Gut microbiota dynamics and fecal SCFAs after colonoscopy: accelerating microbiome stabilization by Clostridium butyricum.

BACKGROUND: Colonoscopy is a classic diagnostic method with possible complications including abdominal pain and diarrhoea. In this study, gut microbiota dynamics and related metabolic products during and after colonoscopy were explored to accelerate gut microbiome balance through probiotics. METHODS: The gut microbiota and fecal short-chain fatty acids (SCFAs) were analyzed in four healthy subjects before and after colonoscopy, along with seven individuals supplemented with Clostridium butyricum. We employed 16S rRNA sequencing and GC-MS to investigate these changes. We also conducted bioinformatic analysis to explore the buk gene, encoding butyrate kinase, across C. butyricum strains from the human gut. RESULTS: The gut microbiota and fecal short-chain fatty acids (SCFAs) of four healthy subjects were recovered on the 7th day after colonoscopy. We found that Clostridium and other bacteria might have efficient butyric acid production through bioinformatic analysis of the buk and assessment of the transcriptional level of the buk. Supplementation of seven healthy subjects with Clostridium butyricum after colonoscopy resulted in a quicker recovery and stabilization of gut microbiota and fecal SCFAs on the third day. CONCLUSION: We suggest that supplementation of Clostridium butyricum after colonoscopy should be considered in future routine clinical practice.

Occurrence of fungi and mycotoxins in herbal medicines and rapid detection of toxin-producing fungi.

Contamination from external hazardous materials may greatly influence the safety and efficacy of herbal medicines. This paper aimed to evaluate the levels of contamination by mycotoxins and toxigenic fungi in herbal medicines and establish a rapid method for detecting toxin-producing fungi. Herein, 62.92%, 36.25%, and 64.17% of herbal medicines were contaminated by aflatoxins (AFs), ochratoxins, and fumonisins, respectively. Aspergillus (43.77%), Fusarium (5.17%), and Cladosporium (4.46%) were the three predominant genera. Spearman's correlation results showed that Aspergillus and Fusarium were significantly and positively correlated with mycotoxin content (R > 0.5, P < 0.05). In addition, 323 fungal strains were isolated from herbal medicines, and 20 species were identified, mainly belonging to Aspergillus and Penicillium. Analysis of potential mycotoxin-producing fungi showed that Aspergillus flavus can produce AFs, and Aspergillus ochraceus and Aspergillus niger can produce ochratoxin A (OTA). Multiplex real-time polymerase chain reaction showed that A. flavus harbored AF synthesis genes (aflR), and A. ochraceus and A. niger harbored OTA synthesis genes (aoksl). With these synthesis genes, 67.07% and 37.20% of 164 herbal medicines were positive for toxigenic genes. Furthermore, an excellent correlation was found between the above gene copies and mycotoxin content (R2 = 0.99). Our results confirmed the high detection rate of mycotoxins in herbal medicines and identified pivotal AF- and OTA-producing fungi. In conclusion, this paper provided the contamination status of fungi and mycotoxins in herbal medicines and established a rapid method for detecting toxigenic fungi.

Biosyntheses characterization of alkaloids and flavonoids in Sophora flavescens by combining metabolome and transcriptome.

Sophora flavescens are widely used for their pharmacological effects. As its main pharmacological components, alkaloids and flavonoids are distributed in the root tissues wherein molecular mechanisms remain elusive. In this study, metabolite profiles are analyzed using metabolomes to obtain biomarkers detected in different root tissues. These biomarkers include alkaloids, phenylpropanoids, and flavonoids. The high-performance liquid chromatography analysis results indicate the differences in principal component contents. Oxymatrine, sophoridine, and matrine contents are the highest in the phloem, whereas trifolirhizin, maackiain, and kushenol I contents are the highest in the xylem. The transcript expression profiles also show tissue specificity in the roots. A total of 52 and 39 transcripts involved in alkaloid and flavonoid syntheses are found, respectively. Among them, the expression levels of LYSA1, LYSA2, AO2, AO6, PMT1, PMT17, PMT34, and PMT35 transcripts are highly and positively correlated with alkaloids contents. The expression levels of 4CL1, 4CL3, 4CL12, CHI5, CHI7, and CHI9 transcripts are markedly and positively correlated with flavonoids contents. Moreover, the quantitative profiles of alkaloids and flavonoids are provided, and the pivotal genes regulating their distribution in S. flavescens are determined. These results contribute to the existing data for the genetic improvement and target breeding of S. flavescens.

