Enterotoxin-related genes PPFIA4 and SCN3B promote colorectal cancer development and progression.

To identify the role of enterotoxin-related genes in colorectal cancer (CRC) development and progression. Upregulated differentially expressed genes shared by three out of five Gene Expression Omnibus (GEO) data sets were included to screen the key enterotoxin-induced oncogenes (EIOGs) according to criteria oncogene definition, enrichment, and protein-protein interaction (PPI) network analysis, followed by prognosis survival, immune infiltration, and protential drugs analyses was performed via integration of RNA-sequencing data and The Cancer Genome Atlas-derived clinical profiles. We screened nine common key EIOGs from at least three GEO data sets. A Cox proportional hazards regression models verified that more alive cases, decreased overall survival, and highest 4-year survival prediction in CRC patients with high-risk score. Protein tyrosine phosphatase receptor type F polypeptide-interacting protein alpha-4 (PPFIA4), STY11, SCN3B, and SPTBN5 were shared in the same PPI network. Immune infiltration results showed that SCN3B and synaptotagmin 11 expression were obviously associated with B cell, macrophage, myeloid dendritic cell, neutrophils, and T cell CD4+ and CD8+ in both colon adenocarcinoma and rectal adenocarcinoma. CHIR-99021, MLN4924, and YK4-279 were identified as the potential drugs for treatment. Finally, upregulated EIOGs genes PPFIA4 and SCN3B were found in colon adenocarcinoma and PPFIA4 and SCN3B were proved to promote cell proliferation and migration in vitro. We demonstrated here that EIOGs promoting a malignancy phenotype was related with poor survival and prognosis in CRC, which might be served as novel therapeutic targets in CRC management.

[Study on the Conformation of Soybean Selenoprotein Solution with Spectroscopy Methods].

The structure characteristic of soybean selenoprotein and soy protein isolate (SPI) were investigated with fluorescence, ultraviolet and Fourier transform infrared (FTIR) spectrum. The unfolding process of two proteins was analyzed with fluorescence phase diagram method. The stability of emulsion properties and the influence of concentration, temperature and pH on the conformation of soy selenoproteins were also determined. The results indicated that the covalent disulfide bond of soybean selenoprotein molecules was damaged; the hydrogen bonding become weak; the hydrophobic interactions were enhanced and the protein chain molecules were extended. Soybean selenoprotein displayed only "folding" and "loose" state in solution, which illustrated soybean selenoprotein more tend to hydrolysis when compared with soybean protein. With temperature increasing, the fluorescence quenching effect occurred and the hydrophobicity of soy selenoproteins was also gradually increased, which reflected the protein molecules tends to be folded. In the range of pH 2.8～8.0, the Trp residue of soybean selenoprotein was mainly distributed in the polarity of the external environment and presented different conformational change on both sides of the isoelectric point under different pH value. In acidic environment, the soybean selenoprotein was easy to appear conformational transition from loose to fold. But it was conducive for soybean selenoprotein to existence in loose structure in alkaline conditions. In addition, the emulsifying properties of soybean selenoprotein were analyzed based on UV spectral data. Results showed that lower temperature helps to enhancement the emulsification but unfavorable the stability of the soybean selenoprotein.

Comparison of soluble dietary fibers from various quinoa microgreens: Structural characteristics and bioactive properties.

Quinoa (Chenopodium quinoa Willd.) microgreens are widely consumed as healthy vegetables around the world. Although soluble dietary fibers exist as the major bioactive macromolecules in quinoa microgreens, their structural characteristics and bioactive properties are still unclear. Therefore, the structural characteristics and bioactive properties of soluble dietary fibers from various quinoa microgreens (QMSDFs) were investigated in this study. The yields of QMSDFs ranged from 38.82 to 52.31 mg/g. Indeed, all QMSDFs were predominantly consisted of complex pectic-polysaccharides, e.g., homogalacturonan (HG) and rhamnogalacturonan I (RG I) pectic domains, with the molecular weights ranged from 2.405 × 104 to 5.538 × 104 Da. In addition, the proportions between RG I and HG pectic domains in all QMSDFs were estimated in the range of 1: 2.34-1: 4.73 (ratio of galacturonic acid/rhamnose). Furthermore, all QMSDFs exhibited marked in vitro antioxidant, antiglycation, prebiotic, and immunoregulatory effects, which may be partially correlated to their low molecular weights and low esterification degrees. These findings are helpful for revealing the structural and biological properties of QMSDFs, which can offer some new insights into further development of quinoa microgreens and related QMSDFs as value-added healthy products.

