Hyperthermia promotes degradation of the acute promyelocytic leukemia driver oncoprotein ZBTB16/RARα.

The acute promyelocytic leukemia (APL) driver ZBTB16/RARα is generated by the t(11;17) (q23;q21) chromosomal translocation, which is resistant to combined treatment of all-trans retinoic acid (ATRA) and arsenic trioxide (ATO) or conventional chemotherapy, resulting in extremely low survival rates. In the current study, we investigated the effects of hyperthermia on the oncogenic fusion ZBTB16/RARα protein to explore a potential therapeutic approach for this variant APL. We showed that Z/R fusion protein expressed in HeLa cells was resistant to ATO, ATRA, and conventional chemotherapeutic agents. However, mild hyperthermia (42 °C) rapidly destabilized the ZBTB16/RARα fusion protein expressed in HeLa, 293T, and OCI-AML3 cells, followed by robust ubiquitination and proteasomal degradation. In contrast, hyperthermia did not affect the normal (i.e., unfused) ZBTB16 and RARα proteins, suggesting a specific thermal sensitivity of the ZBTB16/RARα fusion protein. Importantly, we found that the destabilization of ZBTB16/RARα was the initial step for oncogenic fusion protein degradation by hyperthermia, which could be blocked by deletion of nuclear receptor corepressor (NCoR) binding sites or knockdown of NCoRs. Furthermore, SIAH2 was identified as the E3 ligase participating in hyperthermia-induced ubiquitination of ZBTB16/RARα. In short, these results demonstrate that hyperthermia could effectively destabilize and subsequently degrade the ZBTB16/RARα fusion protein in an NCoR-dependent manner, suggesting a thermal-based therapeutic strategy that may improve the outcome in refractory ZBTB16/RARα-driven APL patients in the clinic.

Diffusion Tensor Imaging Evaluates Effects of Acupoint Injection at Zusanli (ST36) for Type 2 Diabetic Peripheral Neuropathy.

BACKGROUND Acupoint injection is a therapeutic method that combines acupuncture and Western medicine and shows good curative effects for neuropathies. This study aimed to explore the efficacy of acupoint injection for treating diabetic peripheral neuropathy (DPN) by magnetic resonance neuroimaging (MRN). MATERIAL AND METHODS Forty patients with DPN were randomly divided into an acupoint injection group (AI; n=20) and intramuscular injection group (MI; n=20). The AI group received an acupoint injection of mecobalamin at acupoint Zusanli (S36); the MI group received intramuscular injection of mecobalamin. The curative effect was evaluated by the Toronto Clinical Neuropathy Score and diffusion tensor imaging (DTI). RESULTS The neuropathy scores of both groups decreased from baseline (AI 9.31±2.36; MI 9.34±2.54) to after the 2-week treatment (AI 7.12±1.87; MI 7.86±2.11); the differences were not significant. The fractional anisotropy (FA) value showed significant differences on the common peroneal nerve (AI 0.36±0.04; MI 0.31±0.05; P<0.05) and tibial nerve (AI 0.38±0.07; MI 0.34±0.06; P<0.05) after treatment. Likewise, apparent diffusion coefficient (ADC) values between groups showed significant differences for the common peroneal nerve (AI 1.44±0.17×10⁻³ mm²/s; MI 1.61±0.20×10⁻³ mm²/s; P<0.05) and tibial nerve (AI 1.54±0.22×10-3 mm²/s; MI 1.60±0.17 10⁻³ mm²/s; P<0.05). CONCLUSIONS Patients with DPN showed lower nerve FA and higher ADC in DTI-MRN. The acupoint injection of mecobalamin could treat DPN and repair the damaged nerves, which was shown by elevated FA and lowered ADC. Our study provides clinical evidence for the application of acupoint injection therapy and the evaluation of DPN by MRN.

Effects of atorvastatin and Zishen Qingqi granules on immune function and liver function of patients with systemic lupus erythematosus with mild and moderate activity.

