Effects of yeast inoculation methods on caproic acid production and microbial community during anaerobic fermentation of Chinese cabbage waste.

To provide a sufficient supply of electron donors for the synthesis of caproic acid, yeast fermentation was employed to increase ethanol production in the anaerobic fermentation of Chinese cabbage waste (CCW). The results showed that the caproic acid yield of CCW with ethanol pre-fermentation was 7750.3 mg COD/L, accounting for 50.2% of the total volatile fatty acids (TVFAs), which was 32.5% higher than that of the CCW without yeast inoculation. The synchronous fermentation of yeast and seed sludge significantly promoted the growth of butyric acid consuming bacterium Bacteroides, resulting in low yields of butyric acid and caproic acid. With yeast inoculation, substrate competition for the efficient ethanol conversion in the early stage of acidogenic fermentation inhibited the hydrolysis and acidfication. Without yeast inoculation, the rapid accumulation of TVFAs severely inhibited the growth of Bacteroidetes. In the reactor with ethanol pre-fermentation, the key microorganism for caproic acid production, Clostridium_sensu_stricto_12, was selectively enriched.

Regulation of hydraulic retention time on caproic acid production via two-phase anaerobic fermentation of Chinese cabbage waste with autopoietic electron donors.

The effects of hydraulic retention time (HRT) on the performance of two-phase anaerobic fermentation for caproic acid production from Chinese cabbage waste (CCW) were investigated. In the electron donor phase, yeast was inoculated to achieve efficient autopoietic ethanol, providing electron donors for the chain elongation process. Shorter HRT led to drastic fluctuations in microorganisms, thus resulting in lower acid yields at HRT of 6 days. At HRT of 10 days, the balanced collaboration of various key bacteria avoided the accumulation of intermediate by-products, and the caproic acid production reached 4660 mg COD/L, which was 119.5% and 154.8% higher than that at HRTs of 6 and 14 days, respectively. At HRT of 14 days, the low ethanol loading rate resulted in ethanol excessive-oxidation to acetic acid. Acetic acid accounted for 41.5% of the total product, while the selectivity of caproic acid was only 15.3%. The main contributor to the production process of caproic acid was Caproiciproducens, while the Ruminalococcaceae also played a role in the process. This study provided a theoretical basis for the efficient production of caproic acid through continuous fermentation with autopoietic electron donors.

Hexanoic acid production and microbial community in anaerobic fermentation: Effects of inorganic carbon addition.

Carbon dioxide (CO2) plays a crucial role in carbon chain elongation with ethanol serving as an electron donor. In this study, the impacts of various carbonates on CO2 concentration, hexanoic acid production, and microbial communities during ethanol-butyric acid fermentation were explored. The results showed that the addition of MgCO3 provided sustained inorganic carbon and facilitated interspecific electron transfer, thereby increasing hexanoic acid yield by 58%. MgCO3 and NH4HCO3 inhibited the excessive ethanol oxidation and decreased the yield of acetic acid by 51% and 42%, respectively. The yields of hexanoic acid and acetic acid in the CaCO3 group increased by 19% and 15%, respectively. The NaHCO3 group exhibited high headspace CO2 concentration, promoting acetogenic bacteria enrichment while reducing the abundance of Clostridium_sensu_stricto_12. The batch addition of NaHCO3 accelerated the synthesis of hexanoic acid and increased its production by 26%. The relative abundance of Clostridium_sensus_stricto_12 was positively correlated with hexanoic acid production.

Circular RNA CirCHIPK3 promotes cell proliferation and invasion of breast cancer by sponging miR-193a/HMGB1/PI3K/AKT axis.

BACKGROUND: The aim of this study was to explore the potential mechanism of circular RNA (circRNA) CirCHIPK3 on the malignant proliferation and metastasis of breast cancer (BC). METHODS: Human BC samples and their matched normal adjacent tissues were obtained from 50 patients to assess the expression of CirCHIPK3 and its relationship with BC prognosis. A series of in vitro and in vivo functional experiments were carried out to elucidate the role of CirCHIPK3 in BC progression and its underlying molecular mechanisms. Moreover, the interaction of CirCHIPK3, miR-193a, and HMGB1 was examined using bioinformatics, FISH, RIP, RNA-pull down and luciferase reporter assays. Western blot analysis was performed to examine the expression of HMGB1, p-PI3K, total PI3K, p-AKT, and AKT after si-CirCHIPK3 transfection. RESULTS: Upregulation of CirCHIPK3 was identified in BC, which predicted a worse prognosis in BC patients. Furthermore, it was found that CirCHIPK3 facilitated cell proliferation, migration, and invasion in BC by regulating miR-193a/HMGB1/PI3K/AKT signaling. CirCHIPK3 acted as a sponge for miR-193a to facilitate HMGB1 expression. si-CirCHIPK3 also inhibited tumor growth of BC in nude mice. si-CircCHIPK3 decreased HMGB1/PI3K/AKT signal expression in MDA-MB-231 cells, whereas overexpression of CircCHIPK3 enhanced HMGB1/PI3K/AKT signal. CONCLUSIONS: CirCHIPK3 regulated miR-193a/HMGB1/PI3K/AKT signaling to facilitate BC development and progression, providing a novel therapeutic target for BC.