Physicochemical characteristics and biological activities of soluble dietary fibers isolated from the leaves of different quinoa cultivars.

Quinoa leaf is consumed as a promising value-added vegetable in the diet. Although quinoa leaf is rich in soluble dietary fibers, the knowledge regarding their chemical structures and biological activities is still limited, which astricts their application in the functional food industry. Thus, to improve the precise use and application of soluble dietary fibers (SDFs) isolated from quinoa leaves in the food industry, the physicochemical structures and bioactivities of SDFs isolated from different quinoa leaves were systematically investigated. Results indicated that quinoa leaves were rich in SDFs, ranging from 3.30 % to 4.55 % (w/w). Quinoa SDFs were mainly composed of acidic polysaccharides, such as homogalacturonan and rhamnogalacturonan I, which had the molecular weights in the range of 4.228 × 104 -7.059 × 104 Da. Besides, quinoa SDFs exerted potential in vitro antioxidant activities, lipid and bile acid-adsorption capacities, immunoregulatory activities, and prebiotic effects, which might be partially associated with their molecular mass, content of uronic acid, and content of bound polyphenol. Collectively, these findings are beneficial to better understanding the chemical structures and bioactivities of SDFs extracted from different quinoa leaves, which can also provide a scientific basis for developing quinoa SDFs into functional foods in the food industry.

2,2'-(Ethane-1,2-di-yl)bis-(1H-benzimidazole).

The complete mol-ecule of the title compound, C(16)H(14)N(4), is generated by crystallographic inversion symmetry. In the crystal, mol-ecules are linked by N-H⋯N hydrogen bonds, generating (001) sheets. Weak aromatic π-π stacking inter-actions [centroid-centroid distances = 3.7383 (13) and 3.7935 (14) Å] are also observed.

Oosporein from Tremella fuciformis.

THE TITLE COMPOUND [SYSTEMATIC NAME: 3,3',6,6'-tetra-hydroxy-4,4'-dimethyl-1,1'-bi(cyclo-hexa-3,6-diene)-2,2',5,5'-tetra-one], C(14)H(10)O(8), was isolated from Tremella fuciformis. The mol-ecule has 2 symmetry, with the mid-point of the C-C bond linking the cyclo-hexa-dienedione rings located on a twofold rotation axis. In the mol-ecule, the ring is approximately planar, with an r.m.s. deviation of 0.0093 Å, and the two rings make a dihedral angle of 67.89 (5)°. Inter-molecular O-H⋯O hydrogen bonding occurs in the crystal structure.

Poly[[µ-aqua-bis-(µ-4,4'-bipyridine-κN:N')bis-(µ-3-hy-droxy-adamantane-1-carboxyl-ato-κO:O')bis-(3-hy-droxy-adamantane-1-carboxyl-ato-κO)dicobalt(II)] hepta-hydrate].

The title coordination compound, {[Co(C(11)H(15)O(3))(4)(C(10)H(8)N(2))(2)(H(2)O)]·7H(2)O}(n), consists of a pair of Co(II) atoms, four 3-hy-droxy-adamantane-1-carboxyl-ate anions (L), one water mol-ecule, two bridging 4,4'-bipyridine (4,4'-bpy) ligands and seven uncoordinated water mol-ecules. Both of the Co(II) ions are coordinated in a distorted octa-hedral geometry. Four L ligands bind to each pair of Co(II) atoms in a plane, two of which bridge the two Co(II) atoms as bidentate groups while the other two coordinate to a single Co(II) atom in a monodentate mode. Two half-mol-ecules of 4,4'-bipyridine coordinate the Co(II) atoms from the upside and underside. The packing features extensice O-H⋯O hydrogen bonding.

[Establish efficiently transformation system of the Isatis indigotica].

