SLC11A1 promotes kidney renal clear cell carcinoma (KIRC) progression by remodeling the tumor microenvironment.

Kidney renal clear cell carcinoma (KIRC) is a highly immune-infiltrated kidney cancer with the highest mortality rate and the greatest potential for invasion and metastasis. Solute carrier family 11 member1 (SLC11A1) is a phagosomal membrane protein located in monocytes and plays a role in innate immunity, autoimmune diseases, and infection, but its expression and biological role in KIRC is still unknown. In this study, we sought to investigate the potential value of SLC11A1 according to tumor growth and immune response in KIRC. TIMER and UALCAN database was used to analyze the expression feature and prognostic significance of SLC11A1 and its correlation with immune-related biomarkers in KIRC. Proliferation, migration, and invasion were measured using colony formation, EdU, and transwell assays. Role of SLC11A1 on KIRC tumor growth was examined by the xenograft tumor model in vivo. Effects of KIRC cells on macrophage polarization and the proliferation and apoptosis of CD8+ T cells were analyzed using flow cytometry assays. Herein, SLC11A1 was highly expressed in KIRC tissues and cell lines. SLC11A1 downregulation repressed KIRC cell proliferation, migration, invasion, macrophage, and lymphocyte immunity in vitro, as well as hindered tumor growth in vivo. SLC11A1 is significantly correlated with immune cell infiltration and immune-related biomarkers. In KIRC patients, SLC11A1 is highly expressed and positively correlated with the immune-related factors CCL2 and PD-L1. SLC11A1 induced CCL2 and PD-L1 expression, thereby activating the JAK/STAT3 pathway. SLC11A1 deficiency constrained KIRC cell malignant phenotypes and immune response via regulating CCL2 and PD-L1-mediated JAK/STAT3 pathway, providing a promising therapeutic target for KIRC treatment.

Isolation and identification of a salt-tolerant Coelastrum sp. and exploration of its potential for biodiesel production.

Given the escalating demand for renewable biofuels amidst the continual consumption of fossil energy, the exploration and identification of microalgal strains for biodiesel production have become crucial. In this study, a microalgal strain named HDMA-12 was isolated from Lake Chenjiadayuan in China to evaluate its biodiesel potential. Phylogenetic analysis of its internal transcribed spacer sequences revealed HDMA-12 as a new molecular record in the genus Coelastrum. When cultivated in BG11 basal medium, HDMA-12 achieved a biomass of 635.7 mg L-1 and a lipid content of 26.4%. Furthermore, the fatty acid methyl ester profile of HDMA-12 exhibited favorable combustion characteristics. Subjected to 200 mM NaCl stress, HDMA-12 reached its maximum biomass of 751.5 mg L-1 and a lipid content of 28.9%. These findings indicate the promising prospects of HDMA-12 as a promising microalgal strain for further advancements in biodiesel production.

Synergistic benefits of Funneliformis mosseae and Bacillus paramycoides: Enhancing soil health and soybean tolerance to root rot disease.

To explore the response of soil metabolite composition to soybean disease, the effect of the combined inoculation of arbuscular mycorrhizal fungi (AMF) and plant growth-promoting bacteria on soybean root rot caused by Fusarium oxysporum was studied. A factorial completely randomized design with three factors (AMF, Bacillus. paramycoides, and rot disease stress) was conducted, and eight treatments, including normal groups and stress groups, were performed using pot experiments. GC‒MS and enzymatic assays were used to evaluate the soil factors and soybean growth indicators. The results showed that there were significant differences in the composition of metabolites among the different treatment groups, and 23 metabolites were significantly related to soybean biomass. The combined inoculation of Funneliformis mosseae and Bacillus paramycoides resulted in a significant reduction in harmful soil metabolites associated with root rot disease, such as ethylbenzene and styrene. This reduction in metabolites contributed to improving soil health, as evidenced by enhanced soybean defence enzyme activities and microbial activity, and ß-1,3-glucanase, chitinase and phenylalanine ammonia-lyase activities were improved to alleviate plant rhizosphere stress. Furthermore, soybean plants inoculated with the synergistic treatments exhibited reduced root rot disease severity and improved growth indicators compared to control plants. Plant height, root dry weight (RDW), and shoot and root fresh weight (SRFW) were improved by 4.18-53.79%, and the AM fungal colonization rate was also improved under stress. The synergistic application of Funneliformis mosseae and Bacillus paramycoides can effectively enhance soil health by inhibiting the production of harmful soil metabolites and improving soybean tolerance to root rot disease. This approach holds promise for the sustainable management of soil-borne diseases in soybean cultivation.

