Internal Structural Connections in the Popliteus Tendon Complex with Posterior Cruciate Ligament and Clinical Implications / Conexiones Estructurales Internas en el Complejo Tendón Poplíteo con Ligamento Cruzado Posterior e Implicaciones Clínicas

SUMMARY: At present, the anatomical relationship the mid-portion of popliteus tendon complex (PTC) and the surrounding tissues is still unclear, especially its relationship to the posterior cruciate ligament (PCL). It affected the anatomical reconstruction of the posterolateral complex (PLC) injury. A total of 30 cases of the adult human knee joint fixed with formalin were used. Sagittal sections were made in 14 knee joints by the P45 plastination technique and dissection of 16 cases of knee joints. The P45 section revealed that the popliteus muscle fascia ran superiorly over the posterior edge of the tibial intercondylar eminence, and turned forward to be integrated into the PCL. Laterally, near the posterior edge of the lateral tibial plateau, the popliteus tendon penetrates through the articular capsule (AC), where two dense fibrous bundles were given off upwards by the popliteus tendon: one was the ventral fiber bundle, which ran superiorly over the posterior edge of the tibial plateau and then moved forwards to connect with the lateral meniscus; the dorsal fibers bundle ascended directly and participated in the AC. Meanwhile, the popliteus muscle dissection showed that at the posterior edge of the platform of the lateral condyle of the tibia, at the tendon-muscle transition, the PTC and AC were anchored to PCL.

En la actualidad, la relación anatómica entre la porción media del complejo tendinoso poplíteo (CTP) y los tejidos circundantes aún no está clara, especialmente su relación con el ligamento cruzado posterior (LCP). Esto afecta la reconstrucción anatómica de la lesión del complejo posterolateral (LCP). Se utilizaron un total de 30 casos de articulaciones de rodillas humanas de individuos adultos fijadas con formalina. Se realizaron cortes sagitales en 14 articulaciones de rodilla mediante la técnica de plastinación P45 y disección de 16 casos de articulaciones de rodilla. La sección P45 reveló que la fascia del músculo poplíteo discurría superiormente sobre el margen posterior de la eminencia intercondílea tibial y giraba hacia delante para integrarse en el LCP. Lateralmente, cerca del margen posterior de la platillo tibial lateral, el tendón poplíteo penetra a través de la cápsula articular (CA), donde el tendón poplíteo desprendió hacia arriba dos haces fibrosos densos: uno era el haz de fibras ventral, que corría superiormente sobre el margen posterior de la meseta tibial y luego se movió hacia adelante para conectar con el menisco lateral; el haz de fibras dorsales ascendía directamente y participaba en la CA. Por su parte, la disección del músculo poplíteo mostró que en el margen posterior del platillo del cóndilo lateral de la tibia, en la transición tendón-músculo, el CTP y el AC estaban anclados al LCP.

Bony structures at the attachment areas of the cruciate ligaments in human knee joints and their clinical significance-studied using the P45 plastination technique / Estructuras óseas en las áreas de inserción de los ligamentos cruzados en las articulaciones de la rodilla humana y su importancia clínica, estudiadas mediante la técnica de plastinación P45

SUMMARY: For treating cruciate ligament injuries, especially for characterizing the mechanics of the tunnel in cruciate ligament reconstruction, correctly understanding the bony information of the attachment area of the cruciate ligaments is significant. We studied 31 knee joints of middle-aged Chinese adults using the P45 sheet plastination technique, focusing on the attachment areas of the cruciate ligaments, especially the bony structures. The trabeculae at the attachment area were distributed radially and extended deep into the medial wall of the lateral condyle of the femur. However, in the anterior part of the intercondylar eminence, the trabeculae of the anterior group were parallelly arranged along the tendinous fibers of the anterior cruciate ligament, while the trabeculae of the posterior group were parallelly arranged along the perpendicular direction of the anterior cruciate ligament fibers. Similarly, at the attachment area of the lateral wall of the medial condyle of the posterior cruciate ligament, the trabeculae extended radially toward the deep medial condyle. Deep in the posterior part of the intercondylar eminence, the trabeculae were arranged longitudinally. In the anterior part of the intercondylar eminence, the trabeculae were parallelly arranged along the perpendicular directions of ligament fibers. The distribution patterns of the trabecular at the attachment areas of the cruciate ligaments at the ends of the femur and tibia were different. This difference should be considered when orthopedic surgeons reconstruct anterior cruciate ligaments.

