Low-energy shock waves promote the cisplatin chemosensitivity of human osteosarcoma MNNG/HOS cells via the P2X7/Akt/mTOR pathway.

Background: The current dilemma of osteosarcoma treatment is the resistance of chemotherapeutic drugs after long-term usage, which also introduces life-threatening side effects. Methods and results: To minimize chemoresistance in osteosarcoma patients, the authors applied shock waves (SWs) to human osteosarcoma MNNG/HOS cells, then evaluated the cell viability and extracellular ATP levels, and further investigated the effect of SWs on cisplatin (DDP) cytotoxicity in MNNG/HOS cells. The authors' results showed that 400 SW pulses at 0.21 mJ/mm2 exhibited little influence on the MNNG/HOS cell viability. In addition, this SW condition significantly promoted the extracellular ATP release in MNNG/HOS cells. Importantly, low-energy SWs obviously increased Akt and mammalian target of rapamycin (mTOR) phosphorylation and activation in MNNG/HOS cells, which could be partially reversed in the presence of P2X7 siRNA. The authors also found that low-energy SWs strongly increased the DDP sensitivity of MNNG/HOS cells in the absence of P2X7. Conclusions: For the first time, the authors found that SW therapy reduced the DDP resistance of MNNG/HOS osteosarcoma cells when the ATP receptor P2X7 was downregulated. SW therapy may provide a novel treatment strategy for chemoresistant human osteosarcoma.

Stem cell-derived exosomes for chronic wound repair.

Stem cells possess the capability of self-renewal and multipotency, which endows them with great application potential in wound repair fields. Yet, several problems including immune concerns, ethical debates, and oncogenicity impede the broad and deep advance of stem cell-based products. Recently, owing to their abundant resources, excellent biocompatibility, and ease of being engineered, stem cell-derived exosomes were proved to be promising nanomedicine for curing chronic wounds. What is more, stem cell-derived exosomes are almost the mini record of their maternal cells, which even equipped them with the unique characteristics of stem cells. Chronic wound healing efficacy is dominated by several complicated factors, especially the excessive inflammation conditions and impaired vessels. Therefore, this review tries to concentrate on the current advances of stem cell-derived exosomes for reducing inflammation and promoting angiogenesis in chronic wound healing processes. Last but not least, the existing limitations and future perspectives of stem cell-derived exosomes for chronic wound treatment are also outlined.

Treatment of pediatric tibial shaft nonunion using ipsilateral free non-vascularized fibular graft.

BACKGROUND: Although tibial shaft fractures are the third most common long bone fractures in children after the forearm and femur, nonunion of these fractures are rare in the pediatric population. CASE REPORT: Despite seldom seen, tibial nonunion is very complex and it is also a devastating complication of tibial fracture especially when infected. Numerous methods have been employed to treat pediatric tibial nonunion, but there is no consensus. Here, we present a case of a child with right tibial shaft fracture nonunion. We treated this patient with ipsilateral free non-vascularized fibular graft. RESULTS: Both the nonunion site and fibular donor site united well with good function in the injured extremity and no adverse events. CONCLUSION: We recommend the use of ipsilateral free non-vascularized fibular graft for the treatment of pediatric tibial shaft nonunion.

Functional outcomes and adverse effects following three interventions for displaced midshaft clavicular fractures: A Bayesian network meta-analysis of randomized controlled trials.

