Objective To explore the value of immunofluorescent technique for clinical diagnosing Duchenne muscular dystrophy(DMD),Becker muscular dystrophy(BMD) and Limb-girdle muscular dystrophy(LGMD).Methods Immunofluorescent technique was applied,and the expressions of Dys1,Dys2,Dys3 monoclonal antibodies and ?-,?-,?-sarcoglycan(SG) polyclonal antibodies against dystrophin,?-SG,?-SG,?-SG in musculomembranes of frozen section specimens from 25 patients(10 cases of DMD,4 cases of BMD and 11 cases of LGMD) were detected.Results 10 DMD patients had negative staining of dystrophin,and 4 BMD patients had discontinuous or a patchy positive staining pattern.All LGMD patients had positive dystrophin staining.There was one patient presented negative staining of ?-SG and ?-SG,respectively.Conclusions Detecting of dystrophin by immunofluorescent technique is special and helpful in diagnosing and classifying DMD/BMD.At present,SG may not be used in diagnosing the LGMD patients.

Objective To report a rare case of M3r subtype of acute promyelocytic leukemia (APL)with 3'-end of RARα (3'RARα) submicroscopic deletion, and the characters of morphologic, cytogenetic,molecular genetic and molecular biology studies. Methods Chromosomes of bone marrow (BM) cells were prepared with direct method and short-term culture method, and R-banding technique was used for karyotypic analysis. Fluorescence in situ hybridization (FISH) assays were performed on fixed BM cells using the following specific DNA probes: CEP X/Y alpha satellite DNA probe, LSI PML-RARα dual-color dual-fusion and LSI RARα dual-color break apart probes. A quantitative real-time reverse-transcription polymerase chain reaction (RT-PCR) was performed to detect the PML-RARα transcript. A multiplex nested RT-PCR was also performed, which may simultaneously detect the fusion genes derived from 29 chromosomal aberrations in acute leukemia including PML-RARα, PLZF-RARα and NPM-RARα fusion transcripts. Results R-banding analysis revealed a karyotype of 45,X,-Y[6]/46,XY[8], FISH using CEP X/Y probe further confirmed Y-chromosome loss. FISH analysis with RARα dual-color break apart probe demonstrated a deletion of the entire 3'-end of one allele of RARα gene. Cytogenetic, FISH and RT-PCR analyses showed no PML-RARα,PLZF-RARα, NPM-RARα, NuMA-RARα and STAT5b-RARα rearrangements. Conclusion A new RARαrearrangement involving 3'RARα submicroscopic deletion in APL without X-RARα fusion has been identified.FISH analysis with RARα dual-color break apart probe is a useful method for characterization of this abnormality, but its molecular consequences remain to be elucidated.

Objective To study the clinical efficacy of electromyography biofeedback on dystonia after hepatolenticular degeneration.Methods 40 patients with dystonia after hepatolenticular degeneration were divided into treatment group (n=20) and control group (n=20).All patients were treated with copper-cleaning, clonazepam and benzhexol hydrochloride etc., and acupuncture. The treatment group wastreated with electromyographic biofeedback in addition. They were assessed with modified Ashworth scale, modified Barthel index, and ankledorsiflexion active range of motion. Results The lower extremities function of all patients improved after treatment (P<0.05) and thetreatment group improved more than the control group (P<0.05). Conclusion Electromyography biofeedback is more effective to improvethe lower extremities function and activity of daily living in patients with hepatolenticular degeneration following dystonia.

Objective To investigate the effect of electroacupuncture on expression of growth associated protein-43 (GAP-43) and Nogo-A in brain after focal cerebral ischemia/reperfusion in rats. Methods 48 male Sprague-Dawley rats were randomly divided into sham group, model group and electroacupuncture group, and the latter two were modeled as middle cerebral artery occlusion and reperfusion with nylon monofilament suture. Electroacupuncture was performed 90 min after modeling in the electroacupucture group at acupoints of Neiguan (PC06), Shuigou (DU26), Sanyinjiao (SP06), Baihui (DU20), for 30 min, once a day. The sham group and the model group were conventionally fed in cages without any intervention. 8 rats of each group were assessed with Longa's score, and the expressions of GAP-43 and Nogo-A were detected with immunohistochemistry 7 d and 14 d after modeling respectively. Results The sham group presented no neurological symptoms. There was not different in Longa's score between the model group and the electroacupuncture group 7 d after modeling (P>0.05), but was different 14 d after modeling (P<0.05). GAP-43 positive cells was not found in the sham group, but could be found around cerebral ischemia 7 d after modeling, which decreased 14 d after modeling in the model group. GAP-43 positive cells increased significantly in the electroacupuncture group compared with the model group at each time (P<0.01). Nogo-A positive cells was few found in the sham group, and was more in the model group (P<0.01). The expression of Nogo-A decreased significantly in the electroacupuncture group compared with the model group at each time (P<0.01). Conclusion Electroacupuncture can improve neurological function of focal cerebral ischemia/reperfusion rats, which may be associated with the increase of GAP-43 and descrease of Nogo-A in peri-infarct regions

