Tradeoffs in alignment and assembly-based methods for structural variant detection with long-read sequencing data.

Long-read sequencing offers long contiguous DNA fragments, facilitating diploid genome assembly and structural variant (SV) detection. Efficient and robust algorithms for SV identification are crucial with increasing data availability. Alignment-based methods, favored for their computational efficiency and lower coverage requirements, are prominent. Alternative approaches, relying solely on available reads for de novo genome assembly and employing assembly-based tools for SV detection via comparison to a reference genome, demand significantly more computational resources. However, the lack of comprehensive benchmarking constrains our comprehension and hampers further algorithm development. Here we systematically compare 14 read alignment-based SV calling methods (including 4 deep learning-based methods and 1 hybrid method), and 4 assembly-based SV calling methods, alongside 4 upstream aligners and 7 assemblers. Assembly-based tools excel in detecting large SVs, especially insertions, and exhibit robustness to evaluation parameter changes and coverage fluctuations. Conversely, alignment-based tools demonstrate superior genotyping accuracy at low sequencing coverage (5-10×) and excel in detecting complex SVs, like translocations, inversions, and duplications. Our evaluation provides performance insights, highlighting the absence of a universally superior tool. We furnish guidelines across 31 criteria combinations, aiding users in selecting the most suitable tools for diverse scenarios and offering directions for further method development.

Is percutaneous tibial nerve stimulation (PTNS) effective for fecal incontinence (FI) in adults compared with sham electrical stimulation? A meta-analysis.

BACKGROUND: Sacral nerve neuromodulation (SNM) has been considered the optimal second-line treatment for fecal incontinence (FI). However, SNM involves high cost and requires highly skilled operators. Percutaneous tibial nerve stimulation (PTNS) has emerged as an alternative treatment modality for FI, yielding varying clinical outcomes. We conducted this meta-analysis to evaluate the effectiveness and safety of PTNS compared to sham electrical stimulation for FI. METHODS: PubMed, Embase, Web of Science, and the Cochrane Library were searched for studies from May 12, 2012 to May 12, 2022. RESULTS: Four randomized controlled studies were included in this review, involving a total of 439 adult patients with FI (300 in the PTNS group and 194 in the sham electrical stimulation group). Our meta-analysis revealed that PTNS demonstrated superior efficacy in reducing weekly episodes of FI compared to the control groups (MD - 1.6, 95% CI - 2.94 to - 0.26, p = 0.02, I2 = 30%). Furthermore, a greater proportion of patients in the PTNS group reported more than a 50% reduction in FI episodes per week (RR 0.73, 95% CI 0.57-0.94, p = 0.02, I2 = 6%). However, no significant differences were observed in any domains of the FI Quality of Life (QoL) and St Mark's incontinence scores (MD - 2.41, 95% CI - 5.1 to 0.27, p = 0.08, I2 = 67%). Importantly, no severe adverse events related to PTNS were reported in any of the participants. CONCLUSIONS: Our meta-analysis revealed that PTNS was more effective than sham stimulation in reducing FI episodes and led to a higher proportion of patients reporting more than a 50% reduction in weekly FI episodes.

Effects of continuous low-speed biogas agitation on anaerobic digestion of high-solids pig manure: Performance and microbial community.

This study aimed to investigate effects of continuous low-speed biogas agitation on the anaerobic digestion (AD) performance and microbial community of high-solids pig manure (total solids content of 10%). Our results reveal that at a biogas agitation intensity of 1.10 L/g feed VS/d, CH4 production increased by 16.67% compared to the non-agitated condition, the removal efficiency of H2S reached 63.18%, and the abundance of Methanosarcina was the highest. The presence of Hungateiclostridiaceae was associated with H2S concentrations. An increasing biogas agitation intensity led to an elevated pH and a decreased oxidation-reduction potential (ORP). Acetate concentrations, pH, and ORP values indicated changes in H2S concentrations. Sedimentibacter demonstrates the potential to indicate biogas agitation intensity and pH. We demonstrate that continuous low-speed biogas agitation effectively increases CH4 production and reduces H2S concentrations in AD of high-solids pig manure, offering a potential technical pathway for developing AD processes for high-solids pig manure, it also demonstrates that AD process can reduce the risk of pathogen and parasite transmission.

