A quantification method of somatic mutations in normal tissues and their accumulation in pediatric patients with chemotherapy.

Somatic mutations are accumulated in normal human tissues with aging and exposure to carcinogens. If we can accurately count any passenger mutations in any single DNA molecule, since their quantity is much larger than driver mutations, we can sensitively detect mutation accumulation in polyclonal normal tissues. Duplex sequencing, which tags both DNA strands in one DNA molecule, enables accurate count of such mutations, but requires a very large number of sequencing reads for each single sample of human-genome size. Here, we reduced the genome size to 1/90 using the BamHI restriction enzyme and established a cost-effective pipeline. The enzymatically cleaved and optimal sequencing (EcoSeq) method was able to count somatic mutations in a single DNA molecule with a sensitivity of as low as 3 × 10-8 per base pair (bp), as assessed by measuring artificially prepared mutations. Taking advantages of EcoSeq, we analyzed normal peripheral blood cells of pediatric sarcoma patients who received chemotherapy (n = 10) and those who did not (n = 10). The former had a mutation frequency of 31.2 ± 13.4 × 10-8 per base pair while the latter had 9.0 ± 4.5 × 10-8 per base pair (P < 0.001). The increase in mutation frequency was confirmed by analysis of the same patients before and after chemotherapy, and increased mutation frequencies persisted 46 to 64 mo after chemotherapy, indicating that the mutation accumulation constitutes a risk of secondary leukemia. EcoSeq has the potential to reveal accumulation of somatic mutations and exposure to environmental factors in any DNA samples and will contribute to cancer risk estimation.

Precancerous nature of intestinal metaplasia with increased chance of conversion and accelerated DNA methylation.

OBJECTIVE: The presence of intestinal metaplasia (IM) is a risk factor for gastric cancer. However, it is still controversial whether IM itself is precancerous or paracancerous. Here, we aimed to explore the precancerous nature of IM by analysing epigenetic alterations. DESIGN: Genome-wide DNA methylation analysis was conducted by EPIC BeadArray using IM crypts isolated by Alcian blue staining. Chromatin immunoprecipitation sequencing for H3K27ac and single-cell assay for transposase-accessible chromatin by sequencing were conducted using IM mucosa. NOS2 was induced using Tet-on gene expression system in normal cells. RESULTS: IM crypts had a methylation profile unique from non-IM crypts, showing extensive DNA hypermethylation in promoter CpG islands, including those of tumour-suppressor genes. Also, the IM-specific methylation profile, namely epigenetic footprint, was present in a fraction of gastric cancers with a higher frequency than expected, and suggested to be associated with good overall survival. IM organoids had remarkably high NOS2 expression, and NOS2 induction in normal cells led to accelerated induction of aberrant DNA methylation, namely epigenetic instability, by increasing DNA methyltransferase activity. IM mucosa showed dynamic enhancer reprogramming, including the regions involved in higher NOS2 expression. NOS2 had open chromatin in IM cells but not in gastric cells, and IM cells had frequent closed chromatin of tumour-suppressor genes, indicating their methylation-silencing. NOS2 expression in IM-derived organoids was upregulated by interleukin-17A, a cytokine secreted by extracellular bacterial infection. CONCLUSIONS: IM cells were considered to have a precancerous nature potentially with an increased chance of converting into cancer cells, and an accelerated DNA methylation induction due to abnormal NOS2 expression.

Regulating the Interfacial Charge Density by Constructing a Novel Zn Anode-Electrolyte Interface for Highly Reversible Zn Anode.

Aqueous zinc-ion batteries (AZIBs) have attracted considerable attention. However, due to the uneven distribution of charge density at Zn anode-electrolyte interface, severe dendrites and corrosion are generated during cycling. In this work, a facile and scalable strategy to address the above-mentioned issues has been proposed through regulating the charge density at Zn anode-electrolyte interface. As a proof of concept, amidinothiourea (ATU) with abundant lone-pair electrons is employed as an interfacial charge modifier for Zn anode-electrolyte interface. The uniform and increased interfacial charge distribution on Zn anode-electrolyte interface has been obtained. Moreover, the unique Zn-bond constructed between N atoms and Zn2+ as well as the hydrogen bonds are formed among ATU and Ac- anion/active H2 O, which promote the migration and desolvation behavior of Zn2+ at anode-electrolyte interface. Accordingly, at a trace concentration of 0.01 mg mL-1 ATU, these features endow Zn anode with a long cycling life (more than 800 h), and a high average Columbic efficiency (99.52 %) for Zn||Cu batteries. When pairing with I2 cathode, the improved cycling ability (5000 cycles) with capacity retention of 77.9 % is achieved. The fundamental understanding on the regulation of charge density at anode-electrolyte interface can facilitate the development of AZIBs.

