Metabolic flux analysis of coenzyme Q10 synthesized by Rhodobacter sphaeroides under the influence of different pH regulators.

Coenzyme Q10 (CoQ10) is crucial for human beings, especially in the fields of biology and medicine. The aim of this experiment was to investigate the conditions for increasing CoQ10 production. At present, microbial fermentation is the main production method of CoQ10, and the production process of microbial CoQ10 metabolism control fermentation is very critical. Metabolic flux is one of the most important determinants of cell physiology in metabolic engineering. Metabolic flux analysis (MFA) is used to estimate the intracellular flux in metabolic networks. In this experiment, Rhodobacter sphaeroides was used as the research object to analyze the effects of aqueous ammonia (NH3·H2O) and calcium carbonate (CaCO3) on the metabolic flux of CoQ10. When CaCO3 was used to adjust the pH, the yield of CoQ10 was 274.43 mg·L-1 (8.71 mg·g-1 DCW), which was higher than that of NH3·H2O adjustment. The results indicated that when CaCO3 was used to adjust pH, more glucose-6-phosphate (G6P) entered the pentose phosphate (HMP) pathway and produced more NADPH, which enhanced the synthesis of CoQ10. At the chorismic acid node, more metabolic fluxes were involved in the synthesis of p-hydroxybenzoic acid (pHBA; the synthetic precursor of CoQ10), enhancing the anabolic flow of CoQ10. In addition, Ca2+ produced by the reaction of CaCO3 with organic acids promotes the synthesis of CoQ10. In summary, the use of CaCO3 adjustment is more favorable for the synthesis of CoQ10 by R. sphaeroides than NH3·H2O adjustment. The migration of metabolic flux caused by the perturbation of culture conditions was analyzed to compare the changes in the distribution of intracellular metabolic fluxes for the synthesis of CoQ10. Thus, the main nodes of the metabolic network were identified as G6P and chorismic acid. This provides a theoretical basis for the modification of genes related to the CoQ10 synthesis pathway.

The impact of utilizing oyster shell soil conditioner on the growth of tomato plants and the composition of inter-root soil bacterial communities in an acidic soil environment.

Introduction: The objective of this study is to examine the impact of various oyster shell soil conditioners, which are primarily composed of oyster shells, on the growth of tomatoes in acidic soil. Moreover, the aim of this investigation is to analyze the variety and structure of soil bacterial populations in close proximity to tomato roots while also contributing to the understanding of the physical, chemical, and biological mechanisms of oyster shell soil conditioners. Methods: Tomato plants were grown in acidic red soil in three groups: a control group and a treatment group that used two types of oyster shell soil conditioners, OS (oyster shell powder) and OSF (oyster shell powder with organic microbial fertilizer). A range of soil physicochemical properties were measured to study differences in inter-soil physicochemical parameters and the growth of tomato plantings. In addition, this study utilized the CTAB (Cetyltrimethylammonium Bromide) technique to extract DNA from the soil in order to investigate the effects of oyster shell soil conditioner on the composition and diversity of bacterial populations. Utilizing high-throughput sequencing technologies and diversity index analysis, the composition and diversity of bacterial populations in the soil adjacent to plant roots were then evaluated. Ultimately, correlation analysis was used in this study to explore the relationship between environmental factors and the relative abundance of soil bacteria in the inter-root zone of tomato plants. Results: The findings indicated that the oyster shell soil conditioners were capable of modifying the physicochemical characteristics of the soil. This was evidenced by significant increases in soil total nitrogen (16.2 and 59.9%), soil total carbon (25.8 and 27.7%), pH (56.9 and 55.8%), and electrical conductivity (377.5 and 311.7%) in the OS and OSF groups, respectively, compared to the control group (p < 0.05). Additionally, data pertaining to tomato seed germination and seedling growth biomass demonstrated that both oyster shell soil conditioners facilitated the germination of tomato seeds and the growth of seedlings in an acidic red clay soil (p < 0.05). On the other hand, the application of two oyster shell soil conditioners resulted in a modest reduction in the diversity of inter-root soil bacteria in tomato plants. Specifically, the group treated with OSF exhibited the most substantial fall in the diversity index, which was 13.6% lower compared to the control group. The investigation carried out on the soil between tomato plant roots yielded findings about the identification of the ten most abundant phyla. These phyla together represented 91.00-97.64% of the overall abundance. In the inter-root soil of tomatoes, a study identified four major phyla, namely Proteobacteria, Bacteroidetes, Acidobacteria, and Actinobacteria, which collectively accounted for up to 85% of the total abundance. At the general level, the relative abundance of Massilia increased by 2.18 and 7.93%, Brevundimonas by 5.43 and 3.01%, and Lysobacter by 3.12 and 7.49% in the OS and OSF groups, respectively, compared to the control group. However, the pathogenic bacteria unidentified_Burkholderiaceae decreased by 5.76 and 5.05%, respectively. The correlation analysis yielded conclusive evidence indicating that, which involved the use of CCA (Canonical Correlation Analysis) graphs and Spearman correlation coefficients, pH exhibited a positive correlation (p < 0.05) with Shewanella and a negative correlation (p < 0.05) with Bradyrhizobium. The relative abundance of Lysobacter and Massilia exhibited a positive correlation with the levels of total soil nitrogen. Discussion: The utilization of oyster shell soil conditioner on acidic red soil resulted in several positive effects. Firstly, it raised the pH level of the inter-root soil of tomato plants, which is typically acidic. This pH adjustment facilitated the germination of tomato seeds and promoted the growth of seedlings. In addition, the application of oyster shell soil conditioner resulted in changes in the structure of the bacterial community in the inter-root soil, leading to an increase in the relative abundance of Proteobacteria and Bacteroidetes and a decrease in the relative abundance of Acidobacteria. Furthermore, this treatment fostered the proliferation of genera of beneficial bacteria like Massilia, Brevundimonas, and Lysobacter, ultimately enhancing the fertility of the red soil.