ERK1/2-dependent activity of SOX9 is required for sublytic C5b-9-induced expression of FGF1, PDGFα, and TGF-ß1 in rat Thy-1 nephritis.

Mesangial proliferative glomerulonephritis (MsPGN) and its related rat model Thy-1 nephritis (Thy-1N) are associated with C5b-9 deposition and are characterized by proliferation of glomerular mesangial cell (GMC) and expansion of extracellular matrix (ECM) expansion, alongside overexpression of multiple growth factors. Although fibroblast growth factor 1 (FGF1), platelet-derived growth factor alpha (PDGFα), and transforming growth factor beta 1 (TGF-ß1) are well known for their proproliferative and profibrotic roles, the molecular mechanisms responsible for regulating the expression of these growth factors have not been thoroughly elucidated. In this study, we found that sublytic C5b-9 induction of sex-determining region Y-box 9 (SOX9) transactivated FGF1, PDGFα, and TGF-ß1 genes in GMCs, resulting in a significant increase in their mRNA and protein levels. Besides, sublytic C5b-9 induction of activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2) phosphorylated SOX9 at serine 181 and serine 64, which enhanced SOX9's ability to transactivate FGF1, PDGFα, and TGF-ß1 genes in GMCs. Furthermore, we demonstrated that inhibiting ERK1/2 activation or silencing either ERK1/2 or SOX9 gene led to reduced SOX9 phosphorylation, decreased generation of FGF1, PDGFα, and TGF-ß1, and ameliorated glomerular injury in rat Thy-1N. Overall, these findings suggest that expression of FGF1, PDGFα, and TGF-ß1 is promoted by ERK1/2-mediated phosphorylation of SOX9, which may provide a valuable insight into the pathogenesis of MsPGN and offer a potential target for the development of novel treatment strategies for MsPGN.

A large sample-based case-control study of related risk factors of two types of lichenoid vulvar disease (LVD).

PURPOSE: The purpose of this study is to identify the associated factors of two types of lichenoid vulvar disease (LVD) and to compare the differences in related factors between the different pathological types of lichenoid vulvar disease (LVD). METHODS: The study conducted at the West China second Hospital of Sichuan University included a total of 1770 patients with biopsy-confirmed vulvar lichen simplex chronicus (VLSC)and vulvar lichen sclerosus(VLS), along with 1209 patients with normal vulvovagina as control. Further pathological subtype analysis was carried out on 163 cases of vulvar lichen simplex chronicus and 51 cases of vulvar lichen sclerosus. In addition, Univariate chi-square test and multivariate logistic regression were used to analyze the lichenoid vulvar disease group and vulvovaginal normal control group. RESULTS: Univariate analysis revealed that there were statistically significant differences (P < 0.05) in factors between the LVD group and the control group, except for living type, sleep habit, history of drinking, and allergic diseases. There was no significant difference in late sleep, spicy diet, and coffee intake in the factors of life and eating habits and the concomitant disease factors. Furthermore, univariate analysis showed that except for eating seafood, humid living environment, residence, caffeinated drinks, hypertension, and vaginitis, there were statistical differences in the related factors of LVSC. CONCLUSION: The incidence about lichenoid vulvar disease is influenced by various factors such as dietary habits, living environment, mental stress, concomitant diseases, hormone levels and so on, and there were no significant differences in these factors between VLS and VLSC except for income, work stress, systemic immune diseases, and menopause.

Urinary tract infections and intermittent catheterization among patients with spinal cord injury in Chinese community.

