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UNSTRUCTURED The release of a dialogue-based artificial intelligence language model called chatGPT (https://openai.com/blog/chatgpt/) has garnered global attention. An exploratory study published by The Journal of the American Medical Association (JAMA) demonstrated the potential of interactive AI to assist clinical workflows by augmenting patient education and patient-clinician communication. Our study was conducted in February 2023, and we formulated 37 questions that focused on perioperative patient education in the context of thoracic surgery. We made two inquiries to chatGPT for each question, one in English and the other in Chinese, and evaluated the appropriateness and comprehensiveness of the responses separately in both languages. Experienced thoracic surgical clinicians assessed the responses. Thirty-four (91.9%) responses were qualified both in English and Chinese contexts, demonstrating the potential feasibility for perioperative patient education in thoracic surgery.

Background: : The appearance of antibiotic resistance caused by metallo-β-lactamases (MβLs) is a global public health threat. Developing MβLs inhibitor is an effective way to overcome antibiotic resistance. Recently, azolylthioacetamides were reported to be promising MβLs inhibitors. Methods:: Triazolylthioacetamides were synthesized and tested for inhibition activity against the purified MβL IMP-1. Antimicrobial activities of these inhibitors in combination with cefazolin were evaluated. Isothermal Titration Calorimetry (ITC) was employed to characterize the binding of the inhibitor to IMP-1, and their action mechanism was studied by molecular docking. Results: Twenty compounds exhibited specific inhibitory activity against IMP-1 with an IC50 value in the range of 3.1-62.5 μM. Eight of the compounds can restore the antibacterial efficacy of cefazolin against E. coli BL21 strain producing IMP-1 by 2-4 fold. ITC monitoring showed that 1c exhibited dose-dependent inhibition on IMP-1. Docking studies revealed that the triazole group in 1c and 2d played an essential role in the inhibition activity. Cytotoxicity assay showed that 1c and 2d have low toxicity in L929 mouse fibroblastic cells. Conclusion: : The triazolylthioacetamides are efficient inhibitors of IMP-1 in vitro and in vivo.

The d , d -dipeptidase enzyme VanX is the main cause of vancomycin resistance in gram-positive bacteria because of hydrolysis of the D-Ala-D-Ala dipeptide used in cell-wall biosynthesis. Continuous assay of VanX has proven challenging due to lack of a chromophoric substrate. Here, we report a direct approach for continuous assay of VanX in vitro and in vivo from hydrolysis of D-Ala-D-Ala, based on the heat-rate changes measured with isothermal titration calorimetry (ITC). With the ITC approach, determination of kinetic parameters of VanX hydrolyzing D-Ala-D-Ala and the inhibition constant of d -cysteine inhibitor yielded KM of 0.10 mM, kcat of 11.5 s−1, and Ki of 18.8 μM, which are consistent with the data from ninhydrin/Cd(II) assays. Cell-based ITC studies demonstrated that the VanX expressed in E. coli and in clinical strain VRE was inhibited by d -cysteine with IC50 values of 29.8 and 28.6 μM, respectively. Also, the total heat from D-Ala-D-Ala (4 mM) hydrolysis decreases strongly (in absolute value) from 1.26 mJ for VRE to 0.031 mJ for E. faecalis, which is consistent with the large MIC value of vancomycin of 512 μg/mL for VRE and the much smaller value of 4 μg/mL for E. faecalis. The ITC approach proposed here could be applied to screen and evaluate small molecule inhibitors of VanX or to identify drug resistant bacteria.

