OBJECTIVETo observe the relation between Pi deficiency syndrome (PDS) and the configuration and functions of extensor digitorum longus (EDL)and soleus (SOL).

METHODSTotally 36 ICR mice were randomly divided into 3 groups according to weight matching principle, the control group, the exhausted group, and the rhubarb group, 12 in each group. Two PDS models were established by either purgation with rhubarb diarrhea (as Group A) or exhausted swimming plus sleep deprivation (as Group B).The cross sectional area (CSA) of type I and II fibers of extensor digitorum longus (EDL) and soleus (SOL), relative proportions of type I and II fibers were measured by m-ATPase histochemical method. The isotonic contraction and the maximum tetanus contraction of EDL and SOL were detected by PowerLab system.

RESULTSCompared with the control group, the body weight, body temperature, and the general health condition of PDS model rats obviously decreased; the spleen index and the thymus index were also lower; the maximal isotonic contraction and the maximum tetanus contraction obviously decreased; the cross section areas of EDL and SOL were reduced with loosely arranged cells. In EDL, the proportion of type I fibers was added and the proportion of type II fibers was lowered. In SOL, there was no change in the proportion of type I and type II fibers.

CONCLUSIONSEDL and SOL were obviously atrophied in the two PDS model mice. The type I fibers of SOL was more significantly atrophied in Group B.

Animals ; Disease Models, Animal ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred ICR ; Muscle, Skeletal ; physiopathology ; Rats

Objective To clne huma PAIl gene and prepare its monoclonal antibodies(McAbs) for determination of its expression in breast cancer cells.Methods Human PAI1 gene eDNA was amplified by RT-PCR from human breast cancer cell line MDA231 and inserted into the prokaryotic expression vector, which expressed fusion protein of MS2-PAI1 in E.coli.Fusion protein of MS2-PAI1 was purified and used for immunizing BALB/C mouse.Traditional hybridoma technology was used to produce hybridoma cells for preparation of monoclonal antibodies.Western blot and immunohistochemistry were used to detect PAI1 expression in breast cancer.Results The 1209 bp full PAI1 gene was cloned.The two hybridoma cell lines that secreted specific monoclonal antibodies against human PAI1 were identified by ELISA.The immunoglobulin subclasses of the McAbs were IgG1.The McAbs can specifically recognize PAI1 but not other proteins.Western blot showed that the McAbs against PAI1 can specifically react with MS2-PAI1 fusion protein and endogenous proteins in cells.The positive reaction was found in breast cancer cell line MDA231 and breast cancer tissues by immunochemical staining.Conclusions The McAbs against human PAI1 are successfully prepared by hybridoma technology with MS2-PAI1 fusion protein expressed in E.coll.It has been shown that PAI1 can be expressed in MDA231 and breast cancer tissues.The McAbs against PAI1 could be a useful tool for the further study of the human PAI1 functions and detection of clinical tumor samples.

OBJECTIVETo analyze the clinical characteristics of pediatric neuromyelitis optica (NMO) and neuromyelitis optica spectrum disorders (NMOSD).

METHODA retrospective analysis was performed evaluating clinical and laboratory characteristics of ten NMO and NMOSD children who were seen in our hospital from December 2010 to May 2014. Median age at onset was 8.9 years (range 0.8-13.8 years). Seven cases were female and three were male. Median disease duration was 1.5 months (range 1-18.5 months).