Construction of AI Environmental Music Education Application Model Based on Deep Learning.

The art of music, which is a necessary component of daily life and an ideology older than language, reflects the emotions of human reality. Many new elements have been introduced into music as a result of the quick development of technology, gradually altering how people create, perform, and enjoy music. It is incredible to see how actively AI has been used in music applications and music education over the past few years and how significantly it has advanced. AI technology can efficiently pull in the course, stratify complex large-scale music or sections, simplify teaching, improve student understanding of music, solve challenging student problems in class, and simplify the tasks of teachers. The traditional music education model has been modified, and the music education model's audacious innovation has been made possible by reducing the distance between the teacher and the student. A classification algorithm based on spectrogram and NNS is proposed in light of the advantages in image processing. The abstract features on the spectrogram are automatically extracted using the NNS, which completes the end-to-end learning and avoids the tediousness and inaccuracy of manual feature extraction. This study, which uses experimental analysis to support its findings, demonstrates that different music teaching genres can be accurately classified at a rate of over 90%, which has a positive impact on recognition.

Phlebopus roseus, a new edible bolete from China, is associated with insects and plants.

Phlebopus roseus is described as new based on collections from southwest China. Phylogenetic analyses of nuclear rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS) and portions of nuclear 28S rDNA (28S), translation elongation factor 1-alpha (tef1), and the largest and second largest subunits of RNA polymerase II (rpb1, rpb2) support P. roseus as a novel species in the genus Phlebopus (Boletinellaceae, Boletales). The new species resembles P. portentosus but differs from it in that mature basidiomata have a bright rose-red-colored stipe and a radiate tubular hymenophore with nested pores. Despite extensive searching, P. roseus has only been found at four sites within a 24-hectare orchard dominated by Eriobotrya japonica, which is agriculturally important given its fruit production (loquats). Therefore, this species appears to be endemic and geographically restricted. The ecology of this bolete is also unique. In line with the trophic behavior of other species in the Boletinellaceae, our observations indicate that P. roseus forms a symbiotic association with the scale insect Coccus hesperidum, identified through sequence analysis of its mitochondrial cytochrome c oxidase subunit I (COI) region, to form fungus-insect galls that develop on roots of E. japonica trees. Phlebopus roseus is an edible mushroom species and is collected from the type location by farmers and sold commercially in limited quantities at local markets alongside P. portentosus and other fungi.

Smart phase transformation system based on lyotropic liquid crystalline@hard capsules for sustained release of hydrophilic and hydrophobic drugs.