OBJECTIVE: To build up a method of efficiently transforming Isatis indigotica with the Agrobacterium. METHODS: Two kinds of Agrobocterium: ATCC 15834 and RI1601 were used to treat different fraction of Isatis indigotica. Explored the effect of cocultured, different antibiotic concentration and the media on multiply the hairy roots. RESULTS: The explants with out coculture had had higher survival rate rooting rate and earlier sprout time. After ultrasonic treatment of plant, the indacement rate was two times than that of untreated one; The best antibiotic concentration was 400 mg/L; The proportion of the grow speed of the hairy root in the liquid culture media was 2 - 3 times than that of solid culture media, and 37 times of common roots. CONCLUSION: The method is useful for establishing an efficiently transformatiem system of Isatis indigotica by Agrobacterium.

Noni (Morinda citrifolia L.) Fruit Polysaccharides Regulated IBD Mice Via Targeting Gut Microbiota: Association of JNK/ERK/NF-κB Signaling Pathways.

Inflammatory bowel disease (IBD) is a disease characterized by intestinal inflammation with immune dysregulation and intestinal microecological imbalance. In a dextran sulfate sodium salt (DSS)-induced IBD mouse model, noni (Morinda citrifolia L.) fruit polysaccharides (NFP) with homogalacturonan and rhamnogalacturonan-I domain decreased the concentration of serum LPS, TNF-α, and IL-17 by 84, 42, and 65%, respectively. It was abolished when intestinal microbiota were depleted by antibiotics. Sequencing analysis of gut microbiota showed an attenuated disruption of the microbial composition in the DSS+NFP group. Targeted metabolomic analysis revealed that NFP upregulated the content of acetic acid, propionic acid, and butyric acid by onefold but reduced isobutyric acid and isovaleric acid contents. NFP also inhibited JNK, ERK, and NF-κB phosphorylation of IBD mice. Taken together, the mechanism of NFP alleviating IBD is related to the intestinal microecological balance to inhibit inflammatory signaling pathways. This study provides a basis for NFP as a cheap intervention for the prevention and treatment of IBD patients.

Low-grade fever during COVID-19 convalescence: A report of 3 cases.

BACKGROUND: Low-grade fever during convalescence is an atypical symptom of coronavirus disease 2019 (COVID-19). Reports of such cases are rare, and the mechanism and outcome of low-grade fever during COVID-19 convalescence are not completely clear. We report 3 cases with low-grade fever during COVID-19 convalescence and highlight the main clinical, radiographic, and laboratory characteristics, thereby increasing the level of expertise in the clinical management of COVID-19 during convalescence and facilitating individualized decision-making. CASE SUMMARY: We describe 3 patients with COVID-19, two females aged 62 and 66 years and a male 55 years, who had low-grade fever during COVID-19 convalescence. All 3 patients had no other discomfort or comorbidities during low-grade process. Lesions on computed tomography in all 3 patients had resolved during this period. Two patients tested negative on two consecutive severe acute respiratory syndrome coronavirus 2 tests with an interval of at least 24 h between tests. Body temperature in all 3 patients returned to normal after several days without treatment, and fever recurrence was not observed. CONCLUSION: Enhancing the knowledge of low-grade fever during COVID-19 convalescence may increase the expertise in the delivery of optimal healthcare services.

[Experimental study on staphylococcal enterotoxin promoting tendon-bone healing after reconstruction of anterior cruciate ligament in rabbits].