We conducted a cross-sectional study using an online questionnaire to investigate the current status of urinary tract infections (UTIs) and the use of clean intermittent catheterization (CIC) in Chinese community-based SCI patients and to explore the risk factors for UTIs in patients using CIC. Our findings suggest that the prevalence of UTIS is higher in Chinese community-based SCI patients compared with patients in medically developed countries. In addition, we found that CIC had the lower incidence of UTIs than urinary indwelling catheter (UIC) and suprapubic catheter (SPC), and that SCI patients with CIC had low rates of use and poor compliance. Further analysis indicated that most of the risk factors for UTIs in CIC patients were associated with irregular use of CIC. Therefore, we call for not only the need to provide stronger caregiver support and financial assistance to improve CIC adherence in Chinese community SCI patients, but also the establishment of a database of Chinese SCI patients in order to enhance the management of bladder emptying methods and further standardize the CIC operation in such patients, thus reducing the risk of UTIs in Chinese community SCI patients.

In vitro Activity of Cefepime/Avibactam Against Carbapenem Resistant Klebsiella pneumoniae and Integrative Metabolomics-Proteomics Approach for Resistance Mechanism: A Single-Center Study.

Purpose: We aimed to evaluate the in vitro antibacterial effects of combination of cefepime/avibactam against carbapenem-resistant Klebsiella pneumonia (CRKP) and explore the resistance mechanism of FEP/AVI. Patients and Methods: This study explored the in vitro antibacterial activities of ceftazidime/avibactam (CAZ/AVI) and cefepime/avibactam (FEP/AVI) against 40 and 76 CRKP clinical isolates. Proteomics and metabolomics were employed to investigate the resistance mechanisms of CRKP to FEP/AVI. Results: FEP/AVI (MIC50/MIC90 0.5/4-64/4 µg/mL, resistance rate 17.1%) showed better antibacterial activity against CRKP than CAZ/AVI (MIC50/MIC90 4/4-128/4 µg/mL, resistance rate 20%) in vitro. Bioinformatics analysis showed that the differentially expressed proteins (DEPs) were enriched in alanine, aspartate and glutamate metabolism, and ribosome. Remarkably, transcriptional and translational activity-related pathways were inhibited in FEP/AVI resistant CRKP. Overlap analysis suggested that H-NS might play an important role in resistance to FEP/AVI in CRKP. The mRNA levels of DEPs-related genes (adhE, gltB, purA, ftsI and hns) showed the same trends as DEPs in FEP/AVI susceptible and resistant strains. FEP/AVI resistant isolates demonstrated stronger biofilm formation capacity than susceptible isolates. Metabolomics results showed that disturbed metabolites were mainly lipids, and adenine was decreased in FEP/AVI resistant CRKP. Conclusion: These results indicated that H-NS, GltB and SpoT may directly or indirectly promote biofilm formation of CRKP and led to FEP/AVI resistance, but inhibited ribosomal function. Our study provides a mechanistic insight into the acquisition of resistance to FEP/AVI in Klebsiella pneumoniae.

Sublytic C5b-9 induces TIMP3 expression by glomerular mesangial cells via TRAF6-dependent KLF5 K63-linked ubiquitination in rat Thy-1 nephritis.

Rat Thy-1 nephritis (Thy-1N) is an experimental model for studying human mesangioproliferative glomerulonephritis (MsPGN), and its pathological features are glomerular mesangial cell (GMC) proliferation and extracellular matrix (ECM) accumulation. Although we have confirmed that renal lesions of Thy-1N rats are sublytic C5b-9-dependent, and ECM accumulation is related to tissue inhibitor of matrix metalloproteinase (TIMP) inhibiting matrix metalloproteinase (MMP) activity, whether sublytic C5b-9 can induce TIMP production by GMC in Thy-1N rat and the underlying mechanism remains unclear. In the study, we proved that the expressions of TIMP3, krϋppel-like transcription factor 5 (KLF5) and tumor necrosis factor receptor-associated factor 6 (TRAF6) were simultaneously up-regulated both in the renal tissues of Thy-1N rats (in vivo) and in the GMC exposed to sublytic C5b-9 (in vitro). Further mechanism exploration discovered that KLF5 and TRAF6 as two upstream molecules could induce TIMP3 gene transcription through binding to the same region i.e., -1801nt to -1554nt (GGGGAGGGGC) and -228nt to -46nt (GCCCCGCCCC) of TIMP3 promoter. In the process, TRAF6 mediated KLF5 K63-linked ubiquitination at K99 and K100 enhancing KLF5 nuclear localization and binding to TIMP3 promoter, augmenting its gene activation. Furthermore, the experiments in vivo exhibited that silencing KLF5, TRAF6 or TIMP3 gene could markedly lessen renal KLF5 K63-linked ubiquitination or TIMP3 induction, ECM accumulation and other pathological changes of Thy-1N rats. Besides, the positive expressions of above-mentioned these proteins and ECM accumulation and their correlation in the renal tissues of MsPGN patients were also demonstrated. Overall, our findings implicate that KLF5 and TRAF6 play a promoting role in sublytic C5b-9-triggered TIMP3 gene transcription and expression, which might provide a novel mechanistic insight into rat Thy-1N and human MsPGN.

