Objective To clone and express the alkaline protease AprA , one important virulence factor secreted by Pseudomonas aeruginosa(PAE)in Escherichia coli, to clone and express the inhibitor of AprA (AprI) and its substrate flagellin , and to detect the function of AprA and the inhibitory function of AprI .Methods The genes encoding AprA ,AprI and flagellin gene were amplified respectively by PCR using PAE PAO 1 genome DNA as the template .The expression vec-tors (pET-28a-AprA, pET-28a-AprI and pET-28a-Flagellin) were constructed and transformed into E.coli BL21(DE3) respectively.The recombinant AprA protein was expressed by IPTG induction and purified via denaturing and renaturation. The recombinant AprI and flagellin were expressed and purified by Ni 2+affinity chromatography .The cleavage activities of AprA on flagellin were detected by SDS-PAGE.Results Recombinant AprA , AprI and flagellin protein were expressed and purified .It was demonstrated that AprA cleaved flagellin , which was blocked by AprI .Conclusion Recombinant AprA could cleave its substrates as an alkaline protease , and its inhibitor AprI inhibits the activities of AprA .This study will contribute to further investigations on the role of AprA in the pathogenesis of PAE .

Objective To assess the diagnostic predictive value of Wells score and modified Geneva score for acute pulmonary embolism by prospective case series and to explore a more suitable scoring system for Chinese population.Methods All the patients suspected of pulmonary embolism (PE) and received CT pulmonary angiography (CTPA) were enrolled consecutively in Fuxing Hospital,Capital Medical University,China,from June 2009 to August 2011.Before CTPA test or on condition that test results were unknown,clinical scoring was assessed prospectively by the Wells score and the modified Geneva score.The probability of PE in each patient was assessed and the patients were divided into low,moderate and high probability groups according to the clinical scores.The result of CTPA was used as the diagnostic gold standard for PE.Diagnostic accuracy in each group was analyzed.The predictive accuracy of both scores was compared by AUCROC curve.Results A total of 139 patients met our enrollment criteria and 117 eligible patients entered our study at last.PE was diagnosed in 47 patients by CTPA with an overall prevalence of 40.2％.Prevalence of PE in the low,moderate and high pretest probability groups assessed by the Wells score and by the simplified modified Geneva score were 7.1％ (3/42),42.9％ (21/49),88.5％ (23/26)and 10.0％ (3/30),48.1％ (37/77),7/10,respectively.AUCROC curves for the Wells score and the simplified modified Geneva score were 0.872 ( 95％ CI 0.810-0.933 ) and 0.734 ( 95％ CI 0.643-0.825 )respectively,with a significant difference ( P =0.005 ).Conclusion The Wells score is more accurate for clinical predicting acute PE than the modified Geneva score.

BACKGROUNDA prospective 2-night polysomnographic (PSG) study in Chinese snorers was designed to assess the role of the first night effect (FNE) in PSG parameters and the diagnosis of obstructive sleep apnea hypopnea syndrome (OSAHS).

METHODSSeventy-two snorers from two teaching hospitals underwent overnight PSG on two consecutive nights. The night-to-night variability of PSG parameters were recorded and analyzed.

RESULTSSixty-six patients were analyzed. Among all the PSG parameters, only the total time of stage 2 presented a significant difference between two nights: 219.50 (83.50 - 353.50) vs. 215.25 (59.50 - 342.50) (P = 0.000). Subgroup assessment showed a slight night-to-night difference in about 1 - 2 parameters in the group with the apnea-hypopnea index (AHI) ≥ 20 events per hour as well as the group with AHI < 20 events per hour, but there was no night-to-night difference in the AHI in each group. And slighter FNE was found among patients ≥ 40 years old. There was no significant difference in diagnosis of OSAHS. In the decision of severity, a slight difference was found between the two nights with a Kappa value = 0.531.

CONCLUSIONSOnly mild FNE can be found on two consecutive nights of PSG in adult Chinese snorers, but it has no effect on the diagnosis of OSAHS. A single polysomnographic night may be adequate for the diagnosis of OSAHS.

