The microbial populations and community structure characterization technologies of the enhanced biological phosphate removal system were reviewed comprehensively in this paper, and their future research directions were outlined.

The kinase domain of receptor tyrosine kinase(RTK) ErbB2 was expressed fused with GFP in Pichia pastoris. Recombinant expression vector pPIC3.5K was constructed in Escherichia coli TOP10. The right P. pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of recombinant vector, and then induced by methanol in baffled shake bottles. The strain with highest protein yield was scaled up in a 5 L fermentor. Recombinant protein was analyzed with tyrosine kinase assay after Ni2+ affinity chromatograph. Results showed that the 100 kD recombinant protein with tyrosine kinase activity was successfully expressed in P. pastoris.

Objective: It is difficult to repair large-area Ⅲ?burn wound in children.Here we introduce the advantages of the back skin donor site in repairing Ⅲ?burn wound in children.Methods: This clinical study included 15 children with Ⅲ?burn,ranging in age from 18 months to 11 years(mean 7.6 years),with the Ⅲ?burn wound area averaging at 12.6% of the total body surface.The back skin donor site was used to repair the wound.The functional site,such as the joint,was preferentially considered if the skin graft was not enough for all the wounds.Results: Most of the skin grafts survived and the donor sites on the back healed very well.The patients were followed up for six months to three years,which revealed that the skin grafts remained soft without dysfunction and the scar hyperplasia was mild on the donor site.Conclusion: The back donor site supplies larger-area skin with good quality.The donor site heals fast.After healing,patients could sleep on back and compress the scar so as to prevent scar hyperplasia,and the scar does not affect the patient's appearance.With all these advantages,the back donor site is a good choice for repairing Ⅲ?burn wound in children.

Objective To investigate the inhibitory effect of RNA interference on the expression of Aurora A in U251 cells, and the influence on proliferation and apoptosis of U251 cells. Methods The siRNA specific for Aurora A was synthesized and transfected into U251 cells in vitro. Aurora A mRNA expression and protein content were detected by RT-PCR and Western blotting respectively. The cell proliferation and apoptosis were observed by methyl thiazolyl tetrazolium(MTT) and flow cytometry(FCM). Transmission electron microscope was used to observe the ultrastructural changes of U251 cells. Results After transfection, the expression level of Aurora A mRNA was significantly decreased(P<0.01), and the protein content of Aurora A was also obviously reduced. The inhibitory rate of cell proliferation reached up to 67.57% 72 hours after transfection, which was significantly higer than that of normal control group(P<0.01). The apoptosis rate of U251 cells was significantly increased from (3.69±0.87)% to (15.34±2.16)% (P<0.01). Under the transmission electron microscope, it was observed that the U251 cells showed typical morphologic changes of apoptosis after transfection, such as karyopyknosis, chromatin condensation and margination, intracytoplasmic vacuoles formed, and apoptotic bodies formed. Conclusion The expression of Aurora A gene can be inhibited by siRNA successfully, and it results in the suppression of cell growth and induce apoptosis of human glioma cells in vitro. Aurora A may become a new target for the gene therapy of gliomas.

Objective To assess standardized patients (SP) in resident general practitioners training for their consultation skills to explore its role in clinical competence training. Methods SP are employed in standardized training for 52 resident general practitioners in Shanghai during May to July 2008 to assess trainees' clinical consultation skills, including medical history taking and physical examinations, as well as clinical competencies of trainers including medical history taking, physical examinations, clinical thinking (diagnosis, differential diagnosis and treatment plans), health education, medical record writing,and reception time management. Feedback and appraisal for their consultation skills were provided by SP later. Problems encountered in trainees' consultation were discussed in group manner with their trainers.And two mouths later, assessment with SPs was undergone again for their consultation skills to compare their scores in consultation skills and percentages of them passing the assessment before and after training. Results Trainees were enrolled in the training aged (28±1 ) years in average ( with a ratio of men to women of 19/33 ). Percentage of trainees passing the assessment by SP for consultation skills was significantly higher after training (88.5%) than that before it (46. 2% ) ( P < 0. 01 ). Percentage of trainees with scores passing assessment by trainers was also significantly higher after training than that before it (86.5% vs. 51.9%, P < 0.01 ), with a percentage of those qualified for clinical skills and health education after training of 80. 8% and 88.5%, respectively, as compared to those of 46. 2% and 67. 3%,respectively before it, ( P < 0. 01 ), and with a percentage of trainees qualified for clinical thinking, out-patient medical record writing and reception time management not changed after training (71.2%, 100. 0%,100. 0%, respectively) as compared to those before it (51.9%, 100. 0% and 100. 0%, respectively)(P > 0. 05 ). Conclusions Application of SP can effectively enhance consultation skills of the trainees,especially in their medical history taking and physical examinations. However, there is still rooms to improvein their clinical thinking.

