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Background Some clinical researchers have reported that patients with cCR (clinical complete response) status after neoadjuvant chemoradiotherapy (nCRT) could adopt the watch-and-wait (W&W) strategy. Compared with total mesorectal excision (TME) surgery, the W&W strategy could achieve a similar overall survival. Could the W&W strategy replace TME surgery as the main treatment option for the cCR patients? By using the meta-analysis method, we evaluated the safety and efficacy of the W&W strategy and TME surgery for rectal cancer exhibiting cCR after nCRT. Methods We evaluated two treatment strategies for rectal cancer with cCR after nCRT up to July 2021 by searching the Cochrane Library, PubMed, Wanfang, and China National Knowledge Infrastructure (CNKI) databases. Clinical data for primary outcomes (local recurrence, cancer-related death and distant metastasis), and secondary outcomes (disease-free survival (DFS) and overall survival (OS)) were collected to evaluate the efficacy and safety in the two groups. Results We included nine studies with 818 patients in the meta-analysis, and there were five moderate-quality studies and four high-quality studies. A total of 339 patients were in the W&W group and 479 patients were in the TME group. The local recurrence rate in the W&W group was greater than that in the TME group in the fixed-effects model (OR 8.54, 95% CI 3.52 to 20.71, P < 0.001). The results of other outcomes were similar in the two groups. Conclusion The local recurrence rate of the W&W group was greater than that in the TME group, but other results were similar in the two groups. With the help of physical examination and salvage therapy, the W&W strategy could achieve similar treatment effects with the TME approach. Trial registration Protocol registration number: CRD42021244032.

Abnormal RNA m6A methylation is known to lead to the occurrence and progression of multiple cancers including gastric cancer (GC). However, the integrative effects of all m6A methylation regulators on GC prognosis are unclear. Our research aimed to globally analyze the prognosis values of all 33 m6A RNA methylation regulators in GC by univariate and multivariate Cox regression analyses. Among all 33 m6A RNA methylation regulators, fat mass and obesity-associated protein (FTO), an m6A demethylase, was identified as a key prognostic risk factor on overall survival (OS) of GC patients. It was found that FTO could promote GC cell migration and invasion abilities, and we predicted that ITGB1 was a demethylated target of FTO. Knockdown (KD) of FTO significantly down-regulated ITGB1 expression at both mRNA and protein levels and augmented ITGB1 mRNA m6A modification level. Moreover, overexpression (OE) of ITGB1 could partially reverse FTO-KD-inhibited migration and invasion of GC cells. Our study found that FTO was an independent risk factor for overall survival (OS) of GC patients and FTO could promote GC metastasis by upregulating the expression of Integrin β1(ITGB1) via decreasing its m6A level. These results indicated that FTO can be a potent GC biomarker for prognosis prediction as well as a potential target in GC treatment.

Background: Triple negative breast cancer (TNBC) is a tumor characterized by high recurrence and mortality, but without effective targeted therapy. It is urgent to explore new treatment strategy to improve the efficacy of TNBC therapy. Methods: Transcriptomic profiling datasets of TNBC were used for screening TNBC specific gene sets. Drug prediction was performed in Connectivity map (CMap) database. Molecular docking method was used for analyzing drug targets. In vitro and in vivo models of TNBC were constructed to examine the drug efficacy. Results: We screened out Mibefradil, a T-type Ca2+ channel blocker, might be a potential therapeutic drug for TNBC by transcriptomics and bioinformatics analysis, and verified that Mibefradil could inhibit the proliferation of TNBC cells by inducing apoptosis and cell cycle arrest. Furthermore, by network pharmacology and molecular docking analysis, AURKA was predicted as the most possible drug target of Mibefradil. Finally, it was proved that Mibefradil treatment could induce apoptosis by decreasing protein expression and phosphorylation level of AURKA in vitro and in vivo. Conclusions: Mibefradil played anti-cancer role in TNBC cells by targeting to AURKA to induce cell cycle and apoptosis. Our results repurposed Mibefradil as a potential targeted drug of TNBC and provided a fundamental research for a novel strategy TNBC treatment.

