A novel method for gas mixing and distribution in multi-chamber embryo incubators.

BACKGROUND: High-quality control of the gas environment in incubators is crucial for in vitro embryo development, which requires high accuracy, fast recovery, and low gas consumption. OBJECTIVE: In this study, we propose a novel gas mixing and distribution system and method as an alternative solution for multi-chamber embryo incubators. METHODS: The system-based embryo incubator enables a controllable gas circulation process and a quantitative supply of CO2 and N2. To determine the optimal parameters for the mixing time and flow rate of the circulated gases, we conducted contrast experiments on the system-based incubator. To evaluate the performance of the gas system in the incubator, we conducted tests under four different initial conditions, simulating various practical application scenarios. Furthermore, we performed a mouse embryo assay to assess the system's effectiveness. RESULTS: The results show that the system achieved a gas concentration accuracy of ± 0.2% (volume fraction) after stabilization, a minimum recovery time of 5 minutes, an average consumption of 8.9 L/d for N2 and 0.83 L/d for CO2 during routine operation, and a blastocyst rate exceeding 90% observed after 96 hours of culture in the incubator. CONCLUSION: The system and method demonstrate a significant advantage in terms of low gas consumption compared to existing incubators, while still maintaining high accuracy and fast recovery.

Characteristics, predictors and outcomes of new-onset QT prolongation in sepsis: a multicenter retrospective study.

BACKGROUND: Sepsis-induced myocardial injury is a serious complication of sepsis. QT prolongation is a proarrhythmic state which reflects myocardial injury in a group of heterogeneous disorders. However, the study on the clinical value of QT prolongation in sepsis is limited. METHODS: We aimed to investigate the clinical characteristics and predictors of new-onset QT prolongation in sepsis and its impact on the outcome in a multicenter retrospective cohort study. Electrocardiographic and clinical data were collected from patients with sepsis from the wards and intensive care units of four centers after exclusion of QT-influencing medications and electrolyte abnormalities. Clinical outcomes were compared between patients with and without QT prolongation (QTc > 450 ms). Multivariate analysis was performed to ascertain whether QT prolongation was an independent predictor for 30-day mortality. The factors predicting QT prolongation in sepsis were also analyzed. RESULTS: New-onset QT prolongation occurred in 235/1024 (22.9%) patients. The majority demonstrated similar pattern as type 1 long QT syndrome. Patients with QT prolongation had a higher 30-day in-hospital mortality (P < 0.001), which was also associated with increased tachyarrhythmias including paroxysmal atrial fibrillation or tachycardia (P < 0.001) and ventricular arrhythmia (P < 0.001) during hospitalization. QT prolongation independently predicted 30-day mortality (P = 0.044) after multivariate analysis. History of coronary artery disease (P = 0.001), septic shock (P = 0.008), acute respiratory (P < 0.001), heart (P = 0.021) and renal dysfunction (P = 0.013) were independent predictors of QT prolongation in sepsis. CONCLUSIONS: New-onset QT prolongation in sepsis was associated with increased mortality as well as atrial and ventricular arrhythmias, which was predicted by disease severity and organ dysfunction.

Acute epicardial pulmonary vein reconnection: Nondurable transmural lesion or late manifestation of conduction through intercaval bundle.

INTRODUCTION: Acute pulmonary vein reconnection (PVR) via epicardial fibers can be found during observation period after PV isolation, the characteristics and related factors have not been fully studied. We aimed to investigate the prevalence, locations, electrogram characteristics, and ablation parameters related to acute epicardial pulmonary vein reconnection (AEPVR). METHODS: Acute PVR was monitored during observation period after PV isolation. AEPVRs were mapped and distinguished from endocardial conduction gaps. The clinical, electrophysiological characteristics and lesion set parameters were compared between patients with and without PVR. They were also compared among AEPVR, gap-related reconnection, and epicardial PVR in repeat procedures. RESULTS: A total of 56.1% acute PVR were AEPVR, which required a longer waiting period (p < .001) than endocardial gap. The majority of AEPVR were connections from the posterior PV carina to the left atrial posterior wall, followed by late manifestation of intercaval bundle conduction from the right anterior carina to right atrium. AEPVR was similar to epicardial PVR in redo procedures in distribution and electrogram characteristics. Smaller atrium (p < .001), lower impedance drop (p = .039), and ablation index (p = .028) on the posterior wall were independently associated with presence of AEPVR, while lower interlesion distance (p = .043) was the only predictor for AEPVR in acute PVR. An integrated model containing multiple lesion set parameters had the highest predictive ability for AEPVR in receiver operating characteristics analysis. CONCLUSIONS: Epicardial reconduction accounted for the majority of acute PVR. AEPVR was associated with anatomic characteristics and multiple ablation-related parameters, which could be explained by nondurable transmural lesion or late manifestation of conduction through intercaval bundle.

