Objective To establish a multiplex polymerase chain reaction (PCR) method for rapid detection of three kinds of di‐arrheagenic Escherichia coli(EPEC ,EIEC ,EHEC)simultaneously .Methods The eae gene of EPEC ,ipaH gene of EIEC and stx1 gene of EHEC were selected to design primers ;the reaction system and condition were adjusted to optimize the multiplex PCR sys‐tem .Results The target gene fragments were amplified correctly with these primers .The three target bacteria could be detected at the same time by multiplex PCR .Conclusion A rapid multiplex PCR system were successfully established for detection of three di‐arrheagenic Escherichia coli ,and this system could be suitable for rapid screening in food safety .

Objective To put forward a reform assumption of physician payment model based on review of physician payment models abroad. Methods Literature review and expert interview. Results Physician payment model and its reform patterns overseas were discussed. Problems of current physician payment model in Chinese public hospitals were analyzed. Conclusions The physician payment reform in public hospitals should focus on raising physician salary. The investment of resources and technology of medical service projects should be measured reasonably. Setting up responsibility center , saving health care costs and improve the quality of health care are the most important three aspects of physician payment model reform in Chinese public hospitals.

Objective To explore achillis tendon-sliding procedure and subtalar joint capsula release by the same cut and individualization treatment used in the correction of congenital clubfoot.Methods Forty eight cases (76 feet) of congenital clubfoot who were first visit were treated by achillis tendon-sliding procedure and subtalar joint capsula release by the same cut,meantime,footplate fascia release and transfer of the anterior tibial tenden were made by individualization.Results All cases were followed-up,the average time follow-up was 1 year and 6 months,excellent were 71% and good were 15.7%.Five cases were recurrence and its rate was 6.94%.Conclusions Achillis tendon-sliding procedure and subtalar joint capsula release by the same cut can solve rotation of displacement between calcaneus and talus and individualization treatment can also solve high arch deformity and adduction of foot.This technique is a simple,safe and effective treatment for congenital clubfoot.

Objective To evaluate the feasibility and clinical effect of stage Ⅰ posterior vertebral corpectomy filled with titanium mesh cages plus long-segmental instrumentation for treatment of Denis type D and E severe unstable thoracolumbar burst fracture.Methods A retrospectively review was made on 14 patients with Denis type D and E severe unstable thoracolumbar burst fracture,having had posterior vertebral corpectomy,titanium mesh bone grafting and long-segmental pedicle screw fixation.Parameters recorded were operation time,amount of bleeding,bone fusion and reduction,postoperative complications,low back pain,neurological performance,restoration and loss of anterior vertebral height,correction and loss of kyphosis,and intraspinal space occupying lesion.Results Mean operation time was 207.1 min (range,148-306 min) and blood loss was 585 ml (range,300-1,500 ml).Intraoperative fracture reduction was satisfactory.Follow-up ranged from 18 to 54 months (mean,28 months),which showed no complications of infection,screw-rod breakage,loosening or shifting of the internal fixation device,titanium mesh subsidence and pseudarthrosis,and no signs of adjacent segment degeneration.Six months after operation,CT scan showed bony fusion of the bone-implant interface.At the final follow-up,the Denis pain scale improved significantly including 12 patients with completely relieve of pain (P1) and 2 ache slightly but no need of taking painkiller (P2).Neurological functions were improved by 1-3 degree in all patients.Percentage of anterior vertebral height was (41.2 ± 8.9)％ before operation,significantly reduced to (8.3 ± 4.8) ％ one week after operation and to (8.9 ± 5.1) ％ at the final follow-up (P ＜0.01).Mean loss of anterior vertebral height was 0.6％.Sagittal kyphotic angle was (36.9 ± 4.9) °before operation,significantly reduced to (8.1 ± 3.4) ° one week after operation and (8.5 ± 3.8) °at the final follow-up (P ＜0.01).Mean loss of kyphotic angle was 0.9°.Ratio of intraspinal space occupancy was (74.9 ± 11.3) ％ before operation but recovered to (4.1 ± 1.6) ％ one week after operation and (1.8 ± 1.4) ％ at the final follow-up,with significant differences in pair comparison (P ＜ 0.01).Conclusion The use of posterior vertebral corpectomy and fusion with titanium mesh cage fusion in conjunction with long-segment instrumentation are effective for spinal canal decompression,fracture reduction,three-column reconstruction as well as rigid fusion at one stage,lumbar pain relief,neurological function recovery,prevention of correction loss and vertebral height restoration.