Research on Classification Algorithm of Silicon Single-Crystal Growth Temperature Gradient Trend Based on Multi-Level Feature Fusion.

In the process of silicon single-crystal preparation, the timely identification and adjustment of abnormal conditions are crucial. Failure to promptly detect and resolve issues may result in a substandard silicon crystal product quality or even crystal pulling failure. Therefore, the early identification of abnormal furnace conditions is essential for ensuring the preparation of perfect silicon single crystals. Additionally, since the thermal field is the fundamental driving force for stable crystal growth and the primary assurance of crystal quality, this paper proposes a silicon single-crystal growth temperature gradient trend classification algorithm based on multi-level feature fusion. The aim is to accurately identify temperature gradient changes during silicon crystal growth, in order to promptly react to early growth failures and ensure the stable growth of high-quality silicon single crystals to meet industrial production requirements. The algorithm first divides the temperature gradient trend into reasonable categories based on expert knowledge and qualitative analysis methods. Then, it fuses the original features of actual production data, shallow features extracted based on statistical information, and deep features extracted through deep learning. During the fusion process, the algorithm considers the impact of different features on the target variable and calculates mutual information based on the difference between information entropy and conditional entropy, ultimately using mutual information for feature weighting. Subsequently, the fused multi-level feature vectors and their corresponding trend labels are input into a Deep Belief Network (DBN) model to capture process dynamics and classify trend changes. Finally, the experimental results demonstrate that the proposed algorithm can effectively predict the changing trend of thermal field temperature gradients. The introduction of this algorithm will help improve the accuracy of fault trend prediction in silicon single-crystal preparation, thereby minimizing product quality issues and production interruptions caused by abnormal conditions.

Clinical and electromyographic signals analysis about the effect of space-adjustment splint on overerupted maxillary molars.

BACKGROUND: Overerupted maxillary molars is common in adults, which can lead to insufficient intermaxillary vertical space ,great difficulty in prosthetic reconstruction ,and cause occlusal interference in movements.To reconstruct occlusal function, it is necessary to prepare enough space for prostheses. The aim of the present study was to evaluate the effect of space-adjustment occlusal splint on overerupted maxillary molars by clinical and electromyographic signals analysis. METHODS: Eighteen patients with overerupted maxillary molars were selected to wear space-adjustment occlusal splint suppressing overerupted maxillary molars for three months. Satisfaction was assessed by 5-point Likert; intermaxillary vertical space and the teeth transportation distance were measured in models; clinical periodontal status were evaluated by periodontal probing depth (PPT) and bleeding index (BI); electromyographic recordings of the masseter and anterior temporal muscles were monitored by Cranio-Mandibular K7 Evaluation System. RESULTS: All the patients were satisfied with the treatment effect (Likert scale ≧ 4). The intermaxillary space in edentulous areas after treatment showed statistically significant increasing when compared with those before treatment. PPT and BI showed no significant difference. No statistically significant differences were found in electromyographic activity of anterior temporal muscles, while a reduction of muscle activity in masseter in the contralateral side were detected in post-treatment evaluations compared with pre-treatment at mandibular rest position. CONCLUSIONS: Space-adjustment occlusal splint is an efficient treatment option on overerupted maxillary molars by intruding the maxillary molar to obtain adequate intermaxillary space for prostheses.

HSD17B4 methylation enhances glucose dependence of BT-474 breast cancer cells and increases lapatinib sensitivity.

