Search

The widespread of different NDM variants in clinical Enterobacterales isolates poses a serious public health concern, which requires continuous monitoring. In this study, three E. coli strains carrying two novel blaNDM variants of blaNDM-36, -37 were identified from a patient with refractory urinary tract infection (UTI) in China. We conducted antimicrobial susceptibility testing (AST), enzyme kinetics analysis, conjugation experiment, whole-genome sequencing (WGS), and bioinformatics analysis to characterize the blaNDM-36, -37 enzymes and their carrying strains. The blaNDM-36, -37 harboring E. coli isolates belonged to ST227, O9:H10 serotype and exhibited intermediate or resistance to all β-lactams tested except aztreonam and aztreonam/avibactam. The genes of blaNDM-36, -37 were located on a conjugative IncHI2-type plasmid. NDM-37 differed from NDM-5 by a single amino acid substitution (His261Tyr). NDM-36 differed from NDM-37 by an additional missense mutation (Ala233Val). NDM-36 had increased hydrolytic activity toward ampicillin and cefotaxime relative to NDM-37 and NDM-5, while NDM-37 and NDM-36 had lower catalytic activity toward imipenem but higher activity against meropenem in comparison to NDM-5. This is the first report of co-occurrence of two novel blaNDM variants in E. coli isolated from the same patient. The work provides insights into the enzymatic function and demonstrates the ongoing evolution of NDM enzymes.

Background and purpose Pancreatic ductal adenocarcinoma (PDAC) is one of the leading causes of cancer-related deaths. The once highly anticipated antibody-based pathway-targeted therapies have not achieved promising outcomes, due to drugs' low intrinsic anticancer activity and low penetration across the dense physiological barrier of PDAC tumors. Here, an ultra-small-sized (50 kDa), bispecific protein, called Bi-fp50, that can penetrate deep tumor tissue and effectively inhibit PDAC tumor growth is reported. Methods Bi-fp50 was constructed by a typical synthetic biology method and target both EGFR and VEGF of PDAC cells simultaneously. Characteristics for example binding affinity of Bi-fp50 were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), enzyme-linked immunosorbent assay (ELISA) and fourier transform infrared spectra (FTIR). Different cell lines (Bxpc3, Aspc1) were used to test the in vitro targeting effect and anticancer ability of Bi-fp50. The orthotopic PDAC tumor model and subcutaneous PDAC tumor model were used to assess in vivo circulation and antitumor effect of Bi-fp50. Results Bi-fp50 with an ultra-small size of 50 kDa (5 ~ 6 nm) had shown a high target binding capacity and in vitro anticancer effect with significant cell killing for Bxpc3 and Aspc1 human PDAC cells. In vivo imaging had shown that Bi-fp50 could vastly enrich deep tumor tissue and had excellent penetration and accumulation when it was injected into orthotopic Bxpc3 xenograft mice. Bi-fp50 also had a high inhibition effect of tumor growth in vivo, accompanied by vascular normalization. No noticeable side effect of Bi-fp50 was found both in vitro and in vivo. Conclusion Compared with scFv2, anti-EGFR scFv, anti-VEGF scFv and Bi-fp50x group, Bi-fp50 with the ultra-small size had the highest binding affinity to both EGFR and VEGF targets. Since Bi-fp50 could penetrate deep pancreatic tumor tissue and had a high antitumor effect in vivo. Our work demonstrates that Bi-fp50 could be a potential candidate as a PDAC tumor suppressor.

This study aimed to assess the D-dimer level in patients with primary Sjögren syndrome (pSS), uncover its relationship with clinical symptoms, and appraise its predictive value in discriminating disease activity. The laboratory parameters of 101 consecutive patients with pSS and 101 healthy controls were analyzed and compared. Patients were divided into two subgroups according to their D-dimer levels, for the comparison of clinical features. Pearson's correlations were used to measure the relationships between D-dimer levels and other variables. The area under the curve (AUC) was calculated to predict disease activity. The erythrocyte sedimentation rate (ESR), high-sensitivity C-reactive protein (hsCRP) level, and D-dimer level were each higher in patients with pSS than in healthy controls. Compared with the low-D-dimer-level patients, those with elevated D-dimer levels exhibited higher ESRs (p 0.0001) and higher levels of hsCRP (p 0.0001), fibrinogen (p 0.0001), and immunoglobulin A (p = 0.002). Cases with elevated D-dimer levels were prone to be more severe, based on ESSDAI evaluation (p 0.0001). Patients with higher D-dimer levels had more articular involvement (p 0.0001), which was significantly correlated with both the ESR (r = 0.21, p = 0.03) and hsCRP level (r = 0.56, p = 0.001). The D-dimer level may help to discriminate low disease activity from moderate/high disease activity (AUC = 0.754). The D-dimer level was correlated positively with both the ESR and hsCRP level in patients with pSS. The ESR and levels of hsCRP, fibrinogen, and disease activity were higher in the elevated D-dimer level group. The D-dimer level was demonstrated to have predictive value in differentiating pSS disease activity. Key Points •D-Dimer was higher in patients with pSS. •D-Dimer may help for predicting the disease activity in patients with pSS.