Corrigendum: Single-cell RNA sequencing reveals the difference in human normal and degenerative nucleus pulposus tissue profiles and cellular interactions.

[This corrects the article DOI: 10.3389/fcell.2022.910626.].

Potential serum metabolites and long-chain noncoding RNA biomarkers for endometrial cancer tissue.

BACKGROUND: Endometrial carcinoma (EC) is one of the most common tumors in the female reproductive system. There are nearly 200 000 new cases every year. It is the third most common gynecological malignant tumor leading to female death. The incidence rate is closely related to lifestyle, and the incidence rate varies in different regions. The incidence rate of EC is ranking the first in the female reproductive system cancer just second only to breast, lung, and colorectal cancer in North America and Europe and the incidence rate of EC is only second, followed by breast cancer and cervical cancer in China. PURPOSE: The potential metabolic markers of endometrial cancer were screened by liquid chromatograph mass spectrometer (LC-MS), and the tissues of patients with hysteromyoma and endometrial cancer were sequenced to explore the relationship between the disease and change in the content of long-chain noncoding RNA (lncRNA). METHODS: Serum and tissue samples were collected from patients with endometrial dysplasia, endometrial cancer stage I, and endometrial cancer stage III. The metabolites in all serum samples were extracted, and the metabolites in all samples were detected by LC-MS/MS technology. The Pareto-scaling method was used for normalization, and the MetaboAnalyst 4.0 software was used for different analyses. The T test between groups showed that p ≤ 0.05 was regarded as the metabolite with a difference. Further, the function of differential metabolites was determined by metabolite function enrichment and co-expression analysis. Meanwhile, the differentially expressed lncRNA was detected by Illumina second-generation high-throughput sequencing technology, and the expression was analyzed by DEGseq software. Different lncRNA were screened according to p < 0.05. LncRNA with significant differences were screened by p < 0.01, q < 0.001, fold change ≥2, and false discovery rate (FDR) ≤0.001. RESULTS: Through synthesis of T test, cluster heatmap, and ROC curve analysis, five biomarkers with potential diagnostic ability were obtained, including 2,3-Pyridinedicarboxylic acid (area under the curve (AUC) = 0.69), Hematommic acid, ethyl ester (AUC = 0.69), Maltitol (AUC = 0.69), 13(S)-HODE (AUC = 0.88), and D-Mannitol (AUC = 0.69) had potential diagnostic ability between EC phase I versus EC phase III. At the same time, lncRNA sequencing results showed that when endometrial atypical hyperplasia continued to change, including LINC00511, PVT1, and IQCH-AS1 (downregulated), and only changed significantly in the endometrial dysplasia group, including MALAT1, CARMN (downregulated) and LINC00648, BISPR, LINC01534, and LINC00930 (upregulated). Moreover, both differential metabolites and differential lncRNA were annotated to the lipid metabolism pathway, suggesting that this pathway played an important role in the occurrence and development of endometrial carcinoma. CONCLUSIONS: It can combine the results of metabolomics and lncRNA sequencing to assist in the early diagnosis of endometrial precancerous lesions and endometrial cancer patients, to enhance the sensitivity and specificity of diagnosis, which has a certain clinical application prospect.

Long non-coding RNA ERVK13-1 aggravates osteosarcoma through the involvement of microRNA-873-5p/KLF5 axis.