PURPOSE: HER2-positive breast cancer has a high chance of achieving pathological complete response when HSD17B4, responsible for peroxisomal ß-oxidation of very long-chain fatty acids (VLCFA) and estradiol, is methylation-silenced. Here, we aimed to identify the underlying molecular mechanism. METHODS: Using a HER2-positive breast cancer cell line, BT-474, control and knock-out (KO) clones were obtained. Metabolic characteristics were analyzed using a Seahorse Flux analyzer. RESULTS: HSD17B4 KO suppressed cellular proliferation, and enhanced sensitivity to lapatinib approximately tenfold. The KO led to accumulation of VLCFA and a decrease of polyunsaturated fatty acids (PUFAs), such as docosahexaenoic acid (DHA) and arachidonic acid. HSD17B4 KO increased Akt phosphorylation, possibly via decreased DHA, and genes involved in oxidative phosphorylation (OxPhos) and electron transport chain (ETC) were upregulated. Increased mitochondrial ATP production in the KO cells was confirmed by extracellular flux analyzer. Increased OxPhos led to severe dependence of the KO cells on pyruvate from glycolysis. Suppression of glycolysis by lapatinib led to severe delayed suppression of OxPhos in KO cells. CONCLUSION: HSD17B4 KO in BT-474 cells caused a decrease of PUFAs, increased Akt phosphorylation, enhanced glucose dependence of OxPhos, and increased sensitivity to inhibition of HER2, upstream of Akt. This mechanism may be applicable to other HER2-positive glucose-dependent breast cancer cells with HSD17B4 silencing.

Nickel-Catalyzed Three-Component Tandem Radical Cyclization 1,5-Difunctionalization of 1,3-Enynes and Alkyl Bromide.

A nickel-catalyzed three-component tandem radical cyclization reaction of aryl bromides with 1,3-enynes and aryl boric acids to construct γ-lactam-substituted allene derivatives has been described. This protocol provides lactam alkyl radicals through the free radical cyclization process, which can be effectively used to participate in the subsequent multicomponent coupling reaction so that 1,3-enynes could directly convert into corresponding poly-substituted allene compounds. In addition, this efficient method enjoys a broad substrate scope and provides a series of 1,5-difunctionalized allenes in a one-pot reaction.

The methylation level of a single cancer risk marker gene reflects methylation burden in gastric mucosa.

BACKGROUND: Gastric cancer risk can be accurately predicted by measuring the methylation level of a single marker gene in gastric mucosa. However, the mechanism is still uncertain. We hypothesized that the methylation level measured reflects methylation alterations in the entire genome (methylation burden), induced by Helicobacter pylori (H. pylori) infection, and thus cancer risk. METHODS: Gastric mucosa of 15 healthy volunteers without H. pylori infection (G1), 98 people with atrophic gastritis (G2), and 133 patients with gastric cancer (G3) after H. pylori eradication were collected. Methylation burden of an individual was obtained by microarray analysis as an inverse of the correlation coefficient between the methylation levels of 265,552 genomic regions in the person's gastric mucosa and those in an entirely healthy mucosa. RESULTS: The methylation burden significantly increased in the order of G1 (n = 4), G2 (n = 18), and G3 (n = 19) and was well correlated with the methylation level of a single marker gene (r = 0.91 for miR124a-3). The average methylation levels of nine driver genes tended to increase according to the risk levels (P = 0.08 between G2 vs G3) and was also correlated with the methylation level of a single marker gene (r = 0.94). Analysis of more samples (14 G1, 97 G2, and 131 G3 samples) yielded significant increases of the average methylation levels between risk groups. CONCLUSIONS: The methylation level of a single marker gene reflects the methylation burden, which includes driver gene methylation, and thus accurately predicts cancer risk.

Thymus and lung mucosa-associated lymphoid tissue lymphoma with adenocarcinoma of the lung: a case report and literature review.