RESULTEight patients fulfilled diagnostic criteria for NMO and two patients fulfilled diagnostic criteria for NMOSD. The two NMOSD patients had recurrent longitudinally extensive transverse myelitis. Four cases had a monophasic disease course, and six cases had a recurrent course. In eight NMO patients, neuritis was the initial presentation. The two NMOSD patients had no neuritis in the first attack. Nine cases had clinical manifestations of myelitis, one case had asymptomatic spinal cord MRI anomaly. Among the ten patients, seven cases had brain lesions, wherein, four cases had the midbrain involvement and in four cases extensive hemispheric white matter was involved. Three cases had medullary involvement. And two cases had posterior limb of the internal capsule involvement, two cases had thalamus involvement. In one case there was pons, cerebellum or corpus callosum involvement, respectively. One case had accompanied brain symptoms. Of the five patients who had symptomatic brain lesions, four cases had encephalopathy accompanied by large hemispheric lesions on MRI, having a presentation similar to acute disseminated encephalomyelitis. And one case had multiple sclerosis like brain lesion. Of the ten patients tested, nine were seropositive for anti-aquaporin-4 autoantibody. One-patient was complicated with systemic lupus erythematosus. Oligoclonal bands were negative in all cases. All patients received treatment for acute attacks with high-dose intravenous methylprednisolone and intravenous gammaglobulin. The symptoms of 8 cases mitigated. Two cases whose symptoms showed no sign of improvement received plasmapheresis for acute attacks. Seven of the patients were followed up. The median duration of follow-up was 19 months (ranged from 13 months to 30 months). The median Expanded disability status (EDSS) score was 3 (range 1-7).

CONCLUSIONPediatric NMO and(or) NMOSD have a diverse clinical presentation which are more than just optic neuritis and transverse myelitis, including brain symptom. So it may be difficult to distinguish NMO and( or) NMOSD from acute disseminating encephalomyelitis and multiple sclerosis in the early stages of the disease. Antibodies to aquapoin-4 (AQP-Ab) testing is very important for differential diagnosis.

Adolescent ; Anti-Inflammatory Agents ; therapeutic use ; Aquaporin 4 ; Autoantibodies ; Brain ; Brain Diseases ; Child ; Child, Preschool ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Methylprednisolone ; therapeutic use ; Multiple Sclerosis ; etiology ; Neuromyelitis Optica ; complications ; diagnosis ; drug therapy ; Retrospective Studies

The second nonligand binding chain of IL-6 receptor (IL-6R), the membrane glycoprotein with 130 kD mol wt (gp130), is responsible for the signal transduction of IL-6 biological activity. Our experiments indicated that the rat gp130 molecule, which was expressed in the rat acute myeloid leukemia cell line R2, could associate with the complex of rhIL-6 and membrane human IL-6R molecule and transduce the inhibition-inducing signal on R2 cells. In the present study, antisense oligonucleotides of rat gp130 were synthesized and uptaked into the R2 cells. Then the effects of the antisense or sense nucleic acids of rat gp130 on the inhibition induced by rhIL-6 in the R2 cell were investigated. Our results show that the antisense oligonucleotide of rat gp130 blocked the inhibitory effect of rhIL-6 on the R2 cells by (45 +/- 7)% at the optimal concentration
Acute Disease ; Animals ; Antigens, CD ; genetics ; Cell Division ; drug effects ; Cytokine Receptor gp130 ; Drug Interactions ; Interleukin-6 ; antagonists & inhibitors ; pharmacology ; Leukemia, Myeloid ; pathology ; Membrane Glycoproteins ; antagonists & inhibitors ; genetics ; Oligonucleotides, Antisense ; chemical synthesis ; chemistry ; pharmacology ; Rats ; Tumor Cells, Cultured