Pectate lyases (Pels) have a vital function in degradation of the primary plant cell wall and the middle lamella and have been widely used in the industry. In this study, two pectate lyase genes, IDSPel16 and IDSPel17, were cloned from a sheep rumen microbiome. The recombinant enzymes were expressed in Escherichia coli and functionally characterized. Both IDSPel16 and IDSPel17 proteins had an optimal temperature of 60 ℃, and an optimal pH of 10.0. IDSPel16 was relatively stable below 60 °C, maintaining 77.51% residual activity after preincubation at 60 °C for 1 h, whereas IDSPel17 denatured rapidly at 60 °C. IDSPel16 was relatively stable between pH 6.0 and 12.0, after pretreatment for 1 h, retaining over 60% residual activity. IDSPel16 had high activity towards polygalacturonic acid, with a V

Few-shot relation extraction is one of the current research focuses. The key to this research is to fully extract the relation semantic information through very little training data. Intuitively, raising the relation semantics awareness in sentences can improve the efficiency of the model to extract relation features to alleviate the overfitting problem in few-shot learning. Therefore, we propose an enhanced relation semantic feature model based on prototype network to extract relations from few-shot texts. Firstly, we design a multi-level embedding encoder with position information and Transformer, which uses local information in the text to enhance the relation semantics representation. Secondly, the encoded relation features are fed into the novel prototype network, which designs a method that utilizes query prototype-level attention to guide the extraction of supporting prototypes, thereby enhancing the prototypes representation to better classify the relations in query sentences. Finally, through experimental comparison and discussion, we prove and analyze the effectiveness of the proposed multi-level embedding encoder, and prototype-level attention can enhance the stability of the model. Furthermore, our model has substantial improvements over baseline methods.

Cancer stem cells or cancer stemness has been confirmed to a major obstacle for glioma progression and it has also been reported that circRNAs play an important part in cancer progression. This study mainly focuses on revealing the role of circRPPH1 and the underlying mechanisms in glioma cell stemness.In vitro experiment including RT-qPCR, Western blot, sphere-formation analysis, and ALDH1 activity, and in vivo tumorigenesis experiments were performed to evaluate the effects of circRPPH1 on glioma cell stemness. Luciferase reporter, ChIP, and DNA pull-down analysis were used to reveal the underlying mechanisms.It was found that circRPPH1 level was upregulated in glioma cell spheres and facilitated the stemness of glioma cells; C-FOS transcriptionally activated circRPPH1 expression via directly binding to circRPPH1 promoter in glioma cells. Moreover, circRPPH1 promoted the stemness of glioma cells dependent on c-FOS-mediated transcriptional activation.This study indicates that c-Fos-activated circRPPH1 contributes to glioma stemness and provides a potential target for glioma progression based on the c-FOS/circRPPH1 regulatory axis.

The combination of rosuvastatin and ezetimibe has promising clinical benefits with a significant safety and tolerability profile. However, there is a lack of clinical data supporting the drug-drug interaction (DDI) in Chinese population. Thus, the aim of this study is to assess the potential pharmacokinetic DDI between rosuvastatin and ezetimibe in a Chinese population.In this randomized, open-label, phase 1 study, 12 healthy volunteers were randomized to three treatment groups: 10 mg rosuvastatin plus 10 mg ezetimibe, 10 mg rosuvastatin alone, and 10 mg ezetimibe alone under fasting conditions. The plasma concentrations of rosuvastatin and ezetimibe were determined, and the pharmacokinetic parameters were calculated. Primary endpoints were peak plasma concentration (CThe geometric mean ratios (GMRs) of rosuvastatin with 90% confidence intervals (CIs) were 0.94 (0.80-1.12) for CCo-administration of rosuvastatin and ezetimibe showed no clinically significant pharmacokinetic interactions in a healthy Chinese population.

