Objective:To describe the surgical technique and explore clinical and radiological outcomes of Seinsheimer V subtrochanteric femoral fractures treated by clamp-assisted reduction with limited incision and InterTan nailing fixation.Methods:Data of 22 patients with Seinsheimer V subtrochanteric femoral fractures who were treated by clamp-assisted reduction and InterTan nailing fixation with limited incision in our hospital from January 2015 to June 2018 were retrospectively analyzed. There were 14 men and 8 women with an average age of 62.95±12.44 years (range, 36-81 years). Among them, 7 cases with a big butter-fly fragment were fixed by one cable. After the reduction, the fractures were fixed with InterTan nailing. The operative time, intraoperative blood loss and postoperative complication were recorded. The reduction quality was assessed by postoperative radiography according to the criteria proposed by Baumgartner. The function of the hip joint was assessed by the Harris score and VAS score.Results:All patients were followed up for 18±5.33 months (range, 12-30 months). According to the Baumgartner criteria of reduction quality, anatomic reduction was obtained in 20 cases, and satisfactory reduction was gained in 2 cases. All fractures healed in 4.36±1.36 months (range, 3-8 months). At latest follow-up, the mean Harris scores was 89.05±7.75 (range, 71-100 points), including 11 cases of excellent, 8 cases good, and 3 cases fair (satisfactory rate=86.4%). The mean VAS score was 0.64±0.85 (range, 0-3 points). Two patients had limb length discrepancy which was less than 10 mm. There were no significant complications such as infection, deep vein thrombosis, nonunion, nail cut out and implant failure.Conclusion:Clamp-assisted reduction and InterTan nailing with limited incision for Seinsheimer V subtrochanteric femoral fractures is simple and easy to manipulate, which can achieve anatomic reduction and has good clinical effects.

Objective: To investigate the current status and real performance of the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels in China through interlaboratory comparison. Methods: Peking University People’s Hospital (PKUPH) prepared the samples for comparison. That is, the fresh RUNX1-RUNX1T1 positive (+) bone morrow nucleated cells were serially diluted with RUNX1-RUNX1T1 negative (-) nucleated cells from different patients. Totally 23 sets with 14 different samples per set were prepared. TRIzol reagent was added in each tube and thoroughly mixed with cells for homogenization. Each laboratory simultaneously tested RUNX1-RUNX1T1 and WT1 transcript levels of one set of samples by real-time quantitative PCR method. All transcript levels were reported as the percentage of RUNX1-RUNX1T1 or WT1 transcript copies/ABL copies. Spearman correlation coefficient between the reported transcript levels of each participated laboratory and those of PKUPH was calculated. Results: ①RUNX1-RUNX1T1 comparison: 9 samples were (+) and 5 were (-) , the false negative and positive rates of the 20 participated laboratories were 0 (0/180) and 5% (5/100) , respectively. The reported transcript levels of all 9 positive samples were different among laboratories. The median reported transcript levels of 9 positive samples were from 0.060% to 176.7%, which covered 3.5-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.5 to 12.3 (one result which obviously deviated from other laboratories’ results was not included) , 85% (17/20) of the laboratories had correlation coefficient ≥0.98. ②WT1 comparison: The median reported transcript levels of all 14 samples were from 0.17% to 67.6%, which covered 2.6-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.3-13.7, 62% (13/21) of the laboratories had correlation coefficient ≥0.98. ③ The relative relationship of the reported RUNX1-RUNX1T1 transcript levels between the participants and PKUPH was not always consistent with that of WT1 transcript levels. Both RUNX1-RUNX1T1 and WT1 transcript levels from 2 and 7 laboratories were individually lower than and higher than those of PKUPH, whereas for the rest 11 laboratories, one transcript level was higher than and the other was lower than that of PKUPH. Conclusion The reported RUNX1-RUNX1T1 and WT1 transcript levels were different among laboratories for the same sample. Most of the participated laboratories reported highly consistent result with that of PKUPH. The relationship between laboratories of the different transcript levels may not be the same.

AIM:To observe the effects of hawthorn leaf polymeric procyanidins ( PPC) on calcium mobiliza-tion of vascular endothelial cells , and to study the underlying mechanism .METHODS: Free calcium in cultured human umbilical vein endothelial cells (HUVECs) was labeled with Fura-2.HUVECs were treated with ATP, a positive control drug, and PPC at concentrations of 12.5, 25 and 50 mg/L..The intracellular calcium concentrations were measured with a living cell microscope for 30 min.RESULTS:PPC concentration-dependently increased the intracellular calcium concen-tration of HUVECs .The intracellular calcium concentrations in 25 and 50 mg/L PPC groups were significantly higher than that in normal group (P<0.01).The dynamic manner of calcium concentration elevations elicited by PPC was a slow in -crease which happened after a latency time of several minutes , lasted for several minutes , and reached a plateau finally . This manner was quite different from that elicited by ATP , a typical SOC operator , hinting different mechanisms between them .Inhibiting the intracellular calcium release did not influence the effects of PPC;however , deleting extracellular calci-um, inhibiting the reverse mode of Na +-Ca2+exchange, or deleting extracellular sodium , restrained or even abolished the effects of PPC.CONCLUSION:PPC elicits calcium mobilization in vascular endothelial cells , which may be one of the mechanisms of the vascular modulatory activity of hawthorn procyanidins .This effect may be achieved through inducing the influx of sodium and then activating the reverse mode of Na +-Ca2+exchange.

