Extraction-free, immuno-RPA-CRISPR/Cas13a-based one-pot detection of glypican-3 directly from extracellular vesicles.

BACKGROUND: Glypican-3 (GPC3) is a heparan sulfate proteoglycan (HSPG) that binds to the cell membrane via glycosylphosphatidylinositol (GPI). It is not found in healthy adult liver but is overexpressed in human hepatocellular carcinoma (HCC). The protein marker GPC3 on extracellular vesicles (GPC3+ EVs) is also useful for HCC detection. Nevertheless, the absence of practical and dependable quantitative techniques to evaluate EVs proteins prevents their clinical implementation. RESULTS: Here, using an immuno-recombinase polymerase amplification (immuno-RPA) process and dual amplification of clustered regularly interspaced short palindromic repeats (CRISPR)-Cas13a, we firstly create an extraction-free one-pot immuno-RPA-CRISPR (opiCRISPR) for the direct and extremely sensitive detection of EVs proteins. The EVs protein-targeted detection probe is amplified by RPA to generate a long repetitive sequence containing multiple CRISPR RNA (crRNA) targeting barcodes, and the signal is further amplified by the CRISPR-Cas13a side-chain cleavage activity to generate a fluorescent signal. The results show that circulating extracellular vesicle GPC3 (eGPC3) levels are a reliable marker for GPC3 expression in tumor, opening up new avenues for tumor diagnosis. SIGNIFICANCE AND NOVELTY: We created an eGPC3 assay based on the CRISPR-Cas13a system, and successfully study the significance of extracellular vesicle GPC3 markers in hepatocellular carcinoma.

Current methods for the detection of glypican-3.

Glypican-3 (GPC3) is a heparan sulfate proteoglycan (HSPG) that binds to the cell membrane via glycosylphosphatidylinositol (GPI), widely expressed in human embryos, and is undetectable in healthy adult liver but overexpressed in human hepatocellular carcinoma (HCC). Therefore, accurate and sensitive detection of GPC3 is critical for disease diagnosis. In recent years, a series of methods have been developed for the highly sensitive detection of GPC3, but there is a lack of reviews on recent advances in GPC3-related assays. In this review, we provide the recent advances in GPC3 detection and GPC3 concentration detection, mainly in terms of various optical sensor-based assays and electrochemical assays, and also provide new insights into the challenges and future directions of the field.

Superior short-term outcomes of FNS in combination with a cannulated screw in treating femoral neck fractures.

BACKGROUND: This study aimed to evaluate the clinical efficacy of the femoral neck system alone or in combination with a cannulated screw compared with other internal fixation methods for treating femoral neck fractures. We further investigated the predictive effects of tip-apex distance (TAD) on clinical efficacy. METHODS: Data from 129 young adults with femoral neck fractures followed up at The Second Affiliated Hospital of Fujian Medical University between January 2016 and June 2022 were retrospectively collected. The patients were categorized into four groups based on the different internal fixation methods. Analysis and comparisons of the four group were performed according to age, ASA score, operation time, blood loss, fracture classification, fracture healing time, Harris score, TAD value, presence of complications (osteonecrosis of the femoral head, screw failure, and femoral neck shortening), and changes in the neck-shaft angle. RESULTS: All 129 patients were followed up for at least one year. The group who received treatment with the femoral neck system combined with a cannulated screw exhibited the shortest fracture healing time. Differences were observed in the change of neck-shaft angle among the four groups (P < 0.001), with the smallest change observed in the aforementioned group (0.76 ± 0.54°). The femoral neck shortening was also lower in groups with the femoral neck system or combined with a cannulated screw. At the last follow-up surgery, the combined treatment group achieved the highest HHS score. Subgroup analysis revealed that when the TAD was less than 25 and 49 mm for the femoral neck system and combined groups, respectively, there was less femoral neck shortening, less change in the neck-shaft angle, and a higher HHS score. CONCLUSIONS: The femoral neck system alone or combined with a cannulated screw demonstrated better short-term efficacy in the treatment of femoral neck fractures. Furthermore, TAD may serve as a predictive indicator of the potential success of femoral neck fracture treatment.

Roles and regulatory mechanisms of KIN17 in cancers (Review).