OBJECTIVE: To explore highly agglutinative staphylococcin (HAS) pomoting on tendon-bone healing after reconstruction of anterior cruciate ligament(ACL) in rabbits. METHODS: Animal model of ACL reconstruction in 24 mature New Zealand white rabbits(3 months, 2.56 kg on average, either gender) were established using autologous digital long extensor tendon and randomly classified into 2 groups(HAS and control group), 12 rabbits for each group. HAS group was separately injected 0.1ml highly agglutinative staphylococcin immediately into tendon-bone interface during the operation and 2 days after operation. Control group was injected with the same dose of physiological saline for 3 days. All animals were sacrificed at 4, 8, and 12 weeks after operation for histological examinations. The specimens were stained with hematoxylin-eosin, picric acid-sirius red, VEGF immunohistochemistry stain, and toluidine blue to histologically analysis the total pathology changes of the tendon-bone healing tissue, the tendon bone interface morphology classification, hyperplasia and arrangement of collagen fiber, vascularization and new bone formation, respectively. RESULTS: The Yamakado morphological interface results showed that the tissue healing at tendon-bone interface of the HAS group was better than that of the control group. The histological examination revealed that on the 4th week after operation, the tendon-bone interface of HAS group was filled with fibrous connective tissue. The proliferated fibroblasts, chondroblasts and the angiogenesis were rich. On the 8th week after operation, the healing tissue at the bone-tendon interface had developed into dense connective tissue, the neo-vessels were very rich, the collagen fibers were formed abundantly, some Sharpey's fibers spanning parts of the tendon-bone interface. On the 12th week after operation, the transition zones were full of Sharpey's fibers;the neo-vessels were not as much as the 8th weeks, but new bone formation was further increased and immature fibrocartilage appeared. For quantitative histological analysis at 4, 8 and 12 weeks after operation, the proportion of neo-vessel area and the area of now bone formation of the HAS group were all significantly higher than those of the control group(P<0.05). CONCLUSIONS: Under the synergy of staphylococcal enterotoxin C and other active ingredients, Highly Agglutinative Staphylococcin can significantly improve the tendon-bone healing after ACL reconstruction in rabbit knees, which is expected to be a new method to improve the clinical results of ACL reconstruction.

Effects of cyclooxygenase-2 antisense vector on proliferation of human cholangiocarcinoma cells.

OBJECTIVE: To transfect antisense vector of human cyclooxygenase-2 (COX-2) gene into COX-2 highly expressing cholangiocarcinoma cell line QBC939 and explore its biological activities and role in carcinogenesis. METHODS: QBC939 cells were transfected with antisense vector of human COX-2 gene using LipoVec transfecting technique. Transfected cells were selected with G418; COX-2 mRNA was examined using reverse transcription polymerase chain reaction (RT-PCR) and COX-2 protein expression was detected by immunocytochemistry using isozyme selective antibodies. The proliferative status of transfected cells was measured by using methabenzthiazuron (MTT) assay; Cell cycle and apoptosis were analyzed by using flow cytometry. RESULTS: RT-PCR showed a lower COX-2 mRNA level in antisense vector transfected cells and immunocytochemistry showed a weaker COX-2 protein expression in antisense vector transfected cells. The antisense vector transfected cells proliferative index decreased significantly (P < 0.01), the percentage of S phase decreased remarkably (P < 0.05) in antisense vector transfected cells (9.27% +/- 1.91%) compared with that in QBC939 cells without transfection(16.35% +/- 2.87%), and the percentage of G0/G1 phase increased remarkably (P < 0.05) in antisense vector transfected cells (75.16% +/- 4.13%) compared with that in QBC939 cells without transfection (57.31% +/- 10.16%). Transfection with antisense vector of human COX-2 gene had no significant influence on the apoptosis in QBC939 cells (P > 0.05). CONCLUSION: Transfection with antisense vector of human COX-2 gene could inhibit the proliferation of human cholangiocarcinoma QBC939 cells.

High level expression of bikunin in Pichia pastoris by fusion of human serum albumin.

Bikunin is a proteoglycan exhibiting broad-spectrum inhibitory activity against serine proteases and could potentially suppress tumor cell invasion and metastasis. Here, we have successfully expressed recombinant human bikunin (rh-bikunin) in Pichia pastoris and also established the purification procedure. Different fusion genes of h-UTI and domain I, domain I and domain II, domain I, domain II and domain III of human serum albumin (HSA) were inserted into expression vector pPICZαA. After expressed in shake flask, rh-bikunin was produced in an 30-L fermenter and purified by affinity chromatography and cation exchange chromatography. The final expression levels were 200 mg/L and we got totally 1.08 g (3650 IU/mg) of active purified rh-bikunin (purity is 98%) from 20 L of fermentation broth. The rh-bikunin consists of unique form with molecular masses of 25 kDa, and has the same N-terminals sequence as human native bikunin. This study provided a new method for high level expression of active rh-bikunin by using HSA as fusion parter.