Succinylation is a newly discovered and multienzyme-regulated post-translational modification (PTM) that is associated with the initiation and progression of cancer. Currently, no systematic analyses on the role of succinylation regulators in tumors have been reported. In this study, we performed a comprehensive pan-cancer analysis on four well-known succinylation regulators (CPT1A, KAT2A, SIRT5, and SIRT7). We found that these regulators played specific and critical roles in the prognosis of clear cell renal cell carcinoma (ccRCC). We constructed a risk score (RS) based on two independent prognostic prediction factors, CPT1A and KAT2A, and subsequently developed a nomogram model containing the RS, which showed good accuracy in the prediction of overall survival (OS) in ccRCC patients. Furthermore, we used the similar expression pattern of four succinylation regulators according to consensus clustering analysis to divide the patients into three clusters that exhibited prominently different OS as well as clinicopathological characteristics. Differently expressed genes (DEGs) and pathway enrichment analyses of three clusters indicated that succinylation regulators might promote malignant progression of ccRCC by regulating the infiltration of immune cells and RNA N6-methyladenosine (m6A) methylation. Importantly, our data suggest that CPT1A and SIRT5 might up-regulate and down-regulate the expression of LRPPRC and EIF3B, respectively. Our study systematically analyzed the prognostic predictive values of four succinylation regulators and revealed their potential mechanisms in ccRCC aggressiveness. These data provide new insight into the understanding of succinylation modification and present clinical evidence for its role in ccRCC treatments.

Patients with EGFR-mutant non-small-cell lung cancer (NSCLC) greatly benefit from EGFR-tyrosine kinase inhibitors (EGFR-TKIs) while the prognosis of patients who lack EGFR-sensitive mutations (EGFR wild type, EGFR-WT) remains poor due to a lack of effective therapeutic strategies. There is an urgent need to explore the key genes that affect the prognosis and develop potentially effective drugs in EGFR-WT NSCLC patients. In this study, we clustered functional modules related to the survival traits of EGFR-WT patients using weighted gene co-expression network analysis (WGCNA). We used these data to establish a two-gene prognostic signature based on the expression of CYP11B1 and DNALI1 by combining the least absolute shrinkage and selection operator (LASSO) algorithms and Cox proportional hazards regression analysis. Following the calculation of risk score (RS) based on the two-gene signature, patients with high RSs showed a worse prognosis. We further explored targeted drugs that could be effective in patients with a high RS by the connectivity map (CMap). Surprisingly, multiple HDAC inhibitors (HDACis) such as trichostatin A (TSA) and vorinostat (SAHA) that may have efficacy were identified. Also, we proved that HDACis could inhibit the proliferation and metastasis of NSCLC cells in vitro. Taken together, our study identified prognostic biomarkers for patients with EGFR-WT NSCLC and confirmed a novel potential role for HDACis in the clinical management of EGFR-WT patients.

As a vital feature of the microenvironment, hypoxia, especially long-term hypoxia, is known to promote metastasis and lead to poor prognosis in solid tumors. Circular RNAs (circRNAs) participate in important processes of cell proliferation and metastasis in cancers. However, the contribution of circRNAs to metastasis under long-term hypoxia is obscure. In this study, we aim to explore specific functions of circHIPK3 in long-term hypoxia-promoting metastasis of gastric cancer (GC). The hypoxic resistant gastric cancer (HRGC) cell lines we established previously, which were tolerant to 2% O2 conditions, were used as the long-term hypoxia model. We found that circHIPK3 was upregulated by HIF-2α in HRGC cells, and circHIPK3 facilitated the migration and invasion ability of HRGC cells. Further investigation proved that circHIPK3 promoted metastasis of HRGC cells directly by interacting with miR-653-5p and miR-338-3p to relieve the suppression of neuropilin 1 (NRP1), resulting in the activation of downstream ERK and AKT pathways. Our study identified oncogene functions of circHIPK3 under a long-term hypoxic microenvironment and the possibility of using circHIPK3 as a potential biomarker of long-term hypoxia in GC. In conclusion, circHIPK3 could promote GC metastasis via the miR-653-5p/miR-338-3p-NRP1 axis under a long-term hypoxic microenvironment.

The expression of anillin mRNA and protein is regulated in a cell cycle‑dependent manner. However, the mechanism underlying this process is unclear. Previous studies analyzing the sequence of the 5'‑untranslated region of anillin have unveiled several putative p53 binding sites. Therefore, the present study hypothesized that the anillin gene may be repressed by p53 and that the commonly observed mutation (or loss of function) of p53 may serve a role in this phenotype. Bioinformatic analysis of the anillin promoter region revealed potential p53 responsive elements. Of those identified, 2 were able to bind p53 protein, as determined via a chromatin immunoprecipitation assay. Although it was hypothesized that DNA damage and resultant p53 expression would repress anillin expression, the results revealed that anillin mRNA and protein expression levels were negatively regulated by DNA damage in the wild‑type p53 cells, but not in the isogenic p53 null cells. Furthermore, DNA sequences encompassing the p53 binding site downregulated luciferase transgenes in a p53 dependent manner. Taken together, these data indicated that anillin was negatively regulated by p53 and that anillin overexpression observed in cancer may be a p53‑mediated phenomenon. The data from the present study provided further evidence for the role of p53 in the biologically crucial process of cytokinesis.

