Leukemia Inhibitory Factor Facilitates Self-Renewal and Differentiation and Attenuates Oxidative Stress of BMSCs by Activating PI3K/AKT Signaling.

Objective: Transplantation of bone marrow-derived mesenchymal stem cells (BMSCs) remains a hopeful therapeutic approach for bone defect reconstruction. Herein, we investigated the effects and mechanisms of leukemia inhibitory factor (LIF) in the function and viability of hypoxic BMSCs as well as bone defect repair. Methods: The effects of LIF on apoptosis (flow cytometry, TUNEL staining), mitochondrial activity (JC-1 staining), proliferation (colony formation, EdU staining), and differentiation (CD105, CD90, and CD29 via flow sorting) were examined in hypoxic BMSCs. LIF, LIFR, gp130, Keap1, Nrf2, antioxidant enzymes (SOD1, catalase, GPx-3), bone-specific matrix proteins (ALP, BSP, OCN), PI3K, and Akt were detected via immunoblotting or immunofluorescent staining. BMSCs combined with biphasic calcium phosphate scaffolds were implanted into calvarial bone defect mice, and the therapeutic effect of LIF on bone defect was investigated. Results: Hypoxic BMSCs had increased apoptosis and oxidative stress and reduced mitochondrial activity. Additionally, LIF, LIFR, and gp130 were upregulated and PI3K/Akt activity was depressed in hypoxic BMSCs. Upregulated LIF alleviated apoptosis and oxidative stress and heightened mitochondrial activity and PI3K/Akt signaling in hypoxic BMSCs. Additionally, LIF overexpression promoted self-renewal and osteogenic differentiation of BMSCs with hypoxic condition. Mechanically, LIF facilitated self-renewal and differentiation as well as attenuated oxidative stress of BMSCs through enhancing PI3K/AKT signaling activity. Implantation of LIF-overexpressed BMSC-loaded BCP scaffolds promoted osteogenesis as well as alleviated oxidative stress and apoptosis through PI3K/Akt signaling. Conclusion: Our findings demonstrate that LIF facilitates self-renewal and differentiation and attenuates oxidative stress of BMSCs by PI3K/AKT signaling.

Myofascial iliac crest flap for reconstruction of combined oral mucosa-mandibular defects: A single-centre experience.

BACKGROUND: Deep circumflex iliac artery (DCIA) myofascial iliac crest flap has been used for combined oral mucosa-mandibular defects reconstruction. The bone component of this composite flap can reconstruct the mandible with superior contour match, and the muscle fascia which used for repairing the oral mucosa defect will transform into an oral mucosa-like appearance. To explore its scope of clinical application and how the fascia transformed into oral mucosa will give surgeons flexibility to reconstruct the combined oral mucosa-mandibular defects. METHODS: A retrospective review of 18 patients who received combined oral mucosa-mandibular defects reconstruction with DCIA myofascial iliac crest flaps from Dec 2016 to Dec 2020 was performed. The characteristics of the mandibular defects and the flaps were recorded. The postoperative dynamic changes of one graft's fascia were observed from serial photographs. RESULTS: All myofascial iliac crest flaps survived successfully. The bone grafts were from 4.0 to 9.5 cm (mean 7.6 ± 1.5 cm) in length and from 2.0 to 3.5 cm (mean 2.7 ± 0.4 cm) in height. The sizes of fascia were from 13.5 to 48.0 cm2 (mean 27.2 ± 9.4 cm2). The grafted fascia firstly changed into a yellow pseudomembrane-like appearance, and then experienced muscle oedema before finally transformed into an oral mucosa-like appearance at about 60 days after operation. CONCLUSION: Myofascial iliac crest flap is a good option for reconstruction of combined oral mucosa-mandibular defects because of its excellent bone and oral mucosa matches.

Comparison between thermophilic and mesophilic anaerobic digestion of waste activated sludge by combined NaOH-microwave pretreatment.

The purpose of this study was to investigate the performance of the thermophilic and mesophilic anaerobic digestion process (TADP, MADP) fed with NaOH-microwave pretreated waste activated sludge. The experiment was conducted in anaerobic CSTR reactors. During this experiment, the reactors were stable in operation and were not inhibited by ammonia. The methane production and reduction of organic matters from MADP were less than those from TADP. The dewatering performance of mesophilic sludge was better than that of the thermophilic sludge. The experimental results showed that the continuous TADP and MADP were effective, when the reactors were fed with the waste activated sludge pretreated by NaOH-microwave. MADP was more suitable to combine the NaOH-MW pretreatment process than TADP.

A deep learning algorithm for detection of oral cavity squamous cell carcinoma from photographic images: A retrospective study.

