OBJECTIVETo express and purify the protein coded by the TRAF-type zinc finger domain of myasthenia gravis (MG)-related gene P9 (P9-ZFD) and to prepare P9-ZFD antiserum for detecting expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of patient with MG.

METHODSThe cDNA encoding P9-ZFD was amplified by RT-PCR. The cloned P9-ZFD cDNA was ligated into pET24a, and the P9-ZFD recombinant protein was induced via E. coli. BL21 (DE3) and purified by histidine affinity chromatography. P9-ZFD antiserum was prepared and its titer and specificity were determined by ELISA and Western blot. Expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of MG and control were studied.

RESULTSThe molecular weight of purified P9-ZFD protein was about 30 kD. Its purity was more than 95%. Antiserum specific for P9-ZFD was excellent. P9-ZFD protein is fully confined to the cytoplasm membrane of skeletal muscle cell of MG, obvious immunostaining was absent in the A, I, and Z bands of cytoplasm and no immunoreactivity was observed in the skeletal muscle cell of control.

CONCLUSIONP9-ZFD protein is expressed as a cytoplasm membrane-bound protein and has obvious distribution difference in the skeletal muscle cells of patient with MG and normal control.

Adult ; Cell Membrane ; metabolism ; Escherichia coli ; metabolism ; Female ; Humans ; Muscle Proteins ; biosynthesis ; genetics ; Muscle, Skeletal ; metabolism ; pathology ; Myasthenia Gravis ; metabolism ; Peptide Fragments ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Zinc Fingers

OBJECTIVETo observe the effect of salidroside on tic behavior and in vivo dopamine DA) and serotonin (5-HT) levels in Tourette syndrome (TS) model rats.

METHODSForty rats were randomly divided into the blank control group, the TS model group, the haloperidol-treated group (0.5 mg/kg x d(-1)), and the salidroside-treated group (50 mg/kg x d(-1)), 10 in each group. TS rat model was induced by imino-dipropio-nitrile (IDPN). Peritoneal injection of haloperidol and salidroside was started from the 4th day of modeling in the haloperidol-treated group and the salidroside-treated group respectively. Normal saline was peritoneally injected to rats in the blank control group and the TS model group respectively. Stereotyped behavior was scored, and changes of DA and 5-HT levels in blood and striatum were measured before modeling, after modeling, and after intervention.

RESULTSCompared with the blank control group, the score of the tic behavior was elevated (P < 0.01) , levels of DA and 5-HT in plasma and striatum were reduced in the model group (P < 0.01, P < 0.05). Compared with the same group after modeling, the tic behavior score decreased and plasma DA levels increased in the two treated groups after intervention (P < 0.01). 5-HT content increased in the salidroside-treated group (P < 0.01). Compared with the model group after intervention, the tic behavior score was significantly reduced (P < 0.01), and DA levels in plasma and striatum were elevated (P < 0.01, P < 0.05) in the salidroside-treated group and the haloperidol-treated group. Compared with the haloperidol-treated group, the tic behavior score increased (P < 0.01), DA levels in plasma and striatum were lowered (P < 0.01, P < 0.05), the 5-HT level increased in plasma and striatum (P < 0.01, P < 0.05) in the salidroside-treated group.

CONCLUSIONSIn the salidroside-treated group, the tic behavior was significantly reduced, and DA levels in plasma and striatum were elevated. Its mechanism might be related to regulating activities of dopamine neurons in striatum.

Animals ; Corpus Striatum ; Dopamine ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Glucosides ; pharmacology ; therapeutic use ; Haloperidol ; Phenols ; pharmacology ; therapeutic use ; Rats ; Serotonin ; Stereotyped Behavior ; Tics ; drug therapy ; Tourette Syndrome ; drug therapy

Objective To investigate the significance of orthotopic liver transplantation in the management of primary hepatocellular carcinoma (PHC) ; To evaluate the survival and prognostic factors of liver transplantation for PHC.Method The clinical data from 194 consecutive PHC patients who underwent OLT were retrospectively analyzed.Result The 1-,3-and 5-year cumulative survival rate of the 194 patients was 92.78％,78.07％ and 76.80％ respectively.There were 37 cases of recurrence or metastasis of the tumor with the incidence rate being 19.1％ (37/194).The tumor TNM stage (P=0.026 667),tumor diameter (P =0.0045),tumor number (P =0.0318),vascular invasion (P =0.006 059),and AFP level before transplantation (P =0.003 426) were significantly related with tumor recurrence.Multivariate analysis revealed that tumor diameter (P =0.0081,OR =0.789,95％ CI:2.1233-6.124),vascular invasion (P＜0.001,OR =5.928,95 ％ CI:1.173-86.789),and AFP level before transplantation (P =0.019,OR =2.601,95％ CI:2.196-5.658) were independent risk factors for tumor recurrence.There was statistically significant difference in survival rate and tumor-free survival rate between Milan criteria group and beyond-UCSF criteria group (P＜0.01),and between UCSF criteria group and beyond-UCSF criteria group (P＜0.01).There was no significant difference in survival rate and tumor-free survival rate between Milan criteria group and UCSF criteria group (P ＞0.05).Conclusion Liver transplantation is an effective treatment for PHC.The tumor diameter ＞8 cm,vascular invasion and AFP level significantly affect the survival of PHC patients.

