BACKGROUND: The precartilaginous stern cells are limited regarding in vitro proliferative capacity, but the immortalized cell lines can provide a large number of stable immortalized cells, and simian virus 40 large T antigen gene (SV40Tag) is one of gene fragments which are commonly used and effective in vitro immortalized ceils. OBJECTIVE: To construct human immortalized precartilaginous stem cells (IPSCs) using human precartilaginous stem calls induced by SV40LTAg gane. METHODS: The human immortalized precartilaginous stem calls were isolated from aborted fetus and purified with enzyme digestion and immunomagnetic beads screening method. By using liposome-mediated gene transfection technology, plasmid pCMVSV40T/PUR containing SV40Tag was transfected in primary embryonic precartilaginous stem cells, while non-transfected cells sewed as negative controls. Positive clones were cultured to observe the cell morphology and the passage recovery, to calculate cell survival rata and population doubling time, to drew call growth curve. Immunofluorescence cytochemistry was used to detect the expression of IPSCs fibroblast growth factor receptor 3, the expressions of SV40Tag and fibroblast growth factor receptor 3 in the human precartilaginous stem cells were determined by RT-PCR. RESULTS AND CONCLUSION: Morphology of human IPSCs seemed coincidence with primary human precartilaginous stem cells. The survival rate of human IPSCs was not influenced by subculture, freezing and recovery, but the survival rate was descended in the human precartilaginous stem cells at the 6~(th) and 10~(th) passages (P < 0.01). Compared with cells at the 6~(th) and 10~(th) passages, the proliferation of human IPSCs was greater, with short population doubling time and high growth rate (P < 0.01). The immunofluorescence showed that fibroblast growth factor receptor 3 was positive in human IPSCs at the second passage, and the RT-PCR results of fibroblast growth factor receptor 3 revealed a specific amplification band at 400 bp,.while that of SV40Tag revealed at 560 bp. No band was seen in the primary cells. It is indicated that SV40Tag human IPSCs can be constructed successfully using immunomagnatic bead screening technology and liposome transfection technique.

Objective To detect the expressions of HuR and COX-2 in epithelial ovarian carcinoma,and investigate the correlation of HuR and COX-2 expression. In addition, we attempt to seek the pathway to prevent the occurrence and development of epithelial ovarian cancer by combined analysis of various clinicopathologic characteristics. Methods The expressions of HuR and COX-2 in 68 epithelial ovarian carcinoma, 10 borderline ovarian tumors and 5 normal ovarian tissues were examined by S-P immunohistochemical method. The relationship of HuR and COX-2 expressions with clinicopathologic parameterwere evaluated by correlation analysis. Results(1) The expression of HuR in epithelial ovarian cancer tissue (76. 47% ,52/68) was significantly higher than that in borderline epithelial ovarian tumor tissues (30. 00% ,3/10) and normal ovarian tissues (0, x2 = 18. 873, Ps ＜ 0. 05), but there were no significant differences betweenthe expressions of HuR in borderline epithelial ovarian tumor and normal ovarian tissue(P ＞ 0. 05).(2) The positive expression rate of cytoplasmic HuR in epithelial ovarian carcinoma and borderline epithelial ovarian tumor were 45.60% (31/68) and 10. 00% (1/10) respectively,but normal ovarian tissues showed no staining of HuR. We found no significant differences between the expression of cytoplasmic HuR in epithelial ovarian carcinoma and borderline epithelial ovarian tumor or normal ovarian tissue(x2 = 7. 999 ,P =0. 018).(3) The positive expression rate of cytoplasmic HuR in epithelial ovarian carcinoma of FIGO stage Ⅲ - Ⅳ was significantly higher than that of stage Ⅰ - Ⅱ(56. 09% vs. 29. 63%, x2 = 4. 598, P = 0. 032). The positive expression rates of cytoplasmic HuR in epithelial ovarian carcinoma of histological grade 1,2,3 were 10. 00%,46. 67% ,57. 14% respectively, which showed significant difference in the comparison among the three groups (x2 =6. 627 ,P =0. 036). (4) The positive expression rates of COX-2 in epithelial ovarian cancer (67. 64%)and borderline epithelial ovarian tumor tissues (60. 00%) were significantly higher than that in normal ovarian tissues (0, Ps ＜ 0. 05), but we found no significant difference in the comparison between the expression of malignant and borderline ovarian tumors. Statistical analysis showed that the positive expression rate of COX-2 in epithelial ovarian carcinoma was correlated with FIGO stage and lymph node metastasis. (5)There was a significantly positive correlation between cytoplasmic HuR and COX-2 expressions in epithelial ovarian carcinoma. Conclusion The expressions of HuR and COX-2 increased in the epithelial ovarian carcinoma, and the cytoplasmic expression of HuR was significantly correlated with the expression of COX-2. These results suggested that increased cytoplasmic expression of HuR and COX-2 expression might play important roles in the initiation and development of epithelial ovarian carcinoma.

