Objective: To investigate the curative effects of apoptin on the multidrug-resistant model, of human osteosarcoma cell line (R-OS-732). Methods: 363 bpVP 3 gene was cloned into the vector pIRES1 and the recombinant eukaryon expression vector pIRVP3 was transfected into R-OS-732 cells with liposome. The expression of apoptin gene at transcription level was proved by RT-PCR. The pathological changes of the cells was observed by light microscope and electronmicroscope. The transfected R-OS-732 cells were collected after 48 hours. The genomic DNA extracted from the cells was observed by agarose gel electrophoresis. The apoptosis rate of the cells was analysed by flow cytometry. Results: The expression of apoptin gene at transcription level had been proved by RT-PCR. The construction changes of the two groups were obviously different under light microscope: most of R-OS-732 cells in the transfected group existed in form of being away from the bottom of the culture dish after 24 hours, in form of apoptosis after 48 hours and in form of necrosis after 72 hours; but those cells in the controlled group grow luxuriantly. And under electronmicroscope there was much change of cell nucleus between the two groups. DNA ladder of the transfected cells was observed through agarose gel electrophoresis only one band was observed in the controlled group; but bands of fragmented DNA observed in ositive control group. Flow cytometry analysis showed that the apoptosis rate of the cells in the transfected group was relatively higher than that of the controlls( P

Objective To investigate the influences of 5-azacytidine (5-Aza) with different concentrations on mesenchymal stem cells (MSCs) proliferation and differentiation into myoblasts. Methods Bone marrow-derived MSCs of 4 weeks Wistar rats were separated and purified, and then treated with 5-Aza with different concentrations. The growth ability of cell was assayed with methyl thiazolyl tetrazolium (MTT) method. The expression of skeletal muscle actin in MSCs was determined with reverse transcription polymerase chain reaction (RT-PCR) and electrophoresis after MSCs were induced. Results The cell proliferation was not affected by 0 and 1 ?mol ? L-1 5-Aza . There were expressions of skeletal muscle actin treated with 3 -12 ?mol ? L-1 5-Aza. Some cells were obviously enlarged at the 9th day after induction and myotubu-like cells were found at the 12th day when treated with 9 and 12 ?mol ? L-1 5-Aza. 20 - 30 ?mol ? L-1 5-Aza induced the toxic effect on proliferation of MSCs. With the increase of concentration, the proliferation ability of MSCs was weakened. Conclusion 5-Aza affects the expression of regulatory gene to the stem cells and regulate MSCs to orientationally differentiate into myotube-like cells.

Objective To observe the effects of the Dachaihu decoction on intra-abdominal pressure in severe acute pancreatitis. Methods 70 patients with SAP from China-Japan Friendship Hospital were randomly divided into two groups. 35 patients in a conventional treatment group were treated with conventional treatment of severe acute pancreatitis, and 35 patients in a Dachaihu decoction treatment group were treated with the decoction through nasogastric tube based on conventional treatment. The changes of intra-abdominal pressure and the morbidity of IAH and ACS in both groups were observed in acute stage. Results The intra-abdominal pressure in both groups increased. But the pressure in the Dachaihu decoction treatment group (13.31 ± 4.42 mmHg, 13.02 ± 5.23 mmHg, 12.35 ± 3.34 mmHg, 11.26 ± 4.46 mmHg, 9.89 ± 3.44 mmHg) was lower than the conventional treatment group (16.89 ± 5.71 mmHg, 17.52 ± 3.37 mmHg, 16.21 ± 2.15 mmHg, 14.57 ± 5.56 mmHg, 12.11 ± 1.28 mmHg) from the third day on (P<0.05). And the morbidity of intra-abdominal hypertension and abdominal compartment syndrome in the Dachaihu decoction treatment group were lower than the conventional treatment group in the whole acut stage (40%vs. 8.75%).The differences between two groups were statistically significant (P<0.05). Conclusion The Dachaihu decoction may significantly reduce the intra-abdominal pressure and the morbidity of intra-abdominal hypertension and abdominal compartment syndrome among patients with severe acute pancreatitis.