OBJECTIVETo evaluate the feasibility and outcomes of chimeric deep circumflex iliac artery perforator flap (DCIAPF) applied in the simultaneous reconstruction of the oromandibular defect.

METHODSSix patients underwent simultaneous oromandibular reconstruction using DCIAPF following segmental mandibulectomy in Xiangya Hospital from March 2014 to July 2014. The skin paddle was designed to be centered on the pre-operative perforator mapping. Retrograde dissection was performed through the underlying abdominal wall to raise the skin paddle. The pedicle was isolated from the groin, and the iliac crest was cut. The deep iliac circumflex vessels were dissected until the skin paddle was reached. Finally, the donor site was strictly sutured layer by layer to avoid ventral hernia.

RESULTSThe skin paddles ranged from 3.5 cmx5.0 cm to 7.0 cmx 10.0 cm. The length of the bone components was 5.0 cm to 11.0 cm. All donor sites closed primarily without skin grafting. DCIAPF was harvested successfully in five patients, except for one patient whose perforator originated from the superficial iliac circumflex vessels. An additional pair of anastomoses was performed. All iliac flaps survived. However, slight skin-edge necrosis and exfoliation caused by flap thinning occurred in one patient and healed after pruning and dressing change. The heights of all alveolar ridges were significantly restored, and no serious donorsite complication was observed during the three to six months' follow-up.

CONCLUSIONDCIAPF is a reconstructive option for mandibular defects because of its adequate bone tissue and rich blood supply. Satisfactory alveolar ridge restoration greatly facilitates future denture retention. DCIAPF also has a great degree of mobility between the skin paddle and the bone component when appliedin composite oromandibular defect reconstruction.

Humans ; Iliac Artery ; Ilium ; Mandible ; surgery ; Maxillofacial Abnormalities ; surgery ; Perforator Flap ; Reconstructive Surgical Procedures ; methods ; Skin

Objective:To investigate the value of atropine administration before tracheal intubation under general anesthesia in the elderly patients undergoing endobronchial ultrasound-guided transbronchial needle aspiration(EBUS-TBNA)biopsy.Methods:This was a case-control study.A total of 87 elderly hospitalized patients receiving chest enhanced CT test suggesting the risk of lung cancer with enlargement of mediastinal lymph nodes in 7 regions were scheduled to undergo endobronchial ultrasound-guided transbronchial needle aspiration(EBUS-TBNA)biopsy under general anesthesia.The patients were randomly divided into two groups by flipping a coin: the atropine group(n=40)and the control group(n=47). The indicators for evaluating the application values of atropine included preoperative, intra-operative and postoperative systolic blood pressure(SBP), diastolic blood pressure(DBP), heart rate, oropharyngeal and airway secretion volume, oxygen saturation, operation time, positive diagnosis rate and postoperative adverse reactions.Results:SBP and DBP were lower in the atropine group than in the control group before endotracheal intubation(131.7±15.3 mmHg vs.140.7±13.7 mmHg, 79.1±7.6 mmHg vs.85.6±7.4 mmHg, t=2.885 and 4.061, P<0.05). There was no significant difference in SBP and DBP 10 min after endoscopic operation and 10 min after tracheal extubation between the atropine group versus the control group(SBP: 109.1±11.2 mmHg vs.105.0±12.2 mmHg, 136.9±23.0 mmHg vs.129.9±11.2 mmHg, DBP: 66.9±7.5 mmHg vs.68.0±8.3 mmHg, 77.6±10.9 mmHg vs.78.5±6.4 mmHg, t=-1.617, 0.687, -1.751 and 0.448, P>0.05). There was no significant difference in HR between the two groups before endotracheal intubation( t=1.416, P>0.05), while HR was higher in the atropine group than in the control group 10 min after endoscopic operation and 10 min after tracheal extubation( t=-3.323 and -2.181, P<0.01 and P<0.05). The change rates of SBP and DBP were lower in the atropine group than in the control group 10 min after endoscopic operation and 10 min after tracheal extubation( t=7.947, -6.962, -3.187 and -3.232, P<0.01). The change rate of HR was lower in the atropine group 10 min after endoscopic operation and was higher 10 min after tracheal extubation than in the control group( t=-6.467 and -4.131, P<0.01). There were significant differences in the volume of oropharyngeal and airway secretions and fingertip oxygen saturation between the two groups before endotracheal intubation and 10 min after tracheal extubation( t=-2.334, 2.759, -3.314 and -2.767, P<0.01). The endoscopic operation time was less in the atropine group than in the control group with no statistically significant difference[(25.9±5.7)min vs.(26.4±4.7)min, t=0.391, P>0.05]. There was no significant difference in postoperative adverse reactions between the atropine group versus the control group(34 patients or 85.0% vs.43 patients or 91.5%, χ2=1.247, P>0.05). Conclusions:The application of atropine before tracheal intubation under general anesthesia is beneficial to stabilizing the intraoperative blood pressure and heart rate, and can reduce the production of postoperative oropharyngeal and airway secretions in elderly patients undergoing endobronchial ultrasound-guided transbronchial needle aspiration biopsy.