BACKGROUND: Mucosa-associated lymphoid tissue (MALT) lymphoma is a common, low-grade, malignant B-cell lymphoma. However, simultaneous MALT lymphoma in the thymus and lung is extremely rare, and concomitant adenocarcinoma of the lung is even rarer. Herein, we report a rare case of a collision tumor in which MALT lymphoma was found in both the thymus and lung with Sjögren's syndrome (SS) and adenocarcinoma in the lung. CASE PRESENTATION: A physical examination of a 32-year-old woman revealed an anterior superior mediastinal space-occupying lesion, and chest computed tomography (CT) indicated a nodular ground-glass opacity and irregular mixed-density focus in the right lung. All lung cancer-related tumor biomarkers were within normal ranges. The thymus and part of the lung tissue were surgically resected. The histopathology and molecular examinations confirmed MALT lymphoma of the thymus and lung with lung adenocarcinoma. SS was also diagnosed. No special postoperative treatment was performed for the MALT lymphoma, and the patient underwent immunosuppressive therapy for SS after 4 months of follow-up observation. CONCLUSIONS: MALT lymphoma of the thymus and lung tissues has no specific presentation on imaging and is difficult to differentiate from common malignant tumors, and the definite diagnoses of these tumors are highly dependent on histopathological examination in combination with molecular testing and cytogenetics. SS may be an important potential condition for the occurrence of MALT lymphoma in the thymus and lung. Additional similar cases are needed to clarify the biological pathways and potential molecular mechanisms of rare lymphomas and collision tumors.

Neuromuscular and occlusion analysis to evaluate the efficacy of three splints on patients with bruxism.

OBJECTIVE: Occlusal splints are always applied on individuals with bruxism to reduce tooth wear and relieve orofacial symptoms such as myofascial pain. The stomatognathic system is mainly composed of tooth, occlusion, masticatory muscles, and temporomandibular joint. The occlusion and masticatory muscles function are regarded as the important parameters for evaluating the stomatognathic system state objectively. However, the effects of occlusal splints on individuals with bruxism is rarely elucidated from accurate neuromuscular analysis and occlusion evaluation. The aim of the present study was to estimate the effects of three different splints (two clinically common full coverage occlusal splint and an modified anterior splint) on subjects with bruxism using K7-J5 neuromuscular analysis system and Dental Prescale II (DP2) to evaluate occlusion. METHODS: Sixteen subjects claimed to be suffering from nocturnal bruxism,with complete dentition and stable occlusal relationship, were selected for study.The intermaxillary space and the baselines of EMG-activity of the anterior temporalis and masseter were recorded for all the subjects. The participants was treated with three different splints, and outcomes were estimated by comfort index, occlusion and surface electromyography of anterior temporalis and masseter. RESULTS: At teeth clenched position, EMG data were significantly lower in the participants with use of modified anterior splint than with hard, soft occlusal splint or without splint (p < 0.05). The maximum bite force and bite area occur in subjects without use of splint, while the minimal occur in subjects with use of modified anterior splint. Intermaxillary space increased and masticatory muscles presented significant reduction of EMG data at rest position as a result of J5 (p < 0.05). CONCLUSION: Modified anterior splint seems to be more comfortable and effective in reducing occlusion force and electromyographic activity of anterior temporalis and masseter for subjects with bruxism.

Prediction of tissue origin of adenocarcinomas in the esophagogastric junction by DNA methylation.

BACKGROUND: Prediction of tissue origin of esophagogastric junction (EGJ) adenocarcinomas can be important for therapeutic decision, but no molecular marker is available. Here, we aimed to develop such a marker taking advantage of tissue-specific profiles of DNA methylation. METHODS: DNA methylation profiles of gastric adenocarcinomas (GACs) were obtained by an Infinium HumanMethylation450 BeadChip array, and those of esophageal adenocarcinoma (EACs) were obtained from the TCGA database. DNA from formalin-fixed paraffin-embedded (FFPE) samples was analyzed by bisulfite pyrosequencing. RESULTS: In the screening set, 51 of 145,841 CpG sites in CpG islands were methylated at significantly higher levels in 30 GACs compared to those in 30 EACs. Among them, SLC46A3 and cg09177106 were unmethylated in all the 30 EACs. Predictive powers of these two markers were successfully confirmed in an independent validation set (18 GACs and 18 EACs) (SLC46A3, sensitivity = 77.8%, specificity = 100%; cg09177106, sensitivity = 83.3%, specificity = 94.4%), and could be applied to FFPE samples (37 GACs and 18 EACs) (SLC46A3, P = 0.0001; cg09177106, P = 0.0028). On the other hand, EAC-specific markers informative in the FFPE samples could not be isolated. Using these GAC-specific markers, nine of 46 (19.6%) TCGA EGJ adenocarcinomas were predicted to be GACs. CONCLUSIONS: Two GAC-specific markers, SLC46A3 and cg09177106, had a high specificity for identifying the tissue origin of EGJ adenocarcinoma.