Septic liver injury remains a challenge in sepsis treatment. Nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing 3 (NLRP3) inflammasome activation has been suggested to be a major cause of hepatocyte cell death in liver diseases. However, insufficient research has been performed to explore the underlying mechanisms associated with this. In the present study, sophocarpine, a pharmaceutical monomer originally isolated from Sophora flavescens, was suggested to attenuate septic liver injury in a mouse cecal ligation and puncture (CLP) model. By utilizing western blotting, ELISA, H&E staining and immunohistochemistry, the results demonstrated that sophocarpine treatment reversed CLP-induced elevations in serum aspartate transaminase, alanine transaminase, interleukin (IL)-6 and IL-1β levels. Additionally, sophocarpine appeared to have suppressed the activation of the NLRP3 inflammasome, as indicated by observed reductions in liver IL-1β, NLRP3, caspase 1-p20 and gasdermin D-p30 protein levels. Further investigation suggested that sophocarpine-induced autophagy was essential for this suppression of NLRP3 inflammasome activation, the inhibition of which reversed the protective effects of sophocarpine on CLP-induced liver injury. Collectively, results from the present study suggested a protective role for sophocarpine against septic liver injury, where sophocarpine may suppress NLRP3 inflammasome activation by autophagy-mediated degradation.

The X‐linked genetic bleeding disorder caused by deficiency of coagulator factor IX, hemophilia B, is a disease ideally suited for gene therapy with genome editing technology. Here, we identify a family with hemophilia B carrying a novel mutation, Y371D, in the human F9 gene. The CRISPR/Cas9 system was used to generate distinct genetically modified mouse models and confirmed that the novel Y371D mutation resulted in a more severe hemophilia B phenotype than the previously identified Y371S mutation. To develop therapeutic strategies targeting this mutation, we subsequently compared naked DNA constructs versus adenoviral vectors to deliver Cas9 components targeting the F9 Y371D mutation in adult mice. After treatment, hemophilia B mice receiving naked DNA constructs exhibited correction of over 0.56% of F9 alleles in hepatocytes, which was sufficient to restore hemostasis. In contrast, the adenoviral delivery system resulted in a higher corrective efficiency but no therapeutic effects due to severe hepatic toxicity. Our studies suggest that CRISPR/Cas‐mediated in situ genome editing could be a feasible therapeutic strategy for human hereditary diseases, although an efficient and clinically relevant delivery system is required for further clinical studies.

This study developed a high density process using response surface methodology to cultivate Enterococcus faecalis CGMCC1.101 and subsequently investigated this liquid form product in improving the performance of weaned piglets. Exp. I was carried out to use a central composite design (CCD) for the optimization of carbon source, nitrogen sources and pH in the fermentation broth to raise the cell density of E. faecalis CGMCC1.101. Optimum conditions for growth of E. faecalis CGMCC1.101 were (g/l): beef extract, 12.5; yeast extract, 12.5; sucrose, 25; K2HPO4, 2; sodium acetate, 5; di-ammonium hydrogen citrate, 2; MgSO4, 0.2; MnSO4, 0.05; Tween 80, 1 ml/l with pH of 6.7. The ultimate biomass concentration of E. faecalis CGMCC1.101 using the optimized media reached 3.5×109 CFU/ml. In Exp. II, 108 weaned piglets (Duroc×Landrace×Large White), with an initial body weight of 8.9±2.5 kg, were randomly allocated to one of three treatments with six pens per treatment and six pigs per pen. Weaned piglets were fed either a corn-soybean meal based control diet or same diets supplemented with antibiotic (50 mg/kg Kitasamycin) or the liquid fermentation product (3.5×109 CFU/kg E. faecalis CGMCC1.101). The effects of the liquid culture of E. faecalis CGMCC1.101 on performance, intestinal microflora and morphology as well as nutrient digestibility in weaned piglets were investigated. The results of Exp. II demonstrate that the liquid culture of E. faecalis CGMCC1.101 can improve the efficiency of feed utilization and balance the microbial environment in weaned piglets.