Objective To investigate the value of central venous pressure (CVP) combined with visual left ventricular ejection fraction(LVEF) as an indicator of fluid responsiveness in patients with septic shock.Methods A retrospective analysis of 83 patients with septic shock receiving fluid challenge was conducted.The hemodynamic changes were evaluated with the pulse indicator continuous cardiac output (PiCCO) monitor,and all the patients were divided into two groups:the responded group (△CI％ ≥15％)and the unresponded group(△CI ＜ 15％),according to the change in CI(△CI％).Those patients were divided into several subgroups,including low CVP group (CVP ＜ 8 mmHg) and high CVP group (CVP ≥ 8 mmHg),low LVEF group(LVEF ＜ 50％) and high LVEF group(LVEF ≥ 50％),according to the initial value of CVP and LVEF.The changes in hemodynamic variables were compared before and after fluid challenge in each subgroup.The ability of CVP and visual LVEF was evaluated to predict fluid responsiveness with receiver operating characteristic curves.Results Fifty three (63.9％) patients responded to the fluid challenge,and lower CVP and higher LVEF patients were more frequency in responded group.The threshold value of 8 mmHg CVP for prediction of fluid responsiveness revealed the area under the curve (AUC) of 0.646 (P =0.219),and 50％ visual LVEF revealed AUC of 0.729 (P =0.023).CVP in combination with visual LVEF for prediction of fluid responsiveness showed an AUC of 0.817 (P =0.001) with a sensitivity of 76.3％ and a specificity of 88.2％.Conclusions Visual LVEF alone and combined with CVP can be used as an indicator of fluid responsiveness in patients with septic shock.

Objective To investigate the accuracy of careful clinical evaluation in hemodynamic status and guidance of PiCCO monitor in clinical treatment.Methods A total of 96 hemodynamic unstable cases were evaluated prior to the insertion of the PiCCO catheter.The attending physician in charge of the patient was required to complete a questionnaire to predict the range of key hemodynamic variables for CI,GEDI,SVRI and EVLWI.Additionally,the attending was also asked to indicate a plan for therapy based on the predicted hemodynamic profile and decide if the predicted therapy plan was altered after the the first measurement of hemodynamic variables.Results The accurate prediction of hemodynamic variables was CI (55.2％),GEDI(60.4％),SVRI(63.5％) 和 EVLWI (78.1％),among which EVLWI had a higher accuracy(P ＜ 0.05).49％ doctors altered their planned therapy according to the result of the PiCCO information.Doctors had more difficulty in accurately predicting hemodynamic values in critical patients which APACHE Ⅱ scored 15 ～25 (42.3％ vs 67.9％ and 42.3 ％ vs 75.0％,x2 =4.755,5.231,P ＜ 0.05).The prediction of patients with acute myocardial infarction was more accurate than those of without acute myocardial infarction,and less to alter the planned therapy(21.1％ vs 55.8％,x2 =7.382,P =0.007).The patients of impaired oxygenation had less accurate predictions and less therapy alterations(32.3％ vs 56.9％,x2 =5.110,P =0.024).Attending was able to predict the hemodynamic status more accurately(63.9％ vs 40％,x2 =5.152,P =0.023) and alter the predicted therapy less(39.3％ vs 65.7％,x2 =6.189,P =0.013) in patients who were enrolled later.Conclusions Clinical evaluation in hemodynamic status of critically ill patients had a lower accuracy,the information obtained by PiCCO often instruct clinical doctors to choose the optimal treatment.

Objective To study the effects of rehabilitation training on the expression of Nogo-A around the area of cerebral infraction using rats. Methods A total of 60 Wistar rats were randomly divided into a reha- bilitation group and a control group after an experimental cerebral infarction had been established in them. The ani- mals in the rehabilitation group were given exercise with a rotating bar, a balance beam and a rolling cage for one hour daily, while those in the control group were caged without any abnormal exercise. Nogo-A expression in the ar- ea surrounding the infarcts was detected by imunohistochemical techniques at the 3rd, 7th, 14th, 21st and 28th day after infarction. Meanwhile, neurobehavioral evaluations were also conducted. Results The animals in the rehabilitation group scored much lower than the controls in the behavioral evaluations at the 14th, 21st and 28th day. The expression of Nogo-A in tissues around the infracted area increased by the 7th day and peaked at the 21st day in both groups, but the expression of Nogo-A was significantly stronger in the rehabilitation group at the 14th, 21st and 28th days. Conclusion Rehabilitation training decreased the expression of Nogo-A in the brain of rats after infarction. This may have important implications for the functional recovery of the central nervous system.