An enzyme linked immunosorbent assay ( ELISA ) and a flow cytometry assay ( FCA ) based on Wil2-S cells were developed and systematically compared for quantification of recombinant anti-CD20 humanized monoclonal antibody ( rh-anti-CD20zumab) in biological matrix. The specificity, precision and accuracy of each method at correspondingly different linear range showed good results. For ELISA, the precisions of intra-day and inter-day were both <19 . 5%, the relative error was from-18 . 2% to 17 . 6%;For FCA, the precisions of intra-day and inter-day were both <19. 0%, the relative error was from -18. 9% to 18. 4%. The sensitivity of ELISA was significantly higher than that of FCA. The quantitative ranges of ELISA and FCA methods were 0. 04-5. 0 mg/L and 3. 1-200 mg/L, respectively. The concentrations in serum samples and pharmacokinetics analysis were determined by both of two methods after vein drip administration of rh-anti-CD20zumab in rhesus monkeys. Pharmacokinetics data showed that there was excellent consistency between results obtained by two methods at the given dose. We believe that the novel FCA with high speed and high sensitivity can be used to perform PK and PD study of cell surface antigen-targeted antibody derivatives.

Objective To explore the expression levels of RAS-interacting protein 1 (RASIP1) mRNA and protein in hepatocellular carcinoma (HCC) tissues and its cell lines,and to analyze the relationship between RASIP1 and tumorigenesis and progression of hepatocellular carcinoma.Methods The expression levels of RASIP1 mRNA and protein in 29 hepatocellular carcinoma tissues and the corresponding adjacent non-cancer liver tissues (ANLTs),as well as those in the HCC cell lines such as LO2,HEPG2,MHCC97-H and HCCLM3 were detected using real-time PCR and western blot.Results The RASIP1 expression levels decreased significantly in HCC tissues when compared with the corresponding ANLTs; The expression levels of RASIP1 mRNA and protein in LO2 were significantly higher than those in other HCC cell lines (P ＜ 0.05) ; The expression levels of RASIP1 mRNA and protein in MHCC97-H and HCCLM3 were significantly lower than those in HepG2 (P ＜ 0.05).Conclusions HCC tissues had lower expression than those in ANLTs.On analyzing the RASIP1 levels of HCC tissues and its cell lines,we speculated that RASIP1 might suppress recurrence and metastasis of HCC.

BACKGROUND:Damage to nasal ciliated epithelial cels can lead to a severe injury in nasal biological function. Compared with other adult stem cels, human umbilical cord blood stem cels have better differentiation potential.
OBJECTIVE: To explore the feasibility of human umbilical cord blood stem cels differentiating into nasal ciliated epithelial cels through in vitro culture and induction techniques.
METHODS:Normal and healthy umbilical cord blood samples were colected to isolate human umbilical cord blood stem cels, folowed by identification and subculturein vitro. Umbilical cord blood stem cels at passage 3 were infected with recombinant adeno-associated virus carrying enhanced green fluorescent protein and cultured using air liquid interface culture method. Thereafter, PCR assay was employed for detecting MUCS expression in cultured stem cels at 1 and 2 weeks after induction, and immunofluorescent staining for FOXJ1 was performed at 3 weeks.
RESULTS AND CONCLUSION:After subculture, passage 3 umbilical cord blood stem cels that could express stem cel surface markers were visible in a uniform shape and had good refraction. After 3 hours of gene transfection, green fluorescence issued from the passage 3 cels were visible, and the cel positive rate was up to 96.2% until 48 hours, indicating good transfection efficiency. RT-PCR findings showed that MUC8 mRNA had no expression in the umbilical cord blood stem cels, but expressed strongly in the nasal ciliated epithelial cels, whose expression was weak at 1 week of culture and increased at 2 weeks. Additionaly, the positive expression of FOXJ1 red fluorescence was observed under the transfection of green fluorescent protein. These results suggest that human umbilical cord blood stem cels could differentiate into nasal epithelial cels under suitable conditions.