Objective To assess roles of standardized patients (SPs) in training of communication skills for resident general practitioners (GP). Methods Fifty-two resident GPs were trained for communication skills in consultation with SPs during May to July, 2008, including information feedback from SPs, lectures and group discussion coordinated by instructors on problems developed during residents' clinical encounters. Communication skills of the trainees with patients was assessed before and after training, respectively, including building physician-patient relationship, collecting information and providing feedback, understanding complaints from patients' perspective, sharing information and obtaining informed consents and closing consultation. Assessment was scored and proportion of trainees qualified for consultation with SPs were estimated. Results Total score of the trainees was higher after training than that before it (76 ± 13 vs. 49 ± 12, P ＜ 0. 01 ), and 86. 5 percent of them qualified for their communication skills after training, as compared to that of 15.4 percent before it. Proportions of the trainees qualified for building physician-patient relationship, collecting information and providing feedback, understanding complaints from patients' perspective, sharing information and obtaining informed consents during consultation were higher after training than those before it (90. 4% vs. 59. 6%, 65.4% vs. 5.8%, 38.5% vs. 11.5%, 86. 5% vs.17.3% and 65. 4% vs. 3. 9%, respectively, all P ＜0. 01 ). Scores of physician-patient communication in the trainees correlated with those of their consultation skills both before and after training ( r = 0. 55 and r =0. 63, both P ＜ 0. 01 ). Increase in scores of physician-patient communication and consultation skills in trainees after training also associated with that in those of clinical skills ( r = 0. 62, P ＜ 0. 01 ). Conclusions Application of SPs can effectively enhance communication skill of resident general practitioners, thus improve their competencies in clinical consultation.

Objective To evaluate the effect ofcytochrome P450 isoenzyme subfamily 2C19 (CYP2C19)gene polymorphism on the clopidogrel antiplatelet therapy in cirrhosis patients after receiving transjugular intrahepatic portosystemic shunt (TIPS).Methods The clinical data of 171 cirrhosis patients,who were treated with TIPS during the period from January 2013 to December 2014,were retrospectively analyzed.During operation both the portal vein and the elbow vein blood samples were collected and sent for CYP2C19 gene testing.After TIPS,clinical follow-up checkup was made once every 3 months.The gene detection results and clinical follow-up findings were comparatively analyzed.Results A total of 110 patients,who had not received blood transfusion before TIPS and who had regularly taken clopidogrel antiplatelet therapy after TIPS were enrolled in the study.The mean time to take clopidogrel was 192.4 days (31-517 days),and the gene detection results of portal vein and elbow vein were quite consistent.CYP2C19 genotype of *1/*1 was found in 49 patients (44.5％),CYP2C19 genotype of *1/*2 in 27 patients (24.6％),CYP2C19 genotype of *1/*3 in 18patients (16.4％),CYP2C19 genotype of *2/*2 in 11 patients (10.0％),CYP2C19 genotype of *2/*3 in 3patients (2.7％),and CYP2C 19 genotype of *3/*3 in 2 patients (1.8％).Following-up examinations showed that the incidence of shunt dysfunction in patients carrying slow metabolic gene was 87.5％ (14/16),which was significantly higher than that in patients carrying moderate metabolic gene (20.0％,9/45;x2=22.9,P=0.006)as well as in patients carrying fast metabolic gene (8.2％,4/49;x2=37.91,P=O.O00 1).Multivariate analysis of Cox regression model indicated that CYP2C19 slow metabolic gene variation was an important predictive factor for shunt dysfunction (95％CI:1.80-9.03,P=O.O00 7).Conclusion CYP2C19 slow metabolic gene variation,including genotype of *2/*2,*2/*3 and *3/*3,is an important factor that can influence the efficacy of clopidogrel treatment after TIPS.Preoperative CYP2C19 gene detection results can provide useful information,which is very helpful in making an effective and reliable anti-platelet treatment plan for patients after TIPS.