Objective To explore the mechanisms of posterior pilon fractures and evaluate the curative effects of different types of fixation on the treatment of posterior pilon fractures.Methods We included in this retrospective study 20 patients with posterior pilon fracture who had been treated from January 2012 to January 2015 at our department.They were 10 men and 10 women,from 23 to 77 years of age (average,50.6 years).According to the classification by Yu Guangrong,5 cases belonged to type Ⅰ,3 to type Ⅱa,4 to type Ⅱb,and 7 to type Ⅲ.One was not indentified because of lacking CT examination.The mechanisms included ground level fall in 2 cases,motor vehicle accident in 7,fall off stairs in 5,sport injury in 2,fall from a bike in one and fall from a height in 3.More than 25％ of the articular surface was involved in 13 patients.Syndesmosis injury was identified in 6 patients by Cotton test during operation.Internal fixation varied accordingly.We recorded the mechanism,classification,proportion of the articular surface involved (more or less than 25％),and syndesmosis injury to figure out the characteristics of posterior pilon fractures.We used the Burwell-Charnley radiographic criteria to assess the postoperative reduction of the articular surface,and the Olerud-Molander scoring scale and visual analogue scale (VAS) to assess the ankle function.The curative effects of different types of fixation on the treatment of posterior pilon fractures were compared.Results Of the 20 patients,17 were available for follow-up for 6 to 36 months (average,17.8 months).Two patients received reoperation because of implant failure after cannulated screw fixation from anterior to posterior.The Burwell-Charnley radiographic evaluation revealed 12 anatoinical reducations and 8 fair reductions.The mean Olerud-Molander score for the 17 patients at the final follow-ups was 81.5 (range,from 35 to 100) and the mean walking VAS was 1 (from 0 to 3).Conclusions Posterior pilon fractures are mostly caused by medium to high energy violence,resulting from a combination of rotational and vertical forces.Since there is a high risk of implant failure,the cannulated screw fixation from anterior to posterior is not recommended.Good clinical outcomes are observed in the cannulated screw fixation from posterior to anterior and the plate/cannulated screw fixation for posterior pilon fractures.

Objective This study aimed to construct an index system for nurse performance appraisal based on the quantity,quality,efficiency and technology in holistic nursing mode.Methods Expert interviews and Delphi expert enquiry were used to complete the construction of index system,and SPSS17.0 was used to do data statistics and reliability analysis.Results The nurse performance appraisal index system was constructed which contained 4 level-Ⅰ indicators such as level coefficient,nursing workload scores,nursing quality scoring rate and reward amount and 12 level-Ⅱ indicators.The calculation method of each part was also established.The expert positive coefficients were 94.3％ and 100.0％ respectively,the authority coefficients were 0.83 and 0.84,the importance and feasibility mean scores of every index were 4.27-5.00 and 4.26-4.94,and coefficients of variation were 0.00-0.26 and 0.05-0.26 in 2 rounds of enquiry.The importance of level-Ⅰ or level-Ⅱ indicators and operability coordination coefficients were 0.13-0.43,the significance test showed P ＜ 0.05.Conclusions The design of each dimension of the index system gives full consideration to nurse post management,hierarchical using,the nurse in charge of the patient,grading nursing care and many other factors.Representative of key indicators is strong,and it can reflect the distribution concept that more payment for more contributions,best's best reward and rewarding the good and punishing the bad.