Objective To study the clinical effect of acupuncture therapy combining with biofeedback therapy on senile chronic insomnia patients. Methods 150 cases of senile chronic insomnia were divided into 3groups randomly: group A received acupuncture therapy only, group B received biofeedback therapy only, and group C received both therapies. The scores of Pittsburgh Sleep Quality Index (PSQI) ,sleep efficacy,sleep latency, total sleep time and the scores of Hamilton Anxiety Scale (HAMA) and Hamilton Depression Scale (HAMD-17) were assessed before the treatment and at the end of the 4th and 8th week during the treatment.Results After 8 weeks treatment, the scores of PSQI, sleep efficacy, sleep latency, total sleep time and the scores of HAMA and HAMD-17 of group C((7.92 ±2.59)score,(82.52 ±8.93)% ,(24.06 ±8.23)minutes,(413.75± 42.41) minutes, (9.63 ± 3.75) score, (10.10 ± 3.27) score) were better than that of group A (( 9.51 ± 2.92)score, (79.06 ± 10.70) %, (33.16 ± 11.31) minutes, (373.47 ± 40.65) minutes, (15.08 ± 4.20) score, (14.33±± 3.56) score) and group B (( 11.46 ± 3.75) score, (68.85 ± 12.34) %, (33.65 ± 11. 38) minutes, (281.88 ±38.02) minutes, (11.63 ± 4.15) score, (12.08 ± 4.08) score) significantly (P ＜ 0.01). Conclusion The therapy of acupuncture combining with biofeedback is benefit to improve the sleep quality of senile chronic insomnia patients,meanwhile the anxiety and depression associated with insomnia can be improved.

Objective To summarize the experience in the perioperative nursing of one-stage bilateral total hip arthroplaaty (THA) for the systemic lupus erythematosus (SLE) patients with avascular necrosis of the femoral head (ANFH).Methods From June 1998 to April 2007,17 cases of patients who were diagnosed as SLE with bilateral ANFH were treated by one-stage bilateral THA.Perioperative nursing included psychological support,diet control,observation and nursing of various complications,correct functional exercise and health education.Harris score and SF-36 score were evaluated before and after operation.Results There were 2 cases with delayed incision healing,1 with early prosthesis dislocation,1 with thigh pain for 10 months and 1 with acute renal failure postoperatively,which were improved and recovered after proper treatments.Asymptomatic deep vein thrombosis (DVT) in low extremities were detected by color Doppler ultrasonography in 6 cases.There were no pulmonary embolism and no deep infection around prosthesis.There was no Addisonian crisis postoperatively.The pain was relieved and the motion of joint was improved during follow-up.There was no radiological evidence of implant loosening.The Harris hip score and SF-36 score greatly alleviated after operation.All patients were followed up and the mean follow-up time was 28 months.Conclusions One-stage bilateral THA has good results for the patients with SLE and AVNH.Strengthening of perioperative nursing contributed to improvement of success rate of surgery and reduction of complication.

Objective To observe the influence of diabetes educationon the treatment compliance and quality of life (QOL) of diabetics and to find out the influencing factors of diabetics' QOL. Methods This study applied the questionnaire of compliance and the SF-36 to make an investigation on diabetics. Ninety diabetics who participated in the course of diabetes education during the period from October 2008 to March 2009, and 205 diabetics who were seen in the first affiliated hospital of China Medical Universitybut never participated in the course of diabetes education during the period from September to October of 2008. By comparing the data with control group and using self-paired comparison, the influence of diabetes education on the treatment compliance and the QOL of diabetics was studied. The factors influencing the QOL of each dimension were analyzed by means of single or multiple regressions. Results The treatment compliance of patients was improved with the number of times that these patients participated diabetes education was increased (P<0.05). The treatment compliance of the patients in hospital who had not participated in diabetes education was improved temporarily. However, 2 weeks after these patients were discharged the treatment compliance of these patients had began to decline. The treatment compliance would be declined if patients discontinued participating education for a long time. After 24 weeks intevention, the total score of compliance of intervention group (42.44±4.10) was higher than control group (35.60±3.90), and also higher than the intervention group before intervention (34.56 ± 4.34) (all P<0.01). The scores of 5 dimensions were significantly different (P<0.05 or 0.01). Of 8 dimensions of intervention group, the scores of vitality, social functioning, role-emotional and mental health were all increased after intervention, with significant differences except for role-emotional (P<0.05). The mental health index of patients with long-term health education was increased to a certain degree. Conclusion Long-term health education plays an active role in treatment. Insisting on taking part in health education regularly can improve the quantity of mental health.