Erratum: The association between inflammation, the microbiome and urethane-induced pulmonary adenocarcinoma.

[This corrects the article DOI: 10.3892/ol.2018.8167.].

The association between inflammation, the microbiome and urethane-induced pulmonary adenocarcinoma.

Lung cancer is amongst the most common types of cancer throughout the world. The overall 5-year survival rate is ~17%. A number of studies have demonstrated that the microbiome existing within the host may affect the level of inflammation, and consequently contribute to the carcinogenesis of certain types of cancer. To investigate the role of inflammation and the microbiome in the carcinogenesis of lung cancer, an intervention study involving mice, including a control group (C; n=5), a urethane-induced pulmonary adenocarcinoma group (U; n=5) and a prebiotics intervention group (P; n=5) was carried out. This pulmonary adenocarcinoma model was reviewed, and incidences of the disease were identified using histopathology. The levels of the inflammatory cytokines nuclear factor κB (NF-κB), tumor necrosis factor α (TNF-α), interleukin-1ß (IL-1ß) and IL-6 in the sera samples were measured using an ELISA technique. In addition, high-throughput sequencing of the 16S ribosomal RNA gene segment was used to analyze the species present in the microbiome of the lower airways and intestinal tracts of mice. The results demonstrated that groups P and U exhibited altered histopathology and the development of lung adenocarcinoma tumors, but no differences were observed between the groups. The level of inflammation, determined by measuring the levels of NF-κB, TNF-α, IL-1ß and IL-6 inflammatory cytokines, was significantly lower in group P compared with group U (P<0.05), and was significantly higher in group P compared with group C (P<0.05). Overall, the microbiomes of the lower respiratory and intestinal tracts did not change markedly among the 3 groups, in terms of the size of colonies and Shannon diversity indices. However, at a family and operational taxonomic unit (OTU) level, certain microbiota were altered. For example, the abundance of the Clostridiales and Lachnospiraceae families was lower in the lung and intestinal tracts subsequent to urethane-induced treatment compared with in the control group (P<0.05), and the level of abundance of the Clostridiales family increased to similar levels within the control group (P<0.05), when prebiotics were administered. The levels of abundance of the S24-7, Bacteroidales and Firmicutes families were higher in the intestinal tract compared with the control group (P<0.05), and following treatment with prebiotics, the levels of abundance of these families decreased to similar levels observed in the control group (P<0.05). In conclusion, inflammation and the microbiome serve important roles in the carcinogenesis of lung cancer. Additionally, prebiotics may increase the efficacy of lung cancer treatment by modulating levels of inflammation and the composition of the microbiome. The associations between inflammation, the microbiome and lung cancer require attention.

Urine Proteome Profiling Predicts Lung Cancer from Control Cases and Other Tumors.

Development of noninvasive, reliable biomarkers for lung cancer diagnosis has many clinical benefits knowing that most of lung cancer patients are diagnosed at the late stage. For this purpose, we conducted proteomic analyses of 231 human urine samples in healthy individuals (n=33), benign pulmonary diseases (n=40), lung cancer (n=33), bladder cancer (n=17), cervical cancer (n=25), colorectal cancer (n=22), esophageal cancer (n=14), and gastric cancer (n=47) patients collected from multiple medical centers. By random forest modeling, we nominated a list of urine proteins that could separate lung cancers from other cases. With a feature selection algorithm, we selected a panel of five urinary biomarkers (FTL: Ferritin light chain; MAPK1IP1L: Mitogen-Activated Protein Kinase 1 Interacting Protein 1 Like; FGB: Fibrinogen Beta Chain; RAB33B: RAB33B, Member RAS Oncogene Family; RAB15: RAB15, Member RAS Oncogene Family) and established a combinatorial model that can correctly classify the majority of lung cancer cases both in the training set (n=46) and the test sets (n=14-47 per set) with an AUC ranging from 0.8747 to 0.9853. A combination of five urinary biomarkers not only discriminates lung cancer patients from control groups but also differentiates lung cancer from other common tumors. The biomarker panel and the predictive model, when validated by more samples in a multi-center setting, may be used as an auxiliary diagnostic tool along with imaging technology for lung cancer detection.

[Research Progress of Biomakers Proteomics-based in Lung Cancer].

Proteomic technologies can be applied to cancer research to detect differential protein expression that could find cancer biomakers. Lung cancer biomarker discovery is significant due to its anticipated critical role in early diagnosis, therapy guidance, and prognosis monitoring of lung cancer. Therefore, there is an indeed need to identify new biomarkers for early diagnosis and prognosis that could serve to open novel therapeutic means. This article briefly introduces the latest reports in proteomic studies of lung cancer. It contains diagnostic, prognostic, and predictive biomarkers, and a summary based on the most recent literature and our own work.