Up-flow anaerobic sludge blanket treatment of swine wastewater: Effect of heterologous and homologous inocula on anaerobic digestion performance and the microbial community.

The effects of heterogenous (anaerobic sludge from treating distillery sewage, ASDS) and homologous (anaerobic sludge from treating swine wastewater, ASSW) inocula on anaerobic digestion and the microbial community in an up-flow anaerobic sludge blanket treating swine wastewater were compared. The highest chemical oxygen demand removal efficiencies with ASDS (84.8%) and ASSW (83.1%) were obtained with an organic loading rate of 15 kg COD/m3/d. For ASSW compared with ASDS, methane production efficiency was 15.3% higher and excess sludge production was 73.0% lower. The abundance of the cellulose hydrolyzing bacterium Clostridium sensu stricto_1 with ASDS (36.1%) was 1.5 times that with ASSW, while that of Methanosarcina with ASSW (22.9%) was > 100 times that with ASDS. ASDS reduced the content of pathogenic bacteria by 88.0%, while ASSW maintained a low level of pathogenic bacteria. ASSW greatly improved the methane production efficiency of wastewater and is more suitable for treating swine wastewater.

Association of maternal dietary patterns derived by multiple approaches with gestational diabetes mellitus: a prospective cohort study.

We used two a priori diet scores [Mediterranean diet (aMed) and Diet Balance Index (DBI)] and two a posteriori approaches [principal components analysis (PCA) and reduced-rank regression (RRR)] to examine the association of maternal dietary patterns with risk of gestational diabetes mellitus (GDM) and blood glucose among 2202 pregnant women in the Tongji Birth Cohort. Compared to the highest quartile of the aMed and legumes-vegetables-fruits (derived by PCA) scores, the fasting blood glucose (FBG) levels were higher in the lower quartiles (p-trend < 0.05). Lower scores of the meats-eggs-dairy (derived by PCA) and eggs-fish patterns (derived by RRR; characterised by higher intakes of freshwater fish, eggs, and lower intakes of leafy and cruciferous vegetables and fruits) were associated with decreased FBG levels (p-trend < 0.05). Similarities were found across approaches that some dietary patterns were associated with FBG, but not with postprandial glucose and GDM risk.

Pregnancy thiamine and riboflavin intake and the risk of gestational diabetes mellitus: A prospective cohort study.