Objective To demonstrate the carbonic anhydrase Ⅲ (CAⅢ) for 25 000 protein decreased in skeletal muscle of myasthenia gravis (MG). Methods The protein molecular properties responsible to antibodies against 25 000 protein and CAⅢ were analyzed by a combination method of two-dimensional electrophoresis and immuno-Western blot. Competitive binding reactions of the antibodies to the purified 25 000 protein and muscular homogenate were observed by using immuno-Dot blot and immuno-Western blot, respectively. The expression of CAⅢ from normal and MG muscles was detected by immuno-Western blot. Results Combination analysis of two-dimensional electrophoresis and immuno-Western blot showed that the protein of immunological responsible to antibodies against 25 000 protein and CAⅢ had an identical molecular mass and isoelectric point. Competitive binding reactions proved that 25 000 protein and CAⅢ were the same substance, either by immuno-Dot blot or by immuno-Western blot. In addition, a much similar result was obtained when the levels of 25 000 protein from normal and MG muscles were detected by antibodies against 25 000 protein and (CAⅢ) by immuno-Western blot. Conclusion 25 000 protein decreased in the MG skeletal muscle was proved to be just a known protein CAⅢ, which made a basis for further exploring the relationship of CAⅢ deficiency and MG pathogenesis.

Objective To determine the frequency of polymorphism at exon 26 (C3435T) of muhidrug resistance 1 gene (MDR1) in epileptic patients in the southern Chinese and to study the association of this polymorphism with pharmacoresistance.Methods DNA samples were obtained from 134 patients,of whom 72 were resistant to antiepileptic drug treatment and 62 were responsive to the treatment. Genotypes of the C3435T polymorphism were determined by polymerase chain reaction (PCR) followed by restriction digestion and gel electrophoresis.Genotype and allele frequencies in the drug resistant group were compared to those in the response group by Chi-square analysis.Results Of all 134 patients,33 (24.6%) had CC genotype,72 (53.7%) had CT genotype,and 29 (21.6%) had TT genotype.The frequency of CC genotype was significantly higher in the pharmaeoresistance group (33.3%) than that in the responsive group (14.5%,P=0.012).The frequency of the C allele was also significantly higher in the pharmacoresistance group (57.6%) than that in the responsive group (44.4%,P=0.03).When patients were divided by types of seizure into three groups:generalized seizure group,partial seizure group,and undefined seizure group,the CC genotype and C allele were associated with pharmacoresistance in the partial seizure group.Conclusions In the southern Chinese,the CC genotype and C allele are associated with resistance to the antiepileptic treatment.This finding needs to be verified in studies with larger sample size.

OBJECTIVETo observe the effect of glycoprotein receptor blockade tirofiban in acute anterior myocardial infarction patients without ST segment resolution after primary percutaneous coronary intervention (PCI).

METHODSFrom April 2006 to April 2008, 157 acute anterior myocardial infarction patients without ST segment resolution after PCI were randomly allocated to tirofiban (intravenous bolus 10 microg/kg followed by intravenous infusion of 0.15 microgxkg(-1)xmin(-1) for 48 h, n = 80) or equal volume saline (control group, n = 77). Baseline characteristics, PCI features and clinical outcomes during hospitalization, left ventricular ejection fractions (LVEF) and major adverse cardiac events (MACE, including death, re-infarction and target vessel revascularization) at 30 and 180 days after discharge were compared between the two groups.

RESULTSThe baseline clinical characteristics were comparable between the two groups. Compared to control group, the MACE rates and re-infarction rates at 30 days (6.3% vs.18.2%, P < 0.05; 1.3% vs.9.1%, P < 0.05, respectively) and 180 days (10.0% vs.23.4%, P < 0.05; 2.5% vs.10.4%, P < 0.05, respectively) were significantly reduced in tirofiban group. LVEF value was significantly higher in tirofiban group at 30 days and 180 days compared with those in control group [(51 +/- 6)% vs. (46 +/- 8)%, P < 0.05; (57 +/- 7)% vs. (50 +/- 9)%, P < 0.05]. Hemorrhagic complications were similar between the two groups.

