Arthroscopic minimally invasive treatment of tibial intercondylar eminence fractures in children / 中国骨伤

<p><b>OBJECTIVE</b>To analyze the characteristics of children tibial intercondylar eminence fractures, and introduce arthroscopic minimally invasive techniques for the treatment of tibial intercondylar eminence fractures in children.</p><p><b>METHODS</b>From January 2004 to December 2008, 12 children with tibial intercondylar eminence fractures were treated with cross Kirschner wire fixation after arthroscopic reduction. According to Meyers-McKeever classification systems, there were 1 case of type I, 4 cases of type II, and 7 cases of type III. There were 10 fresh and 2 old fractures in all. Among the patients, 10 patients were boy and 2 patients were girl,ranging in age from 8 to 13 years, with an average of 10 years. All the patients underwent arthroscopic exploration, reduction and fixation. During follow-up ranging from 10 to 36 months, the union of fracture, range of motion and stabilization of the knee were assessed. One patient was combined with lesions of the menisci, 1 patient with femoral trochlea cartilage injury, and 5 patients with meniscal entrapment under the bone.</p><p><b>RESULTS</b>The heeling time averaged 5 weeks. No knee laxity or instability and no intercondylar notch impingement was detected in all cases at 3 months postoperatively. At same time, full range of motion of the affected knee returned, and the average Lysholm knee score was (92.7 +/- 2.5), the average Lysholm knee score was (96.4 +/- 1.7) at 6 months postoperatively. The Lachman test and ADT test was negative.</p><p><b>CONCLUSION</b>The type II and type III tibial intercondylar eminence fractures occur frequently in children. Lesions of the menisci and cartilage occur seldom. The method of arthroscopic cross Kirschner wire fixation for the treatment of tibial intercondylar eminence fracture is easy to operate. Simultaneously, this technique is less invasive and allows early recovery. Also it coincidences with the characteristic rapid bone growth of children.</p>

Effects of FMS-like tyrosine kinase 3 targeted RNA interference on proliferation and apoptosis of acute monocytic leukemia cell line THP-1 / 中华儿科杂志

<p><b>OBJECTIVE</b>FMS-like tyrosine kinase 3 (FLT3) is a receptor tyrosine kinase that is constitutively activated in (70-90)% pediatric patients with acute myeloid leukemia (AML) and appears to confer an adverse prognosis. Although several FLT3-selective small molecule inhibitors and antibodies were developed with varied degrees of success, to address the specificity and resistance, new approaches for specifically targeted FLT3 are needed and RNA interference is a promising choice. The aim of the present study was to investigate the efficacy of suppression of FLT3 induced by small hairpin interfering RNA (shRNA) on myeloproliferation and apoptosis in an acute monocytic leukemia (AMOL) cell line THP-1.</p><p><b>METHODS</b>FLT3-targeted small hairpin interfering RNA (FLT3-shRNA) was designed and synthesized by transcription system in vitro was transfected into THP-1 cells. Firstly FLT3 mRNA level was detected by semi-quantitative RT-PCR and FLT3 protein level was detected by flow cytometry (FCM) to verify the efficacy on FLT3-shRNA interference at 48 h after transfection. Cell growth viability was measured at 24 h, 48 h and 72 h after treatment with CCK-8. The distribution of cell cycle was assayed by FCM, and apoptosis was analyzed by DNA Ladder and Annexin V-FITC Staining at 48 h.</p><p><b>RESULTS</b>FLT3 targeted shRNAs was synthesized successfully and the concentration of 15 nmol/L for 48 h could obtain desirable downregulation of FLT3 expression, the inhibitory percentages of FLT3 mRNA and protein were (72.95 +/- 2.07)% and (65.39 +/- 5.57)%, respectively. The suppression of FLT3 induced by FLT3-shRNA resulted in marked inhibition of cell growth and the inhibitory percentages were (36.66 +/- 3.67)% at 48 h, (35.56 +/- 0.73)% at 72 h. FLT3-shRNA induced the inhibition of cell cycle from G(0)/G(1) phase to S phase, the percentage of sub-G(0)/G(1) phase (65.71 +/- 4.47)% was higher than those in the PBS-control group (52.23 +/- 2.98)%, NC-shRNA control group (51.81 +/- 1.44)%, P < 0.01; the percentage of S phase (25.11 +/- 2.70)% was lower than those in the PBS-control group (34.41 +/- 4.07)% and NC-shRNA control group (32.50 +/- 1.46)%, P < 0.05. Furthermore treatment with FLT3-shRNA for 48 h resulted in clear apoptosis ladder, the percentage of early apoptosis detected by Annexin V-FITC was (18.59 +/- 2.07)% which was significantly higher than that in the PBS-control group (4.00 +/- 0.50)% and the NC-shRNA control group (6.06 +/- 0.70)%, P < 0.001.</p><p><b>CONCLUSION</b>The suppression of FLT3 induced by the shRNA can effectively inhibit cell proliferation, and apoptosis induction on THP-1 cells, which indicates that this approach may bear the therapeutic potential on childhood AMOL.</p>

TAp63gamma-induced apoptosis mediated by apoptosis inducing factor in human esophageal squamous carcinoma EC9706 cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the molecular mechanism of TAp63gamma-induced cell apoptosis.</p><p><b>METHODS</b>Transcription and protein expression of apoptosis inducing factor and p63 were investigated by immunohistochemistry and RT-PCR in human esophageal squamous carcinoma cell line EC9706 respectively. Twenty-four hours after transfection with pcDNA3.1-TAp63gamma, the apoptosis and translocation of apoptosis inducing factor in EC9706 cells were studied by flow cytometry, laser confocal microscopy and mitochondrial/cytosol/nuclear extraction analysis respectively. Down-regulation of apoptosis inducing factor protein was achieved by RNAi and pretreatment with caspase inhibitor zVAD.fmk of EC9706 cells.</p><p><b>RESULTS</b>Presence of protein expressions of apoptosis inducing factor and absence of TAp63gamma was observed in the cytoplasm of untransfected cells. RT-PCR verified the subtype of p63 in EC9706 cells was DeltaNp63. After 24 hours of transfection, both nuclear and cytoplasmic expression of apoptosis inducing factor protein were observed in cells transfected with TAp63gamma and p53 expression vectors, but not in cells transfected with control vector. Cell apoptosis rates were 1.37%, 13.64%, 4.52%, 4.03% and 1.91% in the pcDNA3.1 transfection group, pcDNA3.1-TAp63gamma transfection group, apoptosis inducing factor siRNA and pcDNA3.1-TAp63gamma transfection group, zVAD.fmk treatment group, and the group receiving apoptosis inducing factor siRNA, plus zVAD.fmk treatment and pcDNA3.1-TAp63gamma transfection, respectively.</p><p><b>CONCLUSIONS</b>Apoptosis inducing factor of EC9706 cells is released from mitochondria into both the cytoplasm and nucleus during TAp63gamma induced apoptosis. Down-regulation of apoptosis inducing factor inhibits TAp63gamma-induced apoptosis. Overall, TAp63gamma-induced apoptosis is dependent on the expression of apoptosis inducing factor and caspase.</p>