To explore the effect of atorvastatin combined with Zishen Qingqi granules on the immune function and liver function of patients with mild to moderate activity systemic lupus erythematosus. The data of 120 patients with mild to moderate activity systemic lupus erythematosus admitted to our hospital from February 2019 to February 2020 were retrospectively analyzed and they were divided into experimental group (n=60) and the control group (n=60) according to the order of admission; the control group was treated with atorvastatin, and the experimental group was treated with Zishen Qingqi granules plus. The immune function, liver function, TCM syndrome score and systemic lupus erythematosus disease activity index (SLEDAI) were compared between the two groups. The experimental group after treatment was superior to the control group with respect to immune function indexes, liver function indexes, SLEDAI and TCM syndromes (all P<0.001). Atorvastatin combined with Zishen Qingqi granules can improve the liver function of patients with mild to moderate activity systemic lupus erythematosus, enhance their immunity, and relieve their clinical symptoms.

Functional regulation of an ancestral RAG transposon ProtoRAG by a trans-acting factor YY1 in lancelet.

The discovery of ancestral RAG transposons in early deuterostomia reveals the origin of vertebrate V(D)J recombination. Here, we analyze the functional regulation of a RAG transposon, ProtoRAG, in lancelet. We find that a specific interaction between the cis-acting element within the TIR sequences of ProtoRAG and a trans-acting factor, lancelet YY1-like (bbYY1), is important for the transcriptional regulation of lancelet RAG-like genes (bbRAG1L and bbRAG2L). Mechanistically, bbYY1 suppresses the transposition of ProtoRAG; meanwhile, bbYY1 promotes host DNA rejoins (HDJ) and TIR-TIR joints (TTJ) after TIR-dependent excision by facilitating the binding of bbRAG1L/2 L to TIR-containing DNA, and by interacting with the bbRAG1L/2 L complex. Our data thus suggest that bbYY1 has dual functions in fine-tuning the activity of ProtoRAG and maintaining the genome stability of the host.

Effects of Phytosterol Ester on the Fatty Acid Profiles in Rats with Nonalcoholic Fatty Liver Disease.

Both serum and hepatic fatty acid (FA) compositions differ among nonalcoholic hepatic steatosis, nonalcoholic steatohepatitis, and healthy subjects. The severity of the above liver disease is closely associated with the concentration and composition of FAs. Our previous study found that phytosterol ester (PSE) could alleviate hepatic steatosis in nonalcoholic fatty liver disease rats. The aims of this work were to explore the effects of PSE (0.05/100 g·body weight) on FA profiles and the mRNA levels of FA metabolism-related genes. Compared with a high-fat diet alone group, PSE treatment significantly decreased hepatic saturated fatty acid levels (P < .05) and increased monounsaturated fatty acid (especially C16:1 n-7) levels in the liver, serum, and adipose tissue and polyunsaturated fatty acid levels in the serum and liver (P < .05) after 12 weeks of intervention. In particular, PSE treatment increased the level of C22:5 n-3, an FA that was negatively correlated with the degree of hepatic steatosis in the serum, liver, and adipose tissue. The increases in some unsaturated fatty acids are probably related to the upregulation of stearoyl-coenzyme A desaturase-1 and fatty acid desaturase-1.

Geographical origin of Chinese wolfberry (goji) determined by carbon isotope analysis of specific volatile compounds.

Chinese wolfberry or goji berry (Lycium barbarum) is an important traditional Chinese medicine. Its price and function has a close correlation with its geographical provenance. Illegal mislabeling motivated by commercial gains brings serious food safety problems and damages consumer confidence. In this work, a novel analytical strategy combined with chemometrics statistic tools was developed to determine the geographical origin of wolfberries from different provinces in China. Stable carbon isotopic ratios (δ13C) of wolfberry volatile compounds (i.e. limonene, tetramethylpyrazine, safranal, geranylacetone, and ß-ionone) were determined by gas chromatography-combustion-isotope ratio mass spectrometry (GC-IRMS) with headspace-solid phase micro extraction (HS-SPME). Five types of SPME fiber (i.e. DVB/CAR/PDMS, CAR/PDMS, PDMS/DVB + OC, PDMS, and PA), extraction time, temperature and GC-IRMS conditions were comprehensively optimized to obtain the best adsorption of volatile compounds in wolfberry. Method integrity was assessed by comparing volatiles extracted using HS-SPME GC-IRMS with direct injection GC-IRMS and were in good agreement with each other. The geographical variations of volatile compounds using one-way analysis of variance (ANOVA) were explored for individual δ13C values in wolfberry samples from Gansu, Ningxia and Qinghai. Geographical origin of wolfberry was differentiated by linear discrimination analysis (LDA), with an accuracy of 89.16%, 87.77% and 85.87% for these three provinces, respectively. These results showed the combination of SPME and IRMS provides a rapid and valid method to determine the geographical origin of wolfberry.