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Polysomnography ; methods ; Prospective Studies ; Sleep Apnea, Obstructive ; physiopathology ; Snoring ; physiopathology ; Young Adult

China ; epidemiology ; Drug Resistance, Bacterial ; Dysentery, Bacillary ; epidemiology ; microbiology ; Humans ; Serotyping ; Shigella ; classification ; drug effects ; isolation & purification

OBJECTIVETo evaluate speech outcomes in patients of maxillary reconstruction with free fibula composite flap.

METHODSSpeech measurements of 20 patients of maxillary reconstruction with free fibula composite flap were collected. Palatopharyngeal valve, the symmetry and movement of soft palate, hypernasality and nasal emission were examined by nasoendoscopy. At the same time the speech intelligibility was measured.

RESULTSNo palatopharyngeal valve insufficiency was found and all the soft palates had good symmetry and movement. The average speech intelligibility was 98.4%.

CONCLUSIONSFree fibula composite flap can reconstruct the phonatory structure and support the function of soft palate, which can restorate speech function well. It is a good choice for maxillary reconstruction.

Adult ; Female ; Fibula ; Humans ; Male ; Maxilla ; surgery ; Middle Aged ; Speech Intelligibility ; Surgical Flaps

OBJECTIVE: To study the expression levels of glial cell line-derived neurotrophic factor family receptor α-1 (GFRα1) and enhancer of zeste homolog 2 (EZH2) in the intestinal tissue of children with Hirschsprung's disease (HSCR), as well as the role of EZH2 in the regulation of GFRα1 gene expression and the pathogenesis of HSCR. METHODS: The samples of colon tissue with spasm from 24 children with HSCR after radical treatment of HSCR were selected as the experimental group, and the samples of necrotized colon tissue from 18 children with neonatal necrotizing enterocolitis after surgical resection were selected as the control group. Real-time PCR and Western blot were used to measure the expression levels of GFRα1 and EZH2 in colon tissue in both groups. Human neuroblastoma SH-SY5Y cells were divided into an EZH2 over-expression group and a negative control group. The cells in the EZH2 over-expression group were transfected with pCMV6-EZH2 plasmid, and those in the negative control group were transfected with pCMV6 plasmid. The expression levels of EZH2 and GFRα1 were measured after transfection. RESULTS: Compared with the control group, the experimental group had significant reductions in the mRNA and protein expression levels of GFRα1 and EZH2 in colon tissue (P<0.05), and the protein expression of EZH2 was positively correlated with that of GFRα1 (r=0.606, P=0.002). Compared with the negative control group, the EZH2 over-expression group had significant increases in the expression levels of EZH2 and GFRα1 after SH-SY5Y cells were transfected with EZH2 over-expression plasmid (P<0.05). CONCLUSIONS Low expression of EZH2 in the colon tissue of children with HSCR may be one of the causes of inadequate expression of GFRα1 and onset of HSCR.
Child ; Colon ; Enhancer of Zeste Homolog 2 Protein ; genetics ; Glial Cell Line-Derived Neurotrophic Factor Receptors ; genetics ; Hirschsprung Disease ; genetics ; Humans ; Infant, Newborn ; RNA, Messenger

Adult ; Dioxins ; Glutathione Peroxidase ; blood ; Humans ; Lipid Peroxidation ; drug effects ; Male ; Malondialdehyde ; blood ; Middle Aged ; Occupational Exposure ; Oxidative Stress ; drug effects ; Superoxide Dismutase ; blood

OBJECTIVETo investigate the characteristics of lymphatic metastasis in different types of adenocarcinoma of the esophagogastric junction (AEG) and provide guidance for surgical approach adoption.

METHODSClinical data of 228 patients with AEG undergoing surgery were analyzed retrospectively. According to Siewert classification, there were 9 cases of type I (3.9%) who all underwent left thoracoabdominal approach procedures. A total of 121 patients belonged to type II (53.1%), of whom 12 underwent left transthoracic approach, 48 left thoracoabdominal approach, and 61 transabdominal approach. Ninety-eight patients belonged to type III (43%), of whom 22 underwent left thoracoabdominal approach procedures and 76 transabdominal. The pattern of lymph node metastasis was analyzed and the association between surgical approach and oncological clearance was examined.