BACKGROUND: Thiamine and riboflavin deficiencies exist to varying degrees worldwide, especially in developing countries. Evidence regarding the association between thiamine and riboflavin intake and gestational diabetes mellitus (GDM) is scarce. OBJECTIVES: We aimed to evaluate the association of thiamine and riboflavin intake during pregnancy, including dietary source and supplementation, with GDM risk in a prospective cohort study. METHODS: We included 3036 pregnant women (923 in the first trimester and 2113 in the second trimester) from the Tongji Birth Cohort. A validated semi-quantitative food frequency questionnaire and a lifestyle questionnaire were used to assess thiamine and riboflavin intake from dietary source and supplementation, respectively. GDM was diagnosed using the 75 g 2-h oral glucose tolerance test at 24-28 weeks of gestation. A modified Poisson regression or logistic regression model was used to evaluate the association between thiamine and riboflavin intake and GDM risk. RESULTS: Dietary intake of thiamine and riboflavin was at low levels during pregnancy. In the fully adjusted model, compared with participants in quartile 1 (Q1), those who had more total thiamine and riboflavin intake had a lower risk of GDM during the first trimester [thiamine: Q2: RR: 0.58 (95% CI: 0.34, 0.98); Q3: RR: 0.45 (95% CI: 0.24, 0.84); Q4: RR: 0.35 (95% CI: 0.17, 0.72), P for trend = 0.002; riboflavin: Q2: RR: 0.63 (95% CI: 0.37, 1.09); Q3: RR: 0.45 (95% CI: 0.24, 0.87); Q4: RR: 0.39 (95% CI: 0.19, 0.79), P for trend = 0.006]. This association was also observed during the second trimester. Similar results were observed for the association between thiamine and riboflavin supplement use but not dietary intake and GDM risk. CONCLUSIONS: Higher intake of thiamine and riboflavin during pregnancy is associated with a lower incidence of GDM. This trial was registered at http://www.chictr.org.cn as ChiCTR1800016908.

Mapping schistosomiasis risk in Southeast Asia: a systematic review and geospatial analysis.

BACKGROUND: Schistosomiasis is a water-borne parasitic disease estimated to have infected >140 million people globally in 2019, mostly in sub-Saharan Africa. Within the goal of eliminating schistosomiasis as a public health problem by 2030 in the World Health Organization (WHO) Roadmap for neglected tropical diseases, other regions cannot be neglected. Empirical estimates of the disease burden in Southeast Asia largely remain unavailable. METHODS: We undertook a systematic review to identify empirical survey data on schistosomiasis prevalence in Southeast Asia using the Web of Science, ScienceDirect, PubMed and the Global Atlas of Helminth Infections, from inception to 5 February 2021. We then conducted advanced Bayesian geostatistical analysis to assess the geographical distribution of infection risk at a high spatial resolution (5 × 5 km) using the prevalence, number of infected individuals and doses needed for preventive chemotherapy. RESULTS: We identified 494 Schistosoma japonicum surveys in the Philippines and Indonesia, and 285 in Cambodia and Laos for S. mekongi. The latest estimates suggest that 225 [95% credible interval (CrI): 168-285] thousand in the endemic areas of Southeast Asian population were infected in 2018. The highest prevalence of schistosomiasis was 3.86% (95% CrI: 3.40-4.31) in Laos whereas the lowest was 0.29% in Cambodia (95% CrI: 0.22-0.36). The estimated number of praziquantel doses needed per year was 1.99 million (95% CrI: 1.92-2.03 million) for the entire population in endemic areas of Southeast Asia. CONCLUSIONS: The burden of schistosomiasis remains far from the WHO goal and our estimates highlighted areas to target with strengthened interventions against schistosomiasis.

Artificial Nucleobase-Directed Programmable Synthesis and Assembly of Amphiphilic Nucleic Acids as an All-in-One Platform for Cation-Free siRNA Delivery.

Efficient transport of nucleic acid therapeutics into targeted cells is the key step of genetic modulation in disease treatment. Nowadays, delivery systems strongly rely on cationic materials, but how to balance the trade-off between effectiveness and toxicity of these exogenous materials remains incredibly challenging. Here, we take inspiration from nucleic acid chemistry and introduce a new concept of amphiphilic nucleic acids (ANAs), as an all-in-one platform for cation-free nucleic acid delivery, by programmatically conjugating two different artifical nucleobases with sequence-independent activities. Specifically, the hydrophilic artificial nucleobases in ANAs act as both delivery vectors and therapeutic cargos for integrated benefits, while the hydrophobic nucleobases enable molecular self-assembly for improved stability and endosomal membrane oxidation for enhanced endosomal escape. By virtue of these merits, this platform is successfully used for short interference RNA (siRNA) delivery, which demonstrates a high siRNA loading capacity, rapid cellular uptake, and efficient endosomal escape, eliciting remarkable gene silencing and synergistic inhibitory effects on cancer cell proliferation and migration. This work is a case study in exploiting the basis of nucleic acid chemistry to afford new paradigms for advanced cancer theranostics.