Transcriptome and Gene Editing Analyses Reveal MOF1a Defect Alters the Expression of Genes Associated with Tapetum Development and Chromosome Behavior at Meiosis Stage Resulting in Low Pollen Fertility of Tetraploid Rice.

Autotetraploid rice is a useful rice germplasm for polyploid rice breeding. However, low fertility limits its commercial production. A neo-tetraploid rice with high fertility was developed from the progenies of crossing between autotetraploid lines by our research group. Our previous study showed that a myeloblastosis (MYB) transcription factor, MOF1, might be associated with the pollen development in tetraploid rice. However, little information is available about its role in pollen development in tetraploid rice. Here, we identified a new haplotype of MOF1 from neo-tetraploid rice and marked it as MOF1a. Transcriptome and qRT-PCR analysis demonstrated that MOF1a highly expressed in anthers, and displayed differential expression in neo-tetraploid rice compared to tetraploid rice line with low pollen fertility. The mutant (mof1a) of MOF1a, which was generated by CRISPR/Cas9 knockout in neo-tetraploid rice, showed low pollen fertility, and also exhibited abnormal tapetum and middle layer development, and defective chromosome behaviors during meiosis. A total of 13 tapetal related genes were found to be up-regulated in meiotic anthers of MOF1a compared with wild type plants by RNA-seq analysis, including CYP703A3, PTC1, and OsABCG26, which had been demonstrated to affect tapetal development. Moreover, 335 meiosis-related genes displayed differential expression patterns at same stage, including nine important meiosis-related genes, such as metallothionein OsMT1a. These results demonstrated that MOF1a plays an important role in pollen development and provides a foundation for understanding the molecular mechanism underlying MOF1a in reproduction of tetraploid rice.

Antiviral effects of Shuanghuanglian injection powder against influenza A virus H5N1 in vitro and in vivo.

The current study was to identify a protective role of Shuanghuanglian (SHL) injection powder in vitro and in vivo after H5N1 viral infection. Immunofluorescent staining was used to determine the susceptibility of rat intestinal mucosa microvascular endothelial cells (RIM-MVECs) to the H5N1 virus. Viral replication of RIM-MVECs was measured by transmission electron microscopy (TEM) a hemagglutination assay and real-time quantitative PCR. H5N1 virally infected RIM-MVECs, and BALB/c mice were treated with SHL to investigate its therapeutic effect. Animal survival and the weight of H5N1 virally infected BALB/c mice after SHL treatment was noted, and histology and real-time PCR applied to mouse lungs were used to confirm the anti-H5N1 viral effects of SHL. RIM-MVECs supported replication of the H5N1 virus in vitro. SHL treatment reduced viral titers in H5N1 virally infected RIM-MVECs and mouse lungs. SHL -treated mice survived compared to controls. Mild pathological changes, reduced inflammatory cell infiltration and fewer viral antigens were observed in the lungs of SHL-treated mice at days 3 and 6 post-infection. In conclusion, SHL may have the antiviral activity against the H5N1 virus infection by inhibiting viral replication and alleviating lung injury.

Effects of Fe-deficient conditions on Fe uptake and utilization in P-efficient soybean.

Phosphorus (P)-efficient soybean (Glycine max) plants absorb and utilize P with high efficiency. To investigate the effects of iron (Fe)-deficient conditions on the absorption and utilization of Fe in P-efficient soybean plants, two soybean cultivars with different P efficiency, the 03-3 (P-efficient variety) and Bd-2 (P-inefficient variety), were used in this study. The two soybean cultivars were grown in nutrient solution containing Fe concentrations of 0 (Fe0), 20 (Fe20), 40 (Fe40), or 80 (Fe80) µM for 7 days. The Fe reductase activity of roots was higher in 03-3 plants grown under the Fe0, Fe20, and Fe40 treatments than in Bd-2 plants and the total Fe uptake was greater in 03-3 plants under the Fe40 treatment. GmFRD3a was much more highly expressed in the stem of 03-3 than in that of Bd-2, and significantly more iron was transported to 03-3 plant shoots during Fe0 treatment. Chlorosis in young leaves caused by Fe deficiency under the Fe0 and Fe20 treatments was alleviated by increased Fe concentration in shoots. Increased levels of active Fe in young 03-3 leaves under Fe-deprivation conditions (Fe0) and maintenance of stable Fe concentrations in 03-3 shoots subjected to Fe20, Fe40, and Fe80 treatments suggested that the P-efficient 03-3 cultivar is also Fe-efficient. It is suggested that 03-3 soybean cultivar should be a good resource for application to farm field.