Screening of symbiotic Streptomyces spp. and optimization of microalgal growth in a microalgae-actinomycetes co-culture system.

Microalgal biodiesel as a substitute for fossil energy has attracted extensive attention. However, the high cost of microalgae cultivation limits the industrial production of microalgal biodiesel. The co-culture system may offer a means to increase microalgae's biomass production. In this study, Streptomyces strains were selected to construct and optimize co-culture systems with Monoraphidium sp. HDMA-11 and the algal cell biomass, lipid content, phycocyanin content, starch content, and fatty acid composition were determined. The results showed that Streptomyces nojiriensis significantly promoted Monoraphidium sp. HDMA-11 growth and a co-culture system were established. Orthogonal experiments showed that the Monoraphidium sp. HDMA-11 biomass was further increased when the initial culture pH was 7.5, the inoculation time of Streptomyces strain supernatants was 36 h, the volume ratio of microalgal actinomycetes was 1:1, and no additional acetic acid was added. Under these conditions, compared with monocultured Monoraphidium sp. HDMA-11, the cell biomass and lipid productivity of the co-culture system increased by 525.8 and 155.1%, respectively. These results suggest that S. nojiriensis supernatant potentially enhances microalgae biomass and may represent a new method to improve microalgae growth.

Epigallocatechin-3-gallate exerts protective effect on epithelial function via PI3K/AKT signaling in thrombosis.

BACKGROUND: Venous thrombosis (VT) is one of the most frequent cardiovascular diseases, which seriously endangers people's health. Recently, the protective role of epigallocatechin-3-gallate (EGCG) against multiple cardiovascular diseases has been well studied. Nevertheless, whether EGCG is implicated in the progression of VT is still unclear. METHODS: Rat models of VT were established by inferior vena cava (IVC) ligation. Histological characterization of the IVC tissues was examined by hematoxylin-eosin (H&E) staining. TUNEL assay was utilized to detect cell apoptosis in IVC tissues. The concentrations of the oxidative stress biomarkers, malondialdehyde (MDA) and superoxide dismutase (SOD), were estimated by corresponding kits. In addition, the levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-8 in rat plasma were estimated by ELISA. Further, the expression levels of apoptosis markers (Bax, Bcl-2, and Cleaved-caspase 3) as well as key molecules p-PI3K and p-AKT in phosphoinositide 3-kinase (PI3K)/AKT signaling pathway were assessed by western blot. RESULTS: Compared to the sham group, the model group showed obvious thrombus formation in IVC tissues, while the EGCG treatment significantly repressed thrombosis. EGCG inhibited cell apoptosis in IVC tissues of VT rat models. The decreased SOD concentration and increased MDA concentration in the plasma of VT rats were reversed by EGCG treatment. Additionally, the elevated levels of TNF-α, IL-6 and IL-8 in the plasma of VT rats can be partially reduced by the treatment of EGCG. Finally, we also found that EGCG reduced the levels of phosphorylated (p)-PI3K and p-AKT in IVC tissues of VT rat models, indicating that the hyperactivation of the PI3K/AKT signaling pathway was inhibited by EGCG. CONCLUSION: This study proves that EGCG alleviates thrombosis, cell apoptosis, inflammatory response, and oxidative stress injury in VT by inactivating PI3K/AKT signaling pathway.

KCNN4 may weaken anti-tumor immune response via raising Tregs and diminishing resting mast cells in clear cell renal cell carcinoma.