Smart phase transformation systems@hard capsule (SPTS@hard capsule) based on lyotropic liquid crystalline (LLC) were developed for oral sustained release in this study. Doxycycline hydrochloride (DOXY) and meloxicam (MLX) were used as hydrophilic and hydrophobic model drug, respectively. Two systems were added with different additives, that is, gelucire 39/01, PEG 1000 and Tween 80 to adjust their melting point and release profiles. The phase transformation of these systems could be triggered by water as well as temperature. They could spontaneously transform into cubic phase or hexagonal phase when coming across with water, to achieve the 24 h sustained release profile. In addition, the obtained systems could switch between semisolid state and liquid state when temperature changed within room temperature and body temperature, which facilitated the phase transformation in gastrointestinal tract and during their encapsulation into hard capsules. LLC-based SPTS@hard capsule revealed potential for the industrialization of its oral administration on account of its drugs accommodation with different solubility, controllable release profile and simple preparation process.

Tuber borchii Shapes the Ectomycorrhizosphere Microbial Communities of Corylus avellana.

In this study, eight-month-old ectomycorrhizae of Tuber borchii with Corylus avellana were synthesized to explore the influence of T. borchii colonization on the soil properties and the microbial communities associated with C. avellana during the early symbiotic stage. The results showed that the bacterial richness and diversity in the ectomycorrhizae were significantly higher than those in the control roots, whereas the fungal diversity was not changed in response to T. borchii colonization. Tuber was the dominant taxon (82.97%) in ectomycorrhizae. Some pathogenic fungi, including Ilyonectria and Podospora, and other competitive mycorrhizal fungi, such as Hymenochaete, had significantly lower abundance in the T. borchii inoculation treatment. It was found that the ectomycorrhizae of C. avellana contained some more abundant bacterial genera (e.g., Rhizobium, Pedomicrobium, Ilumatobacter, Streptomyces, and Geobacillus) and fungal genera (e.g., Trechispora and Humicola) than the control roots. The properties of rhizosphere soils were also changed by T. borchii colonization, like available nitrogen, available phosphorus and exchangeable magnesium, which indicated a feedback effect of mycorrhizal synthesis on soil properties. Overall, this work highlighted the interactions between the symbionts and the microbes present in the host, which shed light on our understanding of the ecological functions of T. borchii and facilitate its commercial cultivation.

[The effects and mechanisms of BTBD10 on the proliferation of islet beta cell].

OBJECTIVE: To explore the role of BTBD10 overexpression in the proliferation of insulinoma cell line INS-1 and its mechanism. METHODS: The recombined expression plasmid of pcDNA4.0-BTBD10 was constructed by gene cloning technique and was transfected into INS-1 cell by lipofectamine 2000. The stable overexpression BTBD10 of INS-1 cell was selected at 48(th) hour after transfection. INS-1 cell proliferation activity was measured by MTT method. The expression of BTBD10, protein kinase B (Akt), phospho-Akt (p-Akt), mammal target of rapamycin (mTOR) and phospho-mTOR (p-mTOR) were determined by Western blot. RESULTS: The stable overexpression BTBD10 of INS-1 cell was successfully constructed. Overproduction of BTBD10 promoted beta cell proliferation. The phosphorylation of Akt and mTOR was increased and the ratio of p-Akt/Akt and p-mTOR/mTOR was enhanced in the INS-1 overexpressed by BTBD10. But the expression of total Akt and mTOR presented no obvious changes. CONCLUSION: The overexpression BTBD10 of INS-1 cell could activate of Akt/mTOR signalling pathway via stimulating phospho-mTOR and Akt, and enhance overall cell protein translation, so as to promote proliferation of INS-1 cell.

[Diagnostic values of self-assessment tool for Asians for osteoporosis in aged men].