Background The pleth variability index (PVI) has been demonstrated to be a useful, noninvasive indicator of continuous fluid responsiveness. Whether PVI can be used to assess the changes of intravascular volume status remains to be elucidated. Material/Methods Using correlation analysis and receiver operating characteristic (ROC) curves, we sought a correlation between PVI and the initial distribution volume of glucose (IDVG), evaluating PVI as an indicator of the central extracellular fluid volume after anesthesia induction in patients undergoing elective abdominal surgery. Results Strong negative correlations existed between IDVG and PVI (r=−0.72), IDVG, and pulse pressure variation (PPV) (r=−0.73), and between IDVG and systolic pressure variation (SPV) (r=−0.53), P

This study investigated the effects of solid-state fermentation of a compound pig feed on its microbial and nutritional characteristics as well as on pig performance and nutrient digestibility. A mixed culture containing Lactobacillus fermentum, Saccharomyces cerevisae and Bacillus subtilis was used for solid-state fermentation and solid-state fermented feed samples were collected on days 0, 1, 2, 3, 5, 7, 10, 15, 20 and 30 for microbial counts and chemical analysis. Lactic acid bacteria increased rapidly during the first three days of fermentation and then slowly declined until day 10 and, thereafter, the counts were maintained at about 6.7 log cfu/g for the duration of the fermentation period. Enterobacteria also increased during the first two days, and then fell below the detectable level of the analysis (3.0 log cfu/g). The pH of the fermentation substrate declined from 6.1 at the start of fermentation to 5.7 by day 30. The water-soluble protein content increased from 8.2 to 9.2% while the concentration of acetic acid increased from 16.6 to 51.3 mmol/kg over the 30-day fermentation. At the end of the 30-day fermentation, the solid-state fermented feed was used in a pig feeding trial to determine its effects on performance and nutrient digestibility in growing-finishing pigs. Twenty crossbred barrows (14.11+0.77 kg BW) were allotted into two dietary treatments, which comprised a regular dry diet containing antibiotics and a solid-state fermented feed based diet, free of antibiotics. There was no difference due to diet on pig performance or nutrient digestibility. In conclusion, solid-state fermentation resulted in high counts of lactic acid bacteria and low counts of enterobacteria in the substrate. Moreover, feeding a diet containing solid-state fermented feed, free of antibiotics, can result in similar performance and nutrient digestibility in growing-finishing pigs to a regular diet with antibiotics.

Wild type (WT) DNA sequence, which encoded a mature beta-mannanase of Aspergillus sulphureus, composed of 1,152 nucleotides (nt), was amplified from pUCm-T-mann by polymerase chain reaction (PCR). Based on this DNA fragment, mutants designated as E(206)G and E(314)G were constructed by overextension PCR (OE-PCR). Glutamic acids of the 206th and 314th sites in the amino acid sequence of beta-mannanase were separately replaced by glycine in these two mutants. The WT and mutant genes were ligated into prokaryotic vector pET-28a (+) and transformed into the Escherichia coli BL21 strain, respectively. The recombinant enzyme proteins were expressed by IPTG induction and detected by Western blot. The recombinant proteins purified with Ni-NTA column were dialyzed to correctly refold. The WT recombinant beta-mannanase showed optimal activity at 50 degrees C and pH 2.4. The kinetic parameters of K (m) and V (max) for purified beta-mannanase were 1.38 mg/ml and 72.99 U/mg, respectively. However, the mutant proteins did not show any activity. It was demonstrated that E(206) and E(314) were the catalytic residues of beta-mannanase.

Study objective To compare the efficacy of lightwand-guided classic laryngeal mask airway (cLMA) real-time insertion technique with the standard recommended index finger–guided insertion technique. Design Prospective, randomized controlled study. Setting University-affiliated hospital. Patients Three hundred patients undergoing minor gynecological or orthopedic surgeries under general anesthesia using the cLMA as an airway management tool. Interventions Patients were randomly divided into either lightwand-guided group or standard group. Measurements Fiberoptic bronchoscopy was used to determine the cLMA position after a cLMA was inserted. The first attempt and total success rates of the cLMA insertion, insertion time, distances from the end of cLMA pilot tube to the upper central incisors, views of fiberoptic bronchoscopy, blood staining, tidal volume, airway pressure, end-tidal CO 2 , SpO 2 , noninvasive hemodynamic parameters, and others were compared. Main results The cLMA was all successfully inserted within 3 attempts except for 2 patients in the standard group. The success rates of lightwand-guided insertion technique at first attempt were significantly higher than standard insertion technique; the ideal view rates assessed by fiberoptic bronchoscopy in lightwand-guided group patients were also significantly higher than in standard group patients, but the insertion time of first successful attempt was similar; the blood staining rates on the cLMA in lightwand-guided group patients were significantly less than in standard group patients. The depths of cLMA insertion in standard group patients were significantly deeper than those in lightwand-guided group patients. There was no significant difference in end-tidal CO 2 , SpO 2 , airway pressure, and hemodynamic variables. Conclusion Lightwand-guided cLMA insertion technique can provide a more objective indicator for correct cLMA positioning, higher first attempt success rates, better glottic views, and less damage to oropharyngeal or esophagus tissues than standard index finger–guided cLMA insertion technique.