CONCLUSIONUse of tirofiban for acute anterior myocardial infarction patients without ST segment resolution after PCI is safe and can significantly improve 30 and 180 days clinical outcomes after discharge.

Aged ; Angioplasty, Balloon, Coronary ; Anterior Wall Myocardial Infarction ; diagnosis ; drug therapy ; therapy ; Electrocardiography ; Female ; Humans ; Male ; Middle Aged ; Platelet Aggregation Inhibitors ; therapeutic use ; Prognosis ; Treatment Outcome ; Tyrosine ; analogs & derivatives ; therapeutic use

Purpose To describe the clinicopathologic features, diagnosis and differential diagnosis, and prognosis of secretory breast carcinoma (SBC). Methods Clinicopathological and follow-up data of six SBC patients were collected. Histopathologic analysis was performed on hematoxylin and eosinstained (HE) section. Immunohistochemical staining was performed by En Vision two-step method and ETV6 gene detected by fluorescence in situ hybridization (FISH), then relevant literatures were reviewed. Results The ages of the patients ranged from 6 to 76 years with a mean age of 38.7 years, including one male and five female patients. The right breast was involved in 4 cases, and the left, in 2 cases. Five cases showed painless breast mass while one presented with a nipple discharge. The tumor size ranged from 1.0 to 3.1 cm with a mean size of 2.0 cm. Most of the tumors were circumscribed, solid gray white to light brown. Histologically, tumor showed solid nested microcystic, glandular or papillary pattern separating by hyaline fibrous tissue and growed in multiple nodular from. The cytoplasm contains abundant eosinophilic secretions or secretory vesicles. Immunhistochemistry, all cases were positive for CK7, S-100 and CEA, but negative for estrogen and progesterone receptors (ER and PR) and HER-2, and the proliferation index Ki-67 ranged from 10％ to 40％. Molecular testing confirmed the presence of the EVT6 gene translocation in one case. Lumpectomy was performed in 2 cases and modified radical mastectomy in 4 cases, two of them had lymph node metastasis (3/15, 1/16). Five cases were followed up for 6 months to 20 years, 1 case had lung metastasis. Conclusion SBC is a rare breast tumor with relatively indolent clinical and good prognosis. It can be diagnosed according to typical pathological morphology and immunohistochemical characteristics. The characteristic EVT6 gene translocation also has important differential diagnostic value.

OBJECTIVETo summarize characteristics and outcomes of critically ill children with 2009 influenza A (H1N1).

METHODA prospective observational study of 14 critically ill children with 2009 influenza A (H1N1) in pediatric intensive care unit (PICU) in Suzhou between Oct. 1(st) 2009 and Dec. 25(th) 2009. The primary outcome measures included frequency and duration of mechanical ventilation and duration of ICU stay.

RESULTCritical illness occurred in 14 patients with confirmed (n = 14), community-acquired 2009 influenza A virus (H1N1) infection. The mean (SD) age of the 14 patients with confirmed 2009 influenza A (H1N1) was (4.91 ± 4.14) years, 7 were female (50.0%). The median duration from symptom onset to hospital admission was (3.09 ± 1.30) days and from hospitalization to ICU admission was (0.95 ± 0.96) day. All the patients were severely hypoxemic [mean (SD) ratio of PaO2/FiO2 was (191.27 ± 80.58) mm Hg] at ICU admission. ARDS occurred in 11 cases (78.6%). Mechanical ventilation was applied for 10 patients (71.4%). The median duration of ventilation was (12.51 ± 10.03) days and ICU stay was (12.58 ± 10.65) days. The median length of time during which the real-time RT-PCR test results were positive was (17.27 ± 5.57) days; Comorbidities such as iron deficiency anemia, cerebral palsy and congenital heart disease were found in 8 cases (57.1%). The longer length of mechanical ventilation and ICU stay were found in cases with higher admission PRISM III Score and lower Pediatrics Critical Illness Score.

CONCLUSIONCritical illness due to 2009 influenza A (H1N1) in Suzhou occurred rapidly after hospital admission and was associated with severe hypoxemia, ARDS, a condition that required prolonged mechanical ventilation. There were myocardial damages in critically ill children with severe 2009 influenza A (H1N1).

Child ; Child, Preschool ; Critical Illness ; Female ; Humans ; Infant ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; diagnosis ; epidemiology ; virology ; Male ; Prognosis ; Risk Assessment