INTRODUCTION: The efficacy of the most commonly used interventions for clavicle fractures remains controversial. These interventions are: open reduction and plate fixation (ORPF), non-surgical intervention (NSI), and use of an intramedullary nail (IMN). In adult patients with clavicle fractures, choosing which intervention might be best is challenging. MATERIALS AND METHODS: PubMed, Journals@Ovid Full Text, Cochrane Central Register of Controlled Trials (CENTRAL), Ovid MEDLINE, and Embase were performed to search English-language studies from the inception to February 2020. Randomized controlled trials (RCTs) comparing any of these three interventions were included. Patient and baseline characteristics, nonunion, major complications, Constant-Murley score (CMS), and Disabilities of the Arm, Shoulder and Hand score (DASH) were extracted. Then, we evaluated the functional outcomes and adverse effects after use of these three interventions for the management of displaced midshaft clavicle fractures in a Bayesian network meta-analysis. RESULTS: A Bayesian random-effects model was conducted, and nonunion and major complications were evaluated with: risk ratio (RR) and 95% confidential interval (CI); while CMS and DASH were evaluated with mean differences (MD) and the corresponding 95% confidential interval CI. The rank probability of each endpoint was assessed on the basis of the surface area under the cumulative ranking curve (SUCRA). DISCUSSION: ORPF is most likely to be successful in achieving objective functional outcomes as captured by the CMS, and IMN demonstrates significant efficacy for subjective functional outcomes, as captured by DASH scores. Compared with the other interventions examined, IMN was associated with decreased risk for adverse effects. LEVELS OF EVIDENCE: I; meta-analysis.

The applications of research progress of common wheat in teaching genetics.

Common wheat (T. aestivum L.) is also known as allohexaploid wheat. Its genome is composed of A/B/D sub-genomes from three closely related diploid ancestors. The evolutionary history of common wheat is used as a classic example to illustrate the mechanism of species formation and chromosome number variation in the current genetics class. In recent years, with the rapid development and application of research technologies, there have been many breakthroughs in the study of common wheat, at the cytological, molecular and genomic level. Here, we summarize the latest research achievements on common wheat, and discuss our practice in combining them with the genetics teaching. Our approach is not only a supplement to the current genetics textbooks, but also enables students to realize that genetics is a constantly evolving natural science. We aim to enhance students' interests in learning, as well as their systematic learning abilities on genetics and related scientific research frontiers.

Effect of Er on Microstructure and Mechanical Properties of 5052 Aluminum Alloy with Big Width-To-Thickness Ratio.

The effect of Er on microstructure and mechanical properties of the 5052 aluminum alloy with a big width-to-thickness ratio was investigated by a metallurgical microscope, scanning electron microscope and tensile testing machine. The results showed that the precipitates were slightly refined after Er addition and Al3Fe was transformed into Al6Fe and AlEr with/without a small amount of Fe or Si. The effect of Er on grain refinement was related to its content. When Er content was lower or higher than 0.4%, the grain would coarsen. Homogenization could refine the grain by controlling Er content and distribution in the Al matrix. Long time homogenization at high temperature would significantly reduce the strength of the 5052 aluminum alloy and 5052 aluminum alloys with low Er content, but help to improve the plasticity of those with high Er content. The ultimate tensile strength, yield strength and elongation of the as-cast 5052 aluminum alloy were 197 MPa, 117 MPa and 22.5% respectively. The strength was the highest, when Er content was 0.4 wt. % and the elongation was the best at 0.1 wt. % Er content.

An immunomagnetic separation and bifunctional Au nanoparticle probe-based multiamplification electrochemical strategy.

A novel electrochemical magnetoimmunosensor for the rapid and sensitive detection of carcinoembryonic antigen (CEA) was fabricated based on a combination of high-efficiency immunomagnetic separation, bifunctional Au-nanoparticle (bi-AuNP) probes, and enzyme catalytic amplification. The reaction carrier magnetic beads (MBs) effectively reduced the toxicity of the complex sample to the working electrode, and the signal carrier bi-AuNP probes loaded a large amount of signal molecules, both of which enhanced the signal-to-noise ratio and further improved the detection sensitivity. A detection limit as low as 0.11 pg/mL was achieved for CEA detection based on the immunomagnetic separation and bi-AuNP probe-based multiamplification strategy, and the strategy was further successfully applied in human serum samples. The transducer was regenerated via a simple washing procedure, which enabled the detection of all samples on a single electrode with high reproducibility. The proposed strategy, which has the merits of high sensitivity, selectivity, and reproducibility exhibits great potential for detection in complex samples.

Screening of underlying genetic biomarkers for ankylosing spondylitis.