BACKGROUND: The overall prognosis of oral cancer remains poor because over half of patients are diagnosed at advanced-stages. Previously reported screening and earlier detection methods for oral cancer still largely rely on health workers' clinical experience and as yet there is no established method. We aimed to develop a rapid, non-invasive, cost-effective, and easy-to-use deep learning approach for identifying oral cavity squamous cell carcinoma (OCSCC) patients using photographic images. METHODS: We developed an automated deep learning algorithm using cascaded convolutional neural networks to detect OCSCC from photographic images. We included all biopsy-proven OCSCC photographs and normal controls of 44,409 clinical images collected from 11 hospitals around China between April 12, 2006, and Nov 25, 2019. We trained the algorithm on a randomly selected part of this dataset (development dataset) and used the rest for testing (internal validation dataset). Additionally, we curated an external validation dataset comprising clinical photographs from six representative journals in the field of dentistry and oral surgery. We also compared the performance of the algorithm with that of seven oral cancer specialists on a clinical validation dataset. We used the pathological reports as gold standard for OCSCC identification. We evaluated the algorithm performance on the internal, external, and clinical validation datasets by calculating the area under the receiver operating characteristic curves (AUCs), accuracy, sensitivity, and specificity with two-sided 95% CIs. FINDINGS: 1469 intraoral photographic images were used to validate our approach. The deep learning algorithm achieved an AUC of 0·983 (95% CI 0·973-0·991), sensitivity of 94·9% (0·915-0·978), and specificity of 88·7% (0·845-0·926) on the internal validation dataset (n = 401), and an AUC of 0·935 (0·910-0·957), sensitivity of 89·6% (0·847-0·942) and specificity of 80·6% (0·757-0·853) on the external validation dataset (n = 402). For a secondary analysis on the internal validation dataset, the algorithm presented an AUC of 0·995 (0·988-0·999), sensitivity of 97·4% (0·932-1·000) and specificity of 93·5% (0·882-0·979) in detecting early-stage OCSCC. On the clinical validation dataset (n = 666), our algorithm achieved comparable performance to that of the average oral cancer expert in terms of accuracy (92·3% [0·902-0·943] vs 92.4% [0·912-0·936]), sensitivity (91·0% [0·879-0·941] vs 91·7% [0·898-0·934]), and specificity (93·5% [0·909-0·960] vs 93·1% [0·914-0·948]). The algorithm also achieved significantly better performance than that of the average medical student (accuracy of 87·0% [0·855-0·885], sensitivity of 83·1% [0·807-0·854], and specificity of 90·7% [0·889-0·924]) and the average non-medical student (accuracy of 77·2% [0·757-0·787], sensitivity of 76·6% [0·743-0·788], and specificity of 77·9% [0·759-0·797]). INTERPRETATION: Automated detection of OCSCC by deep-learning-powered algorithm is a rapid, non-invasive, low-cost, and convenient method, which yielded comparable performance to that of human specialists and has the potential to be used as a clinical tool for fast screening, earlier detection, and therapeutic efficacy assessment of the cancer.

Transcriptome analysis of Lactobacillus paracasei SMN-LBK under ethanol stress.

Lactobacillus paracasei SMN-LBK (serial number: CCTCC M 2017429) is an ethanol-resistant lactic acid bacteria (LAB) in kumiss. However, the anti-ethanol stress mechanism of L. paracasei SMN-LBK remains unclear. Hence, we performed a transcriptome analysis between L. paracasei SX10 (L. paracasei SMN-LBK under 10% ethanol stress strain, abbreviated as SX10) and L. paracasei SMN-LBK (abbreviated as S10) by RNA sequencing. We performed real-time quantitative PCR (RT-qPCR) to verify the accuracy of the transcription data. The transcriptome data revealed that 315 genes exhibited upregulated expression, and 332 genes were downregulated in the SX10 compared with the S10 group. The PFK, LDH, GPDH, and GK genes were upregulated, with a log2-fold change of 1.10, 0.30, 0.56, and 1.512, respectively. A gene ontology enrichment analysis revealed significant enrichment of ribosomes, ribonucleoprotein complex, non-membrane-bounded organelles, and intracellular non-membrane-bound organelles. Analysis using the Kyoto Encyclopedia of Genes and Genomes database revealed differential genes associated with ribosome function, pyruvate metabolism, biosynthesis of amino acids, fatty acid biosynthesis, fatty acid metabolism, ATP-binding cassette (ABC) transporter, glycolysis, and glycerophospholipid metabolism. The RT-qPCR results were consistent with the transcriptome results. Lactococcus lactis NZ9000 is a typical host bacterium. We performed PFK and GK overexpression to verify the function of the L. paracaseiSX10 resistance gene in Lactococcus lactis NZ9000. Using sodium dodecyl sulfate (SDS)-PAGE electrophoresis, these resistance genes were successfully expressed in Lactococcus lactis NZ9000. The survival rate and key enzyme activity of the recombinant strains were determined under ethanol stress. The survival rate of Lactococcus lactis NZ9000-pNZ8148-PFK and Lactococcus lactis NZ9000-pNZ8148-GK under 10% ethanol stress were 3.43- and 3.80-fold higher compared with the Lactococcus lactis NZ9000-pNZ8148 control, respectively. These results indicate that PFK and GK are important for the ethanol tolerance of LAB and can increase the ethanol tolerance of Lactococcus lactis NZ9000. Hence, PFK and GK were identified as key genes of L. paracasei SX10 with a high ethanol tolerance. Our results provide novel insight for further studies to perform a systematic analysis of the differentially expressed genes and to determine their potential functions in the ethanol tolerance mechanism of LAB.