OBJECTIVE: To explore the effect and mechanism of long noncoding RNA ERVK13-1 on osteosarcoma (OS) cell development by regulation of miR-873-5p/KLF5 axis. METHODS: The expression of ERVK13-1 in the collected tissue was detected by RT-qPCR, and then the relationship between ERVK13-1 expression and clinical characteristics of OS patients was analyzed. After OS cell lines were transfected with miR-873-5p inhibitor, si-ERVK13-1, si-KLF5 or their negative controls, the expression of ERVK13-1, miR-873-5p, and KLF5 in OS cell lines were measured, followed by determination of their effects on cell proliferation, migration, and invasion abilities. Moreover, the binding relationships of ERVK13-1 and miR-873-5p, as well as miR-873-5p and KLF5, were verified by the dual-luciferase reporter gene assay. RESULTS: Highly expressed ERVK13-1 was found in OS tissues, which was closely related to tumor size, tumor node metastasis, and distant metastasis. The overall survival of OS patients with high expression of ERVK13-1 was poorer than those with low expression of ERVK13-1. Elevated ERVK13-1 and KLF5 but suppressed miR-873-5p was observed in the OS cell lines U2OS and MG63. Transfection with miR-873-5p inhibitor enhanced the malignant potentials of OS cells, and transfection with si-ERVK13-1 or si-KLF5 reduced these abilities of OS cells. ERVK13-1 bound to miR-873-5p and KLF5 was a target gene of miR-873-5p. CONCLUSION: The ERVK13-1/miR-873-5p/KLF5 axis confers vital effect on the occurrence and progression of OS, thus providing possible guidance for the clinical treatment of OS.

In situ-prepared poly(ether-ester)-based gel polymer electrolytes for high-performance lithium metal batteries.

Batteries based on an in situ-polymerized amorphous cross-linked poly(ether-ester) GPE exhibit high performance, which is attributed to the synergistic effect of the ether and ester segments of the GPE and the in situ-generated polymer electrolytes.

Corrigendum: Single-cell RNA sequencing reveals the difference in human normal and degenerative nucleus pulposus tissue profiles and cellular interactions.

[This corrects the article DOI: 10.3389/fcell.2022.910626.].

Single-Cell RNA Sequencing Reveals the Difference in Human Normal and Degenerative Nucleus Pulposus Tissue Profiles and Cellular Interactions.

Background: The nucleus pulposus is a constituent structure of the human intervertebral disc, and its degeneration can cause intervertebral disc degeneration (IDD). However, the cellular and molecular mechanisms involved remain elusive. Methods: Through bioinformatics analysis, the single-cell transcriptome sequencing expression profiles of human normal nucleus pulposus (NNP) cells and human degenerative nucleus pulposus (DNP) cells were compared to clarify the transcriptome differential expression profiles of human NNP and DNP. The single-cell sequencing results of the two samples were analyzed using bioinformatics methods to compare the differences in histiocytosis between human NNP and DNP, map the histiocytes of NNP and DNP, perform cell differentiation trajectories for the cell populations of interest and predict cell function, and explore their heterogeneity by pathway analysis and Gene Ontology analysis. Results: Nine cell types were identified, which were chondrocyte 1, chondrocyte 2, chondrocyte 3, chondrocyte 4, chondrocyte 5, endothelial, macrophage, neutrophil, and T cells. Analysis of the proportion of chondrocytes in different tissues revealed that chondrocyte 1 accounted for a higher proportion of NNP cells and highly expressed COL2A1 compared with DNP cells; chondrocyte 2, chondrocyte 3, chondrocyte 4, and chondrocyte 5 accounted for a higher proportion of DNP cells compared with NNP cells. Among them, chondrocyte 2 was an inhibitory calcified chondrocyte with high expression of MGP, chondrocytes 3 were fibrochondrocytes with high expression of COL1A1, chondrocytes 4 were chondrocytes that highly express pain inflammatory genes such as PTGES, and chondrocytes 5 were calcified chondrocytes with high expression of FN1 (chondrocytes 4 and chondrocytes 5 were found for the first time in a study of single-cell transcriptome sequencing of disc tissue). Cell trajectory analysis revealed that chondrocyte 1 was at the beginning of the trajectory and chondrocyte 3 was at the end of the trajectory, while chondrocyte 5 appeared first in the trajectory relative to chondrocyte 2 and chondrocyte 4. Conclusion: After functional identification of the specifically expressed genes in five chondrocytes, it was found that chondrocyte 1 was a chondrocyte with high expression of COL2A1, COL9A2, COL11A2, and CHRDL2 in a high proportion of NNP cells, and chondrocyte 3 was a fibrochondrocyte with high expression of COL1A1, COL6A3, COL1A2, COL3A1, AQP1, and COL15A1 in an increased proportion during nucleus pulposus cell degeneration. Through cell trajectory analysis, it was found that chondrocytes 5 specifically expressing FN1, SESN2, and GDF15 may be the key cells leading to degeneration of nucleus pulposus cells. Chondrocytes 2 expressing MGP, MT1G, and GPX3 may play a role in reversing calcification and degeneration, and chondrocytes 4 expressing PTGES, TREM1, and TIMP1 may play a role in disc degeneration pain and inflammation.