Objective　To observe the effects of Ang-Ⅱ on expressions of c-fos mRNA and pro-α1［Ⅰ］mRNA in fibroblasts (FBC) of rat embryo in the course of collagen synthesis.Methods　The change of free Ca2+(［Ca2+］i) was tested in fluorometry and the expressions of c-fos mRNA and pro-α1［Ⅰ］mRNA were tested by RT-PCR method in incubated FBC.Results　The level of［Ca2+］i in FBC and the expressions of c-fos mRNA and pro-α1［Ⅰ］mRNA were enhanced with the increase of Ang-Ⅱ concentration(10-8～10-5and 10-7～10-6mol/L).Conclusions　The expressions of c-fos mRNA and pro-α1［Ⅰ］mRNA were induced by Ang-Ⅱ activating signal system in FBC.

Objective To determine the effect of mild hypothermia on neonatal piglet cardiac hemodynamic function after hypoxia-ischemia (HI).Method Twenty five 7-day-old piglets were used for hypoxic ischemic brain damage (HIBD) model by the method of temporary occlusion of the bilateral carotid arteries and followed by mechanical ventilation with low concentration of oxygen (FiO_2=6%) for 30 minutes.The piglets were randomly divided into three groups:group A (normothermia with body temperature to 39℃,n=9),group B (body temperature to 36℃for 72 hours,n=8),and group C (body temperature to 34℃for 72 hours,n=8).Mild hypothermia was initiated at 4 hours after HI,the systolic and diastole function were evaluated by Doppler echocardiography at pre-HI,post-Hi 4 hours and post-HI 72 hours.Results There were no significant differences in left ventrieular ejection time/left ventrieular ejection time (LPEP/LVEF),right ventricular ejection acceleration time/right ventricular ejection time (RACT/RVET) and CO at post-HI with hypothermia 72 hours in three groups,but the heart rate decreased in B and group C group.Compared with nonnothermia,mild hypothermia treatment showed no significant differences in MAP,LPEP/LVET,RACT/RVET,CO,SV at post-HI with hypothermia 72 hours.Conclusions Body temperature decreased by 3～5℃for 72 hours will not aggravate hemodynamic abnormity.

Objective To explore the localization of epileptogenic focus and select the appropriate surgical procedures for post-traumatic epilepsy. Methods The clinical data of 21 patients with post-traumatic epilepsy were studied retrospectively. Epileptogenic focus was located by comprehensively analyzing data of electro-neurophysiology, neurological imaging and clinical manifestation. Surgical procedures were performed in all patients, including resection of lesion and peripheral cortex in 12 patients, epileptogenie focus resection plus low power bipolar coagulation in five, anterior temporal iobectomy plus amygdalohippocampectomy in three and corpus callosotomy in one. Results All patients were followed up from 6 months to 3 years, which showed satisfactory outcome in eight patients, marked improvement in six, improvement in five and slight improvement in two. The total effective rate was 90%. Conclusions Surgical procedure is important for intractable post-traumatic epilepsy. The good efficacy depends on precise localization of epileptogenic focus and combined application of various surgical procedures.

Animals ; Lung ; immunology ; pathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; physiopathology ; Spleen ; immunology ; pathology ; fas Receptor ; analysis

OBJECTIVETo evaluate middle and long-term results of total hip arthroplasties (THA) for the treatment of secondary hip traumatic osteoarthritis and femoral head necrosis after acetabular fractures.

