Objective To identify the expression of miRNAs in human bone marrow-derived mesenchymal stem cells (hMSCs). Method Low molecular weight RNA fraction (≤200nt) from human bone marrow-derived mesenchymal stem cells was extracted, and polyadenylated by poly(A) polymerase. Then a 5′RNA adapter was ligated to poly(A)-tailed RNA using T4 RNA ligase. RNAs were reversely transcribed and amplified by RT-PCR. The PCR products with a size of about 109 bp were recovered and cloned into pCR 4-TOPO vector. After sequencing, database searching, and expression profiling, the expression of miRNAs in hMSCs was sieved. The expression of novel and some known miRNAs was examined by Northern blotting with small RNAs (≤200nt) isolated from hMSCs, osteosarcoma cell line SOSP-9607 and UMR-106. Results hMSCs were isolated from bone marrow and purified by centrifuge and in vitro culture. Cell markers CD44 were positive, but CD34 and CD45 were negative. A total of 194 clones were subsequently characterized by DNA sequencing and database searching. 52 clones (correspond to 27species) out of 194 clones from hMSCs were identified as miRNAs. One novel miRNAs (PREDICTED-miR-202) was discovered among other 26 known miRNAs, which were predicted in Nature. Northern blotting confirmed that 1 novel miRNAs and 3 known miRNAs (miR-495, miR-34a, miR-17-5p, PREDICTED-miR-202) were stably expressed in hMSCs. Conclusion These findings indicate that a large diverse population of miRNAs are likely important regulators for hMSCs in maintaining their state of self-renewal, and have potential value in stem cells research.

Objective To evaluate the clinical outcome of alcohol-deactivated autograft-prosthesis composite after resection of bone giant cell tumor in distal femur.Methods From January 2007 to October 2008,5 patients with bone giant cell tumor in distal femur were treated with alcohol-deactivated autograftprosthesis composite,including 3 males and 2 females with an average age of 29.6 years(range,22-40).Three patients were diagnosed with postoperative recurrence,and 2 with pathological fracture.All patients were of Campanacci Ⅲ.Three-dimensional finite element models with 40% bone defect in distal femur were established based on CT images of a healthy volunteer.Three times of body mass load corresponding to the normal walking gait cycle was applied.The influence on stress distribution of femur-cement and prosthesis stem was analyzed.Results All patients were followed up for average 37 months,there was no infection,recurrence,loosening and limb length inequality.The bony healing time was 6 to 11 months.The mean MSTS function score was 25.7(range,25-27).The mean ISOLS graft score was 31.4 (range,28-35).The finite element analysis showed that for the short-term model,the maximum stress was 145.82 MPa in the proximal femur,40.90 MPa in the medial side of 1/4 proximal cement,and 389.24 MPa in the proximal prosthesis stem.The maximum stress was not exceeding the fatigue strength in three sites.For the long-term model,with the bone healing,the maximum stress on three sites decreased to 139.05,36.95,and 253.65 MPa,respectively.Conclusion These results suggest that the alcohol-deactivated autograft-prosthesis composite after resection ot bone giant cell tumor in distal femur can reduce the tumor recurrence and improve the short-term limb function,It is stable in short term and can reduce stress shielding in long term.

Objective To analyze the incidence of remaining symptoms after discectomy with different operation,and investigate its causes and clinical significance.Methods Two hundred and twentyfour patients were followed up.The incidence of remaining symptoms and clinicai situation after discectomy were observed with different operation:discectomy (110 patients,single group),discectomy with bone grafting internal fixation (70 patients,fusion group),and discectomy with dynamic fixation (44 patients,non-fusion group).The SPSS 19.0 software was used to analyze the data.Results One hundred and sixpatients (47.32％,106/224) occurred remaining symptoms in 224 patients.In single group was 69 patients (62.73％,69/110),in fusion group was 26 patients (37.14％,26/70),and in non-fusion group was 11 patients (25.00％,11/44).The incidence of remaining symptoms among three groups was significant difference(x2 =22.177,P =0.000).The incidence of remaining symptoms in single group was significantly higher than that in non-fusion group and fusion group (x2 =17.921,P =0.000;x2 =11.235,P =0.001).The incidence of remaining symptoms in non-fusion group and fusion group had no significant difference (x2 =1.817,P =0.178).Conclusions There is significantly different in the incidence of lumbar disc herniation with different surgery methods.Discectomy with bone grafting internal fixation and with dynamic fixation can effectively reduce the incidence of remaining symptoms.