Expression of Matrix Metalloprotein 13 in Injury Model of Articular Chondrocyte in Rabbits and Analysis of Nano-Drug-Loading System.

This study aimed to analyze the application of a responsive nano-drug-loading system in injury model of articular chondrocyte in rabbits, as well as its effect on expression of matrix metalloprotein 13 (MMP13). The nanoprecipitation method was adopted to prepare camptothecin (CPT)-loaded poly ethylene glycol (PEG)-Poly caprolactone (PCL) and PEG-PCL nanoparticles without CPT. Afterward, the above mentioned nano-drug-loaded system was used to treat an in vitro scratch model of articular chondrocytes. According to different treatment plans, they were divided into groups: G0 (administered CPT-PEG-PCL nanomedicine), G1 (administered PEG-PCL drug), G2 (saline control), and G3 (healthy control). Results showed that the drug-loading capacity and efficiency of CPT-PEG-PCL was higher than that of PEG-PCL. The levels of type II collagen and hyaluronic acid in G0 was higher than that in G1 and G2. The levels of type II collagen and hyaluronic acid in G0 were not obviously different from those in G3. The level of MMP13 in G0 was lower than that in G1 and G2 and the level of tissue inhibitor of metalloproteinases 1 (TIMP1) in G0 was higher than that in G1 and G2. The proliferation activity of cells in G0 was higher than that in G1 and G2, but there was no obvious difference when compared with G3. In conclusion, CPT-PEG-PCL has stronger long-term circulation capacity and drug-loading efficiency. It can effectively up-regulate the levels of type II collagen, hyaluronic acid, and TIMP1, as well as reduce the synthesis and secretion of MMP13 and promote the repair of articular cartilage damage.

Effect of Different Reduction Intramedullary Nails on Spiral Fracture of Middle and Lower Tibia.

Objective: To compare the efficacy of different reduction and intramedullary nailing in the treatment of spiral fracture of middle and lower tibia. Method: A total of 96 patients with spiral fractures of middle and lower tibia treated with intramedullary nails were retrospectively analyzed. The patients were divided into closed functional reduction group, open anatomical reduction group, and closed anatomical reduction group according to different treatment methods. The operation time, intraoperative blood loss, intraoperative fluoroscopy times, fracture healing time, fracture nonunion, wound complications, and healing conditions of the three groups were compared. Results: The operation time and intraoperative fluoroscopy times of patients in the closed anatomical reduction group were significantly increased compared with those in the closed functional reduction group, while the fracture healing time was significantly reduced. However, patients in the open reduction group had significantly more intraoperative blood loss than those in the closed reduction group. The mean follow-up duration of patients was 15.81 ± 3.25 months. Open anatomical reduction was found to have a higher complication rate during follow-up. Specifically, a total of 3 cases recovered after 2 times of surgical treatment. 6 cases showed a small gap at the fracture end which did not affect the function. Conclusion: In the treatment of middle and lower spiral fracture of tibia, closed anatomical reduction and intramedullary nail internal fixation have shorter fracture healing time, less blood loss, and fewer complications, which can act as the first surgical choice. However, open reduction and intramedullary nailing have a high complication rate, which is not recommended.