The superbug infection caused by New Delhi metallo-β-lactamase (NDM-1) has grown into an emerging threat, labelling and inhibition of NDM-1 has proven challenging due to its shuttling between pathogenic bacteria. Here, we report a potent covalent scaffold, ebsulfur, for targeting the protein in vitro and in vivo. Enzymatic kinetic study indicated that eighteen ebsulfurs gained except 1a–b and 1f inhibited NDM-1, exhibiting an IC50 value ranging of 0.16–9 μM, and 1g was found to be the best, dose- and time-dependent inhibitor with an IC50 of 0.16 μM. Also, these ebsulfurs effectively restored the antibacterial activity of cefazolin against E. coli expressing NDM-1, and the best effect was observed to be from 1g, 1i and 1n, resulting in an 256-fold reduction in MIC of the antibiotic at a dose of 16 μg/mL. The equilibrium dialysis study implied that the ebsulfur disrupted the coordination of one Zn(II) ion at active site of NDM-1. Labelling of NDM-1 using a constructed fluorescent ebsulfur Ebs-R suggested that the inhibitor covalently bound to the target through SDS-PAGE analysis in vitro. Also, labelling NDM-1 in living E. coli cells with Ebs-R by confocal microscopic imaging showed the real-time distribution change process of intracellular recombinant protein NDM-1. Moreover, the cytotoxicity of these ebsulfurs against L929 mouse fibroblastic cells was tested, and their capability to restore antibacterial activity of antibiotic against clinical strains E. coli EC08 producing NDM-1 was determined. The ebsulfur scaffold proposed here is valuable for development of the covalent irreversible inhibitors of NDM-1, and also for labelling the target in vitro and in vivo.

Inhibition of &beta, lactamases presents a promising strategy to restore the &beta, lactams antibacterial activity to resistant bacteria. In this work, we found that aromatic carboxyl substituted 2-triazolylthioacetamides 1a&ndash, j inhibited VIM-2, exhibiting an IC50 value in the range of 20.6&ndash, 58.6 &mu, M. The structure-activity relationship study revealed that replacing the aliphatic carboxylic acid with aromatic carboxyl improved the inhibitory activity of 2-triazolylthioacetamides against VIM-2. 1a&ndash, j (16 mg/mL) restored the antibacterial activity of cefazolin against E. coli cell expressing VIM-2, resulting in a 4&ndash, 8-fold reduction in MICs. The isothermal titration calorimetry (ITC) characterization suggested that the primary binding 2-triazolylthioacetamide (1b, 1c, or 1h) to VIM-2 was a combination of entropy and enthalpy contributions. Further, the crystal structure of VIM-2 in complex with 1b was obtained by co-crystallization with a hanging-drop vapour-diffusion method. The crystal structure analysis revealed that 1b bound to two Zn(II) ions of the enzyme active sites, formed H-bound with Asn233 and structure water molecule, and interacted with the hydrophobic pocket of enzyme activity center utilizing hydrophobic moieties, especially for the phenyl of aromatic carboxyl which formed &pi, &pi, stacking with active residue His263. These studies confirmed that aromatic carboxyl substituted 2-triazolylthioacetamides are the potent VIM-2 inhibitors scaffold and provided help to further optimize 2-triazolylthioacetamides as VIM-2 even or broad-spectrum M&beta, Ls inhibitors.

Aim: The discovery and development of novel broad-spectrum MβLs inhibitors are urgent to overcome antibiotic resistance mediated by MβLs. Methods & results: Herein, the synthesized 21 compounds exhibited potent inhibition to the clinically important MβLs (NDM-1, IMP-1 and ImiS) and effectively restored the antibacterial efficacy of cefazolin and imipenem against Escherichia coli harboring MβLs. 5b was first identified to be dual functional broad-spectrum MβLs inhibitor through assemblage of covalent and metal binding scaffold, which irreversibly inhibited B1, B2 MβLs via forming a Se–S covalent bond, and competitively inhibited B3 MβLs by coordinating the metals at active site. Conclusion: The designed compounds can serve as potent broad-spectrum MβLs inhibitors and combat MβLs-producing ‘superbug’ in combination with β-lactams.