Para el tratamiento de lesiones de los ligamentos cruzados, especialmente para caracterizar la mecánica del túnel en su reconstrucción, es importante comprender correctamente la información ósea del área de inserción de estos ligamentos. Estudiamos 31 articulaciones de rodilla de individuos chinos, adultos, de mediana edad, utilizando la técnica de plastinación de láminas P45, centrándonos en las áreas de unión de los ligamentos cruzados, especialmente en las estructuras óseas. Las trabéculas en el área de inserción se distribuyeron radialmente y se extendieron profundamente en la pared medial del cóndilo lateral del fémur. Sin embargo, en la parte anterior de la eminencia intercondílea, las trabéculas del grupo anterior estaban dispuestas paralelamente a lo largo de las fibras tendinosas del ligamento cruzado anterior, mientras que las trabéculas del grupo posterior estaban dispuestas paralelamente a lo largo de la dirección perpendicular de las fibras del ligamento cruzado anterior. De manera similar, en el área de inserción en la cara lateral del cóndilo medial del ligamento cruzado posterior, las trabéculas se extendían radialmente y profundas hacia el cóndilo medial. Profundamente en la parte posterior de la eminencia intercondílea, las trabéculas estaban dispuestas longitudinalmente. En la parte anterior de la eminencia intercondílea, las trabéculas estaban dispuestas paralelamente a lo largo de las direcciones perpendiculares de las fibras del ligamento. Los patrones de distribución del tejido óseo trabecular en las áreas de unión de los ligamentos cruzados en los extremos del fémur y la tibia eran diferentes. Estas diferencias deben tenerse en consideración cuando los cirujanos ortopédicos reconstruyen los ligamentos cruzados anteriores.

Congenital atlanto-occipital fusion and its effect on the myodural bridge: a case report utilizing the P45 plastination technique / Fusión atlanto-occipital congénita y su efecto sobre el puente miodural: reporte de un caso utilizando la técnica de plastinación P45

SUMMARY: The atlanto-occipital joint is composed of the superior fossa of the lateral masses of the atlas (C1) and the occipital condyles. Congenital Atlanto-occipital fusion (AOF) involves the osseous union of the base of the occiput (C0) and the atlas (C1). AOF or atlas occipitalization/assimilation represents a craniovertebral junction malformation (CVJM) which can be accompanied by other cranial or spinal malformations. AOF may be asymptomatic or patients may experience symptoms from neural compression as well as limited neck movement. The myodural bridge (MDB) complex is a dense fibrous structure that connects the suboccipital muscular and its related facia to the cervical spinal dura mater, passing through both the posterior atlanto-occipital and atlanto-axial interspaces. It is not known if atlas occipitilization can induce structural changes in the MDB complex and its associated suboccipital musculature. The suboccipital region of a cadaveric head and neck specimen from an 87-year-old Chinese male having a congenital AOF malformation with resultant changes to the MDB complex was observed. After being treated with the P45 plastination method, multiple slices obtained from the cadaveric head and neck specimen were examined with special attention paid to the suboccipital region and the CVJM. Congenital atlanto-occipital fusion malformations are defined as partial or complete fusion of the base of the occiput (C0) with the atlas (C1). In the present case of CVJM, unilateral fusion of the left occipital condyle with the left lateral mass of C1 was observed, as well as posterior central fusion of the posterior margin of the foramen magnum with the posterior arch of C1. Also noted was a unilateral variation of the course of the vertebral artery due to the narrowed posterior atlanto-occipital interspace. Surprisingly, complete agenesis of the rectus capitis posterior minor (RCPmi) and the obliques capitis superior (OCS) muscles was also observed in the plastinated slices. Interestingly, the MDB, which normally originates in part from the RCPmi muscle, was observed to originate from a superior bifurcation within an aspect of the nuchal ligament. Therefore, the observed changes involving the MDB complex appear to be an effective compensation to the suboccipital malformations.