OBJECTIVE: To evaluate the diagnostic values of osteoporosis self-assessment tool for Asians (OSTA) in aged men. METHODS: Bone mineral density (BMD) was measured by dual energy X-ray absorptiometry (DXA) in 382 male outpatients. Their data were collected. After excluding such influencing factors of BMD as diseases or drugs, they were divided into 3 groups according to the WHO osteoporosis diagnosis value or age. The correlation between OSTA index and BMD was analyzed by Spearman correlation. The concordance of OSTA and BMD were calculated and analyzed by a four-fold table. RESULTS: The prevalence of osteoporosis was 9.42% and osteopenia 42.67%. Age increased gradually in the osteoporosis, osteopenia and normal bone mass groups. But OSTA index, BMD (including lumbar vertebrae L(1-4), femoral neck, trochanter and hip) and weight decreased gradually in those groups. The percent of normal bone mass decreased with advancing age. And the percentages were 60.68%, 51.97% and 33.33% respectively. Osteoporosis increased in the older group (> 80 years old) and the percentage was 24.64% accounting for 94.44% of all osteoporotic subjects. Judging by the OSTA index, the rate of low osteoporotic risk decreased with advancing age. But the rates of mid-risk and high-risk increased. The correlation coefficients of OSTA index and BMD in femoral neck, trochanter, hip and L(1-4) was 0.33, 0.28, 0.29 and 0.06 respectively. And the correlation between OSTA index and BMD had statistical significance except for L(1-4). The sensitivity, specificity, coincidence and Youden index of OSTA index with a T score cutoff of -1 of BMD were 56.28%, 64.48%, 60.21% and 0.21 respectively versus 86.11%, 57.80%, 60.47% and 0.44 with a T score cutoff of -2.5. CONCLUSION: Correlation exists between OSTA index and BMD. The sensitivity, specificity and coincidence of OSTA index are excellent based upon the standard of BMD by DXA. It is a useful and simple diagnostic tool of osteoporosis.

[Preparation, characterization and content determination of cubic phase gel containing capsaicin].

OBJECTIVE: To prepare the cubic phase gel containing capsaicin and characterize its properties. METHOD: The cubic phases gel composed of glycerol monoolein, capsaicin and water was made by self-emulsion technology. The characterization of cubic phase gel was carried out by cross-polarizing light microscopy (CPLM) and Small Angle X-Ray Scattering (SAXS). The capsaicin content was determined by HPLC analysis. RESULT: Under CPLM, cubic phase gel showed dark background. SAXS scattering spectra showed the scattering peaks at 0.1096, 0.1334, 0.1557, 0.1883 A(-1) which was compatible with q1:q2:q3:q4 = mean square root of 2: mean square root of 3: mean square root of 4: mean square root of 6. It was well known that the scattering vector ratio was the characteristic of cubic phase and the internal structure was confirmed to be Pn3m (Q224). The linear range for capsaicin determination was 3.25 x 10(-4) - 2.08 x 10(-2) g x L(-1) (R2 = 1). The average recovery was 97.53% with RSD of 2.9% (n=9). CONCLUSION: CPLM and SAXS technology are suitable to characterize the cubic phase gel The determination of the capsaicin content by HPLC is simple and reproducible.

[The expression of Recombinant adenovirus IkappaBalphaM in human hepatocarcinoma HepG2 and it's inhibitive effect to the activity of NF-kappaB].

OBJECTIVE: To investigate the expression of recombinant IkappaBalphaM (AdIkappaBalphaM) in human hepatocarcinoma cell line HepG(2) and the inhibiting effect to NF-kappaB. METHODS: To test the virus titer in 293 cells and the infective efficiency virus titer in HepG(2) cells with GFP and limited dilution method, then to assay the IkappaBalphaM expression of recombinant adenovirus and the alteration after induction of TNF-alpha in 293 cells and HepG(2) by Western blotting, furthermore, to observe NF-kappaB activity in HepG(2) before and after treatment of TNF-alpha by EMSA. RESULTS: The titer of AdIkappaBalphaM is 2 x 10(8) pfu/L, MOI equals to 20. AdIkappaBalphaM could be expressed stably and efficiently in HepG(2) and will not degrade by induction of TNF-alpha; but IkappaBalpha in the uninfection cell as well as AdIkappaBalpha control was increased at first and then decreased. EMSA demonstrated that the infected cells showed no activation of NF-kappaB before and after the treatment of TNF-alpha, but the cells of uninfected and infected with AdIkappaBalpha appeared excessive activation of NF-kappaB. CONCLUSION: AdIkappaBalphaM could be amplified in 293 cell and effectively infect to target cells HepG(2), and could be expressed stably in cells, and wouldn't be degrade with treatment of TNF-alpha, also it can effectually inhibit the excessive activation of NF-kappaB in HepG(2). The result of our research indicates the theoretical value of IkappaBalphaM as a super inhibitor, inhibition activity of NF-kappaB with IkappaBalphaM super-suppressor aided with routine anti-tumor therapy would become an effective method.