Objective To investigate the clinical value of continuity nursing model in caring patients with malignant obstructive jaundice treated with percutaneous transhepatic biliary drainage (PTBD).Methods A total of 120 patients with malignant obstructive jaundice treated by PTCD were enrolled in this study.The patients were divided into the control group (n=60) and the observation group (n=60).Routine discharge guidance and health education was conducted for the patients of control group,while for the patients of observation group,in addition to conventional discharge education,the continuity nursing was executed by the responsible nurse.Continuity nursing was meant to continue the service,to guide the observation of the wound and dressing change,the observation of the quantity and quality of drainage solution,to teach the knowledge of the management of PTCD catheter as well as its complications,to guide patient's diet and rest,and to establish the continuity nursing records.Results The patients of both groups were followed up for 3 months.The patients' awareness rate of the knowledge related to PTCD tube in the observation group was significantly higher than that in the control group.The wound infection rate,the rate of PTCD tube prolapse or blockage,and the tube-related re-hospitalization rate in the observation group were strikingly lower than those in the control group (P＜0.05).Conclusion The continuity nursing model can significantly improve patients' awareness rate about the knowledge related to PTCD tube,reduce the incidences of biliary tract infection,PTCD tube prolapse or blockage,wound infection,and tube-related re-hospitalization,therefore,the quality of life can be surely improved.

OBJECTIVE To evaluate the commonly encountered pathogens and drng resistance of bacteria causing secondary infection in patients with malignant tumors and provide reference for clinical antimicrobial usage.METHODS Statistical analysis was made retrospectively in the classification and antimicrobial susceptibility testing in 1 204 strains of pathogens isolated from clinical samples of hospitalized patients with malignant tumors from Jan 2003 to Oct 2005.RESULTS Among 1 204 strains,Gram-positive strains accounted for 31.1%;Gram-negative ones accounted for 52.8%;and fungi accounted for 16.1%.The principal strains were Candida albicans,Escherichia coli,Staphylococcus aureus,Klebsiella pneumoniae,and Pseudomonas aeruginosa.The most frequent infection sites were in respiratory tract,urinary tract and skin-soft tissue.Multiple drug resistant rate of Gram-positive and Gram-negative strains had a tendency of elevation.CONCLUSIONS To reduce the coming of drug resistant strains,etiologic examination should be done in treatment of infectious diseases and antimicrobial therapy should be decided according to the results of susceptibility.

Objective:To study the curative efficacy of budesonide combined with huaiqihuang granules in the treatment of children with asthma and its effects on the serum leukotriene D4(LTD4),nerve growth factor (NGF),tissue inhibitor of metalloproteinase-1 (TIMP-1) levels.Methods:90 cases of children with asthma who were treated from March 2014 to March 2015 in our hospital were selected and divided into the observation group (n=45) and the control group (n=45) according to the random number table.Both groups of children were given bronchial antispasmodic agent,oxygen,antibiotics,glucocorticoid and other conventional treatment after admission,the control group was given budesonide atomization inhalation treatment on the basis of conventional treatment,0.5 mg budesonide were add in 3ml saline for continuous inhalation,twice a day,while the observation group was treated with Huaiqihuang Granules on the basis of the control group,twice a day.After 12 weeks of treatment,the curative effect,disappearance time of wheezing,cough relief,pulmonary rales,wheezing and other symptoms,forced vital capacity (FVC),serum LTD4,NGF,TIMP-1 levels,CD3+,CD4+,CD8+ of both groups were compared.Results:After treatment,the total effective rate of observation group was significantly higher than that of the control group [93.33％(42/45) vs 77.77％(35/45)](P＜0.05);disappearance time of breath,cough,moist rales and wheezes were significantly shorter than those of the control group[(4.32± 1.03)d vs (6.08± 1.24)d,(5.60± 1.12)d vs (7.21± 1.30)d,(3.19± 0.98)d vs (4.98± 1.02)d,(3.25± 1.03)d vs (5.89± 1.35)d](P＜0.05);the FEV1,FEV1/FVC were significantly higher than those of the control group [(92.63± 10.01)L/s vs (78.36± 9.19)L/s,(95.37± 11.72)％ vs (80.19± 10.23)％](P＜0.05);the serum LTD4,NGF,TIMP-1 levels were significantly lower than those of the control group [(7.24± 0.86)ng/mL vs (12.68± 1.01vng/mL,(68.18± 9.01)pg/mL vs (80.78± 10.24)pg/mL,(34.16 ± 5.06)ng/mL vs (49.76 ± 5.47)ng/mL] (P ＜0.05);the CD3+,CD4+ were significantly higher than those of the control group [(66.15± 7.20)％ vs (62.03± 6.85)％,(45.13± 7.90)％ vs (37.42± 7.06)％],CD8+ was significantly lower than that of the control group[(34.16± 5.06)％ vs (49.76± 5.47)％] (P＜0.05).Conclusion:Budesonide combined with Huaiqihuang granules was effective for children with asthma,which could enhance the immune function,improve the lung function,reduce the serum levels of LTD4,NGF,TIMP-1.