Genetic biomarkers for the diagnosis of ankylosing spondylitis (AS) remain unreported except for human leukocyte antigen B27 (HLA­B27). Therefore, the aim of the present study was to screen the differentially expressed genes (DEGs), and those that also possess differential single nucleotide polymorphism (SNP) loci in the whole blood of AS patients compared with healthy controls by integrating two mRNA expression profiles (GSE73754 and GSE25101) and SNP microarray data (GSE39428) collected from the Gene Expression Omnibus (GEO). Using the t­test, 1,056 and 1,073 DEGs were identified in the GSE73754 and GSE25101 datasets, respectively. Among them, 234 DEGs were found to be shared in both datasets, which were subsequently overlapped with 122 differential SNPs of genes in the GSE39428 dataset, resulting in identification of two common genes [eukaryotic translation elongation factor 1 epsilon 1 (EEF1E1) and serpin family A member 1 (SERPINA1)]. Their expression levels were significantly upregulated and the average expression log R ratios of SNP sites in these genes were significantly higher in AS patients than those in controls. Function enrichment analysis revealed that EEF1E1 was involved in AS by influencing the aminoacyl­tRNA biosynthesis, while SERPINA1 may be associated with AS by participating in platelet degranulation. However, only the genotype and allele frequencies of SNPs (rs7763907 and rs7751386) in EEF1E1 between AS and controls were significantly different between AS and the controls, but not SERPINA1. These findings suggest that EEF1E1 may be an underlying genetic biomarker for the diagnosis of AS.

Enzymatic gene expression by Pleurotus tuoliensis (Bailinggu): differential regulation under low temperature induction conditions.

Pleurotus tuoliensis is a valuable, rare and edible mushroom that is been commercially cultivated and is rapidly developing in China markets. Low temperatures are required to induces primordia initiation for the successful production of fruiting bodies (basidiomes) during commercial cultivation. In this work, we investigated the enzymatic activities and performed transcription profiling analysis of enzymatic genes under different low temperature conditions. The results suggest that the enzymatic activities and transcription levels decrease or increase significantly at 4 and 13 °C. Lacc10 and mnp6 seems to play a dominant role during nutrition growth. Furthermore, the expression of laccase and peroxidase genes was highly correlated to the detected extracellular enzymatic activity. Cold stress genes expression profiles were upregulated under 4 °C/13 °C (3 days), while only the Hsp70 gene was downregulated (at the stage of fruiting bodies production) at 13 °C (12 days). Our results showed that the transcriptional regulation of laccase and ligninolytic peroxidase genes plays an important role in the fruiting bodies of Bailinggu under low temperature induction (4 °C). Induction at low temperatures was a highly important cultivation condition in Bailinggu.

The Role of Angle in the Evaluation of Ablation Accuracy in Pulmonary Vein Isolation Navigated by Image Integration.

BACKGROUND: The conventional index for ablation accuracy is to compare the distance between mapping points with and without treatment by using image integration. We attempted to quantitatively evaluate the role of angle as an index in the ablation accuracy in patients with atrial fibrillation (AF). METHODS: A total of 48 patients with AF were included in the present study. Virtual fluoroscopy planes were predicted by pulmonary vein (PV) angiography, and the standard image planes were defined on the basis of the computed tomography images. Ablations were performed, guided by image integration; and the ablation planes were defined by the actual ablation rings. The predicted angle (distance) was defined as the angle (distance) between the fluoroscopy (predicted) plane and image (standard) plane, whereas the actual angle (distance) was defined as the angle (distance) between the ablation (actual) planes and the image (standard) planes. RESULTS: We found that all actual angles were significantly smaller than the predicted angles (P <.05), but only the actual distances in the left PV, right inferior PV, right superior PV, and right PV were significantly smaller; the distances in the left inferior PV and left superior PV were not significantly different (P >.05). CONCLUSION: Our finding indicates that both the angle and the distance can be significantly reduced by navigation with image integration, but that the angle exhibited better sensitivity than the conventional index of distance. We suggest that the angle should be considered as a new index for ablation accuracy.

Porcine circovirus type 2 capsid protein induces unfolded protein response with subsequent activation of apoptosis.