The complete chloroplast genome of Annona muricata L.: a tropical fruit with important medicinal properties.

Annona muricata L. (A. muricata) is an important tropical fruit and medicinal plant. It is one of the easily found plants used traditionally in treating cancer. In many tropical countries, especially in Southeast Asia, A. muricata is popular for its edible fruit and medicinal merits. In this study, the complete chloroplast genome of A. muricata was sequenced, assembled, and annotated. The chloroplast genome of A. muricata was found to be a double strand ring structure with the size of 196,038 bp that consists of four regions: a large single-copy region of 75,339 bp, a small single-copy region of 3105 bp, and two inverted repeat regions of 58,797 bp. The GC content of the whole chloroplast genome was 39.92%. It was found that 111 protein-coding genes, one Pseudogene, 38 tRNA genes, and eight rRNA genes were annotated in the chloroplast genome, and the total number of genes was 158. DNA sequences of the chloroplast genomes of 19 species which belonged to three families of Magnoliales order were analyzed and a phylogenetic tree was constructed. The result indicated that A. muricata, Annona cherimola, Uvaria macrophylla, Greenwayodendron suaveolens, and Chieniodendron hainanense had a close phylogenetic relationship. The findings also provided abundant basic data for the genomics study of A. muricata.

Complete chloroplast genome sequence and phylogenetic analysis of Annona reticulata.

Annona reticulata is native to South and Central America which has many phytochemical and pharmacological activities suggesting a wide range of clinical application in lieu of cancer chemotherapy. This study provides abundant genomic data for the genetic relationship study, germplasm resources evaluation and varieties selection of A. reticulata. The complete chloroplast genome of A. reticulata was sequenced, assembled, and annotated in this study. The genome size was 201,906 bp and was divided into four regions: a large single-copy region of 69,650 bp, a small single-copy region of 3,014 bp, and two inverted repeat regions of 64,621 bp. A total number of 164 genes were annotated, including 115 protein-coding genes, one pseudogene, 40 tRNA genes, and eight rRNA genes. In terms of gene function, the 164 genes were divided into four major groups: genes for self-replication, photosynthesis, unknown function, and other genes. A maximum likelihood tree based on the chloroplast genome sequences of 24 plant species was constructed. The result of phylogenetic analysis showed that A. cherimola had the closest relationship with A. reticulate.

The complete chloroplast genome sequence of Manilkara zapota (Linn.) van Royen.

The complete chloroplast genome sequences of Manilkara zapota (Linn.) van Royen in Xishuangbanna, Yunnan province were reported in this study. The length of the sequence was 159,853 bp long with the large single copy (LSC) region of 89,632 bp, the small single copy (SSC) region of 18,747 bp, and two inverted repeat (IR) regions of 27,737 bp. The plastome contained 125 genes, including 84 protein-coding, 8 ribosomal RNA, and 33 transfer RNA genes. The overall GC content was 37.0%. Phylogenetic analysis of 12 representative plastomes within the order Ebenales suggests that M. zapota (Linn.) van Royen is closely related to the species in family Ebenaceae.

Evaluating the microbial ecology and metabolite profile in Kazak artisanal cheeses from Xinjiang, China.

Kazak artisanal cheese is one of the famous fermented food in Uighur Autonomy Region of Xinjiang, China. However, the microbial ecology in Kazak artisanal cheeses across different regions is unclear. In this study, we determined the microbial community composition through amplicon sequencing and measured the flavor profile of 10 cheese samples from different regions of Xinjiang. The associations between microbial communities, flavors and environmental factors were examined by redundancy analysis and Monte Carlo permutation test. Cheeses from different regions had different microbial communities, which was mainly reflected in the relative abundance of Lactobacillus, Streptococcus, Issatchenkia, Debaryomyces and Kluyveromyces. In addition, Pichia and Torulaspora were also the key microbial groups, according to the high relative abundance and large co-occurrence incidence in the correlation network. Using the microbe-metabolites correlation analysis, the major flavor-producing taxa were identified as Kluyveromyces, Anoxybacillus, Torulaspora, Lactobacillus, Streptococcus and Dipodascus. Environmental factors accounted for the majority of the microbial community variations, 88.54% for bacteria and 75.71% for fungi. Compared to physico-chemical factors (temperature, moisture, and pH), geographical factors (longitude, latitude and elevation) had a stronger effect on microbial communities in cheese samples from different regions of Xinjiang.