KIN17, which is known as a DNA and RNA binding protein, is highly expressed in numerous types of human cancers and was discovered to participate in several vital cell behaviors, including DNA replication, damage repair, regulation of cell cycle and RNA processing. Furthermore, KIN17 is associated with cancer cell proliferation, migration, invasion and cell cycle regulation by regulating pathways including the p38 MAPK, NF-κB-Snail and TGF-ß/Smad2 signaling pathways. In addition, knockdown of KIN17 was found to enhance the sensitivity of tumor cells to chemotherapeutic agents. Immunohistochemical analysis revealed that there were significant differences in the expression of KIN17 between cancer tissues and adjacent tissues. Both the Kaplan-Meier survival analysis and multivariate Cox regression analysis indicated that KIN17 is aberrantly high expressed in various tumor tissues and is also associated with poor prognosis in patients with various tumor types. Taken together, KIN17 has key roles in tumorigenesis and cancer development. Investigating the relationship between KIN17 and neoplasms will provide a vital theoretical basis for KIN17 to serve as a diagnostic and prognostic biomarker for cancer patients and as a potential target for cancer therapy.

Long non-coding RNA BC002811 Promotes Gastric Cancer Metastasis by Regulating SOX2 Binding to the PTEN Promoter.

There is increasing evidence that long non-coding RNAs (lncRNAs) are involved in the pathogenesis and progression of gastric cancer (GC), however, the underlying mechanisms remain poorly understood. In this study, we identified lncRNA BC002811 as a critical regulator of GC development and progression. BC002811 was upregulated in GC tissues and cell lines, and that high expression of BC002811 was indicative of a reduction in overall survival of GC patients. Our research reveals that BC002811 promoted GC cell proliferation, migration, invasion, and inhibition of apoptosis in vitro, as well as accelerated tumor growth and metastasis in vivo. We also found that BC002811 upregulated MMP2 and MMP9 and promoted GC cell metastasis partially through downregulating PTEN expression. BC002811 may act as a molecular decoy for the transcription factor SOX2, thereby inhibiting the transcription of PTEN by blocking SOX2 binding to the PTEN promoter. Our study advances the understanding of the role of BC002811 in the pathogenesis of GC and provides new molecular targets for therapeutic intervention against GC metastasis.

The tibial insertion of the hamstring can be considered to be preserved during anterior cruciate ligament reconstruction.

Background: Hamstring as a graft was very common in anterior cruciate ligament reconstruction surgery. Usually the hamstring muscles needed to be taken out and then woven to be used. Aim: In order to investigate whether it was beneficial for patients to preserve the transpedicular insertion of hamstring when using the hamstring as a graft for anterior cruciate ligament reconstruction. Methods: This was a retrospective study. Patients with anterior cruciate ligament injury who underwent surgery in a large hospital from January 2015 to May 2021 were included in the study. These patients underwent anterior cruciate ligament reconstruction assisted by arthroscopic. Autologous hamstring muscles were used as grafts. The tibial insertion of the hamstring were preserved during the operation were included in the observation group. The remaining patients were included in the control group. The knee joint function and operation of the two groups were compared. Results: A total of 97 patients were included in the study. There was no statistical difference between the two groups in general data including gender, age and surgical side. All the patients' operations were successfully completed there was no significant difference in the operation time between the two groups. All patients were followed up for at least 1 year. No patients had complications such as wound infection and graft failure at the last follow-up. There was no significant difference between the two groups in Lysholm score and IKDC score before operation. Similarly, there was no significant difference between the two groups in Lysholm score and IKDC score 3 months after operation. However, the Lysholm score and IKDC score of the two groups 1 year after operation were statistically different, and the patients in the observation group had higher Lysholm score and IKDC score. After comparing the MRI images of the knee of the two groups 3 months after operation through the MRI evaluation system, compared with the patients in the control group, the patients in the observation group have higher scores, and the difference was statistically significant. Conclusion: In the knee arthroscopic assisted anterior cruciate ligament reconstruction using the hamstring as a graft, the tibial insertion of the hamstring can be preserved, which can make the patient have better function after the operation. This kind of operation leads to the increase of operation time and operation risk.

Effect of modified Laprade technique on posterolateral ligament injury of knee.

PURPOSE: To investigate the effect of modified Laprade technique on the reconstruction of posterolateral structure of knee and anterolateral ligament of knee in the treatment of posterolateral injury of knee. METHODS: From December 2013 to June 2020, multiple ligament injury patients who received surgery in our hospital were collected in this research. These patients underwent a modified Laprade technique for posterolateral structural reconstruction of the knee. Lysholm scores of patients pre- and post-operation were recorded. RESULT: The operations of the observation group or the control group patients were completed. There were no significant differences in gender, age, preoperative knee range of motion and preoperative Lysholm score. At the time of follow-up 1 month after operation, there was no significant difference in knee range of motion, dial-up test angle and Lysholm score between the observation and the control group. When followed up 1 year after operation, the Lysholm score of the observation group was higher than that of the control group. The difference was statistically significant. The same situation occurred in the range of motion of the knee in both groups. However, there was still no significant difference between the two groups in the dial-up test 1 year after operation, whether the knee flexion was 30° or 90°. CONCLUSION: For patients with posterolateral structure injury of knee, the modified Laprade technique is a feasible surgical technique.