Soil Microbial Community Structure and Diversity around the Aging Oil Sludge in Yellow River Delta as Determined by High-Throughput Sequencing.

Microorganisms are sensitive indicators of edaphic environmental variation. The Illumina MiSeq sequencing technology was used to analyze soil bacterial community diversity around an aging oil sludge in the Yellow River Delta. The alpha diversity index of soil bacterial community results (Ace, Chao, Shannon, and Simpson) determined that bacterial community diversity sampling within the scope of a 20 cm radius from the center of an aging oil sludge spot showed the most abundant diversity. The level of diversity distributed symmetrically with radial direction from the center of the aging oil sludge spot. Over the distance of 100 m from the center, bacterial community diversity tends to be monotonous, with small differences especially in the horizontal direction underground. The alpha-diversity indicators also showed that the bacterial diversity of samples were close under the aging oil sludge. In addition, the aging oil sludge inhibited the growth of bacteria compared with the referenced unpolluted soil sample and also increased the diversities of soil bacteria. At the phylum level, the Proteobacteria, Chloroflexi, and Actinobacteria existing in the aging oil sludge-contaminated wetland soil constituted a larger proportion of the community, while the proportion of Firmicute was relatively less. On the contrary, Firmicute showed the highest content of 63.8% in the referenced soil. Under the genus level and family level, the corresponding strains that resisted the aging oil sludge were selected. According to the bacterial diversity analysis, the basic structure of the bacterial community which could be used for remediation of aging oil sludge-contaminated soil was also developed.

Changes in soil microbial community structure and function after afforestation depend on species and age: Case study in a subtropical alluvial island.

It is well established that land use change can have a profound impact on soil physicochemical properties but the associated changes in soil microbial communities are poorly understood. We used long-term research sites in a subtropical alluvial island of eastern China to measure changes in soil physicochemical properties and microbial community abundance and composition (via phospholipid fatty acid (PLFA) analysis) and function (via extracellular enzyme activity) across different land use types developed on the same soil matrix, including a camphor (Cinnamomum camphora) plantation, a chronosequence of differently aged dawn redwood (Metasequoia glyptostroboides) plantings, a deforested land and a rice paddy. We hypothesized that afforestation could improve soil quality by enhancing carbon (C) and nitrogen (N) contents, microbial biomass and enzyme activities, but that this effect would vary depending on forest age and tree species. Soil C and N concentrations, PLFA abundances and activities of decomposition enzymes (ß-glucosidase, urease, alkaline phosphatase and catalase) in older plantations all increased significantly compared to cropland. These variables changed little or decreased in deforested land compared to cropland. These variables also increased with planting age in the dawn redwood plantings. Soils under camphor plantations had higher soil nutrient contents, microbial biomass and lower enzyme activities than dawn redwood soils with similar age. We also found some significant relationships between soil chemical and biological properties: PLFA abundances were positively related to soil organic matter (SOM) contents; the fungal-to-bacterial ratio and fungal relative abundance were correlated positively with SOM contents and negatively with C/N ratio; both soil PLFA abundances and enzyme activities were positively linked with soil inorganic N content and potential net N mineralization rate; ratio of specific C, N and P (phosphorus) acquisition activities was limited to 10: 1: 10 across land use types. Our study underscores the fact that land use type can have a profound impact on soil microbial communities; in addition, tree species and planting age also play significant roles in afforestation.

Effect of frying and aluminium on the levels and migration of parent and oxygenated PAHs in a popular Chinese fried bread youtiao.

