Based on the strategy of metabolomics combined with bioinformatics, this study analyzed the potential allergens and mechanism of pseudo-allergic reactions (PARs) induced by the combined use of Reduning injection and penicillin G injection. All animal experiments and welfare are in accordance with the requirements of the First Affiliated Experimental Animal Ethics and Animal Welfare Committee of Henan University of Chinese Medicine (approval number: YFYDW2020002). Based on UPLC-Q-TOF/MS technology combined with UNIFI software, a total of 21 compounds were identified in Reduning and penicillin G mixed injection. Based on molecular docking technology, 10 potential allergens with strong binding activity to MrgprX2 agonist sites were further screened. Metabolomics analysis using UPLC-Q-TOF/MS technology revealed that 34 differential metabolites such as arachidonic acid, phosphatidylcholine, phosphatidylserine, prostaglandins, and leukotrienes were endogenous differential metabolites of PARs caused by combined use of Reduning injection and penicillin G injection. Through the analysis of the "potential allergen-target-endogenous differential metabolite" interaction network, the chlorogenic acids (such as chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, and isochlorogenic acid A) and β-lactam allergens in the combination of the two may be mainly regulated by PLD1, PLA2G12A and CYP1A1. The three upstream signal target proteins mainly activate the arachidonic acid metabolic pathway, promote the degranulation of mast cells, release downstream endogenous inflammatory mediators, and induce PARs.

Lipopolysaccharides (LPS) are major outer membrane components of Gram-negative bacteria. Unlike most Gram-negative bacteria, Acinetobacter baumannii can still survive and acquire polymyxin resistance after complete loss of LPS. Previous studies of LPS-deficient Acinetobacter baumannii mostly focused on LPS-deficient strains induced by polymyxin, whose background was complex and unstable. To investigate LPS loss-mediated polymyxin resistant-Acinetobacter baumannii, this study constructed a stable and clear background LPS-deficient strain by knocking out lpxC gene in Acinetobacter baumannii ATCC 19606 with CIRSPR/Cas9, and then studied the phenotypic changes of the lpxC-deficient strain including morphology, growth rate, antibiotic susceptibility, virulence, membrane permeability, and membrane potential. Animal experiments were approved by the Animal Care and Welfare Committee Institute of Medicinal Biotechnology, CAMS and PUMC (approval number: IMB-20240119D₉). The results indicated that the lpxC gene of Acinetobacter baumannii ATCC 19606 was successfully knocked out. After losing the lpxC, the strain underwent the morphological change from rod-shaped to spherical. Furthermore, it leads to reduced growth rate, enhanced membrane permeability, decreased membrane potential, lower virulence, and increased antibiotic susceptibility to β-lactams, quinolones, aminoglycosides, macrolides, glycopeptides. The lpxC deletion results in significant changes in membrane homeostasis and adaptability of Acinetobacter baumannii. Understanding the phenotypic changes of colistin-resistant Acinetobacter baumannii mediated by LPS loss is useful for exploring the resistance mechanism of Acinetobacter baumannii and developing new therapeutic strategies.

Objective:To observe the changes of optic disc structure in patients with high myopia and the correlation with the morphological markers of the fundus.Methods:A retrospective study. From July 2018 to January 2020, 90 patients (155 eyes) diagnosed as high myopia in Department of Ophthalmology of Beijing Friendship Hospital affiliated to Capital Medical University were included in the study. Among them, there were 31 males (52 eyes) and 59 females (103 eyes), with age of 57.1±14.2 years old and axial length (AL) of 28.5±2.6 mm. According to the classification of myopic macular degeneration, patients were divided into 4 groups based on forms and degree of lesions, including non-pathological myopia group, mild traction lesions group, severe traction lesions group and neovascular lesions group, 35, 58, 41, 21 eyes, respectively. The digitized fundus photographs and an Image J system were used to measure the horizontal, vertical, maximal, and minimal diameter of the optic disc, the horizontal and vertical diameter of the parapapillary δ zone and γ zone, ovality index, distance between the most superior point of the temporal superior arterial arcade and most inferior point of the temporal inferior arterial arcade (VDA), angle between the temporal arterial arcade and optic disc (angle kappa), distance between the optic disc center and the fovea (DFD), angle between the horizontal disc axis and the disc-fovea line (DFA). The correlation between the diameter of the optic disc and other parameters was analyzed. Univariate and multivariate analysis were used to compare differences between groups.Results:The horizontal diameter of the optic disc was positively correlated with the horizontal diameter of the δ zone ( r=0.300, P<0.001), Kappa angle ( r=0.260, P=0.003), and elliptic index ( r=0.650, P<0.001); it was negatively correlated with DFD ( r=-0.190, P=0.030). Optic disc vertical diameter and optic disc horizontal diameter ( r=0.280), δ-zone horizontal diameter ( r=0.330) and vertical diameter ( r=0.460), γ-zone horizontal diameter ( r=0.430) and vertical diameter ( r=0.390), DFD ( r=0.390) was positively correlated ( P<0.001); it was negatively correlated with DFA ( r=-0.210, P=0.001) and Kappa angle ( r=-0.210, P=0.004). Compared with the non-pathological myopia group, there were statistically significant differences in the horizontal and vertical diameters of the optic disc in the severe traction disease group ( P<0.05). Among them, the horizontal diameter difference did not depend on the eye axis and age difference; the vertical diameter difference was caused by the eye axis difference. Compared with the non-pathological myopia group, the difference in the horizontal diameter of the optic disc in the neovascular disease group was statistically significant ( P<0.05), and did not depend on the difference in the axis and age; the difference in the vertical diameter of the optic disc was not statistically significant ( P>0.05). Conclusion:The morphology of optic disc was related to several fundus morphological markers, which was differentiated according to the age, AL and the degree of disease in patients with high myopia.