Objective To evaluate the dynamic contrast-enhanced MRI (DCE-MRI)in detecting experimentally induced testicular ischemia. Methods Thirty healthy male New Zealand rabbits were randomly assigned into 6 groups. There were 5 rabbits in each of the following experimental groups: ( 1 ) Normal control, (2) Sham-operated, (3) ischemia of 3 h group, (4) ischemia of 6 h group, (5) ischemia of 12 h group, (6) ischemia of 24 h group. In all experiment groups, the right testis served as the internal control while the left testis served as the experimental side. DCE-MRI for each animal lasts about 10 minutes. Signal enhanced ratios (SERs) of ROI for both sides of each group were calculated by a computer, and parameters of SERs of 30 s, 75 s, 120 s and maximal SER were used for statistical analysis.Time intensity curves (TICs) were made for two sides of each group via Excel 2003 software and classified into 4 types. Statistical analysis was performed to compare the differences of SERs between left and right testis by two independent Kolmogorov-Smirnov test. Results In group I and 2, significant enhancement was observed on both testes of 10 rabbits. The enhancement decreased gradually with the elongation of ischemia in torsion groups. Three cases of type Ⅰ and 2 cases of type Ⅱ were observed in group 1,5 cases of type Ⅰ in group two, 2 cases of type Ⅰ and 3 cases of type Ⅱ b in group three, 2 cases of type Ⅰ and 2 cases of type Ⅱ b in group four, 5 cases of type Ⅱ b in group five and 5 cases of type Ⅲ in group six were noticed in the left testes. And in TICs of right testes, all cases showed TICs of type Ⅰ except 2 cases of type Ⅱ a in group six. In four torsion groups, the values for SER75 of the left side were 0. 084%, 0. 076%, 0.164% and 0.065%, while the right side were 0.255%, 0.410%, 0.586% and 0.302% (P ＜0.05). The values for SER120 in group three, five and six were 0.221% , 0.158% and 0.059% for the left side, and 0.405%,0.522% and 0.207% for the right side(P ＜0.05). The values for MSER in group three, five and six were 0.217% ,0.164% and 0.072% for the left side, and 0.405%, 0.586% and 0.302% for the right side(P ＜0.05). Conclusion DCE-MRI technique may be useful in the diagnosis of testicular torsion, which shows potential in the clinical application.

OBJECTIVETo investigate the relationship between the frequency of BCL-2/IgH rearrangement in peripheral blood cells of healthy Chinese individuals of Han nationality located in Zhejiang area and the low incidence of follicular lymphoma (FL).

METHODSNested-PCR and direct DNA sequencing were used to detect the Bcl-2/IgH rearrangement in peripheral blood cells of 196 healthy individuals. DNA sequences involved were then searched and aligned in NCBI database to confine the broken points in major breakpoint region and the IgH segments involved.

RESULTSFirst, in this sample the frequency of BCL-2/IgH translocation in Chinese individuals of Han nationality located in Zhejiang area is 9.66%, being much lower than that in North America and Europe countries. Second, the breakpoints tend to fall into 3 clusters: 3055, 3116 and 3165 bp. Usage of J6 segment is most common. Third, There are different subclones of BCL-2/IgH rearrangements in the same individual.

CONCLUSIONThe low frequency of BCL-2/IgH translocation in healthy Chinese individuals of Han nationality located in Zhejiang area may be one of the reasons for the difference in the incidence of FL between China and Western countries.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 18 ; Female ; Gene Rearrangement ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Lymphoma, Follicular ; ethnology ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Polymerase Chain Reaction ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Translocation, Genetic ; Young Adult

OBJECTIVETo investigate COX-2 expression in patients with gastric cancer and its relationship with angiogenesis and clinicopathologic features of gastric cancer.

METHODSCOX-2 expression and CD34-stained microvessel density (MVD) were detected by immunohistochemical methods in specimens from 96 patients with gastric cancer. The correlations among COX-2 expression, MVD and clinicopathologic features were analyzed.

RESULTSThe COX-2 positive rate and MVD in gastric cancer were significantly higher than those in the normal gastric mucosa (80.2% vs. 13.3%; 32.5+/- 8.3 vs. 13.1+/- 2.4, all P< 0.01). The COX-2 positive rate and MVD in the patients with stage III and IV were significantly higher (91.4% and 34.9+/- 8.7 respectively, P< 0.01), than that in the patients with stage I and II. The COX-2 positive rate and MVD in the cases with lymph node metastasis were 87.9% and (35.0+/- 8.5) respectively, higher than those in the cases without lymph node metastasis (P< 0.05). The Spearman rank correlation test showed a significant correlation between COX-2 expression and tumor MVD (r=0.311, P< 0.01).