Genetic and epigenetic profiling indicates the proximal tubule origin of renal cancers in end-stage renal disease.

End-stage renal disease (ESRD) patients on dialysis therapy have a higher incidence of renal cell carcinomas (RCCs), which consist of 2 major histopathological types: clear-cell RCCs (ESRD-ccRCCs) and acquired cystic disease (ACD)-associated RCCs. However, their genetic and epigenetic alterations are still poorly understood. Here, we investigated somatic mutations, copy number alterations (CNAs), and DNA methylation profiles in 9 ESRD-ccRCCs and 7 ACD-associated RCCs to identify their molecular alterations and cellular origins. Targeted sequencing of 409 cancer-related genes, including VHL, PBRM1, SETD2, BAP1, KDM5C, MET, KMT2C (MLL3), and TP53, showed ESRD-ccRCCs harbored frequent VHL mutations, while ACD-associated RCCs did not. CNA analysis showed that ESRD-ccRCCs had a frequent loss of chromosome 3p while ACD-associated RCCs had a gain of chromosome 16. Beadarray methylation analysis showed that ESRD-ccRCCs had methylation profiles similar to those of sporadic ccRCCs, while ACD-associated RCCs had profiles similar to those of papillary RCCs. Expression analysis of genes whose expression levels are characteristic to individual segments of a nephron showed that ESRD-ccRCCs and ACD-associated RCCs had high expression of proximal tubule cell marker genes, while chromophobe RCCs had high expression of distal tubule cell/collecting duct cell marker genes. In conclusion, ESRD-ccRCCs and ACD-associated RCCs had mutation and methylation profiles similar to those of sporadic ccRCCs and papillary RCCs, respectively, and these 2 histopathological types of RCCs were indicated to have originated from proximal tubule cells of the nephron.

Rechargeable Zn-MnO2 batteries: advances, challenges and perspectives.

As one type of advanced alternative batteries, zinc-ion batteries (ZIBs) have attracted increasing attention because of their advantages of cost-effectiveness, high safety and environmentally benign features. However, the performance of cathode materials has become a bottleneck for the future application of ZIBs. In recent years, manganese dioxide (MnO2)-based materials as cathodes for ZIBs have been intensively explored. In this review, recent advances in MnO2-based cathode materials for ZIBs are comprehensively reviewed with a discussion about the reaction mechanisms for the fundamental understanding of the electrochemical processes. Furthermore, several challenges hindering the technology maturity are also analyzed with corresponding strategies to further improve the electrochemical performance of such Zn-MnO2 batteries.

Progress on Range of Motion After Total Knee Replacement by Sensor-Based System.

For total knee replacement (TKR) patients, rehabilitation after the surgery is key toregaining mobility. This study proposes a sensor-based system for effectively monitoringrehabilitation progress after TKR. The system comprises a hardware module consisting of thetriaxial accelerometer and gyroscope, a microcontroller, and a Bluetooth module, and a softwareapp for monitoring the motion of the knee joint. Three indices, namely the number of swings, themaximum knee flexion angle, and the duration of practice each time, were used as metrics tomeasure the knee rehabilitation progress. The proposed sensor device has advantages such asusability without spatiotemporal constraints and accuracy in monitoring the rehabilitation progress.The performance of the proposed system was compared with the measured range of motion of theCybex isokinetic dynamometer (or Cybex) professional rehabilitation equipment, and the resultsrevealed that the average absolute errors of the measured angles were between 1.65° and 3.27° forthe TKR subjects, depending on the swing speed. Experimental results verified that the proposedsystem is effective and comparable with the professional equipment.

Biosynthesis of polyketides by two type III polyketide synthases from Aloe barbadensis.

