULTS: Compared to routine analysis, more differences between HE and noHE were observed in slow-5 and slow-4 bands (p < 0.005, cluster > 12, overall corrected p < 0.05). Sub-frequency band analysis also showed that ReHo abnormalities were frequency-dependent (overall corrected p < 0.05). In addition, ReHo abnormalities in each sub-band were correlated with blood ammonia level and neuropsychological scores, especially in the left inferior parietal lobe (overall corrected p < 0.05 for all frequency bands). Pattern classification analysis demonstrated that ReHo differences in lower slow-5 and slow-4 bands (both p < 0.05) and higher slow-3 band could differentiate the three groups (p < 0.05). Compared to routine analysis, ReHo features in slow-4 band obtained better classification accuracy (89%). CONCLUSION: Cirrhotic patients showed frequency-dependent changes in ReHo. Sub-frequency band analysis is important for understanding HE and clinical monitoring.
Ammonia ; Benchmarking ; Brain* ; Classification ; Healthy Volunteers ; Hepatic Encephalopathy* ; Humans ; Liver Cirrhosis ; Magnetic Resonance Imaging* ; Parietal Lobe

Objective:To investigate the related mechanism of IL-17B in regulating host immune response by studying the role and mechanism of IL-17B in the infection of Listeria monocytogenes in mice. Methods:Eighteen male C57BL/6 mice were randomly divided into three groups with six in each group: control group, PBS group and wild-type (WT) group. The control group was not given any treatment. The mice in the PBS group were injected with 100 μl of sterile PBS, while C57BL/6 mice in the WT group and IL-17B deficient (IL-17B -/-) male mice were injected intravenously with 100 μl of Listeria monocytogenes 19115 (2×10 4 colony forming unit). The mice were sacrificed 48 h after infection and then peripheral blood, spleen and liver samples were collected. Bacterial colonization in mouse spleen and liver was detected by plate count method; HE staining was used to evaluate histopathological damages; flow cytometry was used to detect the immune cells in different tissues. ELISA and qRT-PCR were used to detect the levels of IL-1β, IL-6, IL-12p40, TNF-α, IFN-γ and iNOS in serum and spleen. qRT-PCR were used to detect the expression of IL-17B and IL-17RB. Results:Bacterial colonization in mouse spleen was reduced in the IL-17B -/- group as compared with that in the WT group ( P<0.05). Compared with the PBS group, Listeria monocytogenes infection increased the expression of IL-17B and IL-17RB in mouse spleen ( P<0.05, P<0.01). There was no significant difference in the pathological damages in spleen between WT and IL-17B -/- groups. Moreover, compared with the WT group, the IL-17B -/- group showed increased macrophages, M1 macrophages ( P<0.01) and NK cells ( P<0.05) in spleen, up-regulated macrophages ( P<0.05) and M1 macrophages ( P<0.01) in the peripheral blood, enhanced expression of IL-6 in serum and spleen ( P<0.05), and promoted expression of IL-6, IL-12, IL-1β, TNF-α, IFN-γ and iNOS in spleen. Conclusions:IL-17B might inhibit Listeria monocytogenes clearance by inhibiting macrophage infiltration and the secretion of IL-6.