BACKGROUND: Studies over the past decade have shown that competitive endogenous RNA (ceRNA) plays an essential role in the tumorigenesis and progression of clear cell renal cell carcinoma (ccRCC). Meanwhile, immune checkpoint blocker is gradually moving towards the first-line treatment of ccRCC. Hence, it's urgent to develop a new prediction model for the efficiency of immunotherapy. At present, there is no study to reveal the effect of ceRNA network on the efficiency of immunotherapy for ccRCC. METHODS: To systematically analyze the effect of ceRNA hub genes in ccRCCon immune response, we constructed prognosis models based on ceRNAs and immune cells, respectively. We constructed ceRNA network using hypergeometric distribution test and correlation analysis with R script based on The Cancer Genome Atlas (TCGA) database. We then applied the Cibersort algorithm to simulate the infiltration overview of immune cells in kidney renal clear carcinoma (KIRC) samples. Prognosis-related immune cells were screened and a predictive model of these cells was constructed. Prognosis-related immune cells and ceRNA hub genes were performed with co-expression analysis. Finally, qRT-PCR and immunofluorescence assays were performed to validate the results. RESULTS: The construction of ceRNA related prognosis model contained 8 hub genes, including RELT, MYO9B, KCNN4, SIX1, OTOGL, MALAT1, hsa-miR-130b-3p, and hsa-miR-21-5p. The area under the receiver operating characteristic curve (AUC) was 0.77 at 5 years. For the construction of immune cells prognosis model, 3 immune cells (T cells regulatory, Macrophages, Mast cells resting) were adopted, and the AUC was 0.65 at 5 years. We then merged the two models by correlation analysis and co-expression analysis. Finally, we found that KCNN4 positively correlates with T cells regulatory (Tregs) and negatively correlates with mast cells resting significantly. Furthermore, higher expression of KCNN4 may lead to a higher potential for immune evasion and lower efficiency for immune checkpoint inhibitors (ICIs). CONCLUSIONS: Generally, this is the first study to assess the prognostic value of immune related ceRNA hub genes in ccRCC, and KCNN4 was finally demonstrated to be a key regulatory factor with strong correlation with Tregs and mast cells resting.

Retrospective comparison of clinical outcomes of robotic-assisted laparoscopic partial nephrectomy through transabdominal or retroperitoneal approaches in patients with T1b renal tumor.

BACKGROUND: We compared the intraoperative and postoperative outcomes of robotic-assisted laparoscopic partial nephrectomy (RALPN) via transabdominal or retroperitoneal approaches in patients with stage T1b renal cell carcinoma. METHODS: The medical records for 92 patients who underwent RALPN were retrospectively collected and data on their baseline demographics, duration of operation, duration of renal artery clamping, intraoperative blood loss, recovery time of intestinal functions, surgical margin positive rate, as well as postoperative complications were analyzed. RESULTS: Of the 92 enrolled patients, 43 and 49 patients were subjected to RALPN via the transabdominal and retroperitoneal approaches, respectively. All patients successfully completed the operation. Baseline characteristics for the transabdominal and retroperitoneal groups were comparable. Differences in operative time, renal artery clamping time, intraoperative blood loss, positive rate of surgical margin, and incidences of postoperative complications between the two approaches were insignificant. The recovery time of intestinal function after operation was significantly shorter in patients subjected to the retroperitoneal approach, relative to those subjected to transabdominal approach (p < 0.001). CONCLUSIONS: Application of RALPN via transabdominal or retroperitoneal approaches showed comparable clinical outcomes in patients with stage T1b renal cell carcinoma. The retroperitoneal approach was superior to the transabdominal approach in terms of postoperative intestinal function recovery.

Elaeagnus angustifolia can improve salt-alkali soil and the health level of soil: Emphasizing the driving role of core microbial communities.

Saline-alkali environments are widely distributed in China and significantly hinder the development of agriculture. This study characterizes the long-term effects of planting Elaeagnus angustifolia (E. angustifolia) on the physical and chemical properties, enzyme activities and microbial community characteristics of saline-alkali soil in the Songnen Plain (1, 2 and 3 years). The results showed that planting E. angustifolia reduced soil pH and electrical conductivity (EC) and increased soil total phosphorus (TP), total nitrogen (TN), nitrate nitrogen (Nni), total potassium (TK), dissolved organic C (DOC), dissolved organic matter (DOM) and available potassium (AK) content and catalase, urease, polyphenol oxidase, phosphatase, sucrase and cellulase enzyme activities, and the results peaked in the 3 year. High-throughput sequencing showed that the bacterial abundance and diversity were as follows (from high to low) y3 > y2 > y1 > CK. E. angustifolia resulted in an increase in the relative abundance of the dominant bacteria. Proteobacteria and Pseudomonas were the major phylum and genus, respectively. Redundancy analysis showed that changes in the soil microbial community significantly affect the physical and chemical properties of the soil, with Proteobacteria members being the key microorganisms that reduce soil salinity. Network analysis showed that Pseudomonas (Proteobacteria) participated in the synthesis of key soil enzymes. 16S rRNA sequencing predicted that the expression of genes related to carbon (rbcL, acsA, acsB, Pcc and accA) and nitrogen (amoA/B, nxrA, hao, gdh, ureC and nosZ) transformation increased, and Pseudomonas members were key regulators of carbon and nitrogen dynamics. In conclusion, the planting of E. angustifolia could improve the physical and chemical properties of the soil by releasing root exudates into the soil and increasing the diversity and richness of soil microbial communities to improve saline-alkali soil, providing a theoretical basis for improving saline-alkali soil and promoting the sustainable development of modern agriculture.