METHODSFrom January 2000 to December 2005, 33 patients with secondary hip traumatic osteoarthritis and (or) femoral head necrosis after acetabular fractures were treated with THA. There were 21 males and 12 females, ranging in age from 27 to 69 years old, with an average of 52 years old. Twenty-three patients were performed with open reduction and internal fixation: 5 patients were treated with anterior approach; 12 patients, posterior approach; 6 patients, combined approaches; other 10 patients, conservative treatment in the early stage. All THA were performed with posterior-lateral approach. Bone union was achieved in the all acetabular fractures. Removal of all implants was necessary in 5 patients, and partial removal in 3 patients. Cemented cup was implanted in 6 patients and uncommented cup in 27 patients. Intraoperative and postoperative complications were observed, and Harris hip scores before surgery and 10 years after operation were compared. The prosthetic loosening, osteolysis or revision were used to evaluate 10 years survival rate of prosthesis.

RESULTSAll the patients were followed up,and the duration ranged from 10 to 15 years, with a mean of 12 years. One patient died at the 10th year after operation. The Harris score at the 10th year was higher than the preoperative one. One and two patients were performed with revision total hip arthroplasty caused by aseptic loosening alone and aseptic loosening combined with osteolysis respectively. Osteolysis occurred in 1 patient; deep venous thrombosis in 4 patients; dislocation of prosthesis in 2 patients. One patient had infection of incision and one patient had infection around the prosthesis. Ten years survival rate of implant was 84.8% (28/133).

CONCLUSIONTHA is an effective method to treat secondary hip traumatic osteoarthritis and (or) femoral head necrosis after acetabular fractures in improving hip joint functions with high implant survival rate and good middle and long-term results.

Acetabulum ; injuries ; Adult ; Aged ; Arthroplasty, Replacement, Hip ; methods ; Female ; Femur Head Necrosis ; surgery ; Fractures, Bone ; complications ; Hip Injuries ; complications ; Humans ; Male ; Middle Aged ; Osteoarthritis, Hip ; surgery ; Postoperative Complications ; surgery

Objective:To investigate the effect of lncRNA HOTAIR on the radiosensitivity of glioma cells and its underlying mechanism.Methods:The negative control plasmid, HOTAIR silencing plasmid, miR-NC over expressing plasmid, miR-17-5p over expressing plasmid were transfected into U87R cells, and assigned intothe silencing control, HOTAIR silencing, miR-NC over expressing and miR-17-5 pover expressing groups. Cells in the the above groups were irradiated at a dose of 4Gy, and recorded as silencing control+ 4Gy group, HOTAIRsilencing+ 4Gy group, miR-NC over expressing+ 4Gy group and miR-17-5p over expressing+ 4Gy group. The HOTAIR silencing plasmid, miR-NC suppressing plasmid and miR-17-5p suppressing plasmid were co-transfected into U87R cells and recorded as the HOTAIR silencing+ miR-NC suppressing group and HOTAIR silencing+ miR-17-5p suppressing group. All procedures were transfected by the liposome method. The expression of miR-17-5p and HOTAIR was detected by qRT-PCR. The radio sensitivity of glioma cells was evaluated by cell clone formation assay. The cell apoptosis was assessed by flow cytometry. The fluorescence activity was assessed by dual luciferase reporter assay.Results:HOTAIR was highly expressed in the radiation-resistant glioma cells. Silencing HOTAIR and over-expressing miR-17-5p could increase the radiosensitivity of U87R cells and promote radiation-induced apoptosis of U87R cells. HOTAIR could target and regulate the miR-17-5p expression. Suppressing miR-17-5p reversed the effect of silencing HOTAIR on U87R cell sensitization and promoting radiation-induced U87R cell apoptosis.Conclusions:Silencing lncRNA HOTAIR yields radiation sensitization and promotes radiation-induced apoptosis in glioma cells. The mechanism may be related to the regulation of miR-17-5p.