CONCLUSIONSCOX-2 plays an important role in gastric cancer angiogenesis. COX-2 and angiogenesis induced by COX-2 contribute to tumor invasion and lymph node metastasis.

Adult ; Aged ; Cyclooxygenase 2 ; metabolism ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; metabolism ; pathology ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVESTo establish a primary biliary cirrhosis (PBC) model by AMAM2 autoantigen injection into C57BL/6 mice.

METHODSMice of the model group were immunized intraperitonealy with 200 microl of purified recombinant AMAM2 autoantigen in complete Freund's adjuvant (CFA). Mice immunized with bovine serum albumin and CFA in the same way were used as negative controls. Sixty-six weeks later, mice were sacrificed and their sera were collected. Sera samples were assayed for AMAM2 autoantibody, alkaline phosphatase (ALP), ALT and total bilirubin (TBil). Their liver, stomach, muscle and kidney tissues were sectioned and stained using HE to observe the pathological changes.

RESULTSAntibodies to AMAM2 autoantigen were readily induced in the model group. The mice in the model group had no significant changes in the level of serum ALT and TBil but had an obvious increase of ALP (P<0.05). The stomach, muscle and kidney tissues showed no evident damage while the livers had obvious pathological changes, including bile duct degeneration or proliferation, and mononuclear cell infiltration.

CONCLUSIONThe AMAM2 autoantigen-induced PBC animal model was successfully established in C57BL/6 mice in our experiment and its characteristic biochemical and pathology are quite similar to that in the early stage of human PBC. This model may provide a useful experimental approach for further study of the pathogenesis and clinical treatment of human PBC.

Animals ; Autoantigens ; immunology ; Disease Models, Animal ; Liver Cirrhosis, Biliary ; etiology ; Mice ; Mice, Inbred C57BL ; Mitochondria ; immunology

Adult ; Aged ; Alleles ; Asian Continental Ancestry Group ; Female ; Gene Frequency ; HLA-DQ Antigens ; genetics ; HLA-DQ beta-Chains ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Liver Cirrhosis, Biliary ; genetics ; immunology ; Male ; Middle Aged

OBJECTIVETo analyze the electrophysiological characteristics and efficacy of radiofrequency catheter ablation (RFA) of focal atrial tachycardia (AT) originating from the left atrial appendage (LAA).

METHODSElectrophysiologic study and RFA were performed in 9 patients (4 female) with focal AT originating from the LAA. Atrial appendage angiography was performed to identify the origin of AT. P waves were classified as negative, positive, isoelectric, or biphasic.

RESULTSThe mean age was (21 +/- 9) years. AT occurred spontaneously or was induced by isoproterenol infusion rather than programmed extrastimulation and burst atrial pacing. A characteristic P-wave morphology and endocardial activation pattern were observed. Positive P-wave in inferior leads was seen in all patients, upright or biphasic (+/-) component P wave was observed in lead V1, isoelectric component or an upright component P wave with low amplitude ( < 0.1 mV) was seen in lead V2-V6. Earliest endocardial activity occurred at the distal coronary sinus (CS) in all patients. The earliest endocardial activation at the successful RFA site occurred (36.7 +/- 7.9) ms before the onset of P wave. RFA was successful in all 9 patients immediately post procedure. AT reoccurred in 2 patients within 1 month post RFA and AT disappeared post the 2nd-RFA. AT reoccurred in 1 patient and terminated after the 3rd RFA. At the final follow-up (12 +/ 5) months, all 9 patients were free of arrhythmias without antiarrhythmic drugs.

CONCLUSIONSThe LAA is an uncommon site of origin for focal AT. The characteristic P wave and activation timing are suggestive for focal AT originating from the LAA. LAA focal ablation is safe and effective for patients with focal AT originating from the LAA.

Adolescent ; Adult ; Atrial Appendage ; physiopathology ; Catheter Ablation ; methods ; Child ; Electrophysiological Phenomena ; Female ; Humans ; Male ; Tachycardia, Ectopic Atrial ; physiopathology ; surgery ; Treatment Outcome ; Young Adult