Objective: To explore the association of hypertriglyceridemic waist phenotype ( HTGW ) with impaired fasting glucose ( IFG ) and diabetes, so as to provide reference for the early prevention and control of diabetes. Methods: The survey was conducted among 35 to 75-year-old residents in 8 project sites in Jiangsu Province from 2015 to 2019. The information about demography and lifestyle was collected by the general information questionnaire and the primary screening questionnaire from the National Center for Cardiovascular Diseases; waist circumference, height, weight, triglyceride, high-density lipoprotein cholesterol, total cholesterol and fasting plasma glucose were measured. The multinomial logistic regression model was employed to analyze the association of HTGW with IFG and diabetes. Results: A total of 118 383 subjects were included, among whom 21 851 cases of HTGW, 27 245 cases of IFG and 22 899 cases of diabetes were identified, with the prevalence of 18.46%, 23.01% and 19.34%. The multinomial logistic regression analysis showed HTGW was statistically associated with IFG ( OR=1.414, 95%CI: 1.343-1.489 ) and diabetes ( OR=2.216, 95%CI: 2.098-2.341 ). Conclusion HTGW is associated with IFG and diabetes, which make it possible to be an indicator for screening and assessment of glucose abnormality in middle-aged and elderly population.

Antiviral Oral Liquid is modified on the basis of Baihu Decoction in Treatise on Febrility Diseases by ZHANG Zhongjing and Qingwen Baidu Yin in Qing Dynasty, with effects in clearing toxic heat, repelling dampness and cooling blood. It is widely used in clinical treatment of common colds, influenza and upper respiratory tract infection, mumps, viral conjunctivitis and hand-foot-mouth disease, with a good clinical efficacy and safety. Based on a questionnaire survey of clinicians and a systematic review of study literatures on Antiviral Oral Liquid, the international clinical practice guidelines development method was adopted to analyze the optimal available evidences and expert experiences in the "evidence-based, consensus-based and experience-based" principles. The consensus was jointly reached by more than 30 multidisciplinary experts nationwide, including clinical experts of traditional Chinese and Western medicine in the field of respiratory diseases and infectious diseases, and methodological experts. In the study, literatures were retrieved based on clinical problems in the clinical survey as well as PICO clinical problems. The GRADE system was used for the classification and evaluation of evidence, and fully combined with clinical expert experience, so as to reach expert consensus by the nominal grouping method. This expert consensus recommended or suggested indications, usage and dosage, course of treatment, intervention time for treatment, and the safety and precautions of Antiviral Oral Liquid for treatment of influenza, and can provide reference for the rational use of this drug in clinical practice.
Antiviral Agents/therapeutic use* ; Consensus ; Hand, Foot and Mouth Disease ; Humans ; Influenza, Human/drug therapy* ; Medicine, Chinese Traditional

Objective:To explore the strategies of reducing relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with high-risk myelodysplastic syndrome (MDS) from the perspectives of optimizing the conditioning regimen and pre-transplant cytoreductive therapy.Methods:A total of 84 patients with high-risk MDS undergoing allo-HSCT between January 2013 and September 2019 were retrospectively analyzed. Based upon preparative regimens, they were divided into two groups of decitabine intensified BUCY2 ( n=49) and BUCY2 regimen ( n=35), based upon whether or not pre-treatment prior to allo-HSCT: cytoredutive treatment ( n=34) and none ( n=50). Two groups were compared with regards to hematopoietic reconstitution, graft-versus-host disease (GVHD), relapse rate, transplant-related mortality (TRM) and survival. Results:No significant inter-group differences existed in hematopoietic reconstitution or acute/chronic GVHD. The relapse rate was significantly lower in decitabine intensified group than that in BUCY2 group (18.7% vs 40.0%, P=0.025). Survival was significantly better in decitabine intensified group than that in BUCY2 group (3-year OS: 71.3% vs 51.2%, P=0.038; 3-year DFS: 65.3% vs 45.2%, P=0.033). Moreover, the incidence of recurrence was markedly lower in pre-transplant treatment group than that in non-treatment group (20.7% vs 38.9%, P=0.035). The inter-group incidence of TRM was not different. Three-year OS/DFS of treatment group were remarkably superior to those of non-treatment group (71.2% vs 50.8%, P=0.024; 64.7% vs 45.9%, P=0.044). Conclusions:As an optimal conditioning regimen for high-risk MDS, decitabine intensified BUCY2 regimen could better eliminate tumor burden, remarkably lower relapse rate and improve OS after allo-HSCT. In addition, pre-transplant treatment significantly reduces relapse and offers benefit for OS after allo-HSCT. Therefore intensified conditioning regimen and pre-transplant treatment may be promising strategies of reducing relapse and improving survival for high-risk MDS. However, it still needs further confirmation from prospective randomized controlled trials.