Acetic acid acting as a signaling molecule in the quorum sensing system increases 2,3-butanediol production in Saccharomyces cerevisiae.

2,3-Butanediol (2,3-BD) has been extensively used in chemical syntheses. This study aimed to explore acetic acid as a signaling molecule that activates a quorum sensing (QS) system to promote the production of 2,3-BD. The yield of 2,3-BD is proportional to the cell density. Saccharomyces cerevisiae W141 does not produce 2,3-BD when the cell density is lower than the threshold concentration (OD600 nm = 10 or cell density 4.4 × 108 CFU/mL). When 1.5 g/L acetic acid is added, the yield of 2,3-BD is 3.01 ± 0.04 g/L. Subsequently, S. cerevisiae W141 was cocultured with Acetobacter pasteurianus Huniang 1.01 under the optimal conditions, the acetic acid production was increased by 76.7% and 30.6% compared with the original strain and the strain cultivated with 1.5 g/L acetic acid, and the yield of 2,3-BD was increased by 81.9% and 3.3%, respectively. This difference is due to the activity of acetyl lactic acid synthase (ILV2) and 2,3-BD dehydrogenase (BDH1), as the relative expression of the ilv2 and bdh1 genes is increased. The results showed that the biosynthesis of 2,3-BD was regulated by acetic acid as a signaling molecule. S. cerevisiae is a promising host for producing 2,3-BD for industrial applications.

Enhanced ß-mannanase production by Bacillus licheniformis by optimizing carbon source and feeding regimes.

Bacillus licheniformis HDYM-04 was isolated in flax retting water and showed ß-mannanase activity. Carbon sources for ß-mannanase production, as well as the fermentation conditions and feeding strategy, were optimized in shake flasks. When glucose or konjac powder was used as the carbon source, the ß-mannanase activity was 288.13 ± 21.59 U/mL and 696.35 ± 23.47 U/mL at 24 h, respectively, which was approximately 4.4- to 10.68-fold higher than the values obtained with wheat powder. When 0.5% (w/v) glucose and 1% (w/v) konjac powder were added together, maximum enzyme activities of 789.07 ± 25.82 U/mL were obtained, an increase of 13.35% compared to the unoptimized cultures with only 1% (w/v) konjac powder. The enzyme activity decreased in the presence of 1% (w/v) konjac powder, but the highest enzyme activity was 1,533.26 ± 33.74 U/mL, a 1.2-fold increase compared with that in nonoptimized cultures; when 0.5% (w/v) glucose was used, the highest enzyme activity was 966.53 ± 27.84 U/mL, an increase in ß-mannanase activity of 38.79% compared with control cultures. In this study, by optimizing fed-batch fermentation conditions, the yield of ß-mannanase produced by HDYM-04 was increased, laying the foundation for the industrial application and further research of B. licheniformis HDYM-04.

The response surface optimization of exopolysaccharide produced by Saccharomyces cerevisiae Y3 and its partial characterization.

Response surface methodology (RSM) was used to optimize the conditions of exopolysaccharides (EPSs) by Saccharomyces cerevisiae Y3. The results indicated that the yield of EPS reached 4.52 ± 0.14 g/L with 10.30% (w/v) sucrose, 0.64% (w/v) yeast extract, liquid volume 141.5 mL, which was 2.40 times the original EPS yield. Y3 EPS contained 83.65 ± 0.16% of total sugars, 15.27 ± 0.26% of uronic acid, 0.78 ± 0.02% of protein and 0.30 ± 0.12% of sulfuric acid groups. Y3 EPS maintained a relatively low viscosity, with intrinsic viscosities of 306.58 mL/g (25 °C) and 200.91 mL/g (35 °C), respectively. The EPS had high water solubility index (WSI), high water holding capacity (WHC) and good emulsifying ability (EA). Meanwhile, the EPS could absorb metal ions such as Cu2+, Fe2+ and Zn2+. In addition, Y3 EPS exhibited good antioxidant properties and coagulated skim milk with a concentration-dependent manner. These results indicated that S. cerevisiae Y3 EPS had applicable prospects in medicine, food, especially the dairy industry.