This study investigated the effects of frying conditions on the change, migration and correlation of polycyclic aromatic hydrocarbons (PAHs) and oxygenated PAHs (oxy-PAHs) in youtiao, a typical Chinese fried bread. PAH migration was evaluated using an isotope tracer method. High concentrations of oxy-PAHs, especially 9-fluorenone (1.75µg/kg) and anthracene-9,10-dione (6.41µg/kg), were detected. The results revealed that 1) prolonged continuous frying decreased parent PAH and oxy-PAH concentrations, 2) frying temperature and aluminium content did not have significant effects on parent PAH concentrations, 3) PAH migration rates (2.02-18.48%) varied significantly based on the frying oil type, and 4) the sum of five oxy-PAH concentrations was significantly correlated with the sum of 16 parent PAH concentrations in soybean oil-fried youtiao. Three oxy-PAHs (9-fluorenone, anthracene-9,10-dione, and 9,10-dihydrobenzo[a]pyren-7(8h)-one) were significantly correlated with their respective parent PAHs. The results might be helpful for further study and evaluation on oxy-PAHs and PAHs in fried foods.

A novel alkali extractable polysaccharide from Plantago asiatic L. Seeds and its radical-scavenging and bile acid-binding activities.

A new acidic polysaccharide (PLP) was isolated and characterized from Plantago asiatic L. seeds by hot alkali extraction and chromatographic purification using DEAE cellulose and Sephacryl S-400 columns. PLP has a molecular weight of 1.15 × 10(6) Da, and a monosaccharide composition of xylose (Xyl), arabinose (Ara), glucuronic acid (GlcA), and galactose (Gal) in a molar ratio of 18.8:7.2:6.1:1. The results of methylation analysis, FT-IR, and 1D and 2D NMR indicated that PLP was a highly branched heteroxylan of ß-1,4-linked Xylp backbone with three α-GlcAp-(1→3)-Araf attached to the O-3 position and one α-T-linked-GlcAp and one α-Araf-(1→5)-Araf attached to the O-2 position every eight monosaccharide residues. PLP exhibited scavenging abilities against hydroxyl, peroxyl anion, and DPPH radicals in vitro and showed significant binding capacities against cholic and chenodeoxycholic acids, suggesting its possible cholesterol-lowering activity. The results demonstrated the potential use of PLP in functional foods and nutraceuticals.

Characterization of a novel alkali-soluble heteropolysaccharide from tetraploid Gynostemma pentaphyllum Makino and its potential anti-inflammatory and antioxidant properties.

A novel polysaccharide (GPP-S), with a molecular mass of 1.2 × 10(6) Da, was isolated from the tetraploid Gynostemma pentaphyllum Makino by alkali extraction followed by purifications using DEAE and Sephacryl S-400 column chromatographies. The monosaccharide composition of GPP-S was determined as rhamnose, arabinose, glucose, and galactose with a molar ratio of 1.00:3.72:19.49:7.82. The structural analysis suggested that the backbone of GPP-S is (1→4)-linked-glucose and (1→6)-linked-galactose with a (1→4,6)-linked-glucose branch every six monosaccharide residues. The terminals were 1-)-α-arabinose, glucuronic acid, and other monosaccharides. GPP-S exhibited scavenging capacities against hydroxyl, peroxyl, and DPPH(•) radicals in vitro. GPP-S also had inhibitory activities on IL-1ß, IL-6, and COX-2 gene expressions in RAW 264.7 mouse macrophage cells. These results suggested that GPP-S could be developed as a bioactive ingredient for functional foods and dietary supplements.

Interaction between resveratrol and thrombin and its biological implication.

In view of important implication of resveratrol and thrombin in the process of platelet aggregation and apoptosis, the interaction effect between them and its biological implication was studied. Resveratrol could decrease fluorescence emission intensities of thrombin by a dynamic collision quenching process. The binding process was spontaneous and the resveratrol-thrombin complex formation was an endothermal reaction induced by hydrophobic interaction and hydrogen. Non-radiation energy transfer occurred from thrombin to resveratrol at the molecular level. Kinetic studies showed that the fluorescence intensity of thrombin linearly decreased along with prolonged time, whereas resveratrol intensity reduced continuously with degressive plateau in the form of second exponential decay function. Moreover, the interaction caused a decrease of α-helical structure resulting in thrombin conformation alteration. In addition, the isomerization of resveratrol was not prevented but its stability was improved with the addition of thrombin. This work might give deeper insight into resveratrol as a kind of nutritional factor with the inhibition of platelet aggregation and stimulation of platelet apoptosis in the functional food.

