Treatment of irreducible femoral intertrochanteric fractures using a wire-guided device.

PURPOSE: Treatment of irreducible femoral intertrochanteric fractures often requires open reduction. However, the technique unavoidably causes patients to suffer greater trauma. As such, minimally invasive techniques should be employed to reduce the surgical-related trauma on these patients and maintain a stable reduction of the fractures. Herein, a minimally invasive wire introducer was designed and used for the treatment of femoral intertrochanteric fractures. The effectiveness of using a wire-guided device to treat irreducible femoral intertrochanteric fractures was evaluated. METHODS: Between 2013 and 2018, patients with femoral intertrochanteric fractures who were initially treated by intramedullary nail fixation but had difficult reduction using the traction beds were retrospectively reviewed. Decision for an additional surgery was based on the displacement of the fracture. The patients were then divided into two groups: those in the control group received an open reduction surgery while those in the observation group received a closed reduction surgery using a minimally invasive wire introducer to guide the wire that could assist in fracture reduction. The operation time, blood loss, visual analogue scale scores, angulation, reduction, neck-shaft angle, re-displacement, limb length discrepancy, and union time were then recorded and analyzed to determine the efficiency of the wire introducer technique. Categorical variables were analyzed by using Chi-square test, while continuous variables by independent t-test and the Mann-Whitney test accordingly. RESULTS: There were 92 patients included in this study: 61 in the control group and 31 in the observation group. There were no significant differences in baseline demographic factors between the two groups. All surgeries were successful with no deaths within the perioperative period. The average follow-up time for the patients was 23.8 months. However, the observation group had a significantly shorter operation time, lower visual analogue scale score, less intraoperative bleeding, and shorter fracture healing time. There were no significant differences in the angulation, reduction, neck-shaft angle, and limb length discrepancy between the two groups. CONCLUSION: The minimally invasive wire guide achieved a similar effect to that of open reduction in the treatment of intertrochanteric fractures with difficult reduction. Moreover, the minimally invasive wire introducer is a good technology that accurately guides the wire during reduction. Indeed, it is an effective technique and achieves good clinical outcomes in restoration of irreducible femoral intertrochanteric fractures.

Integrated analysis of microRNA and transcription factors in the bone marrow of patients with acute monocytic leukemia.

Acutemonocytic leukemia (AMoL) is a distinct subtype of acute myeloid leukemia (AML) with poor prognosis. However, the molecular mechanisms and key regulators involved in the global regulation of gene expression levels in AMoL are poorly understood. In order to elucidate the role of microRNAs (miRNAs/miRs) and transcription factors (TFs) in AMoL pathogenesis at the network level, miRNA and TF expression level profiles were systematically analyzed by miRNA sequencing and TF array, respectively; this identified 285 differentially expressed miRNAs and 139 differentially expressed TFs in AMoL samples compared with controls. By combining expression level profile data and bioinformatics tools available for predicting TF and miRNA targets, a comprehensive AMoL-specific miRNA-TF-mediated regulatory network was constructed. A total of 26 miRNAs and 23 TFs were identified as hub nodes in the network. Among these hubs, miR-29b-3p, MYC, TP53 and NFKB1 were determined to be potential AMoL regulators, and were subsequently extracted to construct sub-networks. A hypothetical pathway model was also proposed for miR-29b-3p to reveal the potential co-regulatory mechanisms of miR-29b-3p, MYC, TP53 and NFKB1 in AMoL. The present study provided an effective approach to discover critical regulators via a comprehensive regulatory network in AMoL, in addition to enhancing understanding of the pathogenesis of this disease at the molecular level.

The composition and variation of the BCR CDR3s in gastric cancer.

Gastric cancer (GC) is the fourth most common type of cancer and the second most common cause of cancer-associated mortality worldwide. B cell-associated autoantibodies against tumor-associated antigens are attractive biomarkers for the development of noninvasive serological tests for the early detection of cancer. This is due to their specificity and stability in the sera. In the present study multiplex polymerase chain reaction and Illumina high-throughput sequencing (HTS) was used to study the composition and variation of the B cell receptor (BCR) complimentary-determining region 3 (CDR3) in GC. The peripheral blood, cancer tissues and peri-cancer tissues were included from 7 patients with GC. On average there was a total of 403,959 CDR3 sequences, with 72,367 unique CDR3 nt sequences and 61,709 unique CDR3 aa sequences per sample identified, which are critical for further understanding the BCR repertoire in GC. The details of GC CDR3s may accelerate the screening process for possible new autoantigens and may provide additional information necessary for generating effective B cell targeted diagnosis and therapeutic strategies.