RESUMEN: La articulación atlanto-occipital está compuesta por las caras articulares superiores de las masas laterales del atlas (C1) y los cóndilos occipitales. La fusión atlanto-occipital congénita (FAO) implica la unión ósea de la base del occipucio (C0) y el atlas (C1). La FAO u occipitalización/asimilación del atlas representa una malformación de la unión craneovertebral (MUCV) que puede presentar otras malformaciones craneales o espinales. La FAO puede ser asintomática o los pacientes pueden experimentar síntomas de compresión neural así como movimiento limitado del cuello. El complejo del puente miodural (PMD) es una estructura fibrosa densa que conecta el músculo suboccipital y su fascia relacionada con la duramadre espinal cervical, pasando a través de los espacios intermedios atlanto-occipital posterior y atlanto-axial. No se sabe si la occipitilización del atlas puede inducir cambios estructurales en el complejo PMD y en la musculatura suboccipital. Se observó en la región suboccipital de un espécimen cadavérico, cabeza y cuello de un varón chino de 87 años con una malformación congénita de FAO con los cambios resultantes en el complejo PMD. Se examinaron múltiples cortes obtenidos de la muestra de cabeza y cuello después de ser tratados con el método de plastinación P45, con especial atención a la región suboccipital y la MUCV. Las malformaciones congénitas por fusión atlanto-occipital se definen como la fusión parcial o completa de la base del occipucio (C0) con el atlas (C1). En el presente caso de MUCV se observó la fusión unilateral del cóndilo occipital izquierdo con la masa lateral izquierda de C1, así como fusión posterior central del margen posterior del foramen magnum con el arco posterior de C1. También se observó una variación unilateral del curso de la arteria vertebral por el estrechamiento del espacio interatlanto-occipital posterior. Se observó además agenesia completa de los músculos Rectus capitis posterior minor (RCPmi) y oblicuos capitis superior (OCS) en los cortes plastinados. Curiosamente, se observó que el MDB, que normalmente se origina en parte del músculo RCPmi, se origina en una bifurcación superior dentro de un aspecto del ligamento nucal. Por lo tanto, los cambios observados en el complejo PMD parecen ser una compensación de las malformaciones suboccipitales.

The effectiveness and safety of parathyroid hormone in fracture healing: A meta-analysis

The very large economic and social burdens of fracture-related complications make rapid fracture healing a major public health goal. The role of parathyroid hormone (PTH) in treating osteoporosis is generally accepted, but the effect of PTH on fracture healing is controversial. This meta-analysis was designed to investigate the efficacy and safety of PTH in fracture healing. The EMBASE, PubMed, and Cochrane Library databases were systematically searched from the inception dates to April 26, 2018. The primary randomized clinical trials comparing PTH treatment for fracture healing with placebo or no treatment were identified. We did not gain additional information by contacting the authors of the primary studies. Two reviewers independently extracted the data and evaluated study quality. This meta-analysis was executed to determine the odds ratio, mean difference, standardized mean difference, and 95% confidence intervals with random-effects models. In total, 8 randomized trials including 524 patients met the inclusion criteria. There were significant differences in fracture healing time, pain relief and function improvement. There were no significant differences in the fracture healing rate or adverse events, including light-headedness, hypercalcemia, nausea, sweating and headache, except for slight bruising at the injection site. We determined that the effectiveness and safety of PTH in fracture healing is reasonably well established and credible.

Molecular characteristics of the full-length genome of dengue serotype 4 virus strains isolated from dengue fever cases in Yunnan Province, China / 中国人兽共患病学报

We investigated the molecular characteristics of the full-length genome of 5 dengue serotype 4 virus (DENV-4) strains isolated in Yunnan Province,China,2015 and their molecular epidemiological feature.Isolation of dengue virus was using C6/36 cell culture method.Viral RNA was extracted from virus isolates,then the full-length genome was amplified by RT-PCR.The homology and phylogenetic analysis was made on the nucleotide and deduced amino acid sequences by bioinfor matics softwares including ClastalX1.83 and MEGA6 etc.Results showed that five strains of DENV-4 isolated from dengue fe ver cases in Ruili City of Yunnan Province in 2015,of these,2 strains from indigenous cases,3 from imported cases from Lashio and Nam Can cities of Myanmar to Ruili of China.RT-PCR and sequencing indicated that the full-length genes (10 661 nt) of 5 DENV-4 strains were obtained,and their open reading frame (103-10 264) were coded 3 386 amino acid residues.Phylogenetic tree and homology analysis based on the comeplete genome or structural and non-structural protein genes showed that the 5 DENV 4 isolates were highly homologous and gathered in an evolution as well as they have a closer genetic relationship with the DENV-4 genotype Ⅰ (G-Ⅰ) strains isolated from Thailand.Results indicated that the Yunnan strains belonged to G-Ⅰ.Yunnan strains and Thailand strains compared with DENV 4 prototype strain (H241,Philippines 1956) and Guangzhou strain (B5,1990) of China and showed low homology and evolutionary relationship.When Yunnan strains compared with H241 strain,there were 21 and 45 different sites in amino acid of structural and non-structural proteins,respectively.This is the first time in Yunnan to obtain full-length genomes sequence of DENV-4 and they have closer evolutionary relationship with DENV 4G-Ⅰ strains from Southeast Asia region in recent years.The autochthonous DENV-4 epidemic in Yunnan was detected for the first time,and the virus transmission sources were from neighboring northern Myanmar.It is necessary to further study that change of the amino acid sites of Yunnan strains of DENV-4 is related to its antigenicity and virulence.