BACKGROUND: At present, there are investigations on the expression of cytokines and adhesion molecular in ischemia-reperfusion injury at abroad,but they do not involve in the relative studies on endogenous cytokines and adhesion molecular on microvascular endothelial surface following injury.The expression of endogenous interleukin-1(IL-1) is limited only at mRNA level.OBJECTIVE: To prove into the mechanism of the expression of intercellular adhesion molecular 1 and its regulation factor IL-1 in spinal ischemia-reperfusion injury.DESIGN: A randomized grouping design, animal experiment.SETTING: Department of Sports Medicine, College of Physical Education Affiliated to Jilin UniversityMATERIALS: This experiment was carried out at the Central Laboratory,China-Japan Union Hospital, Jilin University between March 2003 and January 2004. Totally 77 healthy male Wistar rats were randomly divided into normal control group (n=7), simple ischemia group (n=14) and ischemia-reperfusion group (n=56). Among the rats in the simple-ischemia group, 7 rats suffered from blood flow block for 30 minutes and 7 rats for 60 minutes; Rats in the ischemia-reperfusion group were assigned into 8 subgroups according to 8 time phases, respectively at reperfusion for 30,60 minutes, 2, 4, 6,9, 12 and 24 hours following spinal ischemia, with 7 rats at each time phase.METHODS: Spinal ischemia-reperfusion injury animal models were created with Zivin method. The expressions of vascular endothelial intercellular adhesion molecule-1 (ICAM-1) mRNA and IL-1β mRNA following spinal ischemia-reperfusion injury were detected with reverse transcriptionpolymerase chain reaction, immunohistochemistry and immunofluorescent confocal laser scanning microscope technique.MAIN OUTCOME MEASURES: The expression of IL-1β mRNA, activity of IL-1 polypeptide, expression of ICAM-1 mRNA and protein and activity of myeloperoxidase (MPO).RFSULTS: Totally 77 animals were enrolled and all of them entered the stage of result analysis. ① The expression of IL-1β mRNA (A value)was significantly higher in the ischemia-reperfusion group than in the simple ischemia group and normal control group, with significant difference (respectively 1.07±0.33,0.60±0.22,0.57±0.12,t=3.751 7,11.852 6,P ＜ 0.01).② Activity of IL-1 polypeptide (A value )was significantly higher in the ischemia-reperfusion group than in the simple ischemia group and normal control group, with significant difference [respectively (33.7±3.2),(23.8±4.5), (23.1±2.1),t=2.798 8,9.962 7,P ＜ 0.01]. ③ ICAM-1 mRNA(A value)was significantly higher in ischemia-reperfusion group than in simple ischemia group and normal control group, with significant difference[respectively 0.94±0.12,0.52±0.11,0.51±0. 10,t=0.327 0,6.127 4, P＜0.01].④The expression of ICAM-1 protein was significantly higher at ischemiareperfusion for 4,6 and 12 hours than in simple ischemia group and normal control group, with significant difference [Respectively (316.90±26.00),(361.40±18.00),(406.00±23.00),(164.21±2.00),(180.00±32.00) μg/L,t=1.410 3,9.119 3 ,P ＜ 0.01]. ⑤ The activity of MPO was significantly higher at ischemia-reperfusion for 12 hours than in simple ischemia group and normal control group, with significant difference [respectively (15.00±2.00),(7.50±1.67),(6.67±1.00) nkat/g, t=3.012 2,P ＜ 0.01].CONCLUSION: Following reperfusion injury, inflammatory reaction in spinal cord is important molecular basis for causing blood spinal barrier impairment, and plays an important role in the process of secondary spinal cord injury.

Objective To compare ultrasound-and endoscopy-guided methods in nasogastrojejunal tube placement in critical ill patients at bedside.Methods A tot al of 95 critical ill patients planed to perform enteral nutrition (EN) were enrolled.They were divided randomly into ultrasound-guided group (48 patients)and endoscopy-guided group (47 patients).The success rate,the incidence of complications and the time they took were compared between two groups.Results Ultrasound guided group was compared to endoscopy guided group with a lower success rate (81.3％ vs 100％,P =0.003),mainly because of the initial 24-stage having a very low success rate (66.7％ vs 100％,P =0.000),but the success rate of the following 24 cases was significantly improved (95.8％ vs 100％,P =0.338).Ultrasound guided group didn 't cause more complications (2.1％ vs 0,P =1.000),and consumed less time [(13.3 ± 2.8)min vs (15.0 ± 1.4) min,P =0.000].Compared to the following 24 cases,the initial 24 cases of ultrasound guided group in nasogastrojejunal tube placement had the lower success rate (66.7％ vs 95.8％,P =0.023),the longer time-consuming [(15.4 ±2.1)rin vs (11.2 ± 1.4) min,P =0.000],and but didnt cause more complications (4.2％ vs 0,P =1.000).The success rate of different diseases [severe acute pancreatitis(SAP) vs cerebrovascular disease] hadn't statistically significant difference (94.9％ vs 87.5 ％,P =0.300),but the SAP group consumed less time [(12.6 ± 1.9)min vs (15.5 ± 2.0)min,P =0.000].Conclusions The placement of nasogastrojejunal tube under ultrasound guidance represented a safe,quick and effective method to provide enteral nutrition.