Superbug infection caused by metallo-β-lactamases (MβLs) is a global public health threat. Previous studies reported that the thioesters specifically inhibited the B3 subclass MβL L1. In this work, nine amino acid thioesters 1-9 were synthesized, the activity evaluation revealed that all of these molecules exhibited broad-spectrum inhibitory efficacy against ImiS, IMP-1, NDM-1, and L1, with IC50 values range of 0.02-54.9 µM (except 5 and 7 on NDM-1), and 1 was found to be the best inhibitor with IC50 range of 0.02-16.63 µM. Minimal inhibitory concentration (MIC) assays showed that thioesters 1, 5 and 9 restored 2-32-fold antibacterial activity of cefazolin and/or imipenem against both Escherichia coli BL21 and DH10B strain expressing ImiS, L1, IMP-1 and NDM-1 (except 5 on NDM-1), and also, thioester 1 increased 2-4-fold antimicrobial activity of cefazolin on two clinical strains Pseudomonas aeruginosa and Klebsiella pneumoniae producing NDM-1. Stability evaluation indicated that thioester 1 was partially hydrolysed by MβLs to be converted into the mercaptoacetic acid, revealing that the thioester and its hydrolysate mercaptoacetic acid jointly inhibit MβLs. Isothermal titration calorimetry (ITC) monitoring showed that thioester 1 exhibited dose-dependent inhibition on four MβLs tested, and the binding of 1/L1 showed mainly enthalpy driven, while 1/NDM-1 was found to be more entropy driven. Docking studies suggested that 1 bound to Zn(II) ion(s) preferentially via its carboxylate group, while other moieties interacted mostly with the conserved active site residues.

We discovered a promising metallo-β-lactamase inhibitor, a DNA nanoribbon, by enzymatic kinetics and isothermal titration calorimetry evaluations. Atomic force microscopy, gel electrophoresis, competitive binding experiments, circular dichroic and thermal denaturation studies suggested that the DNA nanoribbon could bind to the enzyme through a minor groove.

// Chen Yu 1, * , Peng Li 2, * , Dong Qi 2, * , Lei Wang 3, * , Hong-lin Qu 1 , Yue-juan Zhang 4 , Xue-kai Wang 5 , Hua-Ying Fan 5 1 School of Pharmacy, Binzhou Medical University, 264003 Yantai, Shandong, P.R. China 2 Department of Nephrology, Yu-Huang-Ding Hospital/Qingdao University, 264000 Yantai, Shandong, P.R. China 3 Yantai Food and Drug Inspection Center, 264000 Yantai, Shandong, P.R. China 4 Yantai Yan-Tai-Shan Hospital, Yantai, Shandong, P.R. China 5 School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation (Yantai University), Ministry of Education, Collaborative Innovation Center of Advanced Drug Delivery System and Biotech Drugs in Universities of Shandong, Yantai University, Yantai, P.R. China * These authors have contributed equally to this work Correspondence to: Chen Yu, email: yuchen810624@163.com Hua-Ying Fan, email: katieflydong@sina.com Keywords: osthole, sepsis, CLP, NF-κB signal pathway, acute kidney injury Received: July 20, 2016 Accepted: November 08, 2016 Published: November 25, 2016 ABSTRACT BACKGROUND AND PURPOSE: As a natural coumarin derivative from the Cnidium monnieri(L)Cusson fruit, osthole consists of 7-methoxy-8-isopentenoxy-coumarin. The purpose of this research is to study the mechanism and effect of osthole on sepsis-induced acute kidney injury. EXPERIMENTAL APPROACH: The protective effect of osthole on mouse macrophage RAW 264.7 and HK-2 cells induced by LPS in vitro and on acute kidney injury model induced by sepsis and established by puncture and cecal ligation (CLP) in vivo were tested. KEY RESULTS: Osthole (20, 40 mg·kg -1 ) group can greatly attenuate the changes of the score and kidney histopathology damage and enhance the survival time of septic mice. After the CLP surgery, degrees of SCr and BUN related to kidney injury were upregulated. The concentrations of SCr and BUN can be greatly reduced by treatment with osthole. Furthermore, osthole could increase bacterial killing activity and phagocytic activities of macrophages impaired after CLP partly and attenuate blood bacterial counts and leukocyte infiltration markedly. Furthermore, osthole can suppress NF-κB signal pathway through the inhibition of the nuclear translocation by regulating phosphorylation of IκBα and IKKβ and hinder the production of chemoattractant (MCP-1 and IL-8) and proinflammatory cytokines (TNF-α, IL-1β and IL-6). CONCLUSION AND IMPLICATIONS: Mainly because of its immunomodulatory properties and anti-inflammatory activity, which might be closely associated with suppression of the stimulation of the NF-κB signal pathway, osthole has protective effect on sepsis-induced kidney injury. It can be seen from such evidence that osthole can be potentially applied in the treatment of acute kidney injury.