Expression of Gal-3 and BCL-2 in patients with ulcerative colitis and their significance / 临床与实验病理学杂志

Purpose To study the expression and significance of Gal-3 and BCL-2 in ulcerative colitis (UC).Methods 64 cases of patients diagnosed as UC in the Department of Digestive Dseases were selected as the observation group in our hospital from August 2014 to December 2015.And 50 healthy volunteers were selected as control group.The expression levels of BCL-2 and Gal-3 in the two groups were detected,and the correlation between and UC and between BCL-2 and Gal-3 was studied.Results The positive rate of Gal-3 in the control group was 100％,higher than 37.5％ in the observation group(x2 =48.142,P ＜0.001).The positive rate of BCL-2 in the control group was 20％,which was lower than that in the observation group of 90.62％ (x2 =58.171,P ＜ 0.001).The Gal-3 positive rate of UC Grade Ⅰ was higher than UC Grade Ⅱ (x2 =5.539,P =0.019),The Gal-3 positive rate of UC Grade Ⅰ was higher than UC Grade Ⅲ (x2 =4.532,P =0.033),The Gal-3 positive rate of UC Grade Ⅲ was lower than UC Grade Ⅱ (x2 =0.183,P=0.669).The BCLo2 positive rate of UC Grade Ⅰ was lower than UC Grade Ⅱ (x2 =5.539,P=0.019).The BCL-2 positive rate of UC Grade Ⅰ was lower than UC Grade Ⅲ (x2 =6.098,P=0.014).The BCL-2 positive rate of UC Grade Ⅲ was higher than UC Grade Ⅱ (x2 =0.511,P =0.475).BCL-2 and Gal-3 expression was not related to gender,age and disease duration (P ＞ 0.05).The expression of BCL-2 and Gal-3 was not correlated.Conclusion The expression level of BCL-2 in UC is higher than that in normal colorectal mucosa.The expression level of Gal-3 is lower than that of normal colorectal mucosa.The expression of UC has no effect on sex,age and course of disease,and it could be used as the prediction standard of UC.

Prognostic significance of coagulation disorders in children with hemophagocytic syndrome / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the prognostic significance of coagulation disorders in children with hemophagocytic syndrome (HPS).</p><p><b>METHODS</b>Thirty-five children with HPS were retrospectively studied to analyze the etiology, clinical characteristics, laboratory results and treatment outcomes.</p><p><b>RESULTS</b>After treatment, 27 of the 35 HPS patients survived, and the other 8 cases died. All cases were treated according to the HLH-2004 protocol, but etoposide (VP-16) was not used in 10 of them. The response rate in patients who received VP-16 (22/25, 88%) was significantly higher than that in those not receiving VP-16 (5/10, 50%) (P<0.05). Compared with the survival group, the dead group had significantly lower platelet count, fibrinogen level, and VP-16 utilization rate (P<0.05) but significantly longer activated partial thromboplastin time and prothrombin time (P<0.05).</p><p><b>CONCLUSIONS</b>Coagulation function can be used as an indicator of disease outcome. It is essential for improving the clinical outcome of HPS to monitor the coagulation function during treatment, detect and correct abnormalities in time, and provide treatment strictly according to the HLH-2004 protocol.</p>

Ultrastructural observation of Birbeck granules in 8 cases of Langerhans cell histiocytosis / 中华实用儿科临床杂志

Objective To study the ultrastructural features of Langerhans cell(LC),especially Birbeck granules in Langerhans cell histiocytosis(LCH) and provide some ultrastructural evidence to clarify the origin,function and stereoscopic structure of Birbeck granules.Methods Pathologic tissues of 8 LCH patients were treated with routine transmission electron microscope (TEM) procedures,fixture,dehydration,permeation,embedding,polymerization,ultramicrocut,staining and examined by TEM.Results Under the TEM,8 cases of LCH revealed typical LC cell,and LC cell in 6 cases revealed typical rod,tennnis-racquet like Birbeck granules.Some double membrane structures revealed irregular shape,sometimes accompanied with vacuole-like structure.Most of Birbeck granules locate inside the cytoplasmic membrane,in continuity with the cell membrane.There was Birbeck granule-like structure outside the cytoplasmic membrane in one case of multisystem LCH.Conclusions The ultrastructure of Birbeck granules could be various and irregular.The irregular Birbeck granule-like structure can also be useful for the diagnosis of LCH.Birbeck granules may arise from the cell membrane.The stereoscopic structure of Birbeck granules could be irregular.