Objective: To study the protective effect of vitamin E in preventing contrast-induced nephropathy (CIN) in patient with coronary artery disease (CAD) after percutaneous coronary intervention (PCI).
Methods: We prospectively studied 206 CAD patients with elective PCI in our hospital and divided them in 2 groups: Treatment group, the patients received oral vitamin E combining vinous hydration,n=102 and Control group, the patients received vinous hydration only,n=104. CIN was deifned by at 48h after contrast media injection, serum cretinin increased up to 25% from the baseline, or reached 44.2 μmol/L. Excluding the other kidney injury factors, the renal functions at 48 h before and after PCI were compared, the occurrence rate of CIN were also compared between 2 groups.
Results:①Overall, there were 19/206 (9.22%) patients suffered from CIN, the occurrence rate in Treatment group (4.90%) was lower than Control group (13.46%), χ2=4.506,P=0.034. For patients with hypertension, diabetes, chronic kidney disease, anemia and mehran risk score<10, the occurrence rate of CIN in Treatment group was lower than Control group,P<0.05.②Compared with pre-operative condition, at 48 h post-operation,Control group showed increased serum creatinine (Scr), blood urea nitrogen (Bun) and decreased creatinine clearance rate (Ccr), allP<0.05.③At 48 h post-operation, compared with Control group, Treatment group presented decreased Scr (86.72 ± 17.73) μmol/L vs (95.13 ± 21.67) μmol/L and increased Ccr (96.75 ± 27.23) ml/min vs (90.70 ± 17.85) ml/min, allP<0.05.④Multivariate regression analysis revealed that elder than 75 years of age (OR=7.278, 95% CI 5.158-11.480), diabetes (OR=3.919, 95% CI 1.330-8.200), chronic kidney disease (OR=6.325, 95% CI 2.137-16.816) and mehran risk score>10 (OR= 4.461, 95% CI 1.589-14.724) were the independent risk factors for CIN occurrence, allP<0.05.
Conclusion: Short-term application of vitamin E may reduce the risk of CIN occurrence at certain degree in CAD patients after PCI.

Objective To observe the effects of PTD-hepatitis B core antigen (HBcAg) induced murine bone marrow-derived dendritic cells (DCs) maturation on T lymphocyte proliferation in vitro.Methods Bone marrow derived DCs isolated from BALB/c mice were cultured with recombinant granu|ocyte-macrophage colony-stimulating factor (rGM-CSF) and recombinant interleutin-4 (rIL-4)for 5 days.Tumor necrosis factor (TNF)-a,HBcAg and PTD-HBcAg were added to induce DCs maturation.The distribution and localization of intracellular immunofluorescence were observed by confocal microscopy,and DCs phenotypes were analyzed by flow cytometry.The level of IL-12 p70 in the supernatant was detected by enzyme linked immunosorhent assay (ELISA).The proliferation of T lymphocytes was performed by using cell counting kit-8 (CCK-8).All data were analyzed using t test.Results DCs were cultured and identified successfully.Recombinant PTD-HBcAg could penetrate into DCs cytoplasm while recombinant HBcAg was detected on the surface of cells.DCs surface molecules,such as CD80,CD86 and major histocompability complex (MHC) II were upregulated by PTDHBcAg;IL-12 p70 levels induced by 50 mg/L and 100 mg/L recombinant PTD-HBcAg were (142.50±18.31) ng/L and (124.30±15.12) ng/L,respectively,which were significantly higher than those induced by recombinant HBcAg [(42.31±4.21 ) ng/L,t = 9.234 and 9.045,respectively,P＜0.05].The proliferation of T lymphocytes induced by PTD-HBcAg was much higher than that in HBcAg group or positive control TNF-a group.Conclusions PTD-HBcAg could penetrate membrane of DCs and promote the differentiation and maturation of DCs.PTD-HBcAg could up-regulate the expressions of costimulatory molecules on cell surface of DCs,and enhance the ability of DCs on stimulating T lymphocytes proliferation and IL-12 p70 production.