CircRNA PVT1 promotes proliferation and chemoresistance of osteosarcoma cells via the miR-24-3p/KLF8 axis.

OBJECTIVE: To investigate the regulatory effect and mechanism of circular RNA PVT1 (circPVT1) in proliferation and chemoresistance of osteosarcoma (OS) cells. METHODS: The expression of circPVT1 in human OS and adjacent normal tissues was detected. The correlation between circPVT1 expression and clinical features of OS was analyzed. The expressions of circPVT1 and miR-24-3p in OS cells resistant to cisplatin, doxorubicin or methotrexate and parental OS cells were detected after cell transfection. CCK-8 and colony formation assay assessed the viability and proliferative ability of OS cells. qRT-PCR and Western blotting measured the expression of KLF8. Dual-luciferase reporter and RNA pull-down assays verified the targeting relationships of circPVT1/miR-24-3p and miR-24-3p/KLF8. RESULTS: CircPVT1 was over-expressed in OS tissues and cells, and correlated with clinical features of OS. Over-expressed circPVT1 reduced the survival of OS patients. CircPVT1 was up-regulated in chemoresistant OS cells compared to their parental cells. CircPVT1 inhibition suppressed the proliferation and chemoresistance of OS cells. MiR-24-3p was under-expressed in OS cells and further down-regulated in chemoresistant cells. CircPVT1 could bind and down-regulate miR-24-3p. MiR-24-3p overexpression inhibited the proliferation and chemoresistance of OS cells. KLF8 was over-expressed in OS cells and further up-regulated in chemoresistant cells. MiR-24-3p negatively regulated the expression of KLF8. CONCLUSION: CircPVT1 mediates proliferation and chemoresistance of OS cells via the miR-24-3p/KLF8 axis. The findings may provide guidance for clinical treatment of OS.

Single versus dual elastic nails for closed reduction and antegrade intramedullary nailing of displaced fifth metacarpal neck fractures.

Closed reduction and internal fixation with antegrade intramedullary nails is a feasible and effective treatment for displaced fifth metacarpal neck fractures (FMNFs). The present study aimed to compare clinical and radiological outcomes in patients with displaced FMNFs after treatment with single or dual antegrade elastic intramedullary nails (AEIMNs). Thirty-three patients were treated with a single 2.0 mm AEIMN and 34 patients were treated with two 1.5 mm AEIMNs. Clinical and radiological outcomes included grip strength, active range of motion (ROM), active flexion and extension of the fifth metacarpophalangeal (MCP) joint, dorsal angulation loss, and metacarpal shortening of the fifth metacarpal at 12 months after treatment. No significant difference was observed between the two groups with respect to grip strength, ROM or flexion of the fifth MCP joint. The average values of dorsal angulation loss, metacarpal shortening, and extension of the fifth MCP joint of the dual nails group were better than those of the single nail group (dorsal angulation loss, 2.79 ± 1.93° vs. 4.05 ± 1.59°, P = 0.009; metacarpal shortening, 1.66 ± 0.80 mm vs. 2.12 ± 0.88 mm, P = 0.028; extension of the fifth MCP joint, 7.71 ± 4.43° vs. 4.82 ± 4.09°, P = 0.012). In conclusion, dual AEIMNs fixation provided better MCP extension and radiological outcomes than single AEIMN fixation.

A new low-profile anatomic locking plate for fixation of comminuted, displaced greater tuberosity fractures of the proximal humerus.