Objective To observe the effect of needle pricking therapy on rheological indices and efficacy for infertile patients with varicocele (VC). Methods A total of 215 infertility patients with VC were assigned to the treatment group (109 cases, treated by needle pricking therapy) and the control group (106 cases, treated by drug therapy). Patients in the treatment group received CHEN's needle pricking therapy, once per week for a total of 10 times. Those in the control group took Guizhi Fuling Cap- sule (GFC), 0. 31g/pill, 3 pills each time, three times per day for 70 successive days. The diameter of VC, whole blood apparent viscosity, and plasma viscosity value were detected in the two groups after 75- day treatment. Sperm count, sperm motility rate, sperm motility, teratopermia rate, and other parame- ters were determined to evaluate the efficacy. Routines of blood, urine, and stool were detected. Renal and liver functions were examined to evaluate the safety. Results Compared with before treatment in the same group, the diameter of VC was narrowed in grade I and II patients of the two groups after treatment (P0.05). The diameter of VC was obviously narrower in grade II patients of the treatment group after treatment (P0. 01). Compared with before treatment in the same group, semen quality was improved in the two groups after treatment (P0. 05,P0.01). Besides, sperm density was reduced, sperm motility rate was elevated, spermatozoa with normal morphology was increased with longer fluidized time in the treatment group after treatment, which were all better than those of the control group (P0.05). Compared with before treatment, whole blood apparent viscosity and plasma viscosity value both obviously decreased in the treatment group after treatment (P0. 05). The total effective rate was better in the treatment group than in the control group (P0. 05). Conclusion Needle pricking therapy could obviously improve blood flow and vasomotic elasticity of infertile patients with VC, lower plasma viscosity, thus promoting blood circulation and lowering VC.

The emergence of antibiotic resistance caused by β-lactamases, including serine β-lactamases (SβLs) and metallo-β-lactamases (MβLs), is a global public health threat. L1, a B3 subclass MβL, hydrolyzes almost all of known β-lactam antibiotics. We report a simple and straightforward UV-Vis approach for real-time activity assays of β-lactamases inside living bacterial cells, and this method has been exemplified by choosing antibiotics, L1 enzyme, Escherichia coli expressing L1 (L1 E. coli), Escherichia coli expressing extended-spectrum β-lactamases (ESBL-E. coli), clinical bacterial strains, and reported MβL and SβL inhibitors. The cell-based studies demonstrated that cefazolin was hydrolyzed by L1 E. coli and clinical strains, and confirmed the hydrolysis to be inhibited by two known L1 inhibitors EDTA and azolylthioacetamide (ATAA), with an IC50 value of 1.6 and 18.9 μM, respectively. Also, it has been confirmed that the breakdown of cefazolin caused by ESBL-E. coli was inhibited by clavulanic acid, the first SβL inhibitor approved by FDA. The data gained through this approach are closely related to the biological function of the target enzyme in its physiological environment. The UV-Vis method proposed here can be applied to target-based whole-cell screening to search for potent β-lactamase inhibitors, and to assays of reactions in complex biological systems, for instance in medical assays.

We report an UV-Vis method for monitoring the hydrolysis of the β-lactam antibiotics inside living bacterial cells. Cell-based studies demonstrated that the hydrolysis of cefazolin was inhibited by three known NDM-1 inhibitors. This approach can be applied to the monitoring of reactions in a complex biological system, for instance in medical testing.