The aim was to analyze the influence of fasting plasma glucose-lowering rate (FPGLR) on plasma BNP levels in type 2 diabetes mellitus (T2DM) patients with coronary microcirculation dysfunction (CMD) and to determine the optimal FPGLR for these patients.A total of 170 T2DM patients who received intensive glucose-lowering therapy during hospitalization in the First Affiliated Hospital of Harbin Medical University were enrolled. Ninety-two patients with CMD and 78 patients without CMD were assigned to a study and a control group, respectively. The study group was stratified as S1 (4.1 ~ 6.0 mmol·LIn the study and the control group, the BNP level in S1 was significantly higher than that in C1 (87 vs. 12 pg/ml, P 0.001), although there was no significant difference between S2 and C2, S3 and C3. In the study group, the BNP level in S1 was significantly higher than that in S2 (87 vs. 22 pg/ml, P 0.001) and S3 (87 vs. 15 pg/ml, P 0.001), but there was no significant difference between S2 and S3.Rapid intensive glucose-lowering may lead to increased plasma BNP levels in T2DM patients with CMD. Optimal FPGLR for these patients was determined to be no more than 4.0 mmol·L

The emerging SARS-CoV-2 variants, commonly with many mutations in S1 subunit of spike (S) protein are weakening the efficacy of the current vaccines and antibody therapeutics. This calls for the variant-proof SARS-CoV-2 vaccines targeting the more conserved regions in S protein. Here, we designed a recombinant subunit vaccine, HR121, targeting the conserved HR1 domain in S2 subunit of S protein. HR121 consisting of HR1–linker1–HR2–linker2–HR1, is conformationally and functionally analogous to the HR1 domain present in the fusion intermediate conformation of S2 subunit. Immunization with HR121 in rabbits and rhesus macaques elicited highly potent cross-neutralizing antibodies against SARS-CoV-2 and its variants, particularly Omicron sublineages. Vaccination with HR121 achieved near-full protections against prototype SARS-CoV-2 infection in hACE2 transgenic mice, Syrian golden hamsters and rhesus macaques, and effective protection against Omicron BA.2 infection in Syrian golden hamsters. This study demonstrates that HR121 is a promising candidate of variant-proof SARS-CoV-2 vaccine with a novel conserved target in the S2 subunit for application against current and future SARS-CoV-2 variants.

Myocardial infarction (MI) caused by ischemic cardiomyocyte necrosis induces inflammatory responses that strongly affect ventricular remodeling. Tolerogenic dendritic cells (tDCs) can suppress this effect on inflammatory responses. However, the precise role of atorvastatin-induced tDCs in ventricular remodeling after MI remains unclear.To explore the effect of necrotic cardiomyocytes (SNC) and/or atorvastatin on DC function, the expression of CD40, CD80, CD86, and MHC-II was determined using flow cytometry. The protein levels of TLR-4/NF-κB-related molecules were evaluated using western blotting. The infarct area after MI was determined via 2,3,5-triphenyltetrazolium chloride staining. The TUNEL assay was employed to evaluate the apoptosis of cardiomyocytes in heart sections. Masson's trichrome method was used to determine the extent of fibrosis.Compared to the DCs co-cultured with PBS (control), cells co-cultured with Supernatant-IM or Supernatant-NH produced higher levels of inflammatory cytokines, including TNF-α, IL-1, IL-6, IL-12P40, and IL-8. This cytokine production was impaired by atorvastatin treatment. SNC treatment induced DC maturation and enhanced inflammatory cytokine secretion and oxidative stress through TLR-4/NF-κB pathway activation. Compared to that in the PBS-treated group, the left ventricular ejection fraction was significantly improved after tDC treatment. Additionally, compared to that in the PBS-treated group, tDC treatment reduced the left ventricular end-diastolic and end-systolic diameters in mice. Furthermore, treatment with tDCs improved the left ventricular systolic function, attenuated inflammatory cell infiltration, and reduced cardiomyocyte apoptosis, myocardial fibrosis, and infarct size compared to those in the control group.Adoptive transfer of atorvastatin-induced tDCs alleviated post-infarction cardiomyocyte apoptosis and myocardial fibrosis in association with decreased inflammatory cell infiltration and inhibited oxidative stress, likely by suppressing TLR-4/NF-κB activation after myocardial infarction.