Objective:To explore the influencing factors of physicians' professionalism,to understand the cognitive status and changes of professional responsibility and put forward the corresponding strategy.Methods:The satisfaction and cognitive status of physicians'professionalism were investigated using the self-designed questionnaire that adapted to medical staff,and the data was analyzed using statistical software SPSS19.0.Results:It showed that the satisfaction with the job itself was quite high.But they thought that the work risk was high,their own value was difficult to realized,and the professionalism was lack of cultivation.Conclusions:It suggests to put forward measures and suggestions from the perspective of system reform,vocational education,hospital culture,social atmosphere,decision-making of doctors and patients together.

Objective To observe the effects of hawthorn leaf procyanidins (HLP) on over expressions of ICAM-1 and E-selectin in human umbilical vein endothelial cells (HUVEC) induced by TNF-α,and clarify the mechanism of HLP's anti-inflammation effect. Methods HUVEC were cultured in vitro. MTT assay was used to detect cell viabilities. The expressions of ICAM-1 and E-selectin in HUVEC were detected by flowcytometry. Results Up to 200 mg/L, HLP showed no significant decrease in cell viabilities; the expression levels of ICAM-1 and E-selectin in the model group significantly increased, compared with that in the normal group; 10, 20, 30, 40, 50 mg/L HLP inhibited the expression elevations of ICAM-1 and E-selectin in concentration-dependent manner; and there were statistical significances in 40, 50 mg/L HLP groups, compared with the model group. Conclusion HLP can inhibit the over expressions of ICAM-1 and E-selectin of vascular endothelial cells induced by TNF-α, which possibly underlies HLP's anti-inflammation effect.

Objective To explore clinical and radiological outcomes of treating displaced fractures of proximal clavicle by open reduction and internal fixation with an inverted anatomic locking plate for distal clavicle.Methods From August 2013 to August 2015,12 patients with displaced fracture of proximal clavicle were treated in our hospital by open reduction and internal fixation with an inverted anatomic locking plate for distal clavicle.They were 11 men and one woman,with an average age of 43.5 years (range,25 to 62 years).There were 9 fresh and 2 old fractures.According to the Edinburgh classification,10 fractures were classified as type 1B1 and 2 as type 1B2.After fixation,the 180° inverted plate on the ipsilateral side was placed on the superior aspect of proximal clavicle.The medial fragment was fixed with 2 to 4 pieces of 2.7 mm multidirectional locking screw and the lateral fragment with 2 to 3 pieces of 3.5 mm locking screw.X-ray and CT were performed to assess union,delayed union,nonunion,and hardware failure.Functional outcomes were assessed by Constant-Murley scores and Disabilities of the Arm,Shoulder and Hand (DASH) scores at final follow-ups.Results There were no significant neurovascular injuries intraoperatively.All patients were followed up for an average of 15.6 months (range,12 to 24 months).All fractures healed after an average of 14.3 weeks (range,8 to 24 weeks).At final follow-ups,the mean Constant-Murley score was 96.0 points (range,84 to 100 points) and the mean DASH score 1.9 points (range,0 to 14.8 points).There were no such significant complications as infection,reduction loss or implant failure.Conclusion Displaced fractures of proximal clavicle may be treated with an inverted anatomic locking plate for distal clavicle on the ipsilateral side because of rigid fixation,fine stability and good chance for early rehabilitation.

Objective: To observe the correlation between Nε-carboxymethyl-Lysine (CML), the main component of advanced glycation end products and the calcification of the anterior tibial artery plaque in patients with diabetic foot post foot amputation. Methods: Sixty patients hospitalized for foot amputation operation due to diabetic foot from June 2012 to June 2016 in the Department of Orthopedics, Affiliated Hospital of Jiangsu University were prospectively recruited.The patients were categorized into mild stenosis (0n=20), moderate stenosis (50%≤stenosis<70%, n=20) and severe stenosis (70%≤stenosis≤100%, n=20) based on the color Doppler ultrasound assessed severity of anterior tibial artery stenosis.The baseline clinical data of patients were collected and anterior tibial artery was isolated.Then, HE staining, O-Cresolphthalein Complexone method, enzymic method and ELASA analysis were then performed to detect the evolution of calcification, arterial calcium content, alkaline phosphatase activity and serum CML concentration, respectively. Results: The results from both color Doppler ultrasound scan before amputation and HE staining after amputation showed that echo intensity as well as spotty blue calcium particles of anterior tibial artery plaque increased significantly in proportion to degree of stenosis and destructed elastic plate of the arterial wall was evidenced in patients with severest stenosis.The content of calcium ((2.3±0.9), (3.9±1.3), (6.6±1.7) μmol/mg, respectively, P<0.001), ALP activity ((102.4±39.4), (202.3±73.4), (483.7±117.9) U/mg, respectively, P<0.001) and serum CML level ((28.9±4.4), (37.9±5.3), (57.3±7.1)μg/L, respectively, P<0.001) increased significantly in proportion to stenosis severity.Pearson correlation analysis showed that serum CML level was positively correlated with the content of calcium (r=0.749, P<0.001) and ALP activity (r=0.923, P<0.001), respectively. Conclusions Serum CML level is positively correlated with the calcification of anterior tibial arterial plaque in patients with diabetic foot and could be used to evaluate the calcification of anterior tibial arterial plaque and stenosis degree of anterior tibial arterial in these patients.