Heat shock protein 90 inhibition results in altered downstream signaling of mutant KIT and exerts synergistic effects on Kasumi-1 cells when combining with histone deacetylase inhibitor.

KIT mutations may be associated with a poor prognosis in t(8;21) AML. Heat shock protein 90 (Hsp90) is a molecular chaperone frequently used by cancer cells to stabilize mutant oncoproteins. Inhibition of Hsp90 by 17-allylamino-17-demethoxygeldanamycin (17-AAG) disrupted downstream signaling pathways of mutant KIT in Kasumi-1 cells. AML1-ETO fusion gene and mutated KIT act as "two-hit" factors in Kasumi-1 cells. Histone deacetylation (HDAC) inhibitors sodium phenylbutyrate (PB) and valproic acid (VPA) block AML1-ETO. Co-treatment with 17-AAG and PB or 17-AAG and VPA resulted in a synergistic effect in Kasumi-1 cells. Our results confirmed that Hsp90 and mutated KIT were valid molecular targets in the therapy of AML.

Differential metabolic responses of perennial grass Cynodon transvaalensis×Cynodon dactylon (C4) and Poa Pratensis (C3) to heat stress.

Differential metabolic responses to heat stress may be associated with variations in heat tolerance between cool-season (C3) and warm-season (C4) perennial grass species. The main objective of this study was to identify metabolites associated with differential heat tolerance between C4 bermudagrass and C3 Kentucky bluegrass by performing metabolite profile analysis using gas chromatography-mass spectrometry. Plants of Kentucky bluegrass (Poa Pratensis'Midnight') and hybrid bermudagrass (Cynodon transvaalensis x Cynodon dactylon'Tifdwarf') were grown under optimum temperature conditions (20/15 °C for Kentucky bluegrass and 30/25 °C for bermudagrass) or heat stress (35/30 °C for Kentucky bluegrass and 45/40 °C for bermudagrass). Physiological responses to heat stress were evaluated by visual rating of grass quality, measuring photochemical efficiency (variable fluorescence to maximal fluorescence) and electrolyte leakage. All of these parameters indicated that bermudagrass exhibited better heat tolerance than Kentucky bluegrass. The metabolite analysis of leaf polar extracts revealed 36 heat-responsive metabolites identified in both grass species, mainly consisting of organic acids, amino acids, sugars and sugar alcohols. Most metabolites showed higher accumulation in bermudagrass compared with Kentucky bluegrass, especially following long-term (18 days) heat stress. The differentially accumulated metabolites included seven sugars (sucrose, fructose, galactose, floridoside, melibiose, maltose and xylose), a sugar alcohol (inositol), six organic acids (malic acid, citric acid, threonic acid, galacturonic acid, isocitric acid and methyl malonic acid) and nine amino acids (Asn, Ala, Val, Thr, γ-aminobutyric acid, IIe, Gly, Lys and Met). The differential accumulation of those metabolites could be associated with the differential heat tolerance between C3 Kentucky bluegrass and C4 bermudagrass.

[The effect of electro-acupuncture on endogenous EPCs and serum cytokines in cerebral ischemia-reperfusion rat].

In this research project, rats were made into animal models of acute focal cerebral ischemia and reperfusion (IR) by occlusion of their middle cerebral artery (MCAO). We observed the effect of endogenous endothelial progenitor cells (EPCs) and serum cytokines on cerebral ischemia rats treated by electro-acupuncture(EA). The results showed: MCAO model had high stability after EA treatment which was delivered via the acupuncture needles inserted into "quchi" and "zusanli" points, the nervous functions of cerebral IR rats recovered faster than those of rats not treated; EPCs in rats' blood increased after acute focal cerebral ischemia and reperfusion; and the growth rate was obvious in IR group. This phenomenon might be related to the inflammation elicited by injury of ischemia and self-repair. Besides, EA treatment could decrease induced nitric oxide synthase (iNOS) activity, alleviate injury after cerebral ischemia, and regulate the quantity of EPCs in blood. The quantity of EPCs in blood increased in IR-24hr. In IR-48 hr, the rise of EPCs quantity was significant (P < 0.01). The level of vascular endothelium growth factor (VEGF) in serum of rats after cerebral ischemia was escalated, which indicated to a certain extent that cerebral ischemia could stimulate stress reaction. EA treatment could raise VEGF level, which suggested that high expression of VEGF could accelerate mobilization, chemotaxis and homing of EPCs. At the same time, the levels of matrix metalloproteinase-9 (MMP-9) and basic fibroblast growth factor (bFGF) also changed. In conclusion, EA treatment could promote neovascularization after cerebral ischemia by mobilizing EPCs, decreasing iNOS activity and increasing VEGF level. This may be one of the ways by which EA could treat cerebral ischemia.