Integrated analysis of microRNA and transcription factor reveals important regulators and regulatory motifs in adult B-cell acute lymphoblastic leukemia.

B-cell acute lymphoblastic leukemia (B­ALL) is an aggressive hematological malignancy and a leading cause of cancer-related mortality in children and young adults. The molecular mechanisms involved in the regulation of its gene expression has yet to be fully elucidated. In the present study, we performed large scale expression profiling of microRNA (miRNA) and transcription factor (TF) by Illumina deep­sequencing and TF array technology, respectively, and identified 291 differentially expressed miRNAs and 201 differentially expressed TFs in adult B­ALL samples relative to their controls. After integrating expression profile data with computational prediction of miRNA and TF targets from different databases, we construct a comprehensive miRNA­TF regulatory network specifically for adult B­ALL. Network function analysis revealed 25 significantly enriched pathways, four pathways are well­known to be involved in B­ALL, such as PI3K­Akt signaling pathway, Jak­STAT signaling pathway, Ras signaling pathway and cell cycle pathway. By analyzing the network topology, we identified 28 hub miRNAs and 19 hub TFs in the network, and found nine potential B­ALL regulators among these hub nodes. We also constructed a Jak­STAT signaling sub­network for B­ALL. Based on the sub­network analysis and literature survey, we proposed a cellular model to discuss MYC/miR­15a­5p/FLT3 feed-forward loop (FFL) with Jak­STAT signaling pathway in B­ALL. These findings enhance our understanding of this disease at the molecular level, as well as provide putative therapeutic targets for B-ALL.

Molecular dysfunctions in acute myeloid leukemia revealed by integrated analysis of microRNA and transcription factor.

Acute myeloid leukemia (AML) is a heterogenic hematological malignancy with pathogenesis that has yet to be elucidated. MicroRNAs (miRNAs) and transcription factors (TFs) are two major regulators of gene expression, which may play important roles in the etiology of AML. However, the global regulation of gene expression in AML, involving miRNAs and TFs, still remains elusive. To characterize the global role of miRNAs and TFs in AML pathogenesis, large scale expression profiling of miRNA and TF was performed using miRNA sequencing and TF array technology, respectively, and validated by qPCR. In the present study, 308 miRNAs and 84 TFs were identified to be differentially expressed (fold-change ≥2.0) in AML samples relative to their controls. After integrating the expression profiling data into bioinformatic analysis, we identified 1,462 miRNA-gene pairs, 982 TF-gene pairs and 296 TF-miRNA pairs. By merging these regulatory relations together, we constructed a comprehensive AML-specific miRNA-TF regulatory network. In this network, we identified 22 hub miRNAs and 11 hub TFs. KEGG pathway analysis showed that the network nodes were significantly enriched in 33 different pathways, of which the AML pathway was the most significant. After analyzing the topology of the subnetwork, we propose that TCF3 was a potential key regulator in this regulatory network. In conclusion, this is the first study perform on global expression profiling of miRNAs and TFs relating to AML. These results may enhance our understanding of the molecular mechanisms underlying AML and provide potential targets for future therapeutics.

Identification of microRNAs and their target genes in Alport syndrome using deep sequencing of iPSCs samples.

MicroRNAs (miRNAs) are a class of small RNA molecules that are implicated in post-transcriptional regulation of gene expression during development. The discovery and understanding of miRNAs has revolutionized the traditional view of gene expression. Alport syndrome (AS) is an inherited disorder of type IV collagen, which most commonly leads to glomerulonephritis and kidney failure. Patients with AS inevitably reach end-stage renal disease and require renal replacement therapy, starting in young adulthood. In this study, Solexa sequencing was used to identify and quantitatively profile small RNAs from an AS family. We identified 30 known miRNAs that showed a significant change in expression between two individuals. Nineteen miRNAs were up-regulated and eleven were down-regulated. Forty-nine novel miRNAs showed significantly different levels of expression between two individuals. Gene target predictions for the miRNAs revealed that high ranking target genes were implicated in cell, cell part and cellular process categories. The purine metabolism pathway and mitogen-activated protein kinase (MAPK) signaling pathway were enriched by the largest number of target genes. These results strengthen the notion that miRNAs and their target genes are involved in AS and the data advance our understanding of miRNA function in the pathogenesis of AS.

Integrated analysis of long non-coding RNAs and mRNA expression profiles reveals the potential role of lncRNAs in gastric cancer pathogenesis.