BACKGROUND: Although various implants exist for the fixation of isolated greater tuberosity fractures, few implants are specifically designed for such fractures. The purpose of this study was to investigate the clinical and radiologic outcomes of open reduction-internal fixation with a low-profile anatomic locking plate for comminuted greater tuberosity fractures of the proximal humerus. METHODS: From November 2012 to February 2018, 24 patients with displaced and comminuted isolated greater tuberosity fractures were treated with the new low-profile anatomic locking plate. To determine clinical outcomes, we evaluated active range of motion; the visual analog scale pain score; the Constant-Murley score; the Disabilities of the Arm, Shoulder and Hand score; radiographs; and complications. RESULTS: In all cases, a mean follow-up period of 29.3 months (range, 18-48 months) was completed. All patients achieved bone union with a mean healing time of 11.3 weeks (range, 8-16 weeks). The mean Constant-Murley score was 91.1 points (range, 69-100 points), with a rate of good to excellent results of 95.8%. The average Disabilities of the Arm, Shoulder and Hand score was 9.9 points (range, 2-25 points), and the mean visual analog scale pain score was 1.1 points (range, 0-4 points). Mean active forward flexion, abduction, external rotation, and internal rotation (level) were 157°, 152°, and 40°, and T11, respectively. Postoperatively, 1 patient had persistent shoulder stiffness, and 1 patient had recurrence of shoulder dislocation because of a falling injury during badminton. No serious complications such as subacromial impingement, malunion, nonunion, loss of reduction, or implant failure occurred. CONCLUSIONS: The new low-profile anatomic locking plate was useful for the treatment of comminuted isolated greater tuberosity fractures as it provided reliable stability and satisfactory radiographic and functional results. The described technique is a simple and effective method and provides a new reliable option for the treatment of isolated greater tuberosity fractures.

miR-129-3p alleviates chondrocyte apoptosis in knee joint fracture-induced osteoarthritis through CPEB1.

BACKGROUND: Osteoarthritis (OA), a refractory disease, is one of the leading contributors for disability worldwide. Since chondrocyte is the only resident cell in cartilage, this study aims to explore the roles of miR-129-3p and CPEB1 in chondrocyte apoptosis in knee joint fracture-induced OA. METHODS: Cartilage was collected from 20 OA patients who underwent total knee replacement (OA group) and 20 patients with knee contusion (normal group). Then, miR-129-3p and CPEB1 levels in the cartilage were quantified by qRT-PCR. Primary rat chondrocytes in the knee were isolated and identified by toluidine blue staining and immunofluorescent staining of type II collagen. OA cellular models were induced by TNF-α treatment, in which miR-129-3p and CPEB1 expressions were assessed. Subsequently, cell viability, apoptosis, and the expression levels of apoptotic protein and caspase-3 were measured. Dual luciferase reporter assay identified the interaction between miR-129-3p and CPEB1. RESULTS: Patients in the OA group had decreased miR-129-3p expression and increased CPEB1 expression than those in the normal group. TNF-α treatment successfully induced the OA cellular model. Downregulated miR-129-3p and upregulated CPEB1 expressions were found in OA-treated chondrocytes. miR-129-3p overexpression or CPEB1 knockdown improved chondrocyte viability and attenuated apoptosis, and vice versa. miR-129-3p negatively regulated CPEB1, thus ameliorating apoptosis and enhancing cell viability. CONCLUSION: miR-129-3p negatively targeted CPEB1 to facilitate chondrocyte viability and hamper apoptosis.

Linical efficacy of percutaneous endoscopic lumbar discectomy for the treatment of lumbar spinal stenosis in elderly patients: a retrospective study.

BACKGROUND: Although percutaneous endoscopic lumbar discectomy (PELD) is increasingly being used to treat lumbar degenerative disease, the treatment of elderly patients with lumbar spinal stenosis (LSS) involves considerable uncertainty. The purpose of this study was to evaluate the safety and effectiveness of PELD for the treatment of LSS in elderly patients aged 65 years or older. METHODS: In this retrospective review, 136 patients aged 65 years or older who underwent PELD to treat LSS were evaluated. The patients were divided into two groups, group A (ages 65-74) and group B (age ≥ 75), and perioperative data were analyzed. The Japanese Orthopaedic Association (JOA) score, visual analog scale (VAS) score, and MacNab classification were used to evaluate postoperative clinical efficacy. RESULTS: All patients successfully underwent the operation with satisfactory treatment outcomes. Compared to preoperative scores, the self-reported scores or pain while performing daily activities were significantly improved in both treatment groups (P < 0.05). No statistically significant between-group differences were observed in operation time, intraoperative blood loss, postoperative bed rest, and postoperative hospital stay (P > 0.05). The overall postoperative complication rate was similar between the two groups. Moreover, no statistically significant differences in VAS-back pain scores, VAS-leg pain scores, JOA scores, and MacNab classification were found between the groups at the 3-month and 1.5-year follow-up examinations (P > 0.05). CONCLUSION: PELD is safe and effective for the treatment of LSS in elderly patients. Age is not a contraindication for decompressive lumbar spine surgery. PELD has advantages such as reduced trauma, fewer anesthesia-related complications, and a fast postoperative recovery. Elderly patients should be considered good candidates for lumbar decompression surgery using minimally invasive techniques.

