Objective To define the role of HCV-specific cytotoxic T lymphocytes (CTL) in chro-nic hepatitis C virus (HCV) infection. Methods HCV-specific CTL activity (HCV-CTL) was assessed in the liver and peripheral blood cells in 62 patients with chronic hepatitis C and 8 non-HCV-infection controls by using bulk-expanded liver/peripheral blood mononuclear lymphocytes (PBMC)-derived CD8+ cells as effector cells, EBV-transformed B cells as autologous target cells and recombinant vaccinia vectors expressing various regions of the HCV genome as transduction vector, in a standard chromium release assay. Results HCV-CTL activity was detected from the liver in 28 of the 60 patients (46.7%), but not from PBMC. CTL activity could not be detected from the liver and PBMC in all non-HCV-infection controls. Five patients with non-type 1 HCV infection were found to have HCV-specific CTL activity against HCV type 1 epitope. Compared with the patients without detectable HCV-specific CTL activity based on our assay, those exhibiting CTL activity had higher serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, higher histologic activity indices and lower levels of HCV RNA (all P

Objective:To investigate the accuracy of 3.0 T MR in the diagnosis of full thickness tear and partial thickness tear of the supraspinatus-infraspinatus tendon, and to analyze the causes of missed diagnosis and misdiagnosis of the supraspinatus-infraspinatus tendon tear.Methods:The MRI diagnosis report and arthroscopic surgery records of 210 patients with shoulder joint injuries in Peking University Third Hospital were analyzed retrospectively. Taking the results of arthroscopic surgery as the gold standard, the diagnostic accuracy, sensitivity, specificity, positive predictive value and negative predictive values of 3.0 T MR for full thickness tear and partial thickness tear of the supraspinatus-infraspinatus tendon were calculated, and the reasons of missed diagnosis and misdiagnosis were analyzed.Results:In 210 patients with rotator cuff injury, the diagnostic accuracy, sensitivity, specificity, positive predictive value and negative predictive value of MRI for full thickness tear of the supraspinatus-infraspinatus tendon were 81.4% (171/210), 83.1% (98/118), 79.3% (73/92), 83.7% (98/117) and 78.4% (73/93) respectively. The diagnostic accuracy, sensitivity, specificity, positive predictive value and negative predictive value of MRI for partial tears of the supraspinatus-infraspinatus tendons were 77.6% (163/210), 65.7% (44/67), 83.2% (119/143), 64.7% (44/68) and 83.8% (119/142) respectively. Among the 44 cases of missed diagnosis and misdiagnosis of the supraspinatus-infraspinatus tendon tear, 21 cases (47.7%) occurred in the junction supraspinatus and infraspinatus tendon-osseous (stop). Among the 7 cases of intra-tendon tear of supraspinatus and infraspinatus muscle, 6 cases were misdiagnosed cases, which was the most easily missed diagnosed type of partial tear. The reason for missed diagnosis was that the signal of partial tear was not high (no fluid signal was shown on T 2WI). Complex laceration was also a type of missed diagnosis in partial tears. Among the 4 cases of superior and inferior surface tendons combined with intra-tendon lacerations, 3 cases were missed diagnosed. Conclusion:3.0 T MR has higher accuracy for the diagnosis of full thickness tear and partial thickness tear of supraspinatus-infraspinatus tendon, but there is still a certain rate of misdiagnosis and missed diagnosis. A full understanding of the causes of misdiagnosis and missed diagnosis of MR in the diagnosis of rotator cuff tears of supraspinatus and infraspinatus tendons is helpful to further improve the accuracy of MR in the diagnosis of supraspinatus and infraspinatus tendon injuries of shoulder joint.

Aim To evaluate the vasorelaxant effect of two new chemical entities, J35242 and J35243, on iso-lated rat thoracic aorta rings as Rho-kinase inhibitors, and further to explore the underlying mechanisms of these two compounds. Methods Isolated rat thoracic aorta rings pre-contracted by KCl or norepinephrine ( NE) were used to evaluate the vasodilatory effect of J35242 and J35243 . Through the interventions of sev-eral tool drugs, the mechanisms of compounds concern-ing endothelium, K+ channels and Ca2+ were studied. Results J35242 and J35243 showed potent relaxant effect on both KCl and NE pre-contracted vessels, and exhibited partial endothelium dependency. L-NAME and Methylene Blue( MB) could influence the relaxant effect of these compounds. Meanwhile, the compounds could inhibit intracellular Ca2+ release and extracellu-lar Ca2+ influx, which indicated that the compounds might block the calcium channels to relax the vessels. In addition, the two compounds probably did not dilate the aorta rings through opening potassium channels. Conclusions J35242 and J35243 have vasorelaxant effects on vessels in vitro and the potency of J35242 is stronger than that of J35243 . The underlying mecha-nisms might be endothelium-dependent. Also the com-pounds might block Ca2+ channels, lowering intracel-lular Ca2+ concentration to relax the vessels.