Long non-coding RNAs (lncRNAs) have been shown to play a critical role in cancer biology and are frequently aberrantly expressed. Despite their important role in pathology, little is known mechanistically regarding their role in gastric cancer (GC) pathogenesis. To characterize the role of lncRNAs in GC pathogenesis, 8 paired human GC tissue samples and matched adjacent normal tissue were examined. Large scale expression profiling of lncRNA and mRNA was performed utilizing microarray technology and validated by qPCR. Differentially expressed lncRNAs were subjected to bioinformatic analysis to predict target genes, followed by the integration of differentially expressed mRNA data and GO and network analysis to further characterize potential interactions. In our study, 2,621 lncRNAs and 3,121 mRNAs were identified to be differentially expressed (≥2.0-fold change) in GC samples relative to their matched counterparts. lncRNA target prediction revealed the presence of 221 potential lncRNA-mRNA target pairs for the 75 differentially expressed lncRNAs and 60 differentially expressed genes. KEGG pathway analysis showed that these target genes were significantly enriched in 7 different pathways, of which the p53 signaling pathway was the most significant and has been previously implicated in GC pathogenesis. Construction of a lncRNA-mRNA correlation network revealed 10 differentially expressed lncRNAs potentially regulating the p53 signaling pathway. Overall, this is the first study perform global expression profiling of lncRNAs and mRNAs relating to GC. These results may provide important information for further insights into the pathogenesis of GC and provide potential targets for future therapeutics.

Pelvic step: the contribution of horizontal pelvis rotation to step length in young healthy adults walking on a treadmill.

Transverse plane pelvis rotations during walking may be regarded as the "first determinant of gait". This would assume that pelvis rotations increase step length, and thereby reduce the vertical movements of the centre of mass-"the pelvic step". We analysed the pelvic step using 20 healthy young male subjects, walking on a treadmill at 1-5 km/h, with normal or big steps. Step length, pelvis rotation amplitude, leg-pelvis relative phase, and the contribution of pelvis rotation to step length were calculated. When speed increased in normal walking, pelvis rotation changed from more out-of-phase to in-phase with the upper leg. Consequently, the contribution of pelvis rotation to step length was negative at lower speeds, switching to positive at 3 km/h. With big steps, leg and pelvis were more in-phase, and the contribution of pelvis rotation to step length was always positive, and relatively large. Still, the overall contribution of pelvis rotations to step length was small, less than 3%. Regression analysis revealed that leg-pelvis relative phase predicted about 60% of the variance of this contribution. The results of the present study suggest that, during normal slow walking, pelvis rotations increase, rather than decrease, the vertical movements of the centre of mass. With large steps, this does not happen, because leg and pelvis are in-phase at all speeds. Finally, it has been suggested that patients with hip flexion limitation may use larger pelvis rotations to increase step length. This, however, may only work as long as the pelvis rotates in-phase with the leg.

Effects of experimentally increased trunk stiffness on thorax and pelvis rotations during walking.

Patients with non-specific low back pain, or a similar disorder, may stiffen their trunk, which probably alters their walking coordination. To study the direct effects of increasing trunk stiffness, we experimentally increased trunk stiffness during walking, and compared the results with what is known from the literature about gait coordination with, e.g., low back pain. Healthy subjects walked on a treadmill at 3 speeds (0.5, 1.0 and 1.5m/s), in three conditions (normal, while contracting their abdominal muscles, or wearing an orthopedic brace that limits trunk motions). Kinematics of the legs, thorax and pelvis were recorded, and relative Fourier phases and amplitudes of segment motions were calculated. Increasing trunk stiffness led to a lower thorax-pelvis relative phase, with both a decrease in thorax-leg relative phase, and an increase in pelvis-leg relative phase, as well as reduced rotational amplitude of thorax relative to pelvis. While lower thorax-pelvis relative phase was also found in patients with low back pain, higher pelvis-leg relative phase has never been reported in patients with low back pain or related disorders. These results suggest that increasing trunk stiffness in healthy subjects causes short-term gait coordination changes which are different from those seen in patients with back pain.

Gait adaptations in low back pain patients with lumbar disc herniation: trunk coordination and arm swing.

Patients with chronic non-specific low back pain (LBP) walk with more synchronous (in-phase) horizontal pelvis and thorax rotations than controls. Low thorax-pelvis relative phase in these patients appears to result from in-phase motion of the thorax with the legs, which was hypothesized to affect arm swing. In the present study, gait kinematics were compared between LBP patients with lumbar disc herniation and healthy controls during treadmill walking at different speeds and with different step lengths. Movements of legs, arms, and trunk were recorded. The patients walked with larger pelvis rotations than healthy controls, and with lower relative phase between pelvis and thorax horizontal rotations, specifically when taking large steps. They did so by rotating the thorax more in-phase with the pendular movements of the legs, thereby limiting the amplitudes of spine rotation. In the patients, arm swing was out-of phase with the leg, as in controls. Consequently, the phase relationship between thorax rotations and arm swing was altered in the patients.