Objective To investigate the changes of cardiac structure and function in patients with hepatolenticular degeneration (HLD).Methods From March 2010 to February 2011,90 HLD patients who did not receive formal treatment (observation group) and 30 healthy people (control group) were collected and analyzed with color Doppler echocardiography in the Institute of Neurology,Affiliated Hospital of Anhui University of Traditional Chinese Medicine.Based on the clinical manifestations,cases were divided into hepatic type group (n =31),brain type group (n =42) and brain-visceral type group (n =17).According to the Child-Pugh classification,patients were divided into Child A group (n =71),Child B group (n =10) and Child C group (n =9).Results (1) The left atrial diameter ((28.00 ± 3.11),(29.62 ± 3.44) mm) and left ventricular diameter ((45.69 ± 3.75),(47.10-± 4.73) mm) of HLD patients in brain-visceral type group and hepatic type group were larger than that of the control group ((24.86 ± 2.63),(41.93 ±3.56) mm;t =3.143,4.761,P=0.018,0.000;t=3.764,5.167,P=0.018,0.000).The left atrial diameter and left ventricular diameter of hepatic type group were significantly larger than the brain type group (26.06 ± 3.68,43.34 ± 3.88;t =3.557,P =0.000;t =3.751,P =0.001).The value of E/A in the hepatic type group (1.57 ± 0.37) was significantly lower than the control group (1.93 ± 0.20;t =-0.352,P =0.006).(2) The left atrial diameter ((31.29 ± 1.70),(34.67 ± 1.97) mm) and left ventricular diameter ((48.29 ± 2.81),(53.67 ± 2.67) mm) of Child B and C groups were significantly larger than the control group ((24.86 ± 2.63),(41.93 ± 3.56) rm;t =6.429,9.810,P =0.000,0.000;t =6.357,10.738,P =0.000,0.000),and the Child A group ((26.42 ± 3.05),(43.89 ± 3.76) rm;t=4.871,8.252,P=0.000,0.000;t =4.399,8.780,P=0.003,0.000).The value of E/A of Child B and C groups (1.58 ± 0.32,1.26 ± 0.39) was lower than that of the control group (t =-0.347,0.662,P=0.020,0.000);At the same time,the value of E/A of Child C group was significantly lower than that of Child A group (1.80 ± 0.33;t =-0.530,P =0.000).Conclusions The HLD patients may have cardiac structural and functional changes,mainly manifested as left atrial and left ventricular enlargement and cardiac diastolic dysfunction,whereas a serious impact has not yet been found in systolic function.And the changes of cardiac structure and function were related to the degree of liver cirrhosis in patients with HLD.

in 30minutes.Antibody expression located accurately.Background stained clearly and no interfering signal.The back-ground was better than conventional immunohistochemistry of frozen remaining tissue.The positive expression rate of p40,34βE12,p504s with rapid immunohistochemistry in prostatic hyperplasia was 97.4%(77 /79),93.7%(74 /79),0%(0 /79),and in prostatic carcinoma was 0%(0 /39),0%(0 /39),97.4%(38 /39).The positive expression rate of p40,34βE12,p504s with conventional immunohistochemistry in prostatic hyperplasia was 96.2%(76 /79), 93.7%(74 /79),2.5%(2 /79),and in prostatic carcinoma was 0%(0 /39),0%(0 /39),92.3%(36 /39).The difference of expression between prostatic hyperplasia and prostatic carcinoma with rapid immunohistochemical detec-tion p40 group:χ2 =109.402,P =0.000,34βE12 group:χ2 =97.971,P =0.000,p504s group:χ2 =113.537,P =0.000;The difference of expression between prostatic hyperplasia and prostatic carcinoma with conventional immuno-histochemical detection p40 group:χ2 =105.410,P =0.000,34βE12 group:χ2 =97.971,P =0.000,p504s group:χ2 =96.388,P =0.000;The expression of prostatic hyperplasia with between rapid immunohistochemical detection and conventional immunohistochemical detection 34βE12 group was identical,p40 group:χ2 =0.207,P =0.649, p504s group:χ2 =2.026,P =0.155;The expression of conventional immunohistochemical detection with between rap-id immunohistochemical detection and conventional immunohisto -chemical detection p40 group and 34βE12 group were identical,p504s group:χ2 =1.054,P =0.305.The expression of three markers between prostatic hyperplasia and prostatic carcinoma had statistical significance(P <0.05).The expression results between rapid immunohisto-chemistry and conventional immunohistochemistry had no statistically significant difference (P >0.05).Conclusion MaxVision rapid immunohisto -chemical staining technique has the advantages of rapid,accurate,timely.It could be used to rapid diagnosis of prostate biopsy tissue.The combined detection of p40,34βE12,p504s has very high practi-cal value in the differential diagnosis of benign and malignant lesions of the prostate.

Objective To investigate the feasibility and safety of transumbilical laparoscopic single-port choledocholithotomy in the treatment of common bile duct stones. Methods The clinical data of four patients who underwent transumbilical laparoscopic single-port choledocholithotomy at Shengjing Hospital of China Medical University from January to June, 2010 were retrospectively analyzed. The operation method and postoperative effects were reviewed. Results All the operations were successfully carried out. The mean operation time was 100 minutes and the mean blood loss was 62. 5 ml. All patients were cured without the occurrence ofcomplications except for one patient who had a peritoneal infection and was cured after surgical drainage. Conclusion Transumbilical laparoscopic single-port choledocholithotomy is safe and feasible, but indications should be strictly followed.