The changes of carotid intima media thickness (cIMT) in type 2 diabetic patients with vitamin D insufficiency and subclinical macrovascular disease after 12 months' intervention therapy with calcitriol were observed.86 patients with vitamin D insufficiency [25 (OH) D＜30 ng/ml] and subclinical macrovascular disease were randomized to group A (n =44,treated with 0.5 μg/day calcitriol) or group B (n =42,blank control) for 12 months.40 patients in each group completed the follow up.After 12 months of therapy with calcitriol (0.5 μg.qd.po.),levels of high sensitive-Creactive protein,sCD36,and cIMT were significantly decreased,while 25 (OH) D and ankle brachial index were significantly increased (P＜0.05) in group A,suggesting that calcitriol could improve cIMT in patients with vitamin D insufficiency and subclinical macrovascular disease.

Aim To investigate the in vitro vasorelax-ant effect of DL0805-0, a Rho kinase inhibitor, on iso-lated rat thoracic aorta and explore its underlying mechanism. Methods Tension was measured to eval-uate the vasorelaxant effect of DL0805-0 on rat endo-thelium-intact and endothelium-denuded thoracic aorta rings. Rho kinase inhibitor fasudil, nitric oxide syn-thase inhibitor Nω-nitro-L-arginine methyl ester ( L-NAME), guanylate cyclase inhibitor methylene blue, cyclooxygenase inhibitor indomethacin, calcium-activa-ted potassium channel blocker tetraethyl ammonium ( TEA ) , ATP-sensitive potassium channel blocker glibenclamide and voltage-dependent potassium chan-nel blocker 4-aminopyridine ( 4-AP ) were used to il-lustrate the mechanisms of vasorelaxant effect of DL0805-0 . Results DL0805-0 exerted vasorelaxation in a dose-dependent manner in KCl (60 mmol·L-1 ) or NE ( 0. 1 μmol · L-1 ) -induced contraction. DL0805-0-induced vasorelaxation was significantly re-duced by L-NAME. However, methylene blue and in-domethacin did not significantly affect vasorelaxation of DL0805-0. In endothelium-denuded rings, TEA re-markably attenuated the vasorelaxant effect of DL0805-0 , while glibenclamide and 4-AP did not affect vasore laxation of DL0805-0 significantly. DL0805-0 also re-duced NE-induced transient contraction and inhibited contraction induced by increasing extracellular calci-um. Conclusion These results suggest that DL0805-0 induces vasorelaxation through an endothelium-depend-ent pathway. The opening of calcium-activated K+channels and blocking of Ca2+ channels in vascular smooth muscle cells may be one of the mechanisms of DL0805-0-induced vasorelaxation.

Aim ToestablishthemethodofHighper-formance liquid chromatography ( HPLC ) for detecting plasma concentration of indazole compound DL0805-1 , a Rho kinase inhibitor, and to investigate its pharma-cokinetics in rats with intravenous injection. Methods ThedetectingsystemwasAgilent1200-DAD;chro-matographic column was Agilent TC-C18 ( 4. 6 mm × 250 mm, 5 μm); the ultraviolet detection wavelength was 235 nm; the column temperature was 35 ℃; the flow rate was 1 ml·min-1;the mobile phase was ace-tonitrile-0. 05% H3 PO4 gradient elute. Rat blood sam-ples were collected at different intervals after intrave-nous injection of a single dose of DL0805-1 , and the concentration of DL0805-1 in rat plasma were deter-mined by HPLC method for estimating pharmacokinetic parameters.Results Afterintravenousinjectionof DL0805-1 in rats, prototype and its metabolite were detected in plasma. T1/2 of DL0805-1=(2. 34 ± 1. 42) h, Cmax=(3. 51 ± 0. 44) mg·L-1, T1/2 of metabolite of DL0805-1 = ( 1. 27 ± 0. 45 ) h, Cmax = ( 3. 55 ± 0.22)mg·L-1.Conclusion Theseresultssuggest that DL0805-1 may be metabolized into another sub-stance in vivo and play biological functions. The meth-od is sensitive, simple, and accurate, and can be used for the determination of DL0805-1 in rat plasma and pharmacokinetic studies.

BACKGROUNDTo determine the relationship between diversity of hepatitis C virus quasispecies and viremia, activity of liver disease and response to interferon therapy.

METHODSHCV quasispecies heterogeneity in 68 patients with chronic hepatitis C were detected by single stranded conformational polymorphism (SSCP) analysis of the HCV E2 hypervaribale region 1 (HVR1); of these, 48 were subsequently treated with interferon-alpha for 6 months.