Various bioactive polyketides have been found in Aloe barbadensis. However, the polyketide synthases (PKSs), which participate in biosynthesis of polyketides in A. barbadensis remain unknown. In this study, two type III PKSs (AbPKS1 and AbPKS2) were identified from A. barbadensis. AbPKS1 and AbPKS2 were able to utilize malonyl-CoA to yield heptaketides (TW93a and aloesone) and octaketides (SEK4 and SEK4b), respectively. AbPKS1 also exhibited catalytic promiscuity in recognizing CoA thioesters of aromatics to produce unusual polyketides. What Is more, a whole cell biocatalysis system with the capability of producing 26.4 mg/L of SEK4/SEK4b and 2.1 mg/L of aloesone was successfully established.

Photophysical Identification of Three Kinds of Low-Energy Green Band Defects in Wide-Bandgap Polyfluorenes.

Blue-light-emitting semiconductors based on polyfluorenes often exhibit an undesired green emission band. In this report, three well-defined oligofluorenes corresponding to three types of "defects" attributed to aggregation, keto formation, and chain entanglement, respectively, are systemically investigated to unveil the origins of the green emission band in fluorene-based materials. First, the optical properties of defect molecules in different states are studied. The defect associated with aggregation is absent in dilute solutions and in films doped at 0.01 wt % with poly(methyl methacrylate). Second, the dependence of the emission spectra on the solvent was monitored to compare the effects of the "keto-" and "chain-entanglement defect" molecules. The green emission of keto defects exhibited a strong dependence on solvent polarity, whereas this cannot be observed in case of chain-entanglement defect. Third, energy transfer between poly[4-(octyloxy)-9,9-diphenylfluoren-2,7-diyl]- co-[5-(octyloxy)-9,9-diphenyl-fluoren-2,7-diyl] and the keto or chain-entanglement defect molecules is illustrated. Compared to those of the chain-entanglement defect, the spectra of the keto defect molecule (1:10-3) show signs of defect emission at lower proportions. These investigations not only provide insight into the photophysics of oligofluorenes but also supply a new strategy to explore defects in semiconductor polymers, which will aid in the development of effective approaches to obtain stable, pure blue organic light-emitting diodes based on polyfluorenes.

Estrogen deficiency compromised the ß2AR-Gs/Gi coupling: implications for arrhythmia and cardiac injury.

Estrogen and ß2-adrenergic receptors (ß2AR) play important roles in the processes that protect the heart. Here, we investigated how ovariectomy influenced the ß2AR downstream pathways in the context of catecholaminergic stress. In vivo and in vitro stress models were developed in female Sprague-Dawley (SD) rats by epinephrine (Epi) treatments. The cardiac function was evaluated at in vivo and in vitro levels in terms of contraction, rhythm, and injury. We found that myocardial contractility was not significantly different between Sham and ovariectomized (OVX) group rats in the normal state. However, Epi pretreatment decreased the contractility and increased abnormal rhythms especially in OVX group, which were attributed to lack of estrogen. Inhibition of the ß2AR-Gi-PI3K/p38MAPK pathway with ICI118,551, PTX or LY294002 increased contractility and aggravated Epi-induced injury on cardiomyocytes, decreased p38MAPK phosphorylation, and only increased arrhythmia in Sham group. These results indicated that OVX exacerbated cardiac injury and abnormal rhythms through ß2AR-Gi-PI3K and ß2AR-Gi-p38MAPK pathways, respectively. In normal state, the levels of activated Gi were similar in both groups, but those of cAMP and activated Gs were higher in OVX group. Epi treatment increased activated Gi (especially in Sham group) and activated Gs and cAMP in Sham group but decreased it in OVX group. These results suggested that estrogen increased the Gi activity in normal and stress states and Gs activity in stress state. These results indicated that lack of estrogen impaired the ß2AR-Gs/Gi coupling during stress which compromised cardiac contractility and increased abnormal rhythms.

Sperm-inherited organelle clearance in C. elegans relies on LC3-dependent autophagosome targeting to the pericentrosomal area.