Phthalate acid esters and polycyclic aromatic hydrocarbons concentrations with their determining factors among Chinese pregnant women: A focus on dietary patterns.

BACKGROUND: Pregnant women are susceptible to adverse health effects associated with phthalate acid esters (PAEs) and polycyclic aromatic hydrocarbons (PAHs), and diet is a significant exposure source. Little is known about the contributions of dietary patterns during pregnancy to the exposure variability of these environmental contaminants. OBJECTIVES: To identify dietary patterns in relation to PAEs and PAHs exposure in the Chinese pregnant population. METHODS: Dietary data and urinary concentrations of environmental pollutants were obtained from 1190 pregnant women in the Tongji Birth Cohort (TJBC). PAEs and PAHs were measured in spot urine samples. Food intake was assessed using a food-frequency questionnaire. Dietary patterns were constructed by principal component analysis (PCA). Through PCA, we also extracted three chemical mixture scores that represent different co-exposure patterns of PAEs and PAHs. Multiple linear regression models were adopted to identify predictors of PAEs and PAHs exposure. RESULTS: Four dietary patterns were identified by PCA that explained 44.9 % of the total variance of food intake. We found egg-dairy products pattern, whole grain-tuber crop pattern, and meat-aquatic products pattern were positively associated with specific pollutants exposure. In contrast, fruit-nut-vegetable pattern was negatively correlated with PAEs and PAHs exposure. Every SD increase in this pattern score was associated with 14.36 % reduced mono(2-ethyl-5-oxohexyl) phthalate (MEOHP) (95 % CI: -24.50 ~ -2.96, p-trend = 0.01), 10.86 % reduced 2-hydroxynaphthalene (2-OHNap) (95 % CI: -20.07 ~ -0.60, p-trend = 0.04), 19.35 % reduced 9-hydroxyphenanthrene (9-OHPhe) (95 % CI: -34.49 ~ -0.70, p-trend = 0.01), and 8.33 % reduced scores of PAHs group (95 % CI: -15.97 ~ -0.10, p-trend = 0.02). In addition, disposable tableware usage and passive smoking were suggested as potentially modifiable sources of PAEs and PAHs exposure, respectively. CONCLUSION: Adhering to egg-dairy products pattern, whole grain-tuber crop pattern, and meat-aquatic products pattern may be related to increased PAEs and PAHs exposure, while following fruit-nut-vegetable pattern seems to correlate with a lower burden of such exposure.

A prospective study of specimen eversion to lateral rectum and valgus resection for low rectal cancer.

Purpose: To investigate the safety and efficacy of a reverse puncture device (RPD) and specimen eversion of the rectum for resection in total laparoscopic proctectomy. Methods: In a prospective study from August 2019 to March 2021, 40 patients underwent a procedure with an RPD and specimen eversion of the rectum for total laparoscopic low rectal cancer resection, that is natural orifice specimen extraction surgery (NOSES), were included in the NOSES group. Forty patients in the control group underwent conventional laparoscopic radical resection for low rectal cancer and were included in the LAP group. Intraoperative- and postoperative-related indicators, recovery and inflammatory factors, quality of life (QOL) and mental health were compared. Results: All operations were successfully completed. Compared with the LAP group, the NOSES group showed better short-term outcomes, such as time to eating, postoperative pain, and especially postoperative incision-related complications. At the same time, postoperative inflammatory factor levels, psychological trauma, life-related anxiety and depression scores, and QOL were better in the NOSES group than in the LAP group. Conclusions: The application of an RPD and specimen eversion of the rectum for total laparoscopic low rectal cancer resection is a technically feasible and safe approach with a short-term curative effect.