Objective To design a novel electronic device for measuring the pressure in the cuff of the artificial airway; and to study the advantage of this device on continuous and intermittent cuff pressure monitoring. Methods ① a portable electronic device for cuff pressure measurement was invented, which could turn pressure signal into electrical signal through a pressure transducer. Meantime, it was possible to avoid pressure leak from the joint and the inside of the apparatus by modified Luer taper and sophisticated design. If the cuff pressure was out of the normal range, the apparatus could release a sound and light alarm. ② Six traditional mechanical manometers were used to determine the cuff pressure in 6 tracheal tubes. The cuff pressure was maintain at 30 cmH2O (1 cmH2O =0.098 kPa) by the manometer first, and repeated every 30 seconds for 4 times. ③ Study of continuous cuff pressure monitoring: We used a random number generator to randomize 6 tracheal tubes, 6 mechanical manometers and 6 our products by number 1-6, which has the same number of a group. Every group was further randomized into two balanced groups, one group used the mechanical manometer first, and the other used our product first. The baseline pressure was 30 cmH2O, measurement was performed every 4 hours for 6 times. Results When traditional mechanical manometer was used for cuff pressure monitoring, cuff pressure was decreased by an average of 2.9 cmH2O for each measurement (F = 728.2, P = 0.000). In study of continually monitoring, at each monitoring point, the pressure measured by electronic manometer was higher than the mechanical manometer. All the pressures measured by mechanical manometer were dropped below 20 cmH2O at 8th hour, and there was no pressure decrease below 20 cmH2O measured by electronic manometer in 24 hours by contrast. In study of intermittent monitoring, the same result was found. The pressure was dropped significantly with time when measured by mechanical manometer (F = 61.795, P = 0.000), the drops below 20 cmH2O began at 8th hour; but when measured by electronic manometer, all the value stayed unchanged around the baseline in 24 hours (F = 0.511, P = 0.796). Conclusions Compared with traditional mechanical manometer, cuff pressures monitored by our novel electronic manometer were steadier in both continuous and intermittent monitoring. The device is compact and convenient, and can provide a good solution for continuously monitor of the tracheal cuff pressure.

Objective To investigate the clinical outcomes in the treatment of complex acetabular fractures through a single supra-ilioinguinal approach after the application of preoperative 3D printing and rapid prototyping simulating the operative procedure.Methods From January 2013 to January 2016,24 cases of complex pelvic and acetabular fractures in our hospital were retrospectively analyzed.Among them 18 cases were male and 6 were female.The average age was 39.2+±13.8 years (range,24-68 years).According to Letoumel-Judet's classification,there were 6 cases of transverse,4 cases of T shaped,4 cases of combined anterior column and posterior hemi-transverse (ACPHT) and 10 cases of 2-column fractures.The time from injured to operation averaged 9 days (range,4-18 days).Before operation,pelvic AP view,three-dimensional CT as well as CTA of iliac blood vessels was taken routinely in all cases.The CT and CTA scan data were then imported into Mimics software for 3D reconstruction.Surgical simulation and optimal placement of internal fixations were performed on computer.After that,the individual simulated model of the pelvis was printed by the use of 3D printing and rapid prototyping technique.The order of reduction,selection and optimal position of the implants were determined.The plates were bent before operation and the length and direction of the screws were recorded.A uniform supra-ilioinguinal approach was used in all cases.Results The length of incision averaged 9 cm (range,8-13 cm).The operative time average 130 min (range,90-220 min).The blood loss averaged 600 ml (range,300-1 500 ml).Once a satisfactory reduction was obtained,single placement of the plates was achieved.In accordance with the preoperative plan,the plates were perfectly fitted to the bone surface.The 24 patients were followed for an average time of 18.3 months (range,15-30 months).The fractures healed in all cases.The average healing time was 4.2 months (range,3-6 months).According to Matta criteria,there were 16 cases of excellent,6 cases of good,2 cases of poor and the excellent rate was 91.7％ (22/24).The average modified Merle d'Aubigne score was 16.5 (range,13-18).The functional outcome was excellent in 13 cases,good in 6 cases and poor in 5 cases.The excellent rate of hip functions was 79.2％ (19/24).There were 2 cases of femoral lateral cutaneous nerve injury (recovered one month later) and another 1 case of fat liquefaction of the incision happened three days postoperatively (the wound healed completely after several dressing changes).No inguinal hernia or abdominal wall hemias was found.No other postoperative complications was found.Conclusion The application of 3D printing and rapid prototyping technique allows preoperatively simulating reduction in vitro,determining the optimal position of the implants,bending the plates and measuring the length of the screws in advance,which could largely reduce the operative time,intraoperative blood loss and occurrence of postoperative complications.Adopting supra-ilioinguinal approach for the treatment of complex acetabular fractures with a small incision and less trauma,a full exposure could be obtained,so the reduction and fixation of the anterior,posterior and middle column (also named pubo-ischiatic column) could be completed in a direct visualization,resulting in a satisfactory clinical outcomes.