Clinical efficacy of desmopressin in the treatment of mild hemophilia A in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of desmopressin (DDAVP) on coagulation factor Ⅷ (FⅧ) and activated partial thromboplastin time (APTT) in children with mild hemophilia A.</p><p><b>METHODS</b>Eighteen children with mild hemophilia A were enrolled. DDAVP (0.3 μg/kg•d) was injected intravenously for 5 days. Plasma FⅧ levels and APTT were measured before and after DDAVP treatment.</p><p><b>RESULTS</b>In 16 of 18 children with mild hemophilia A, the bleeding symptoms, including the articular or musclar hematoma, were significantly alleviated as a result of DDAVP treatment. The plasma FⅧ levels increased significantly to (27±4)% and APTT was shortened to (66±10)s 60 minutes after the first dose of DDAVP treatment. The plasma FⅧ remained at the levels of 25%-30% during 3-4 days of DDAVP treatment. Five days after DDAVP treatment, the plasma FⅧ levels decreased [(21±3)%], and APTT was prolonged when compared with 1-4 days of DDAVP treatment.</p><p><b>CONCLUSIONS</b>DDAVP treatment can increase plasma FⅧ levels and shorten APTT in children with mild hemophilia A. DDAVP is effective in the treatment of mild hemophilia A. The duration of DDAVP therapy for mild hemophilia A is recommended as 3 to 4 days.</p>

Levels of Toll-like receptors-2,-4 on platelets in children with idiopathic thrombocytopenic purpura / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the changes and significance of Toll-like receptor-2 (TLR2) and Toll-like receptor-4 (TLR4) on platelets, CD86 on lymphocytes and concentrations of IL-2, IFN-gamma, IL-4 and IL-10 in serum in children with idiopathic thrombocytopenic purpura (ITP).</p><p><b>METHODS</b>Peripheral blood samples were collected from 24 children with acute idiopathic thrombocytopenic purpura (AITP), 21 children with chronic idiopathic thrombocytopenic purpura (CITP) and 20 normal children (control group). Expression of TLR2 and TLR4 on platelets and CD86 on lymphocytes were detected by flow cytometry. Serum concentrations of IL-2, IL-4, IL-10 and IFN-gamma were measured using ABC-ELISA.</p><p><b>RESULTS</b>The expression of CD41+TLR2+ and CD61+TLR4+ in the AITP and the CITP groups were significantly lower than those in the control group (p<0.01). The AITP group had lower expression of CD41+TLR2+ and CD61+TLR4+ than the CITP group (p<0.01). The expression of CD86+ in the AITP and the CITP groups was significantly higher than that in the control group (p<0.01). The serum concentrations of IL-2, IL-4, IL-10 and IFN-gamma in the AITP and the CITP groups were significantly higher than those in the control group (p<0.05). There was a positive correlation between CD41+TLR2+ and CD61+TLR4+ expression. CD41+TLR2+ and CD61+TLR4+ expression were negatively correlated with CD86+ expression and serum concentrations of IL-2, IL-4 and IL-10.</p><p><b>CONCLUSIONS</b>The detections of TLR2 and TLR4 on platelets, CD86 on lymphocytes and serum concentrations of IL-2, IFN-gamma, IL-4 and IL-10 are of great value in understanding the pathogenesis and predicting types of ITP in children.</p>

In vitro transcription synthesis and effects of FLT3 targeted short hairpin RNA / 中国实验血液学杂志