Reduced porin expression with EnvZ-OmpR, PhoPQ, BaeSR two-component system down-regulation in carbapenem resistance of Klebsiella Pneumoniae based on proteomic analysis.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) has proven to be an urgent threat to human health. Proteomics (TMT/LC-MS/MS) and bioinformatics approaches were employed to explore the potential mechanisms underlying carbapenem resistance. Proteomic profiling of CRKP and susceptible KP (sKP) isolates revealed the involvement of outer membrane, beta-lactam resistance pathway, and two-component systems (TCSs) in carbapenem resistance. 27 CRKP strains and 27 susceptible Klebsiella pneumoniae strains were isolated from inpatients at the Second Xiangya Hospital, China to verify the mechanisms. Modified carbapenem inactivation method (mCIM) and PCR of common carbapenem resistance genes confirmed that 77.8% (21/27) of CRKP isolates were carbapenemase-producing. Porin decrease in CRKP isolates was found by SDS-PAGE and mRNA levels of major porins (OmpK35 and OmpK36). RT-qPCR detection of two-component systems (envZ, ompR, phoP, phoQ, baeS and baeR) revealed down-regulation of EnvZ-OmpR, PhoPQ, BaeSR TCSs. Expression of the TCSs, except ompR, were closely correlated with OMPs with the R-value >0.7. Together, this study reaffirmed the significance of the ß-lactam resistance pathway in CRKP based on proteomic analysis. OmpK35/36 porin reduction and the controversial downregulation of EnvZ-OmpR, PhoPQ, and BaeSR TCSs were confirmed in carbapenem resistance of Klebsiella pneumoniae.

Analysis of Hypoxia Inducible Factor-1α Expression and Its Effects on Glycolysis of Esophageal Carcinoma.

We investigated the regulatory effects of hypoxia-inducible factor-1a (HIF-1α) on glycolysis metabolism in esophageal carcinoma (ESCA) cells. A series of bioinformatics databases and tools were used to investigate the expression and role of HIF-1α in ESCA. The expression of HIF-1a in ESCA tissues and adjacent tissues was validated by real-time PCR. Small interfering RNA (siRNA) was used to inhibit HIF-1α-related genes in human ESCA cells (Eca109 and KYSE150). Cell proliferation was detected by the CCK-8 assay. The expression of HIF-1α and glycolytic enzymes were investigated by real-time PCR and Western blot. HIF-1α is highly expressed in ESCA and is involved in many biological processes such as cell hypoxia reaction, glucose metabolic process. Further in vitro experiments showed that expression of HIF-1α in Eca109 and KYSE150 significantly increased under hypoxia compared with normoxia conditions. Also, the glucose uptake and lactate production under hypoxia were higher. The expression levels of hexokinase 2 (HK2) and pyruvate dehydrogenase kinase 1 (PDK1), glycolysis-related genes, were significantly increased under hypoxia. After siRNA knockdown of HIF-1a in Eca109 and KYSE150, the glucose uptake and lactate production, as well as cell proliferation were significantly decreased under hypoxia, and HK2 and PDK1 were significantly downregulated. HIF-1α promotes glycolysis of ESCA cells by upregulating the expression of HK2 and PDK1 under hypoxia.

Konjac glucomannan improves the gel properties of low salt myofibrillar protein through modifying protein conformation.

Improving the characteristics of low salt proteins is the key to the gel properties of low-salt meat products which are demanded by people nowadays. The present study focused on the effects of KGM concentrations on the changes in structure and gelling properties of low-salt myofibrillar protein (MP). KGM addition (≤0.75 %) irrespective of salt concentration modified secondary and tertiary structures of MPs, enhanced the binding capacity of Troponin-T and Tropomyosin, augmented the gelling behavior of proteins, and remarkably improved the storage modulus (G') and gel strength of heat-induced MP gels. Interestingly, KGM addition in low salt condition showed the transformation of the all-gauche SS conformation into gauche-gauche-trans and trans-gauche-trans, and the partial transformation of α-helices into ß-sheets. overall, KGM modified the structure of low salt MPs and thus improved the gel properties of low salt MPs. Therefore, KGM is recommended for low-salt meat processing to enhance the MP gelling potential.