Effects of pyruvate decarboxylase (pdc1, pdc5) gene knockout on the production of metabolites in two haploid Saccharomyces cerevisiae strains.

Saccharomyces cerevisiae has good reproductive ability in both haploid and diploid forms, a pyruvate decarboxylase plays an important role in S. cerevisiae cell metabolism. In this study, pdc1 and pdc5 double knockout strains of S. cerevisiae H14-02 (MATa type) and S. cerevisiae H5-02 (MATα type) were obtained by the Cre/loxP technique. The effects of the deletion of pdc1 and pdc5 on the metabolites of the two haploid S. cerevisiae strains were consistent. In S. cerevisiae H14-02, the ethanol conversion decreased by 30.19%, the conversion of glycerol increased by 40.005%, the concentration of acetic acid decreased by 43.54%, the concentration of acetoin increased by 12.79 times, and the activity of pyruvate decarboxylase decreased by 40.91% compared to those in the original H14 strain. The original S. cerevisiae haploid strain H14 produced a small amount of acetoin but produced very little 2,3-butanediol. However, S. cerevisiae H14-02 produced 1.420 ± 0.063 g/L 2,3-BD. This study not only provides strain selection for obtaining haploid strains with a high yield of 2,3-BD but also lays a foundation for haploid S. cerevisiae to be used as a new tool for genetic research and breeding programs.

Optimization of cultivation strategy and medium for bacteriocin activity of Enterococcus faecium HDX-2.

A lactic acid bacteria (LAB) isolated from pickled Chinese cucumber was screened for bacteriocin production. The strain was identified to be Enterococcus faecium HDX-2. Based on the Plackett-Burman (PB) experiment, glucose, Ca2+, and initial pH were found to be the most significant parameters of bacteriocin production. Afterward, effects of the three main parameters on bacteriocin activity were further investigated by central composite design (CCD) and the optimum composition was glucose 22 g/L, Ca2+ 0.6 mM, and initial pH 7.2. Optimum results showed that bacteriocin activity was increased to 1337.60 ± 36.71 AU/mL, 2.23-fold higher than in MRS medium without parameters optimization. The bacteriocin also showed significant antimicrobial activity against Listeria monocytogenes in meat and cheese model system.

Bacteriocin production and inhibition of Bacillus subtilis by Lactobacillus paracasei HD1.7 in an indirect coculture system.

A broad-spectrum antimicrobial peptide named Paracin 1.7 was produced by Lactobacillus paracasei HD1.7, which was isolated from Chinese sauerkraut juice. In this study, the influence of cocultivation on the communication mechanism of L. paracasei HD1.7 and Bacillus subtilis was investigated. The two bacterial strains were grown in monoculture and indirect coculture, and the growth of both bacteria and bacteriocin production as well as the transcriptional level of luxS in L. paracasei HD1.7 and spo0A in B. subtilis were monitored. Bacteriocin production and cell numbers were increased significantly when L. paracasei HD1.7 cells were indirectly cocultured with B. subtilis, and bacteriocin-producing L. paracasei HD1.7 can prevent the growth and sporulation of B. subtilis. After indirect coculture with B. subtilis, the expression of luxS in L. paracasei HD1.7 increased in the exponential growth phase and decreased in the stationary phase compared to monoculture. The expression of spo0A in B. subtilis dropped in the indirect coculture compared to the monoculture. It indicate that the upregulation of luxS is due to a response to a secreted compound produced by B. subtilis. The results show L. paracasei HD1.7 has an amensalism on B. subtilis, while B. subtilis has a commensalism on L. paracasei HD1.7.

The response surface optimization of ß-mannanase produced by Weissella cibaria F1 and its potential in juice clarification.