Porcine circovirus type 2 (PCV2) has recently been reported to elicit the unfolded protein response (UPR) via activation of the PERK/eIF2α (RNA-activated protein kinase-like endoplasmic reticulum (ER) kinase/eukaryotic initiation factor 2α) pathway. This study attempted to examine which viral protein might be involved in inducing UPR and whether this cellular event would lead to apoptosis of the cells expressing the viral protein. By transient expression, we found that both replicase (Rep) and capsid (Cap) proteins of PCV2 could induce ER stress as shown by increased phosphorylation of PERK with subsequent activation of the eIF2α-ATF4 (activating transcription factor 4)-CHOP (CCAAT/enhancer-binding protein homologous protein) axis. Cap expression, but not Rep, significantly reduced anti-apoptotic B-cell lymphoma-2 (Bcl-2) and increased caspase-3 cleavage, possibly due to increased expression of CHOP. Since knockdown of PERK by RNA interference clearly reduced Cap-induced CHOP expression, caspase-3 cleavage, and apoptotic cell death possibly by partially rescuing Bcl-2 expression, we propose that there is connection between Cap-induced UPR and apoptosis via the PERK/eIF2α/ATF4/CHOP/Bcl-2 pathway. This study, together with our earlier studies, provides insight into the mechanisms underlying PCV2 pathogenesis.

The Effects of Pre-Operative Enteral Nutrition from Nasal Feeding Tubes on Gastric Outlet Obstruction.

We examined gastric outlet obstruction (GOO) patients who received two weeks of strengthening pre-operative enteral nutrition therapy (pre-EN) through a nasal-jejenal feeding tube placed under a gastroscope to evaluate the feasibility and potential benefit of pre-EN compared to parenteral nutrition (PN). In this study, 68 patients confirmed to have GOO with upper-gastrointestinal contrast and who accepted the operation were randomized into an EN group and a PN group. The differences in nutritional status, immune function, post-operative complications, weight of patients, first bowel sound and first flatus time, pull tube time, length of hospital stay (LOH), and cost of hospitalization between pre-operation and post-operation were all recorded. Statistical analyses were performed using the chi square test and t-test; statistical significance was defined as p < 0.05. The success rate of the placement was 91.18% (three out of 31 cases). After pre-EN, the levels of weight, albumin (ALB), prealbumin (PA), and transferrin (TNF) in the EN group were significantly increased by pre-operation day compared to admission day, but were not significantly increased in the PN group; the weights in the EN group were significantly increased compared to the PN group by pre-operation day and day of discharge; total protein (TP), ALB, PA, and TNF of the EN group were significantly increased compared to the PN group on pre-operation and post-operative days one and three. The levels of CD3+, CD4+/CD8+, IgA, and IgM in the EN group were higher than those of the PN group at pre-operation and post-operation; the EN group had a significantly lower incidence of poor wound healing, peritoneal cavity infection, pneumonia, and a shorter first bowel sound time, first flatus time, and post-operation hospital stay than the PN group. Pre-EN through a nasal-jejunum feeding tube and placed under a gastroscope in GOO patients was safe, feasible, and beneficial to the nutrition status, immune function, and gastrointestinal function, and sped up recovery, while not increasing the cost of hospitalization.

Methylation of the suppressor of cytokine signaling 3 gene (SOCS3) in bladder cancer.

BACKGROUND: It has been identified consequences of dysregulation of JAK-STAT signalling, particularly in regard to JAK-STAT signalling that has been shown to have roles in the oncogenesis of several cell types. SOCS3 protein, the negative regulatory protein of JAK-STAT signaling pathway, may also plays critical regulatory roles in cancer initiation and progression. SOCS3 promoter hypermethylation has often been identified in human cancers; however, the precise role of SOCS3 in bladder cancer is unclear. METHODS: The methylation status of the SOCS3 was analyzed in an age (±5 years) and sex-matched case-control study, including 112 bladder cancer cases and 118 normal controls, using the MassARRAY EpiTYPER system. RESULTS: Methylation rate of JAK2, SOCS3 and STAT3 gene were shown to vary among different CpG island. The methylation rate of SOCS3 gene was also much higher in BCa than in normal control participants, but the methylation rate of JAK2, STAT3 gene weren't different in Bca and normal control participants. CONCLUSIONS: Our study demonstrates that promoter hypermethylation of SOCS3 gene is associated with BCa and thus, may serve as an independent prognostic biomarker.