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.

Arthroscopic fixation of humeral greater tuberosity fracture using a W-shaped suture.

BACKGROUND: Patients with greater tuberosity fractures of the humerus often require surgery. Therefore, there is a need to find a minimally invasive and effective surgical procedure with great patient outcomes. AIM: To evaluate the clinical outcomes of the W-shaped suture technique under shoulder arthroscopy in the treatment of greater tuberosity fractures of the humerus. METHODS: In this retrospective study, a total of 17 patients were included. The fractures were closed, and there was no neurovascular injury. These patients underwent arthroscopically assisted reduction and internal fixation of the greater tuberosity fractures. Fixation was performed using sighting nails combined with a W-shaped suture. The imaging data of the patients were collected, and the ASES score, Constant-Murley score, and VAS score were used to evaluate the patient's outcome. At the last follow-up (at least 1 year), the range of motion in the affected shoulder was compared with that of the contralateral side. RESULTS: The operation was successful in all the patients. The average follow-up time was 13 months. There were no reported complications such as fracture displacement, nonunion, and internal fixation failure during the follow-up period. Post-operative X-ray examinations revealed good function recovery, with a healing time of between 10 and 12 weeks, and an average healing time of 11.5 weeks. Following the operation, patients reported reduced shoulder joint pain that no longer influenced their activity or caused discomfort in their daily life. The patient's VAS score ranged from 0 to 3, with an average of 0.52 ± 0.73, while at the last follow-up, the Constant-Murley score ranged from 83 to 97, with an average of 92.33 ± 7.55. The ASES score ranged from 81 to 98, with an average of 93.15 ± 6.93. At the last follow-up, there was no significant difference in the overall range of motion with the unaffected limb. CONCLUSION: This study demonstrates that the W-shaped suture can be used to effectively fix the fractures of the greater tuberosity of the humerus, by increasing the fixed area to promote healing.

Do we need to suture the pronator quadratus muscle when we do open reduction and internal fixation for fracture of the distal radius.

BACKGROUND: Open reduction and internal fixation is often used for the treatment of distal radius fracture. Opening the pronator quadratus muscle during the process of open reduction and internal fixation is necessary to achieve sufficient exposure. Therefore, knowledge on how to suture the pronator quadratus muscle will be of essence. AIM: The aim of the present study was to determine if suturing the pronator quadratus during the treatment of the distal radius fracture can enhance limb function . METHODS: A total of 126 patients were enrolled for the study. The patients underwent open reduction and internal fixation. During the procedure, the pronator quadratus was cut open to allow insertion of the plate. The pronator quadratus muscles of the patients were stitched together before the surgery was completed. After the fracture healed, the patients underwent surgery to remove the internal fixations. Patients received wrist function scores prior to removal of the internal fixations. Healing of the pronator quadratus was during surgery. Patients were grouped according to the healing of the pronator quadratus. Functional scores between the two groups were compared. RESULTS: Muscle healing was observed in 23 patients during surgery. However, the PQ muscles of these patients were remarkably atrophic, with scar hyperplasia and fibrosis. The muscle fibers were loose, thin, and had decreased in number. The remaining muscle fibers presented different degrees of adhesion with radial carpal flexor muscles, steel plates and interosseous membrane. A total of 23 patients were included in group A and 103 patients in group B based on the intraoperative condition. No statistically significant differences was observed in age and type of fracture between group A and group B. In addition, no statistically significant differences was observed in the isokinetic forearm pronation strength and clinical outcomes including grip strength, wrist ROM, and PRWE scores between the two groups. CONCLUSION: This study demonstrates that healing of the PQ muscle does not affect the outcomes of volar plating for distal radius fractures with reference to the isokinetic forearm rotation strength, grip strength, wrist ROM, and PRWE scores. The results of this study support our current practice of PQ muscle incision.

[Corrigendum] Oncogenic function of Plac1 on the proliferation and metastasis in hepatocellular carcinoma cells.

Following the publication of this article, the authors have realized that Table I contained some important errors. The data shown for the positive or negative HBsAg patient characteristic in terms of the no. of patients, Plac1 (+) and Plac1 (­) were incorrect. A corrected version of the Table is shown below (the corrected data are highlighted in bold). The authors sincerely apologize for the errors that were introduced during the preparation of this Table. Furthermore, they regret any inconvenience that this mistake has caused. [the original article was published in Oncology Reports 37: 465­473, 2017; DOI: 10.3892/or.2016.5272].