OBJECTIVE:To analyze bacterial infection of chronic bacterial prostatitis and to provide reference for clinical therapy. METHODS: Prostatic fluid was collected in sterile condition. Positive results of bacterial culture in 261 cases of chronic bacterial prostatitis were analyzed. RESULTS: Of total 261 cases, gram-postive bacteria accounted for 56.7%, which were mainly staphylococcus bacteria (41.9%) and gram-negative bacteria (43.3%) among which Escherichia coli accounted for 42.5%. CONCLUSION: Culture and drug sensitivity analysis of prostatic fluid and rational use of antibiotics based on results of drug sensitivity test has significance for drug use in the clinic.

Objective:To improve the sensitivity and the linear range of electrochemical immunosensor to detect Schistosoma japonicum (S.japonicum) antibody.Methods:Carbon inks and silver/silver chloride inks were printed on a polyethylene terephthalate (PET) board to make a two-electrode test strip,where carbon was the working electrode and S.japonicum soluble egg antigen (SEA) was fixed at one end of working electrode by different methods; silver/silver chloride electrode was used as control.We tested the valency of the antibody by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in an electrochemistry workstation,and conducted comparison with the results of ELISA.Two new immunosensing electrodes have been developed,based on glutaraldehyde cross-linked (GA) or chitosan-glutaraldehyde cross-linked (Chit-GA) transducer fixing S.japonicum antigen.We tested the titer of the antibody by means of CV and DPV.Results:Our experimental S.japonicum antigen (50 μg/L) is the optimal test concentration for the GA sensor,and 10 μg/L for Chit-GA sensors.The immune reaction time of both electrodes is all essentially complete in 1 minute.The linear range for S.japonicura antibody in human positive serum sample detection by the glutaraldehyde cross-linked immunosensor is 1∶1000 to 1∶400,and by the chitosan-glutaraldehyde cross-linked immunosensor is 1∶1000 to 1∶500.As the concentration of dilution ratio of S.japonicum antibody in human positive serum sample increased,the test value of DPV increased proportionally.Conclusion:GA sensor and Chit-GA cross-linked S.japonicum sensors have high sensitivity and broad linear range response,and both exhibited a good linear relationship between the DPV signal and the test antibody titer.

Objective:To investigate the value of the three-dimensional CT reconstruction by computer photo process system for laryngeal carcinoma and control study with pathologic feature. Methods: Fifty patients with laryngeal carcinoma were scanned by helical CT which photos were three-dimensionally reconstructed with a photo processing system,and the images were compared with pathologic feature.Results:Clear three-dimensional photo of the laryngeal was obtained through the reconstruction of the helical CT.The images could be circular fully around the axis of X,Y and Z in order to find the best view of the diagnosis.Ninety-six percentage of the TNM stage of patients according to the images of three-dimensional reconstruction were consistent with the TNM stage after operation.The safe edge was obtained in 98% of the surgery for patients.Good correlations were showed in the images of the three-dimensional reconstruction of the larynx and pathologic specimens.Conclusion:Good consistent of the three-dimensional images of the larynx with pathologic specimen were useful for diagnosis,operation design and education.