With the improvement of industrialization, numerous rural laborers migrate to urban areas in search of off-farm jobs. Farmers change agricultural production decisions to adapt to the change of labor force, which will inevitably affect the crop planting structure. However, few studies have explored the sustainability of crop planting structure. Based on the calculation of the multiple cropping index (MCI), grain crops planting rate (GCR), economic crops planting rate (ECR), and ecological sustainability index (ESI) of crop planting structure, this study analyzes the impact of labor transfer rate (LTR) and labor cost (LC) on the sustainability of crop planting structure using a geographically and temporally weighted regression (GTWR) model. The results show that the scale of rural labor transfer and labor cost in China remains on the rise, but the growth rate has slowed down. The total carbon absorption of crops in China shows a U-shape trend, and the rice and maize have the largest carbon absorption. The impact of LTR on MCI is mainly positive, especially in the North China Plain in the early stage and some provinces in the Southwest China in the later stage. The impact of LTR on ECR and ESI is negative in most provinces. And the negative influence of LC on MCI is increasing, showing the spatial distribution characteristics of large influence in the southeast and small influence in the northwest. The impact of LC on ESI shows a negative effect in most provinces in the early stage, and the negative effect is more concentrated in some provinces in the southwest in the later stage.

The acute promyelocytic leukemia (APL) driver ZBTB16/RARα is generated by the t(11;17) (q23;q21) chromosomal translocation, which is resistant to combined treatment of all-trans retinoic acid (ATRA) and arsenic trioxide (ATO) or conventional chemotherapy, resulting in extremely low survival rates. In the current study, we investigated the effects of hyperthermia on the oncogenic fusion ZBTB16/RARα protein to explore a potential therapeutic approach for this variant APL. We showed that Z/R fusion protein expressed in HeLa cells was resistant to ATO, ATRA, and conventional chemotherapeutic agents. However, mild hyperthermia (42 °C) rapidly destabilized the ZBTB16/RARα fusion protein expressed in HeLa, 293T, and OCI-AML3 cells, followed by robust ubiquitination and proteasomal degradation. In contrast, hyperthermia did not affect the normal (i.e., unfused) ZBTB16 and RARα proteins, suggesting a specific thermal sensitivity of the ZBTB16/RARα fusion protein. Importantly, we found that the destabilization of ZBTB16/RARα was the initial step for oncogenic fusion protein degradation by hyperthermia, which could be blocked by deletion of nuclear receptor corepressor (NCoR) binding sites or knockdown of NCoRs. Furthermore, SIAH2 was identified as the E3 ligase participating in hyperthermia-induced ubiquitination of ZBTB16/RARα. In short, these results demonstrate that hyperthermia could effectively destabilize and subsequently degrade the ZBTB16/RARα fusion protein in an NCoR-dependent manner, suggesting a thermal-based therapeutic strategy that may improve the outcome in refractory ZBTB16/RARα-driven APL patients in the clinic.

Purpose To compare the three-dimensional (3D) heads-up surgery with the traditional microscopic (TM) surgery for various vitreoretinal diseases. Methods A medical record review of patients that underwent 3D heads-up or TM vitreoretinal surgeries was performed from May 2020 to October 2021 in this retrospective case–control study. Main outcome measures included surgery-related characteristics, efficacy, safety, and satisfaction feedback from the surgical team. Results A total of 220 (47.6%) and 242 (52.4%) eyes were included in the 3D and TM groups, respectively. The 3D heads-up system significantly benefits delicate surgical steps, like the epiretinal membrane (ERM) peeling for ERM and internal limiting membrane peeling for idiopathic macular holes (P P = 0.049), while no difference by TM surgery (P = 0.45). For the satisfaction feedback, the 3D heads-up system was rated significantly higher in most subscales and the overall score (P P P > 0.05). Conclusion The efficacy and safety of the 3D heads-up surgery were generally comparable to the TM surgery. The 3D heads-up system could significantly benefit delicate surgical steps and achieve better surgical team satisfaction.

Due to the multiple influences of natural and anthropogenic factors, stormwater runoff from urban roads generally presents heterogeneous pollution among cities. The identification of regional heterogeneity and related driving factors of road runoff pollution is of significance for the optimal management of road runoff pollution according to the local circumstances. In this study, the regional heterogeneity of urban road runoff pollution from fourteen representative cities in China is analyzed for four typical pollutants including total suspended solids (TSS), chemical oxygen demand (COD), total nitrogen (TN) and total phosphorus (TP). The results show wide variations in TSS, COD, TN and TP pollution among cities, with the average event mean concentrations ranging from 77.0 to 1347.9, 31.4 to 488.1, 0.81 to 8.46, 0.139 to 1.930 mg/L, respectively. One-way ANOVA analyses demonstrate significant differences in road runoff pollution among cities. The TSS pollution is significantly heavier for northern and northwestern inland cities than that for eastern and southern cities. Pearson correlation analysis and Stepwise linear regression analysis are performed to identify and rank the influence of climate, population, economy, industry structure, traffic and environmental quality. Direct relationships of road runoff pollution are detected with PM