[Effect of seven kinds of flavonoids on recombinant human protein tyrosine phosphatase].

OBJECTIVE: To observe the effects of seven kinds of flavonoids on recombinant human phosphatase of regenerating liver-3 activity. METHODS: The inhibitory effect of flavonoids was tested by DiFMUP assay. Calculation of IC50 values was performed according to the law of semi-effect-probit. RESULTS: Myricetin and gossypin could significantly inhibit recombinant PRL-3 activity in a concentration dependent manner with IC50 values of 55.54 and 68.86 micromol/L, respectively. Quercetin, luteolin and 7,8-Dihydroxyflavone had a weak inhibitory effect with IC50 values of 113.38, 151.56 and 249.49 micromol/L. respectively. While 3-hydroxyflavone and 6-hydroxyflavone had no significant effect on PRL-3 activity. Structure activity study indicated that C4 and C7 hydroxyls on the flavone skeleton were key functional groups to influencing PRL-3 inhibotory activity. The inhibitory effect of flavonoids on PRL-3 was increased with the number of hydroxyl group. CONCLUSION: Myricetin and gossypin were two strong inhibitors on recombinant human protein tyrosine phosphatase PRL-3.

[Effects of mitoxantrone on the activity of human protein kinase CK2 holoenzyme].

BACKGROUND & OBJECTIVE: Protein kinase CK2, a highly conserved protein serine/threonine kinase that is ubiquitously distributed in eukaryotes, has a close relationship with human leukemia. Mitoxantrone is an effective drug used for acute leukemias. This study was to observe the effects of mitoxantrone on the activity of recombinant holoenzyme of human protein kinase CK2 and proliferation in human leukemia cell line HL-60. METHODS: The CK2 holoenzyme composed of alphao and beta subunits was recombined in vitro. Subsequently, CK2 was treated with mitoxantrone at various concentrations, followed by addition of reacting liquid containing [gamma-32P] ATP. CK2 activity was measured by detecting the radioactivity of 32P transferred onto the substrates of CK2. The effect of mitoxantrone on the proliferation of HL-60 cells was detected by the trypan blue dye exclusion assay; the changes in the cell cycle were analyzed by flow cytometry (FCM); apoptosis was analyzed by FCM and DNA agarose gel electrophoresis; effects of the drugs on the intrinsic CK2 activity were measured by a specific CK2 peptide substrate. RESULTS: Mitoxantrone strongly inhibited the holoenzyme activity of recombinant human protein kinase CK2 (IC50=0.66 micromol/L). The inhibition of CKZ by mitoxantrone was competitive with respect to ATP (Ki 0.25 micromol/L) and mostly non-competitive with respect to casein (Ki 0.66 micromol/L). Mitoxantrone exerted strong cytotoxicity to HL-60 cells. When treated with 0.1 micromol/L mitoxantrone for 12 h, the apoptotic rate of HL-60 cells was 9.3%. Mitoxantrone did not affect intracellular CK2 activity. CONCLUSIONS: Mitoxantrone is a strong inhibitor of recombinant human protein kinase CK2 in vitro. Apoptosis induced by mitoxantrone in HL-60 cells has no correlation to intracellular CK2 activity.

[Inhibitory effect and its kinetic analysis of baicalein on recombinant human protein kinase CK2 holoenzyme].

BACKGROUND & OBJECTIVE: Protein kinase CK2 is a ubiquitous and pleiotropic Ser/Thr protein kinase in eukaryotic cells. CK2 activity has been shown to be markedly elevated in solid tumors and leukemia cells. Its alpha or alpha' gene is a protooncogene. CK2 is attracting increasing attention as a potential target for anti-neoplastic. This study was to search specific CK2 inhibitors in tumor cells through observation of the inhibitory effects of baicalein on recombinant human protein kinase CK2 holoenzyme and its kinetics in vitro. METHODS: Recombinant human protein kinase CK2 alpha' and beta subunits were cloned and expressed by gene engineering, and purified to homogeneous. These 2 subunits were mixed at equal molar ratio to reconstitute CK2 holoenzyme. The CK2 activity was evaluated by detecting radioactivity of 32P of [gamma-32P]ATP which was incorporated into the substrate in various conditions. RESULTS: Baicalein was shown to strongly inhibit the holoenzyme activity of CK2 with IC50 of 2.54 micromol/L. Kinetic studies of baicalein on CK2 showed that baicalein acted as an inhibitor of noncompetitive with ATP(KI=7.73 micromol/L) and mixed types with casein(KI=3.07 micromol/L). CONCLUSION: Baicalein is an effective inhibitor of protein kinase CK2 in vitro.