Simultaneous quantification of phencynonate and its active metabolite N-demethyl phencynonate in human plasma using liquid chromatography and isotope-dilution mass spectrometry.

A sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed to simultaneously quantify phencynonate (PCN) and its major metabolite N-demethyl phencynonate (DM-PCN) in human plasma. Following one-step liquid-liquid extraction, the analytes were separated on a reversed-phase C18 column. Methanol and 0.02% formic acid in 10 mM ammonium acetate (62:38, v/v) was used as isocratic mobile phase at a flow-rate of 0.3 mL/min. An API 5000 tandem mass spectrometer equipped with a Turbo IonSpray ionization source was used as the detector and was operated in the positive ion mode. Multiple reaction monitoring using the transition of m/z 358.4 → m/z 156.2, m/z 344.4 → m/z 142.2, and m/z 361.3 → m/z 159.2 was performed to quantify PCN, DM-PCN, and the internal standard (D3 -PCN), respectively. This approach showed a lower limit of quantification of 10 pg/mL and 25 pg/mL for PCN and DM-PCN in plasma, respectively. This sensitivity was at least 50-fold superior to previously reported ones and thus enabled the approach well applicable to low-dose pharmacokinetic studies. The intra- and inter-day precisions were less than 14.2 % at each QC level for both PCN and DM-PCN. The inter-day relative errors ranged from -1.9% to -4.9% for PCN, and from 0.6% to 6.4% for DM-PCN. As a proof of principle, the validated method was successfully applied to simultaneous quantification of circulating PCN and DM-PCN in healthy subjects after a single oral administration of 2 mg phencynonate hydrochloride pellet.

[Analysis of differentially expressed proteome in urine 
from non-small cell lung cancer patients].

BACKGROUND: Screen differentially expressed proteins in patients with non-small cell lung cancer (NSCLC), and aim to identify biomarkers for early screening, monitoring prognosis and evaluating therapy of NSCLC. METHODS: Urinary samples were collected from 40 newly diagnosed NSCLC patients, 8 patients with lung benign disorders and 22 healthy people. 0.9% sodium dodecylsulfate- polyacrylamide gel electrophoresis (1D SDS-PAGE) and MS-Thermo-Orbitrap-Velos were applied to separate, extract and identify proteins in urinary samples from non-neoplastic groups and NSCLC patients, in order to find out differentially expressed proteins in patients with NSCLC. Then, sensitivity and specificity of candidate proteins were tested by certain experiments. Finally, biomarkers related to NSCLC could be determined. RESULTS: The differences of urinary proteins between non-neoplastic groups and NSCLC patients mainly focused on 90 kDa, 60 kDa and 20 kDa-30 kDa stripes. Four differently expressed proteins were found in urinary proteins in NSCLC group, including LRG1, CA1 (up-regulating proteins) and VPS4B, YWHAZ (down-regulating proteins). The sensitivity of these four proteins for biomarker of NSCLC was relatively low when they were used to screen or diagnose independently. The sensitivity and specificity of LRG1 was 83.0% (25/30) and 90.0% (18/20), respectively; 60.0% (18/30) and 90.0% (18/20) for CA1; 73.3% (22/30) and 90.0% (18/20) for VPS4B; 60.0% (18/30) and 95.0% (19/20) for YWHAZ. However, the sensitivity and specificity would increase to 96.7% (29/30) and 85% (17/20) after the four biomarkers were combined. CONCLUSIONS: LRG1 and CA1 are abundant in urine in patients with NSCLC, while VPS4B and YWHAZ are low-abundance proteins. They could be regarded as biomarkers for early screening, monitoring prognosis and evaluating therapy of patients with NSCLC because of differential expression. The sensitivity of the four biomarkers of NSCLC is relatively low when they are used to screen or diagnose independently, while significantly improvement if they were in combined pattern, which will be of excellent applications to clinical diagnosis and treatment.

Serum cyfra21-1 as a biomarker in patients with nonsmall cell lung cancer.

OBJECTIVE: The purpose of this retrospective study was to evaluate the clinical value of serum cytokeratin-19-fragment (cyfra21-1) as a biomarker in nonsmall cell lung cancer (NSCLC). METHODS: Sixty-six patients with NSCLC and 48 cases with benign lung disease were retrospectively analyzed in the department of thoracic surgery in our hospital. The serum level of cyfra21-1 was detected in the above patients. The diagnosis sensitivity, specificity and the receiver operating characteristic curve were calculated by using the stata11.0 statistical software to evaluate the clinical diagnosis value of serum Cyfra21-1 as NSCLC serologic biomarker. RESULTS: The mean of serum cyfra21-1 were 8.95 eat. 01 µ/L and 4.28 eat. 89 µ/L in NSCLC patient and control groups respectively, which indicated that the NSCLC group were much higher (P < 0.05). The diagnosis sensitivity and specificity were 77.08% and 63.64% at the threshold of 6.32 µ/L respectively. Moreover, the area under the curve was 0.78 (95% confidence interval: 0.70-0.87). CONCLUSION: Serum cyfra21-1 can be a potential serologic biomarker in evaluation of NSCLC.