A total of 124 samples were collected from the intestine of broiler chickens, piglet faeces, fermented foods, soils and Chinese herbs. More than 750 strains of aerobic, spore-forming bacteria were isolated from these samples. The inhibitory activity of these spore-forming strains against Escherichia coli K88, E. coli K99, Salmonella typhimurium and Staphylococcus aureus was assessed using a disc plate diffusion assay. The six bacilli with the largest inhibition zones against the four indicator bacteria were chosen and assessed for their resistance to unfavorable conditions within simulated gut environments. The strain Bacillus subtilis MA139 showed full resistance to pH 2, 0.3% bile salts and exhibited the highest antimicrobial activity. Based on these results, B. subtilis MA139 was selected as a potential probiotic and fed to piglets at concentrations of 2.2 x 10(5), 2.2 x 10(6) or 2.2 x 10(7) CFU/g of feed during a 28-day feeding trial. A negative control consisting of the basal diet with no additives and a positive control consisting of the basal diet supplemented with 16 g/ton flavomycin were also included. Ninety piglets between 35 and 40 days old were used in the in vivo animal trials. B. subtilis MA139 enhanced daily gain (P = 0.10) and feed conversion (P = 0.03) compared with the negative control. The performance of pigs fed B. subtilis MA139 supplemented diets did not differ from that of pigs fed the antibiotic diet. There was a significant increase in Lactobacilli cell counts (P = 0.02) and a numerical decrease in E. coli counts (P = 0.05) in the faecal samples of pigs fed B. subtilis MA139 with 2.2 x 10(5) CFU/g of feed. The overall results of this study show that the use of initial co-culture with indicator pathogens, a disc plate diffusion assay and simulated gut environment tolerance tests is one of effective methods of screening Bacillus for probiotic use and that B. subtilis MA139 is a promising alternative to antibiotics for use as a feed additive in piglet diets.

We compared extracts and toxicities of organic compounds (OCs) in drinking water concentrated by composite XAD-2/8 resin (mixed with an equal volume of XAD-2 and XAD-8 resins) with those extracted by single XAD-2 (non-polar) and XAD-8 (polar) resins. Drinking water was processed from raw water of the Han River and the Yangtze River in Wuhan section, China. The extraction efficiency of all resins was controlled at 30%. The types of extracted OCs were detected by gas chromatography–mass spectrometry, and the cytotoxicity and genotoxicity were assessed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and comet assays, respectively, in human hepatoma HepG2 cells. Our results showed that XAD-2/8 extracted a larger variety of OCs, compared with XAD-8 and XAD-2. The cytotoxicity and genotoxicity of extracted OCs were in the order of XAD-8> XAD-2/8> XAD-2 at almost all tested concentrations after 24 h treatment (P < 0.05). Our findings suggest that single XAD resin selectively extracts either polar or non-polar OCs, which would lead to over- or under-estimation of the toxicity of drinking water. Nevertheless, composite resin extracts both polar and non-polar OCs, and could be utilized as a useful extraction technique to evaluate the level and toxicity of OCs in drinking water.

A monoclonal antibody (Mab), 4B2, against soybean glycinin was prepared using the preliminary extracted natural glycinin as the immunogen in our previous study. Herein, we established a novel method for the purification of glycinin by Mab 4B2-based immunoaffinity chromatography. The characteristics of the purified glycinin were identified by SDS-PAGE, Western blot, and histamine release assay. Glycinin was successfully isolated from soybeans with a yield of 16.8% and a purity of 93.8%, which were significantly higher than those produced using other traditional procedures. The acidic polypeptides of the purified glycinin can be recognized by the Mab 4B2, but not the basic polypeptides. In addition, the histamine release ratio of the purified glycinin was similar to that of natural glycinin, which indicated that the purified glycinin maintained its biological activities. Further study revealed that the Mab/gel ratios ranging from 6.0 to 12.0 mg/mL were suitable for the isolation of glycinin using immunoaffinity chromatography. Taken together, this new method based on immunoaffinity chromatography could be used for high-yield and high-purity natural glycinin production and would facilitate future study on the mechanism of soybean-induced food allergy.