The effect of NaOH pretreatment on coal structure and biomethane production.

Biogenic CBM is an important component of detected CBM, which is formed by coal biodegradation and can be regenerated by anaerobic microorganisms. One of the rate-limiting factors for microbial degradation is the bioavailability of coal molecules, especially for anthracite which is more condense and has higher aromaticity compared with low-rank coal. In this paper, NaOH solution with different concentrations and treating time was employed to pretreat anthracite from Qinshui Basin to alter the coal structure and facilitate the biodegradation. The results showed that the optimal pretreatment conditions were 1.5 M NaOH treating for 12 h, under which the biomethane production was increased by 17.65% compared with untreated coal. The results of FTIR and XRD showed that NaOH pretreatment mainly reduced the multi-substituted aromatics, increased the C-O in alcohols and aromatic ethers and the branching degree of aliphatic chain, and decreased the aromatic ring structure, resulting in the improvement of coal bioavailability and enhancement of biomethane yield. And some organics with potential to generate methane were released to filtrate as revealed by GC-MS. Our results suggested that NaOH was an effective solution for pretreating coal to enhance biogenic methane production, and anthracite after treating with NaOH could be the better substrate for methanogenesis.

Clinical significance of tumor miR-21, miR-221, miR-143, and miR-106a as biomarkers in patients with osteosarcoma.

OBJECTIVE: To investigate the expression of miR-21, miR-221, miR-143, and miR-106a in patients' osteosarcoma samples, and to explore the correlation between these microRNAs (miRNAs) and the clinical stage of osteosarcoma. METHODS: RNA was extracted from tumor and tumor-adjacent normal bone tissues from 94 patients with osteosarcoma. RNA reverse-transcription was carried out using an miRNA reverse transcription kit. The levels of miR-21, miR-221, miR-143, and miR-106a in osteosarcoma and normal bone tissues were analyzed by real-time polymerase chain reaction using SYBR Premix Ex Taq™II. RESULTS: The expression levels of miR-21, mirR-221, and miR-106a were significantly higher in 90.42%, 84.04%, and 92.55 % of the osteosarcoma samples compared to the adjacent normal tissues (P<0.05), respectively. While the expression of miR-143 was significantly lower compared to the adjacent normal tissues (P <0.05). Moreover, the expression levels of miR-21 and miR-221 were positively correlated with the Enneking clinical stage and the presence of lung metastasis (P <0.05), while the expression levels of miR-143 and miR-106a showed a significant inverse and direct correlation respectively, with the tumor grade. CONCLUSIONS: The upregulation of miR-21, miR-221, and miR-106a, as well as the down-regulation of miR-143 were correlated with the pathological stage, tumor grade, and lung metastasis. Therefore, the levels of these miRNAs can serve as potential biomarkers for the early diagnosis of osteosarcoma, and can be used as potential therapeutic targets.

Screening, Identification, and Characterization of an Affinity Peptide Specific to MT1-MMP and Its Application in Tumor Imaging.