Veratric Acid Inhibits LPS-Induced IL-6 and IL-8 Production in Human Gingival Fibroblasts.

Veratric acid, one of the major benzoic acid isolated from vegetables and fruits, has been reported to have anti-inflammatory activity. The purpose of this study was to investigate the anti-inflammatory effects of veratric acid on lipopolysaccharide (LPS)-induced inflammatory response in human gingival fibroblasts (HGFs). HGFs were pretreated with veratric acid 1 h before LPS stimulation. The productions of IL-6 and IL-8 were detected by ELISA. The expression of iNOS, COX-2, PI3K, AKT, and NF-κB were detected by western blotting. The results showed that veratric acid inhibited LPS-induced IL-6 and IL-8 production, as well as iNOS and COX-2 expression. Veratric acid also inhibited LPS-induced NF-κB activation. In addition, veratric acid was found to suppress LPS-induced PI3K and AKT phosphorylation. In conclusion, the anti-inflammatory mechanism of veratric acid is due to its ability to inhibit PI3K/Akt/NF-κB signaling pathway.

Posterior dislocation of the hip with bilateral femoral fractures: An unusual combination.

Posterior hip dislocation with concomitant femoral fracture is very rare. Here, we report a rare case of a 43-year-old man who was injured in a car accident. The patient sustained right posterior hip dislocation with concomitant right acetabular transverse and posterior wall fracture, ipsilateral femoral shaft fracture, and contralateral proximal femoral fracture (AO type 31-A3). Closed reduction of the hip was attempted, but failed. The acetabular fracture and posterior hip dislocation were reduced and acetabular fracture was fixed using plates through the Kocher-Langenbeck approach. The ipsilateral femoral fracture was treated with closed reduction and intramedullary nailing. The contralateral femoral fracture was treated with closed reduction and Gamma 3 nailing. Postoperative X-rays revealed reduction of the fractures. The patient achieved bone union and recovered function of the hip 4 months after surgery.

A novel technique to prevent guide wire related complications while inserting the 4.0 mm cannulated screws.

BACKGROUND: Cannulated screws (4.0 mm) provide inter-fragmentary compression and stability to fractures. A guide wire is used to define the screw trajectory and hold the fracture fragment while the screw is being inserted. The cannulated shaft typically accommodates a 1.25 mm guide pin. Since the guide pin is very slender and undergoes elastic deformation during insertion, there is a high probability of pin breakage. METHODS: The authors have devised a new way to place the 4.0 mm cannulated screws in a manner that prevents the intraoperative complication of guide wire breakage. For this technique, predrilling was achieved using a 2.0 mm K-wire which was subsequently replaced with a 1.25 mm guide pin under the protection of sleeve. 4.0 mm cannulated screws were then inserted into a defined trajectory over the guide pin. RESULTS: Using the technique, over 20 patients were managed in our department over a period of two years without any complications. CONCLUSION: We have observed that patients treated with this method experience short operation time, combined with good clinical outcome and we recommend its use in cases where cannulated screw use is warranted.

KISS1 methylation and expression as predictors of disease progression in colorectal cancer patients.