Given the clinical importance of metallo-β-lactamases (MβLs), a new scaffold, N-substituted carbamylmethyl mercaptoacetate thioether, was constructed. The obtained molecules 1–16 inhibited MβLs from all three subclasses, but preferentially L1 from subclass B3. Compound 9 with a p-carboxyphenyl substituent exhibited the broadest spectrum with at least 70% inhibition of enzymes from all subclasses at 100 μM, while compound 5 with a p-methylphenyl substituent was the most potent inhibitor of any individual enzyme, with 97% inhibition at 100 μM and an IC50 value of 0.41 μM against L1. Isothermal titration calorimetry assays corroborate findings from UV–vis spectrophotometric assays that the inhibition of L1 by 5 is dose-dependent. Docking studies suggest that the carboxyl group, the sulfide atom, and the carbonyl group of the carbamyl coordinate Zn2 in a chelating fashion. Using E. coli cells expressing L1, 6 and 8 were able to decrease cefazolin minimum inhibitory concentration 8-fold.

Background The management of breakthrough pain in cancer patients is always a challenge for medical professions. Occurring in 80% of cancer patients with advanced disease, breakthrough pain significantly decreases both patient’s and caregiver’s quality of life. The aim of this study is to assess the analgesic efficacy of a fixed inhaled nitrous oxide/oxygen mixture for adult cancer patients with breakthrough pain. Methods/design This is a randomized, placebo-controlled, double-blind study; it will be conducted in the General Hospital of Ningxia Medical University. The target study subjects are at least 18 years old, and are hospitalized cancer patients who are receiving routine opioids to control cancer-related pain but still experience breakthrough pain. A total of 240 patients will be recruited and randomly allocated between three treatment groups (A, B, C) and a control group (group D) in a ratio of 3:1. All treatment groups (A, B, C) will receive standard pain treatment (oral immediate-release morphine) plus a pre-prepared nitrous oxide/oxygen mixture, and the control group (D) will receive the standard pain treatment plus oxygen. Patients, doctors, nurses, and data collectors are all blind to the experiment. Assessments will be taken before treatment (T0), at 5 min (T1) and 15 min (T2) during treatment, and at 5 min after treatment (T3). The primary endpoint measures will be the percentage of patients whose pain is relieved at T1, T2, and T3. Secondary outcome measures will include the safety of treatment, adverse events, and satisfaction from both health professionals and patients. Discussion This study aims to provide an effective and practical intervention for a fast breakthrough pain relief and to improve cancer patients’ quality of life significantly. The Evidence-Based Medicine Working Group claim that a randomized, double-blind, placebo-controlled experimental intervention is the most appropriate design to demonstrate its efficacy, so this study could give a new approach to controlling breakthrough pain episodes. Trial registration ChiCTR-INC-16008075. Registered on 8 March 2016. Electronic supplementary material The online version of this article (doi:10.1186/s13063-016-1739-9) contains supplementary material, which is available to authorized users.

The emergence of antibiotic resistance caused by metallo-β-lactamases (MβLs) is a global public health problem. Recently, we found amino acid thioesters to be a highly promising scaffold for inhibitors of the MβL L1. In order to optimize this series of inhibitors, nine new amino acid thioesters were developed by modifying the substituents on the N-terminus of the thioesters and the groups representing the amino acid side chain. Biological activity assays indicate that all of them are very potent inhibitors of L1 with an IC50 value range of 20–600 nM, lower than those of most of the previously reported inhibitors of this scaffold. Analysis of structure–activity relationship reveals that big hydrophobic substituents on the N-terminus and a methionine amino acid side chain improves inhibitory activity of the thioesters. All these inhibitors are able to restore antibacterial activity of a β-lactam antibiotic against Escherichia coli BL21(DE3) cells producing L1 to that against E. coli cells lacking a β-lactamase. Docking studies reveal that a large N-terminal hydrophobic group results in a slightly different binding mode than smaller hydrophobic groups at the same position.