Dynamic correlations between microbiota succession and flavor development involved in the ripening of Kazak artisanal cheese.

Cheese is a typical handcrafted fermented food in Kazak minority from the Uighur Autonomy Region in China. The ripening process of the cheese is crucial for quality and flavor. The aim of this study was to gain a deeper knowledge on the bacterial and fungal community diversity at different time points during the post-ripening of the cheese and to understand the relationship between bacterial and fungal profiles and the chemical components including amino acids, fatty acids and volatile compounds related the cheese flavor. Cheese samples were collected from days 5, 10, 15, 20 and 30 after the starting point of post-ripening. The bacterial and fungal compositions were analyzed with next generation sequencing targeting the 16S rDNA loci for bacteria and ITS loci for fungi. The amino acids contents were analyzed by reversed phase high performance liquid chromatography combined with UV detection. The fatty acids and the volatile components were analyzed by Solid Phase Micro Extraction followed by Gas Chromatography/Mass spectrometry. We found that Lactobacillus, Streptococcus, Kluyveromyces and Torulaspora were the dominant cheese's population. Bidirectional orthogonal partial least squares (O2PLS) based correlation analysis between microbiota succession and flavor dynamics showed that bacteria made more contributions to flavor formation than fungi. Eight bacteria genera and seven fungi genera were determined as functional core microbiota for the flavor production based on their dominance and functionality in microbial community. This study provided a comprehensive picture of the dynamic changes of microbiota profiles through the post-ripening process. The elucidation of the causal relationship between microbiota and the flavor components has advanced our understanding of the mechanism underlying the cheese development.

Potential characterization of yeasts isolated from Kazak artisanal cheese to produce flavoring compounds.

Cheese is a typical handcrafted fermented food in Kazak minority from the Uighur Autonomy Region in China and Central Asia. Among the microbial community that is responsible for Kazak cheese fermentation, yeasts play important role in flavor formation during ripening. To develop ripening cultures, we isolated 123 yeasts from 25 cheese products in Kazak, and identified 87 isolates by the D1/D2 domain of the large subunit rRNA gene sequence. Pichia kudriavzevii was the dominant yeast in Kazak cheese, followed by Kluyveromyces marxianus and Kluyveromyces lactis. Of these, the ability to exhibit enzyme of dominant isolates and contribution to the typical flavor of cheeses was assessed. Enzyme producing yeast strains were inoculated in Hazak cheese-like medium and volatile compounds were identified by head space solid phase micro extraction coupled to gas chromatography and mass spectroscopy. Pichia kudriavzevii N-X displayed the strongest extracellular proteolytic and activity on skim milk agar and produced a range of aroma compounds (ethanol, ethyl acetate, 3-methylbutanol, and acetic acid) for Kazak cheese flavor, could be explored as ripening cultures in commercial production of Kazak cheeses.

Analysis of components and study on antioxidant and antimicrobial activities of oil in apple seeds.

In order to improve the comprehensive utilization of major by-products in apple-juice processing, the components, antioxidant and antimicrobial activities of oil in two species apple seeds, Fuji and New Red Star, were investigated. The Soxhlet extracted oil content of apple seeds raged from 20.69 to 24.32 g/100 g. The protein, fiber and ash contents were found to be 38.85-49.55 g/100 g, 3.92-4.32 g/100 g and 4.31-5.20 g/100 g, respectively; the extracted oils exhibited an iodine value of 94.14-101.15 g I/100 g oil; refractive index (40 degrees C) was 1.465-1.466; density (25 degrees C) was 0.902-0.903 mg/ml; saponification value was 179.01-197.25 mg KOH/g oil; and the acid value was 4.036-4.323 mg KOH/g oil. The apple seed oils mainly consisted of linoleic acid (50.7-51.4 g/100 g) and oleic acid (37.49-38.55 g/100 g). Other prominent fatty acids were palmitic acid (6.51-6.60 g/100 g), stearic acid (1.75-1.96 g/100 g) and arachidic acid (1.49-1.54 g/100 g). Apple seed oil was proven to possess interesting properties, emerging from its chemical composition and from the evaluation of its in vitro biological activities. The apple seed oil was almost completely active against bacteria, mildews were less sensitive to apple seed oil than yeasts, and the minimum inhibitory concentration (MIC) of apple seed oil ranged from 0.3 to 0.6 mg/ml. The observed biological activities showed that the oil had a good potential for use in the food industry and pharmacy.