FMS-like tyrosine kinase 3 (FLT3) is a receptor of tyrosine kinase that is constitutively activated in most of acute myeloid leukemia patients and seems to give an adverse prognosis. In order to explore the silencing effect of FLT3 targeted short hairpin RNA (FLT3-shRNA) on acute leukaemia cell line THP-1, three FLT3-shRNAs (shRNA1, shRNA2, shRNA3) were designed and synthesized by transcription system in vitro and then transfected into THP-1 cells. FLT3 mRNA was analyzed by semi-quantitative RT-PCR, FLT3 protein was detected by Flow cytometry and immunofluorescence. The results indicated that FLT3 expression was downregulated by shRNA1 and shRNA3, and shRNA1 showed stronger inhibitory effect. At 48 hours following transfection, the inhibitory rate of 25 nmol/L shRNA1 was 72.95 +/- 2.07%, lasting 72 hours. The 5 nmol/L and more concentration of FLT3 shRNA1 could downregulate FLT3 mRNA level, which displayed a quantity-effect relation; the inhibitory rate of 15 nmol/L shRNA1 was 67.53 +/- 0.66%. FLT3 protein was located on THP-1 cell membrance, its expression was downregulated obviously by shRNA1, at 72 hours following transfection the inhibitory rate of shRNA1 was 79.67 +/- 0.66%. shRNA1 showed the best inhibitory effect on FLT3 protein, the optimal time of which was 72 hours with an inhibitory rate of 79.67%. It is concluded that FLT3-shRNA1 shows a desireable FLT3-targeted inhibitory effect, which can be used for further investigation of FLT3 mechanism or FLT3 targeting treatment.

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>

Expression levels of CXCR4 on megakaryocytes and its ligand in bone marrow in children with acute idiopathic thrombocytopenic purpura / 中华儿科杂志

<p><b>OBJECTIVE</b>Chemokine receptor CXCR4 and its ligand stromal-derived factor 1 alpha (SDF-1alpha) have been paid increasing attention for their involvement in megakaryocytic hematopoiesis. It has been revealed in recent years that they can induce mature and immature megakaryocytes (MKs) to migrate through bone marrow endothelial cells (BMEC) by increasing the affinity of MKs for BMEC. Thus MKs maturity and eventual release of platelet from MKs ensues. While maturity disturbance of MKs and impaired production of platelets have been regarded as the main pathogenesis of ITP, the mechanism of which still remains unclear. Therefore, a clear understanding of the levels of CXCR4 and SDF-1alpha within bone marrow in children with ITP will help us to elucidate further the mechanism of ITP as well as to provide direct theoretical evidence for predicting treatment effect and evaluating prognosis.</p><p><b>METHODS</b>Bone marrow were aspirated from 28 children with AITP and 12 normal children. Percoll density gradient and immunomagnetic beads method were used to purify megakaryocytes from the bone marrow. The immune cytochemistry was used to detect CXCR4 on megakaryocytes. The levels of SDF-1alpha were detected by ELISA. SPSS10.0 statistical software was used to deal with the experimental data.</p><p><b>RESULTS</b>Before the treatment in children with AITP, both the CXCR4 expression on megakaryocytes and the SDF-1alpha level in bone marrow plasma were markedly decreased compared with the normal controls (P < 0.05). As to the cases who were sensitive to the high-dose intravenous immunoglobulin (HDIVIgG), the CXCR4 and SDF-1alpha levels were much higher in children after the treatment than those before the treatment (P < 0.05). In 6 cases insensitive to HDIVIgG, before the treatment the CXCR4 level was much lower than the children sensitive to HDIVIgG (P < 0.05).</p><p><b>CONCLUSIONS</b>The low levels of CXCR4/SDF-1alpha system in bone marrow may be one of the factors which contribute to the maturity disturbance of megakaryocytes and disturbance of platelets production in AITP, while decreased CXCR4/SDF-1alpha system may be caused by the effect of autoantibody against platelet. The mechanism of HDIVIgG in the treatment of AITP may involve in the increasing expression of CXCR4/SDF-1alpha system. The level of CXCR4 on megakaryocytes may play a certain role in predicting the treatment effect of immunoglobulin.</p>

Level of Transforming Growth Factor - ?_1 in Bone Marrow and Expression of Its Receptor III on Megakaryocytes in Children with Idiopathic Thrombocytopenic Purpura / 实用儿科临床杂志

Objective To study changes and effects of transforming growth factor-?1(TGF- ?1) and its receptorIII (TGF-?1 R III) in children with idiopathic thrombocytopenic purpura (ITP). Methods Bone marrow were respectively collected from 28 children with acute idiopathic thrombocytopenic pupura(AITP),16 children with chronic idiopathic thrombocytopenic purpura(CITP) and 20 comparably normal children; Percoll density gradient and immunomagnetic beads methods were used to purify megakaryocytes from bone marrow; ABC- ELISA was used to detect TGF - ?1 in bone marrow; in situ hybridization was used to detect TGF-?1 RIIImR-NA expression on megakaryocytes.Results In AITP and CITP group, the levels of TGF-?1 and TGF-?1 RIIImRNA were significant higher than those in control group(P