Objective:To investigate the effect of miR-125b targeting Sema4C on STAT3 signaling pathway on invasion and metastasis of non-Hodgkin′s lymphoma.Methods: Expression of miR-125b in non-Hodgkin′s lymphoma tissues and cell lines was detected by QT-PCR.Expression of Sema4C in normal lymphocyte tissues and non-Hodgkin′s lymphoma tissues was detected by immunohistochemistry.Dual luciferase effect of miR-125b on the transcriptional activity of Sema4C was examined by the reporter gene system.Transwell invasion assay was used to detect the expression of miR-125b in the non-Hodgkin′s lymphoma cell line NK-92.Scratch test Western blot was used to detect the expression of Sema4C.Western blot was used to detect the protein expression of STAT3 signal pathway after silencing Sema4C.The protein expression of Sema4C was detected by Western blot.Results: Expression of miR-125b was significantly lower in non-Hodgkin′s lymphoma tissues than that in normal tissues[(0.48±0.05)% vs (1.59±0.38)%,P<0.05].Sema4C was highly expressed in non-Hodgkin′s lymphoma[(326.25±7.75)% vs (58.75±5.76)%].After overexpression of miR-125b,non-Hodgkin′s lymphoma cells expression level of Sema4C was down-regulated after silencing Sema4C[(326.25±7.75)% vs (58.75±5.76)%],and the expression of JAK and STAT3 protein were down-regulated after miR-125b overexpression[(85.26±6.94)% vs (12.61±4.32)%,P<0.05].The dual luciferase reporter gene system showed that miR-125b could directly regulate the transcriptional activity of Sema4C.Conclusion: miR-125b can regulate the invasion and migration of non-Hodgkin′s lymphoma cells by targets the expression of Sema4C.

Objective To investigate the efficacy of gelatin sponge particles(GSP)or polyvinyl alcohol particles (PVA) for hemoptysis secondary to bronchiectasis or pulmonary tuberculosis. Methods The clinical data on 271 patients with bronchiectasis- or tuberculosis-induced hemoptysis were retrospectively analyzed. The efficacy and rates of recurrence and complications were analyzed. Results A total 271 patients were included in this study, 176 of whom suffered from bronchiectasis and the rest 95 had tuberculosis. One-week cure rate was signifi-cantly higher in bronchiectasis group than in tuberculosis group(73.3%vs. 46.3%,P<0.05),and one-year recur-rence rate was significantly lower in bronchiectasis group(17.6%vs. 26.3%,P<0.05). One-year recurrence rate was slightly higher in patients receiving GSP than in those undergoing PVA(bronchiectasis group:22.2%vs. 10.3%, P<0.05;tuberculosis group:28.8%vs. 22.2%,P>0.05). No severe complications occurred. Conclusions In-terventional artery embolization therapy for hemoptysis secondary to bronchiectasis is better than tuberculosis-induced hemoptysis,and PVA is more effective than GSP. Recurrence of massive hemoptysis mostly occurrs within one month ,and most of the patients are complicated with blood supply and have a history of hemoptysis.

Objective:Dipeptidyl peptidase I(DPPI),a lysosomal cysteine protease for serine proteases activation,highly expressed in granule immune cells.This study used collagen induced arthritis(CIA) rat model to investigate the effects of Triperygium Wilfordii Polyglucoside(TWP) on DPPI activity and the pharmacological mechanism in RA treatment.Methods:Rats were divided into four groups randomly,the blank control group,the CIA model group,the high dose (5.0 mg/100 g body-weight) and low dose(2.5 mg/100 g body-weight)treatment group.Bovine collagen-Ⅱ plus complete Freund′s adjuvant injected twice in rats.Physical assessments were carried out.12 days post-injections,the rats of treatment group were intragastric administered with TWP every day.The rats were killed after two week administrations.Serum and synovial membrane homogenates were collected and DPPI activity was detected by fluorescence substrate.Joint HE staining and cell counting were carried out,Zymography was used to detect the MMP-2/9 activity in synovial fluids.Total protein in synovial membrane homogenates were measured by BCA method.Results:TWP could reduce the number of CIA synovial tissue mast cells,inhibited DPPI activity in the synovial fluids and in serum.The expression levels of MMP-2/9 activity and synovium total protein content were also reduced by TWP.Conclusion:Triperygium Wilfordii Polyglucoside has inhibitory effects on DPPI activity on CIA rats,which might be the one of the pharmacological mechanisms in RA treatment.