Membrane type-1 matrix metalloproteinase (MT1-MMP) plays a crucial role in many physiological and pathological processes, especially in tumor invasion and metastasis. Bioimaging of this key molecule may find wide usage in various applications. MT-loop is a unique sequence of MT1-MMP and locates in the surface of the protein. In our previous studies, AF7p, an affinity peptide that targeting the MT-loop domain of MT1-MMP, was identified by screening a phage display (Ph.D.) peptide library. However, the target of AF7p is a synthetic sequence which lacked native conformation of the MT-loop region; thus, the binding affinity and specificity in reality may not be optimal. In this study, we considered the 3-dimensional (3-D) conformation of the MT-loop area in the MT1-MMP molecule and designed a novel strategy to screen the Ph.D. peptide library. The peptide we obtained showed a better binding affinity to WT-MT1-MMP than AF7p as observed through enzyme-linked immunosorbent assay (ELISA) and biolayer interferometry (BLI). The new peptide labeled and attached MT1-MMP expression cell lines HT1080 and did not show any toxicity to cells. Furthermore, for in vivo imaging, HT1080 tumor-bearing mice with higher MT1-MMP expression accumulated more Cy5.5-HS7 than mice with MT1-MMP low-expression cell lines A549 at tumor sites, and the half-life of HS7 was longer than that of AF7p, as confirmed by ex vivo imaging of the main organs. These results suggest the feasibility of using the subtraction biopanning strategy to screen the affinity peptide targeting MT-loop regions and HS7 is a superior probe for noninvasively imaging MT1-MMP expression in MT1-MMP-positive tumor models. It provides impetus for further studies to use HS7 in early diagnosis of tumors and in peptide-mediated drugs.

[Mid-term effectiveness of Coflex interspinous dynamic internal fixation combined with spinal fusion for lumbar disc degeneration].

Objective: To evaluate the effectiveness of Coflex interspinous dynamic internal fixation combined with spinal fusion for lumbar disc degeneration. Methods: The clinical data of 39 patients with two-level lumbar disc degeneration who met the selection criteria between June 2010 and December 2011 was retrospectively analyzed. They were divided into group A (20 cases, simple lumbar decompression and fusion) and group B (19 cases, Coflex interspinous dynamic internal fixation combined with spinal fusion) according to different surgical methods. There was no significant difference in age, gender, disease diagnosis, lesion segment, disease duration, Oswestry disability index (ODI), visual analogue scale (VAS) score, and the intervertebral height, foramen intervertebral height (FIH), and range of motion (ROM) of upper operative segment and adjacent segment between the two groups ( P>0.05). ODI and VAS score were used to evaluate the effectiveness before operation and at last follow-up, and the improvement rates were calculated. The intervertebral height [anterior disc height (ADH), middle disc height (MDH), and posterior disc height (PDH)], FIH, and ROM were measured and compared between the two groups. Results: The operation time and intraoperative blood loss in group A were significantly more than those in group B ( P<0.05), and there was no significant difference in hospitalization time between the two groups ( t=0.992, P=0.328). All patients were followed up; the follow-up time was 33-50 months (mean, 40.5 months) in group A and 39-51 months (mean, 42.6 months) in group B. No complication such as displacement, loosening, or rupture of internal fixator was found in both groups. At last follow-up, ODI and VAS score of the two groups significantly improved when compared with preoperative scores ( P<0.05). At last follow-up, there was no significant difference in ODI, VAS score, and improvement rate of ODI between the two groups ( P>0.05); the improvement rate of VAS score in group B was significantly higher than that in group A ( t=2.245, P=0.031). There was no significant difference in the intervertebral height and FIH of the upper operative segment at last follow-up between the two groups and between preoperation and last follow-up in the two groups ( P>0.05). At last follow-up, the ADH of adjacent segment in group B was significantly higher than that in group A, and MDH, PDH, and FIH were significantly lower than those in group A ( P<0.05). Compared with preoperation, the ADH of adjacent segment in group A decreased and MDH, PDH, and FIH increased at last follow-up ( P<0.05), while all indexes in group B did not change significantly ( P>0.05). The ROM of adjacent segment in group A increased significantly at last follow-up ( t=2.318, P=0.026). There was significant difference in ROM of adjacent segment between the two groups ( P<0.05). Conclusion: The mid-term effectiveness of Coflex interspinous dynamic internal fixation combined with spinal fusion is similar to that of simple decompression fusion. For those patients whose adjacent segments of the responsible segments have degeneration but have no symptoms or mild symptoms, this treatment can slow down the adjacent segment degeneration.