[Effect of electroacupuncture on plasma endogenous endothelial progenitor cell counts in cerebral ischemia-reperfusion rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on peripheral blood endothelial progenitor cell (EPC) counts, vascular endothelial growth factor (VEGF) level and total nitric oxide synthase (TNOS) and inducible nitric oxide synthase (iNOS) activity in cerebral ischemia-reperfusion injury (CI/RI) rats. METHODS: A total of 72 male rats were randomly and evenly assigned to normal control, sham-operation (sham), model and EA groups which were further divided into 24 h, 48 h and 72 h subgroups, with 6 cases in each. Acute focal cerebral ischemia model was established by occlusion of the middle cerebral artery (MCAO, 120 min) and reperfusion. EA (2/15 Hz, 1 mA) was applied to "Quchi" (LI 11) and "Zusanli" (ST 36) for 30 min, once daily. Peripheral blood was collected from abdominal aorta for detecting EPC count by using flow cytometry, serum VEGF level by using enzyme-linked immunosorbent assay (ELISA), and serum TNOS and iNOS activity by spectrophotometry, respectively. RESULTS: Compared with the corresponding normal control subgroups, blood EPC counts, serum TNOS and INOS activity and serum VEGF content at 24 h, INOS activity and VEGF level at 48 h, and EPCs and INOS at 72 h in model subgroups all increased significantly (P < 0.01, P < 0.05). In comparison with the corresponding model subgroups, EPC count at 24 h and 72 h, and TNOS activity at 24 h in EA subgroups decreased considerably (P < 0.01, P < 0.05); while EPC and VEGF levels at 48 h in EA subgroup increased evidently (P < 0.05, P < 0.01). No significant differences were found among normal, sham, model and EA subgroups in serum TNOS activity at 48 h and 72 h (P > 0.05). CONCLUSION: EA of LI 11 and ST 36 can suppress CI/ RI induced increase of blood EPC count and serum TNOS activity, and upregulate serum VEGF level, which may contribute to its effect in relieving CI/RI.

Tapetum degeneration retardation is critical for aliphatic metabolism and gene regulation during rice pollen development.

As a complex wall system in flowering plants, the pollen outer wall mainly contains aliphatic sporopollenin; however, the mechanism for synthesizing these lipidic precursors during pollen development remains less well understood. Here, we report on the function of the rice tapetum-expressing TDR (Tapetum Degeneration Retardation) gene in aliphatic metabolism and its regulatory role during rice pollen development. The observations of transmission electron microscopy (TEM) and scanning electron microscopy (SEM) analyses suggested that pollen wall formation was significantly altered in the tdr mutant. The contents of aliphatic compositions of anther were greatly changed in the tdr mutant revealed by GC-MS (gas chromatography-mass spectrometry) testing, particularly less accumulated in fatty acids, primary alcohols, alkanes and alkenes, and an abnormal increase in secondary alcohols with carbon lengths from C29 to C35 in tdr. Microarray data revealed that a group of genes putatively involved in lipid transport and metabolism were significantly altered in the tdr mutant, indicating the critical role of TDR in the formation of the pollen wall. Also, a wide range of genes (236 in total-154 up-regulated and 82 down-regulated) exhibited statistically significant expressional differences between wild-type and tdr. In addition to its function in promoting tapetum PCD, TDR possibly plays crucial regulatory roles in several basic biological processes during rice pollen development.