AIM: To examine the effect of aberrant methylation of the KISS1 promoter on the development of colorectal cancer (CRC) and to investigate reversing aberrant methylation of the KISS1 promoter as a potential therapeutic target. METHODS: KISS1 promoter methylation, mRNA expression and protein expression were detected by methylation-specific polymerase chain reaction (PCR), real-time quantitative PCR and Western blotting, respectively, in 126 CRC tissues and 142 normal colorectal tissues. Human CRC cells with KISS1 promoter hypermethylation and poor KISS1 expression were treated in vitro with 5-aza-2'-deoxycytidine (5-Aza-CdR). After treatment, KISS1 promoter methylation, KISS1 mRNA and protein expression and cell migration and invasion were evaluated. RESULTS: Hypermethylation of KISS1 occurred frequently in CRC samples (83.1%, 105/126), but was infrequent in normal colorectal tissues (6.34%, 9/142). Moreover, KISS1 methylation was associated with tumor differentiation, the depth of invasion, lymph node metastasis and distant metastasis (P < 0.001). KISS1 methylation was also associated with low KISS1 expression (P < 0.001). Furthermore, we observed re-expression of the KISS1 gene and decreased cell migration after 5-Aza-CdR treatment in a CRC cell line. CONCLUSION: These data suggest that KISS1 is down-regulated in cancer tissues via promoter hypermethylation and therefore may represent a candidate target for treating metastatic CRC.

Blending of soybean oil with selected vegetable oils: impact on oxidative stability and radical scavenging activity.

BACKGROUND: Soybean oil may protect against cancer of the breast and prostate. It may also exert beneficial influence in combination with other oils. Here, blends (20%, v/v) of sea buckthorn oil (SEBO), camellia oil (CAO), rice bran oil (RBO), sesame oil (SEO) and peanut oil (PEO) with soybean oil (SBO) were formulated. MATERIALS AND METHODS: Oxidative stability (OS) and radical scavenging activity (RSA) of SBO and blends stored under oxidative conditions (60°C) for 24 days were studied. By blending with different kinds oils, levels of polyunsaturated fatty acids (PUFA) decreased, while monounsaturated fatty acid (MUFA) content increased. Progression of oxidation was followed by measuring peroxide value (PV), p-anisidine (PAV), conjugated dienes (CD) and conjugated trienes (CT). RESULTS: Inverse relationships were noted between PV and OS at termination of storage. Levels of CD and CT in SBO, and blends, increased with increase in time. The impact of SEO as additives on SBO oxidation was the strongest followed by RBO, CAO, SEBO and PNO. CONCLUSIONS: Oxidative stability of oil blends was better than SBO, most likely as a consequence of changes in fatty acids and tocopherols' profile, and minor bioactive lipids found in selected oils. The results suggest that these oil blends could contribute as sources of important antioxidant related to the prevention of chronic diseases associated to oxidative stress, such as in cancer and coronary artery disease.

Intrinsic karyotype stability and gene copy number variations may have laid the foundation for tetraploid wheat formation.

Polyploidy or whole-genome duplication is recurrent in plant evolution, yet only a small fraction of whole-genome duplications has led to successful speciation. A major challenge in the establishment of nascent polyploids is sustained karyotype instability, which compromises fitness. The three putative diploid progenitors of bread wheat, with AA, SS (S ∼ B), and DD genomes occurred sympatrically, and their cross-fertilization in different combinations may have resulted in fertile allotetraploids with various genomic constitutions. However, only SSAA or closely related genome combinations have led to the speciation of tetraploid wheats like Triticum turgidum and Triticum timopheevii. We analyzed early generations of four newly synthesized allotetraploid wheats with genome compositions S(sh)S(sh)A(m)A(m), S(l)S(l)AA, S(b)S(b)DD, and AADD by combined fluorescence and genomic in situ hybridization-based karyotyping. Results of karyotype analyses showed that although S(sh)S(sh)A(m)A(m) and S(l)S(l)AA are characterized by immediate and persistent karyotype stability, massive aneuploidy and extensive chromosome restructuring are associated with S(b)S(b)DD and AADD in which parental subgenomes showed markedly different propensities for chromosome gain/loss and rearrangements. Although compensating aneuploidy and reciprocal translocation between homeologs prevailed, reproductive fitness was substantially compromised due to chromosome instability. Strikingly, localized genomic changes in repetitive DNA and copy-number variations in gene homologs occurred in both chromosome stable lines, S(sh)S(sh)A(m)A(m) and S(l)S(l)AA. Our data demonstrated that immediate and persistent karyotype stability is intrinsic to newly formed allotetraploid wheat with genome combinations analogous to natural tetraploid wheats. This property, coupled with rapid gene copy-number variations, may have laid the foundation of tetraploid wheat establishment.