A ß-mannanase-producing lactic acid bacteria (LAB) was identified as Weissella cibaria F1 according to physiological and biochemical properties, morphological observations, partial sequence of 16S rRNA gene and API 50 CHL test. In order to improve the yield of ß-mannanase, the response surface methodology (RSM) was originally used to optimize the fermentation conditions. The optimization results showed that when the konjac powder, glucose, and initial pH were 9.46 g/L, 14.47 g/L and 5.67, respectively, the ß-mannanase activity increased to 38.81 ± 0.33 U/mL, which was 1.33 times compared to initial yield (29.28 ± 0.26 U/mL). This result was also supported by larger clearance on the konjac powder-MRS agar plate through Congo Red dyeing. The W. cibaria F1 ß-mannanase could improve the clarity of five fruits juice, i.e., apple, orange, peach, persimmon and blue honeysuckle. Among these, peach juice was the most obvious, clarity increasing by 12.8%. These results collectively indicated that W. cibaria F1 ß-mannanase had an applicable potential in food-level fields.

[Extragonadal germ cell tumor: A case report and review of the literature].

OBJECTIVE: To discuss the clinical diagnosis and treatment of extragonadal germ cell tumor. METHODS: We analyzed the clinical data on a case of extragonadal germ cell tumor diagnosed and treated in the General Hospital of Eastern Theater Command and reviewed the relevant literature. RESULTS: The patient was initially diagnosed with retroperitoneal tumor and treated by resection of the tumor together with the left kidney due to the large volume of the tumor, which was complicated by pancreatic injury. Postoperative pathology showed it to be extragonadal germ cell malignancy. Postoperative examination revealed space-occupying lesion in the left testis, with serum alpha fetoprotein (AFP), human chorionicgonadotropin (hCG) and lactate dehydrogenase (LDH) negative, followed by stage-two resection of the left testis, which was pathologically shown with testicular seminoma. The patient received 7 courses of cisplatin, etoposide bleomycin (PEB) regimen and was followed up for 8 years, which found no recurrence or metastasis, and the patient fathered no child during the postoperative follow-up. CONCLUSION: For patients with a history of cryptorchidism and tumors located in the central axis, special attention should be paid to physical examination of the testes, testicular ultrasonography, and determination of AFP and other indicators to identify gonadal tumor metastasis. And if so, radiotherapy and chemotherapy can be considered first to reduce surgical complications and achieve accurate management.

[Long-term effect of finasteride on high-risk patients with benign prostatic hyperplasia after implantation of thermo-expandable spiral prostatic stent].

OBJECTIVE: To investigate the long-term effect of finasteride (FS) on high-risk BPH patients after treated by implantation of thermo-expandable spiral prostatic stent (TESPS). METHODS: We retrospectively analyzed the clinical data on 63 cases of BPH treated by implantation of TESPS in our Department of Urology from January 2017 to January 2019. All the patients received oral FS after operation except two cases of stent removal because of infection, 37 for more than 12 months (the long-term FS group) and the other 24 for less than 12 months (the control group). We followed up the patients at 3, 6, 12, 24, 36 and 48 months postoperatively, recorded the incidence of hematuria and infection, IPSS, maximum urinary flow rate (Qmax) and residual urine volume (PVR), and compared them between the two groups of patients. RESULTS: At 48 months after operation, the incidence rates of postoperative hematuria and infection were significantly lower in the long-term FS group than in the control (P < 0.05), but evidently increasing with the prolonging of medication time. The total effectiveness rate was as high as 95.1% at 3 months, but only 63.6% at 48 months, significantly higher, however, in the long-term FS than in the control group (69.2% vs 55.6%, P < 0.05), and the IPSS, Qmax and PVR were also remarkably higher in the former than in the latter group (P < 0.05). CONCLUSION: The long-term effect of TESPS implantation is definite in the treatment of BPH-induced dysuria, and it can be used as a first-choice method for the patients at high risk and unsuitable for surgery. Finasteride has an evident advantage in preventing hematuria and infection after prostatic stent implantation, and long-term medication of finasteride improves long-term outcomes.

Nobiletin inhibits viability of human renal carcinoma cells via the JAK2/STAT3 and PI3K/Akt pathway.