[Disease of root-knot nematode and control strategy in medicinal plants].

Root-knot nematode (Meloidogyne spp.) is one of the important disease of medicinal plant cultivation and seriously hinders the sustainable development of traditional Chinese medicine industry. We introduced the main species, identification methods and control strategies of root-knot nematode diseases in the medicinal plants in this study. Identifications of morphology and molecular were the main tools for the distinction of root-knot nematodes at present. This study stated that integrated system was established for root-knot nematode control, including that integrated control technique was the first step, disease-resistant varieties with high yield were the basis, and normalized patterns of cultivation and management were the measure. These strategies would improve the sustainable development of medicinal plants and environmental protection.

Baicalin protects sertoli cells from heat stress-induced apoptosis via activation of the Fas/FasL pathway and Hsp72 expression.

Certain Chinese herbal medicines have antipyretic effects in both animal and human clinical practice. However, no report indicates their antipyretic effects on heat-stressed cells. The present study aimed to identify the protective effects of baicalin on the apoptosis of primary cultured bovine sertoli cells (SCs) subjected to heat stress (HS). The results demonstrated that HS induced apoptosis in the SCs exposed to 43°C for 1h as Fas/FasL was activated and caspase-3 was cleaved, the cells apoptotic rate was decreased. Moreover, the mRNA and protein levels of Hsp72 increased, whereas the cells apoptotic rate and expression of Fas, FasL, caspases 8 and 3 decreased in the SCs pretreated with various concentrations (0.1, 1, 10, 20µg/mL) of baicalin prior to HS. In conclusion, baicalin ameliorates heat stress-induced cell apoptosis via the modulation of the cell survival rate through Fas/FasL pathway activation and the upregulation of Hsp72 expression in bovine SCs.

Saikosaponin-D attenuates heat stress-induced oxidative damage in LLC-PK1 cells by increasing the expression of anti-oxidant enzymes and HSP72.

Heat stress stimulates the production of reactive oxygen species (ROS), which cause oxidative damage in the kidney. This study clarifies the mechanism by which saikosaponin-d (SSd), which is extracted from the roots of Bupleurum falcatum L, protects heat-stressed pig kidney proximal tubular (LLC-PK1) cells against oxidative damage. SSd alone is not cytotoxic at concentrations of 1 or 3 µg/mL as demonstrated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To assess the effects of SSd on heat stress-induced cellular damage, LLC-PK1 cells were pretreated with various concentrations of SSd, heat stressed at 42°C for 1 h, and then returned to 37°C for 9 h. DNA ladder and MTT assays demonstrated that SSd helped to prevent heat stress-induced cellular damage when compared to untreated cells. Additionally, pretreatment with SSd increased the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) but decreased the concentration of malondialdehyde (MDA) in a dose-dependent manner when compared to controls. Furthermore, real-time PCR and Western blot analysis demonstrated that SSd significantly increased the expression of copper and zinc superoxide dismutase (SOD-1), CAT, GPx-1 and heat shock protein 72 (HSP72) at both the mRNA and protein levels. In conclusion, these results are the first to demonstrate that SSd ameliorates heat stress-induced oxidative damage by modulating the activity of anti-oxidant enzymes and HSP72 in LLC-PK1 cells.

[Clinical research advancement on male breast cancer].

Male breast cancer is a rare disease and the incidence has increased over the past 25 years. Current knowledge regarding its biology, natural history, and treatment strategies is mainly based on the research findings on female breast cancer. Genetic risk factors include BRCA2 mutations. Occupational risk factors include high temperature environments and exhaust fumes. Hormonal imbalances, such as gonadal dysfunction, obesity, and radiation exposure also contribute to the occurrence of male breast cancer. It may be indolent or progress slowly, or invade rapidly. Most cases are ductal tumors and 10% of the cases are ductal carcinoma in situ. The surgical operation is usually mastectomy with axillary clearance or sentinel node biopsy. Indications for radiotherapy, as well as the steps and methods, are similar to that for female breast cancer. Because 90% of the patients are estrogen receptor-positive, tamoxifen is a standard adjuvant therapy, but some individuals could also benefit from chemotherapy. In this article, the latest information on the epidemiology, biology, and treatment of male breast cancer is reviewed.