Macroautophagic degradation of sperm-inherited organelles prevents paternal mitochondrial DNA transmission in C. elegans. The recruitment of autophagy markers around sperm mitochondria has also been observed in mouse and fly embryos but their role in degradation is debated. Both worm Atg8 ubiquitin-like proteins, LGG-1/GABARAP and LGG-2/LC3, are recruited around sperm organelles after fertilization. Whereas LGG-1 depletion affects autophagosome function, stabilizes the substrates and is lethal, we demonstrate that LGG-2 is dispensable for autophagosome formation but participates in their microtubule-dependent transport toward the pericentrosomal area prior to acidification. In the absence of LGG-2, autophagosomes and their substrates remain clustered at the cell cortex, away from the centrosomes and their associated lysosomes. Thus, the clearance of sperm organelles is delayed and their segregation between blastomeres prevented. This allowed us to reveal a role of the RAB-5/RAB-7 GTPases in autophagosome formation. In conclusion, the major contribution of LGG-2 in sperm-inherited organelle clearance resides in its capacity to mediate the retrograde transport of autophagosomes rather than their fusion with acidic compartments: a potential key function of LC3 in controlling the fate of sperm mitochondria in other species.

Tanshinol alleviates impaired bone formation by inhibiting adipogenesis via KLF15/PPARγ2 signaling in GIO rats.

Glucocorticoid (GC)-induced osteoporosis (GIO) is characterized by impaired bone formation, which can be alleviated by tanshinol, an aqueous polyphenol isolated from Salvia miltiorrhiza Bunge. In this study we investigated the molecular mechanisms underlying GC-induced modulation of osteogenesis as well as the possibility of using tanshinol to interfere with GIO. Female SD rats aged 4 months were orally administered distilled water (Con), prednisone (GC, 5 mg·kg-1·d-1), GC plus tanshinol (Tan, 16 mg·kg-1·d-1) or GC plus resveratrol (Res, 5 mg·kg-1·d-1) for 14 weeks. After the rats were sacrificed, samples of bone tissues were collected. The changes in bone formation were assessed using Micro-CT, histomorphometry, and biomechanical assays. Expression of Kruppel-like factor 15 (KLF15), peroxisome proliferator-activated receptor γ 2 (PPARγ 2) and other signaling proteins in skeletal tissue was measured with Western blotting and quantitative RT-PCR. GC treatment markedly increased the expression of KLF15, PPARγ2, C/EBPα and aP2, which were related to adipogenesis, upregulated FoxO3a pathway proteins (FoxO3a and Gadd45a), and suppressed the canonical Wnt signaling (ß-catenin and Axin2), which was required for osteogenesis. Thus, GC significantly decreased bone mass and bone quality. Co-treatment with Tan or Res effectively counteracted GC-impaired bone formation, suppressed GC-induced adipogenesis, and restored abnormal expression of the signaling molecules in GIO rats. We conclude that tanshinol counteracts GC-decreased bone formation by inhibiting marrow adiposity via the KLF15/PPARγ2/FoxO3a/Wnt pathway.

Cardioprotection by Low-dose of Estrogen and Testosterone at the Physiological Ratio on Ovariectomized Rats During Ischemia/Reperfusion Injury.

Although estrogen and testosterone deficiency have often been associated with the development of cardiac diseases in postmenopausal women, the benefits of estrogen or testosterone therapy are controversial. Supplementation with high dose of estrogen or testosterone alone has been associated with many side effects, especially estrogen. This study was aimed to investigate whether supplementation of testosterone in combination with low-dose estrogen conferred stronger cardioprotective effects on ovariectomized rats subjected to ischemia/reperfusion injury. Female Sprague Dawley rats were subjected to sham operation (Sham) or bilateral ovariectomy (OVX). Two weeks after ovariectomy, OVX rats were treated with one of the following: (1) vehicle (OVX), (2) testosterone (100 µg·kg·d) (OVX+T), (3) estrogen (20 µg·kg·d) (OVX+E), (4) testosterone (100 µg·kg·d) + estrogen (20 µg·kg·d) (OVX+T+E) for 4 weeks. The hearts were mounted on the Langendorff apparatus and subjected to ischemia/reperfusion injury subsequent to the determination of hemodynamic parameters. We examined the release of lactate dehydrogenase, serum estrogen, and testosterone levels and the expression of pAkt/Akt and bax/bcl-2. Testosterone supplementation alone improved the heart function, increased p-Akt/Akt and bcl-2 expression, and decreased the release of lactate dehydrogenase. Accordingly, these effects of testosterone were more pronounced when low-dose estrogen was administered simultaneously, whereas estrogen alone at the dose of the experiment had no significant effects. These effects might be partially orchestrated by the Akt signaling pathway.