To study the effect of PKC signalling pathway and aldose reductase (AR) on the expression of fibronectin (FN) induced by transforming growth factor-β1 (TGF-β1).Human mesangial cells (HMCs) were cultured and transfected with pcDNA3-AR, and subject to AR gene silencing with small interfering RNA (siRNA) and then the cell was treated with recombinant human TGF-β1. The AR mRNA expression in the HMCs was examined using real time RT-PCR and protein expression of AR and FN was detected by Western blotting.The cultured HMC treated with TGF-β1 showed increased expression of AR and FN, the normal HMC showed not reduced expression of FN after incubation with single inhibitors of AR.Pre-incubation of cells with inhibitors of AR and PKC, then the different groups of cells were treated with TGF-β1, and the induction effect on FN expression was suppressed (34%) in HMC. HMCs transfected with AR showed a strong protein expression of FN, which was increased by 3.6-fold after treatment with TGF-β1 (P0.05), and the induction effect on FN expression was suppressed by GÖ6983 (42%) in HMCs (P0.05). The HMC with AR gene knock-down by siRNA showed a decreased expression of AR and 90% decrease of FN protein in HMCs (P0.01), and TGF-β1-induced up-regulation of FN was significantly suppressed by siRNA (12%) in HMCs (P0.01).AR is capable of regulating FN expression only in the presence of TGF-β1, and this reaction is possibly accomplished through the activation of PKC signalling pathway.

To observe the effect of needle-pricking bleeding combined with pulling-rotating manipulation and simple manipulation on blood rheology in vertebral artery type cervical spondylosis (VATCS) patients, so as to analyze their mechanisms in relieving VATCS.A total of 198 VATCS patients were randomly divided into treatment group (n=101) which was treated with needle-pricking plus pulling-rotating manipulation, and control group (n=97) which was treated with simple pulling-rotating manipulation, according to the random number table. The treatment was given once every 7 days, 9 times altogether. The peak systolic blood flow velocity (Vpeak), end-diastolic blood velocity (Vmin), pulsatility index (PI) and resistent index (RI) of bilateral vertebral arteries (VA) and basilar artery (BA) were detected by transcranial doppler sonography (TCD). Whole blood apparent viscosity and the plasma viscosity in the treatment group were determined by using a blood viscosimeter.Of the 101 and 97 VATCS cases in the treatment and control groups, 62 (61.38%) and 12 (12.37%) were cured basically, 23 (22.77%) and 26 (26.80%) experienced marked improvement, 14 (13.86%) and 41 (42.27%) were improved. 2 (1.98%) and 18 (18.55%) failed in the treatment, with the total effective rates being 98.01% and 81.44% separately. The effective rate of the treatment group was significantly higher than that of control group (P0.05). Compared with pretreatment, Vpeak and Vmin of the bilateral VA and BA in the treatment group, and Vpeak of the right VA in the control group increased significantly (P0.01, P0.05), PI and RI of the bilateral VA and BA in the treatment group, and PI and RI of the right VA in the control group decreased significantly (P0.01, P0.05), suggesting a marked reduction of the vascular resistance and an apparent increase of the cerebral blood supply after the treatment. The therapeutic effects of the above-mentioned indexes of treatment group were significantly superior to those of the control group (P0.05). In comparison with pre-treatment, the whole blood apparent viscosity (high, medium and low shear rates) and plasma viscosity of the treatment group post-treatment were obviously reduced (P0.01, P0.05).Needle-pricking therapy combined with pulling-rotating manipulation can significantly improve VATCS patients' clinical symptoms, which may be closely related to its effects in lowering vascular blood resistance and blood viscosity and increasing cerebral blood supply.

To search for the best method for increasing clinical therapeutic effect on primary abnormality of sperm.One hundred and sixty-eight cases of infertility were randomly divided into a treatment group of 85 cases and a control group of 83 cases. The treatment group were treated with needle-picking at bilateral Shengzhi points, Dicong Shenjing points and L2 Shenjing points as main. The control group were treated with oral administration of Wuzi Yanzong Pills [symbol: see text]. Their therapeutic effects were observed in 3 hospitals.The total effective rate of 83.5% and the pregnancy rate of the patient's wife of 78.8% in the treatment group, and corresponding figures were 54.2% and 43.4% in the control group, with a very significant difference between the two groups (P0.01); and reproductive hormones improved significantly after treatment in the two groups (P0.01); after treatment, superoxide dismulase (SOD) activity and Zn content in semen were elevated and cadmium level decreased significantly in the treatment group (P0.05).Needle picking therapy can significantly improve and regulate endocrine function, increase quality of semen and elevate pregnancy rate of the patient's wife for the patient of primary abnormal sperm.