RESULTSHVR1 was amplified in 61 patients. The average number of SSCP bands was 6.2+/-2.4. Quasispecies heterogeneity significantly correlated with serum HCV RNA levels (P<0.01), but not with serum ALT, AST levels and histological activity index (P>0.05). Of the patients who received interferon therapy, 43 were HVR1 positive. Patients who gained sustained response (n=11) had lower pre-treatement quasispecies heterogeneity (3.3+/-1.2) compared to those who had complete end-of-treatment response (ETR) with relapse (6.3+/-2.2, n=12, P<0.5) or no response (8.0+/-3.3, n=20, P<0.01). At the end of treatment, HVR1 could still be detected in 16 patients. The number of quasispecies heterogeneity in these patients decreased to 3.4+/-1.2, which was significantly lower than that in the patients who didn't receive interferon therapy (6.8+/-2.5, P<0.01). Of these 16 patients, 10 had change in quasispecies patterns.

CONCLUSIONSIncreased quasispecies heterogeneity can cause high HCV viremia, but it is not related to severity of liver disease. Quasispecies heterogeneity is another marker to predict the response to interferon-alpha in patients with chronic hepatitis C.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Female ; Genetic Heterogeneity ; Hepacivirus ; drug effects ; genetics ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Middle Aged ; Polymorphism, Single-Stranded Conformational ; RNA, Viral ; blood ; Viremia ; virology

OBJECTIVETo investigate the cytogenetic differences between children and adults with acute lymphoblastic leukemia (ALL) using eight-probe fluorescence in situ hybridization and karyotype analysis.

METHODSEight-probe (MYC, P16, E2A, TEL/AML1, BCR/ABL , MLL , IGH, and hyperdiploidy) fluorescence in situ hybridization and karyotype analysis were performed for 86 adults and 39 children with acute lymphoblastic leukemia.

RESULTSEight-probe fluorescence in situ hybridization showed significant differences in the positivity rate of TEL/AML1, BCR/ABL, and hyperdiploidy between adult patients and children with ALL. By karyotype analysis, the positivity rate of t(9;22) and hyperdiploidy differed significantly between the children and adult patients (P<0.05).

CONCLUSIONAdults and children with ALL have different expression profiles of the fusion genes. Eight-probe fluorescence in situ hybridization is time-saving, accurate and efficient in detecting common genetic abnormalities in ALL patients, and can be well complementary to karyotype analysis in clinical diagnosis of ALL.

Adolescent ; Adult ; Child ; Child, Preschool ; Cytogenetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infant ; Karyotype ; Karyotyping ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics ; Young Adult

Objective To investigate the immediate brain effect of acupuncture at Fengchi using amplitude of low-frequency fluctuation(ALFF)and functional connectivity by the resting-state functional magnetic resonance imaging(rs-fMRI)in patients with posterior circulation ischemia vertigo(PCIV).Methods Twenty patients with PCIV were enrolled.The dizziness handicap inventory(DHI)was used to evaluate the severity of vertigo.The patients were randomly divided into acupuncture group and sham acupoint acupuncture group.Rs-fMRI scan was performed before and after acupuncture.MATLAB-based DPABI 6.1 software was used to analyze rs-fMRI data.Correlation analysis was used between the altered ALFF values and DHI scores.The regions of altered ALFF were taken as seeds to analyze functional connectivity.Results Compared with the sham acupoint acupuncture group,the increased ALFF values were mainly located on the left precuneus,left superior frontal gyrus and left caudate nucleus after acupuncture in the acupuncture group;the decreased ALFF values were mainly located on the left cerebellum and right inferior occipital gyrus.The ALFF value of the left superior frontal gyrus was negatively correlated with the DHI score(P=0.04).The increased functional connectivity was mainly found between left precuneus and the right middle frontal gyrus,the right superior frontal gyrus,the decreased functional connectivity was mainly found between left precuneus and the bilateral paracentral lobule and right cerebellum.Conclusion The ALFF value and functional connectivity are different before and after acupuncture,indicating that the vestibular network,visual and motor brain regions functional activities are changed after needling at Fengchi,which may be the brain functional basis of Fengchi for vertigo in PCIV.

Conventional IgG is composed of heavy and light chains. The light chain has one variable region (VL) and one constant region (CL) domain, whereas the heavy chain has one variable region (VH) and three constant region domains (CH1, CH2 and CH3). Single domain antibody (sdAb) is a kind of antibody that is composed of a variable domain of heavy chain and devoid of the light chain completely. Due to its small size, it is also called as nanobody. Although the sdAb has a simple structure, it can exhibit a comparable even better antigen-binding affinity than conventional antibody. Compared with conventional antibody, the small size, high stability and simplicity in recombinant expression are representative advantages of sdAb. In recent years, scientists are becoming increasingly interested in the roles of sdAb in fundamental biomedical research and clinical application. In this review, we summarized the structural features, physicochemical properties, screening strategies and recent advances in application of sdAb.
Antibodies ; immunology