To investigate the diagnostic efficiency of Raman spectroscopy for the diagnosis of breast cancer, we searched PubMed, Web of Science, Cochrane Library, and Embase for articles published from the database establishment to May 20, 2022. Pooled sensitivity, specificity, diagnostic odds ratio, and area under the receiver pooled operating characteristic curve were derived for the included studies as outcome measures. The methodological quality was assessed according to the questionnaires and criteria suggested by the Diagnostic Accuracy Research Quality Assessment-2 tool. Sixteen studies were included in this meta-analysis. The pooled sensitivity and specificity of Raman spectroscopy for breast cancer diagnosis were 0.97 (95% CI, [0.92-0.99]) and 0.96 (95% CI, [0.91-0.98]). The diagnostic odds ratio was 720.89 (95% CI, [135.73-3828.88]) and the area under the curve of summary receiver operating characteristic curves was 0.99 (95% CI, [0.98-1]). Subgroup analysis revealed that all subgroup types in our analysis, including different races, sample types, diagnostic algorithms, number of spectra, instrument types, and laser wavelengths, turned out to have a sensitivity and specificity greater than 0.9. Significant heterogeneity was found between studies. Deeks' funnel plot demonstrated that publication bias was acceptable. This meta-analysis suggests that Raman spectroscopy may be an effective and accurate tool to differentiate breast cancer from normal breast tissue, which will help us diagnose and treat breast cancer.

Glycosylation change is one of the landmark events of tumor occurrence and development, and tumor cells may be inhibited by regulating the aberrant expression of glycosyltransferases. Currently, fucosyltransferase VI (FUT6), which is involved in the synthesis of α-1, 3 fucosyl bond, has been detected to be closely associated with multiple tumors, but its function and mechanism in head and neck squamous cell carcinoma (HNSCC) still need further research. In this study, FUT6 knockdown and overexpression strategies were used to investigate the effects of FUT6 on cell proliferation, migration, and invasion, as well as the growth and metastasis of HNSCC in a xenografts mouse model. The protein expression levels of epidermal growth factor receptor (EGFR), extracellular signal-regulated kinase (ERK), Signal Transducer and Activator of Transcription (STAT), protein kinase B (AKT), c-Myc, and epithelial-mesenchymal transition (EMT) markers were determined by western blot analysis. Our research found that the mRNA expression of FUT6 was lower in HNSCC tissues than in normal mucosal epithelial tissues. In Cal-27 and FaDu cells, FUT6 overexpression inhibited cell proliferation, migration and invasion, causing upregulation of ZO-1 and E-cadherin, downregulation of N-cadherin and Vimentin, and finally decreased the phosphorylation levels of EGFR, ERK, STAT, and c-Myc. In HSC-3 cells, knockdown of FUT6 promoted cell proliferation, migration and invasion, downregulating ZO-1 and E-cadherin, upregulating N-cadherin and Vimentin, and increased the phosphorylation levels of EGFR, ERK, STAT, and c-Myc. In the HNSCC xenografts mouse, FUT6 overexpression inhibited tumor growth and metastasis. In summary, FUT6 controls the proliferation, migration, invasion, and EGF-induced EMT of HNSCC by regulating EGFR/ERK/STAT signaling pathway, indicating its potential future therapeutic application for HNSCC.

Artesunate (ART) has been indicated as a candidate drug for hepatocellular carcinoma (HCC). Glucosylceramidase (GBA) is required for autophagic degradation. Whether ART regulates autophagic flux by targeting GBA in HCC remains to be defined. Herein, our data demonstrated that the dramatic overexpression of GBA was significantly associated with aggressive progression and short overall survival times in HCC. Subsequent experiments revealed an association between autophagic activity and GBA expression in clinical HCC samples, tumor tissues from a rat model of inflammation-induced HCC and an orthotopic mouse model, and human HCC cell lines. Interestingly, probe labeling identified GBA as an ART target, which was further verified by both a glutathione-S-transferase pulldown assay and surface plasmon resonance analysis. The elevated protein expression of LC3B, the increased numbers of GFP-LC3B puncta and double-membrane vacuoles, and the enhanced expression of SQSTM1/p62 indicated that the degradation of autophagosomes in HCC cells was inhibited by ART treatment. Both the in vitro and in vivo data revealed that autophagosome accumulation through targeting of GBA was responsible for the anti-HCC effects of ART. In summary, this preclinical study identified GBA as one of the direct targets of ART, which may have promising potential to inhibit lysosomal autophagy for HCC therapy.