RESULTSThe resection margin was positive in 20(8.8%) patients, including 10 with type II (8.3%) and 10 with type III (10.2%), and the difference was not statistically significant (P>0.05). The rate of positive resection margin was 12.4%(17/137) in the transabdominal group and 16.7%(2/12) in the left transthoracic group, both significantly higher than the left thoracoabdominal group (1.1%, 1/88) (both P<0.05). Lymph node metastasis was found in 159(69.7%) patients. The metastasis was found in 4 of 9 patients with type I cancer and two were thoracic metastasis, no metastasis was found in the upper mediastinum. For type II cancer, the rate of lymph node metastasis was 66.9%(81/121), including thoracic metastasis ( n=32, 26.4%) and abdominal metastasis (n=81, 66.9%). For type III cancer, the rate of lymph node metastasis was 66.9%(81/121), including thoracic metastasis (n=15, 15.3%) and abdominal metastasis (n=69, 70.4%).

CONCLUSIONSFor type I AEG, left thoracoabdominal approach should be used because the pattern of lymph node metastasis is similar to that of the distal esophageal carcinoma. For type II , left thoracoabdominal approach should be used to ensure adequate resection of the tumor and clearance of lymph node in the lower esophagus and upper mediastinum because of high rate of intrathoracic lymph node metastasis. For type III cancer, transabdominal incision offers better benefit with less impact on respiratory function. However, thoracic incision should be used to ensure adequate clearance for tumors of larger size and significant external invasion.

Adenocarcinoma ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Esophagectomy ; methods ; Esophagogastric Junction ; pathology ; surgery ; Female ; Humans ; Lymph Node Excision ; methods ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Objective To compare the clinical efficacy and safety of pegylated interferon α-2a (Peg IFN α-2a) or adefovir dipivoxil(ADV) monotherapy and their combination therapy in HBeAg positive chronic hepatitis B (CHB) patients. Methods An open randomized controlled multicenter clinical trial was performed. One hundred and twenty cases with CHB were divided into 3 groups: Peg IFN α-2a monotherapy (group A), ADV monotherapy (group B) and Peg IFN α-2a plus ADV combination therapy (group C). The virological response (VR), serological response (HBeAg, HBsAg clearance and seroconversion), biochemical response (BR) and sustained response (SR) were tested at week 24 and 48 of therapy and week 48 of follow-up after end of treatment (EOT) for'evaluation of therapeutic effects, safety and drug resistance. The efficacy was compared using X2 test. Results At week 48 of treatment, the VR (HBV DNA ≤500 copy/mL) rates were 36. 8%(14/38), 37. 5%(15/40) and 62. 9% (22/35), respectively in groups A, B and C; that in group C was higher than those in groups A and B (X2 = 4. 933, 4. 801, respectively; both P < 0. 05); HBeAg seroconversion rates in three groups were 44. 7% (17/38), 17. 5% (7/40) and 51. 4% (18/35), respectively. At week 48 of follow-up,SR rates in three groups were 34. 2%(13/38), 15. 0%(6/40) and 48. 6% (17/35), respectively; those in groups C and A were higher than that in group B (X2 = 9. 894,P<0. 01;X2 =3. 903, P<0. 05, respectively). Conclusions VRs at week 24 and 48 of Peg IFN α-2a plus ADV combination therapy are better than Peg IFN α-2a or ADV monotherapy. SRs at week 48 of follow-up after Peg IFN α-2a monotherapy and combination therapy are both better than ADV monotherapy.

OBJECTIVETo evaluate the clinic characteristic and treatment of senile chronic submandibular sialadenitis.

METHODSThe clinical records of the patients over 50 years old whose final diagnosis was chronic submandibular sialadenitis were reviewed. The patients were diagnosed as neoplasm of submandibular gland before operation, There was no radiographic sign of sialolithiasis, no clinical and laboratory signs of Sjögren syndrome or related autoimmune disease.

RESULTSThirty-five patients were presented in masses in the submandibular space, which was not related to eating. Histopathologic study showed destruction of acinus, infiltration of inflammatory cell, and micro-calcification in the intercalated duct.

CONCLUSIONSThe diagnosis of senile chronic submandibular sialadenitis should be considered if a mass in the submandibular space occurs in the old patients and radiographic study shows no sialolithiasis and neoplasm. Resection of the submandibular gland is not necessary for these patients.

Aged ; Aged, 80 and over ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Sialadenitis ; diagnosis ; therapy ; Submandibular Gland Diseases ; diagnosis ; therapy