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.

Oncogenic function of Plac1 on the proliferation and metastasis in hepatocellular carcinoma cells.

Placenta-specific protein 1 (Plac1), which is selectively expressed in the placental syncytiotrophoblast in adult normal tissues, plays an essential role in normal placental and embryonic development. Accumulating evidence suggests that enhanced Plac1 expression is closely associated with the progression of human cancer. Whether Plac1 contributes to the pathophysiology of hepatocellular carcinoma (HCC) remains unclear. In the present study, our data revealed that the expression of Plac1 in human HCC tissues was upregulated, which significantly correlated with metastasis of HCC. Knockdown of Plac1 by small interfering RNA (siRNA) in Bel-7402 and HepG2 cells resulted in decreasing tumor cell proliferation and increasing apoptosis, which implied the oncogenic potential of Plac1. Moreover, silencing of Plac1 induced G1 cell cycle arrest through suppression of cyclin D1 and CDK4 expression. Furthermore, depletion of Plac1 repressed epithelial-mesenchymal transition (EMT), with decreased cell migration and invasion, supporting upregulated E-cadherin expression and downregulated vimentin, twist and snail expression that characterize EMT. Further study suggested that decreased Plac1 expression attenuated the phosphorylation of Akt. These findings have uncovered that Plac1 plays a pivotal role in the progression of HCC, and may serve as a novel therapeutic target for HCC.

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.

Generation of induced pluripotent stem cells from renal tubular cells of a patient with Alport syndrome.

Alport syndrome (AS) is a hereditary disease that leads to kidney failure and is caused by mutations in the COL4A3, COL4A4, and COL4A5 genes that lead to the absence of collagen α3α4α5 (IV) networks in the mature kidney glomerular basement membrane. Approximately 80% of AS is X-linked because of mutations in COL4A5, the gene encoding the alpha 5 chain of type IV collagen. To investigate the pathogenesis of AS at the genetic level, we generated induced pluripotent stem cells (iPSCs) from renal tubular cells of a patient with AS. The successful iPSC generation laid the foundation to master the repair of the COL4A5 gene and to evaluate the differentiation of iPSC into Sertoli cells and the accompanying epigenetic changes at each stage. The generation of iPSCs from AS patients not only confirms that iPSCs could be generated from renal tubular cells, but also provides a novel type of genetic therapy for AS patients. In this study, we generated iPSCs from renal tubular cells via ectopic expression of four transcription factors (Oct4, Sox2, c-myc, and Klf4). According to the human embryonic stem cell (hESC) charter, iPSC formation was confirmed by comparatively analyzing hESC markers via colony morphology, immunohistochemistry, qRT-PCR, flow cytometry, gene expression profiling of the three germ layers, and karyotyping. Our results demonstrated that iPSCs were similar to hESCs with regard to morphology, proliferation, hESC-specific surface marker expression, and differentiation into the cell types of the three germ layers. The efficient generation of iPSCs from the renal tubular cells of an AS patient would provide a novel model to investigate the mechanisms underlying AS and to develop new treatments for AS.

Frontal plane kinematics in walking with moderate hip osteoarthritis: Stability and fall risk.

BACKGROUND: Hip abductor weakness and unilateral pain in patients with moderate hip osteoarthritis may induce changes in frontal plane kinematics during walking that could affect stability and fall risk. METHODS: In 12 fall-prone patients with moderate hip osteoarthritis, 12 healthy peers, and 12 young controls, we assessed the number of falls in the preceding year, hip abductor strength, fear of falling, Harris Hip Score, and pain. Subjects walked on a treadmill with increasing speeds, and kinematics were measured opto-electronically. Parameters reflecting gait stability and regressions of frontal plane center of mass movements on foot placement were calculated. We analyzed the effects of, and interactions with group, and regression of all variables on number of falls. FINDINGS: Patients walked with quicker and wider steps, stood shorter on their affected leg, and had larger peak speeds of frontal plane movements of the center of mass, especially toward their unaffected side. Patients' static margins of stability were larger, but the unaffected dynamic margin of stability was similar between groups. Frontal plane position and acceleration of the center of mass predicted subsequent step width. The peak speed of frontal plane movements toward unaffected had 55% common variance with number of falls, and adding the Harris Hip Score into bivariate regression led to 83% "explained" variance. INTERPRETATION: Quickening and widening steps probably increase stability. Shorter affected side stance time to avoid pain, and/or weakened affected side hip abductors, may lead to faster frontal plane trunk movements toward the unaffected side, which could contribute to fall risk.

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.