Nobiletin, a flavonoid found chiefly in oranges and lemons, has exhibited potential anti-proliferative and pro-apoptotic activities in various types of cancers. However, the inhibitory effect and mechanisms of nobiletin on renal cancer cells are unclear.  CCK8 and plate clone formation assay were used to determine the effect of nobiletin on the proliferation of renal cancer cells, while scratch healing test was used to assay its effect on migration ability. The effect of nobiletin on the invasion of renal cancer cells was determined using Trans well chamber assay. Flow cytometry was used to determine the effect of nobiletin on apoptosis of renal cancer cells, while Western blotting assay was used to determine its effect on the expressions of JAK2/STAT3 and PI3K/Akt pathway proteins, and apoptosis-related proteins. Nobiletin inhibited the proliferation of renal carcinoma cells in a time- and dose-dependent manner. It inhibited the migration and invasion of renal cancer cells, and promoted their apoptosis. Western blot results showed that nobiletin inhibited the phosphorylations of JAK2, STAT3, PI3K, and Akt, and promoted the expressions of apoptosis-related proteins. Nobiletin inhibits the proliferation, invasion and migration of renal cell carcinoma by inhibiting JAK2/STAT3 and PI3K/Akt pathways, and promotes their apoptosis. These findings provide a new experimental basis for the application of nobiletin in the treatment of renal cancer.

Disruption of the lactate dehydrogenase and acetate kinase genes in Klebsiella pneumoniae HD79 to enhance 2,3-butanediol production, and related transcriptomics analysis.

OBJECTIVES: 2,3-Butanediol (2,3-BD) is widely used in several chemical syntheses as well as the manufacture of plastics, solvents, and antifreeze formulations, and can be manufactured by microbial glucose fermentation. Conventional (2,3-BD) fermentation typically has low productivity, yield, and purity, and is expensive for commercial applications. We aimed to delete the lactate dehydrogenase and acetate kinase (ldhA and ack) genes in Klebsiella pneumoniae HD79 by using λRed homologous recombination technology, to eliminate by-products and thereby improve (2,3-BD) production. We also analyzed the resulting gene changes by using transcriptomics. RESULTS: The yield of (2,3-BD) from the mutant Klebsiella strain was 46.21 g/L, the conversion rate was 0.47 g/g, and the productivity was 0.64 g/L·h, which represented increases of 54.9%, 20.5%, and 106.5% respectively, compared to (WT) strains. Lactate and acetate decreased by 48.2% and 62.8%, respectively. Transcriptomics analysis showed that 4628 genes were differentially expressed (404 significantly up-regulated and 162 significantly down-regulated). Moreover, the (2,3-BD) operon genes were differentially expressed. CONCLUSION: Our data showed that the biosynthesis of (2,3-BD) was regulated by inducers (lactate and acetate), a regulator (BudR), and carbon flux. Elimination of acidic by-products by ldhA and ack knockdown significantly improved (2,3-BD) production. Our results provide a deeper understanding of the mechanisms underlying (2,3-BD) production, and form a molecular basis for the improvement this process by genetic modification in the future.

The response surface optimization of exopolysaccharide produced by Weissella confusa XG-3 and its rheological property.

The response surface methodology (RSM) was used to optimize the exopolysaccharide (EPS) production by a lactic acid bacteria (LAB) Weissella confusa XG-3. Two-level factorial design screened three significantly influencing factors sucrose, initial pH and sodium acetate. Central composite design (CCD) predicted under the condition of sucrose 80.1 g L-1, initial pH 5.8 and sodium acetate 3.7 g L-1, the maximal EPS yield obtained a 2.9-fold increase, reaching 97.5 ± 1.1 g L-1. This maximal value was far exceeding EPS production by other W. confusa species strains reported so far. The results suggested that W. confusa XG-3 had a potential for large-scale EPS production. The rheological properties of XG-3 EPS was further investigated. It was a typical non-Newtonian fluid, exhibiting pseudo-plastic behavior. The EPS concentration and temperature exerted positive and negative impact on apparent viscosity, respectively. The XG-3 EPS maintained relatively higher viscosity at moderate pH (6-8). The intrinsic viscosity [η] was 409.7 (25 °C) and 201.7 (35 °C), which was relevant to temperature but irrelevant to EPS concentration. This EPS efficiently coagulated sucrose-supplemented milk in a concentration-dependent manner. These results indicated that XG-3 EPS had an applicable potential in food processing fields especially dairy products.