Objective To observe the effects of matrix metalloproteinases (MMP)-2 and MMP-9 secreted by leukemic cells on tight junction proteins ZO-1,claudin-5 and occluding and the permeability of the blood-brain barrier (BBB) and explore the mechanisms of MMP-2 and MMP-9 in leukemic cell infiltration of the central nervous system (CNS).Methods The rnRNA expressions of MMP-2 and MMP-9 in leukemic cell lines SHI-1,HL-60 and U937 were detected by quantitative RT-PCR.The MMP inhibitor GM6001 was used to inhibit the secretion of MMP-2 and MMP-9.RNA interference (RNAi) was used to knock down the expression of MMP-2 and MMP-9.Zymography was used to analyze the secretion of MMP-2 and MMP-9 in the supematant of different leukemia cell lines treated or untreated with drugs,as well as the RNAi-treated cells.An in vitro BBB model composed of human brain microvascular endothelial cells (BMVECs) was developed on a Matrigel-based insert.Cell invasion through a barrier of Matrigelbased human basement membrane and the BMVECs-based human BBB barrier was assayed to measure the invasive capacity and the capacity to breakdown the BBB of different leukemia cell lines treated or untreated with drugs,as well as the RNAi-treated cells.The morphologic changes of BMVECs after co-culture with different leukemia cell lines treated or untreated with drugs,as well as the RNAi-treated cells in vitro BBB models were observed by invert microscopy and tight junction proteins in these BMVECs were analyzed with a laser-scanning confocal microscope.Results (①)The mRNA expression in different leukemic cell lines shown a pronounced transcription of MMP-2 and-9,and the transcriptional level in SHI-1 cells was the highest among all leukemic cell lines tested (P＜0.01).The data of activities of MMP-2 and-9 were consistent with the results of mRNA expression and SHI-l displayed higher capacity of invasion (P＜0.01).(②)After incubation 24h with different leukemic cells,the BMVECs disrupted to loss cell-cell contacts and grew in single cell.Confocal imaging showed down-regulations of ZO-1,claudin-5 and occluding accompanied by the disruption of BBB in vitro models.SHI-1 cells had stronger alterations to BMVECs,tight junction proteins and the permeability of the BBB than HL-60 and U937 cells.However,GM6001 and the knock-down of MMP-2 and MMP-9 altered the responses of BBB.They reduced the degradation of three tight junction proteins with a decreased permeability of BBB.Conclusion MMP-2 and MMP-9 secreted by leukemic cells could disrupt the BBB by degrading the tight junction proteins ZO-1,claudin-5 and occluding,which contributed the infiltration of leukemic cell into CNS.

OBJECTIVETo investigate the effect of up-regulation of Rap1GAP on the invasion ability of leukemic HL-60 cells in vitro, and to establish leukemia mouse model to verify the effects in vivo.

METHODSQuantitative RT-PCR and Western blot methods were used to detect the expression of Rap1GAP in Venus/HL-60 (vehicle control) and Rap1GAP/HL-60 cells (R1 andR2). Transwell method was used to examine the invasion ability in vitro. Quantitative RT-PCR and gelatin zymograph were used to study the expression of MMP-2 and MMP-9. Four-week-old BALB/c nu/nu mice were pre-treated and inoculated with leukemic cells from different groups, several index including survival time were then monitored.

RESULTSRap1GAP mRNA level of R1 and R2 increased about 16-17 folds as compared to the control cells. The invasion rate of R1 and R2 are (55 ± 5)% and (59 ± 4)%, which are significantly higher than (14 ± 4)% of the control cells. The mRNA level of MMP-9 was up-regulated about 12.0 folds in R1 and R2 cells compared to the corresponding control cells. The median survival times of R1 and R2 mice are (32.00 ± 1.85) d and (33.37 ± 2.50) d, respectively, which are shorter than (43.62 ± 2.32) d of the control group. Three mice of R1 and R2 groups showed leukemic cells infiltration in meninges tissue, and the genes of Rap1GAP and MMP-9 were amplified by PCR method.

CONCLUSIONUp-regulated expression of Rap1GAP increased the invasion ability of HL-60 cells accompanied with enhancement of MMP-9 expression in vitro, and the experiment in mouse model also confirmed that Rap1GAP enhanced the invasion of HL-60 cells in vivo.

Animals ; GTPase-Activating Proteins ; metabolism ; HL-60 Cells ; Humans ; Leukemia ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; RNA, Messenger ; Transcriptional Activation ; Up-Regulation