Objective To explore the molecular pathogenesis of 3 Glanzmann's thrombasthenia pedigree by using bioinformatics software and provide evidence for in vitro experiments. Methods The genetic analysis of 3 pedigree diagnosed as Glanzmann's thrombasthenia was carried out. Clustalx-2.1 win software was used to analyze the conservatism of mutant sites in homologous sequences. Bioinformatics software such as PolyPhen-2, PROVEAN, SIFT and Mutationtaster was used to analyze the biological effect of mutation. SPDBV software constructed the molecular structure model of mutant protein and evaluated the influence of mutation on protein structure. Results The "new mutations" found in 3 Glanzmann's thrombasthenia pedigree were ITGA2B:c. 814G>C (p. Val272Leu), ITGA2B:c. 432G>A (p. Trp144Ter) and ACTN1:c. 2458A>G (p. Ile820Val). All three mutations were highly conserved among homologous species. Mutationtaster software showed that 3 new mutations were likely pathogenic. PolyPhen-2 and PROVEAN software showed ITGA2B p.Val272Leu and ACTN1 p.Ile820Val were benign and SIFT software showed that ITGA2B p. Val272Leu were likely pathogenic, while ACTN1 p. Ile820Val is benign. The result of SPDBV software showed that the Val272 of ITGA2B was transformed to Leu, neutralizing all the original hydrogen bond. The Trp144 of ITGA2B is transformed to Ter, resulting in the truncated proteins with only 113 amino acid residues. All these mutations affected the molecular structure of GPⅡb, resulting in a decrease ofGPⅡb/Ⅲa expression. When the Ile820 of ACTN1 is transformed to Val, onlyretained the hydrogen bond of Ile820 and Asp822, neutralized the rest hydrogen bond, whichaffected the molecular structure and protein function of ACTN1. Conclusion The mutations of ITGA2B:c.814G>C (p.VAL272LEU), ITGA2B:c.432G>A (p.Trp144Ter) and ACTN1:c.2458A>G (p.Ile820Val) are pathogenic.

Objective: To investigate the molecular pathogenesis for a patient with Glanzmann thrombasthenia (GT). Methods: The peripheral blood of a patient with Glanzmann′s thrombasthenia was collected, and the genetic mutations were detected by gene sequencing technology. The mutant plasmids were prepared by PCR site-directed mutagenesis and transfected into CHO-K1 cells of Chinese hamster ovary to construct in vitro eukaryotic expression system. The expressions of αⅡb and β3 protein subunits in CHO-K1 cells were detected by western blot. The expression levels of αⅡb and β3 in cellular membrane and cytoplasm of CHO-K1 cells were detected by flow cytometry. The expression and distribution of αⅡb and β3 in CHO-K1 cells were observed by immunofluorescent labeling under microscope. Results: This patient was diagnosed with type Ⅱ GT. Gene sequencing revealed two mutations in ITGB3 gene which has not been reported in the literature. ITGB3 c.1495 T＞C missense mutation resulted in replacement of cysteine no.499 by arginine (p.C499R). ITGB3 c.1728 delC code shift mutation resulted in a change in the amino acid synthesis initiated by the β3 protein subunit serine no.577 and terminated by the 92nd amino acid following these changes. The results of western blotting showed that the synthesis and expression of primary structures of αⅡb and β3 were detectable in the lysates of mutant CHO-K1 cells. The results of flow cytometry showed that no expression of β3 on the surface and intracellular of mutant CHO-K1 cells was observed. Under fluorescence microscopy no distribution of β3 protein subunit was displayed in mutant CHO-K1 cells. Conclusion The mutation of ITGB3 c.1728 del C or ITGB3 c.1495 T＞C should be relevant to the cause of GT in this patient. The mutation of ITGB3 c.1728 del C and ITGB3 c.1495 T＞C seems not to affect the formation of the primary structure of β3 protein subunit, but did affect the formation of its high-level structure.

UHPLC-QTOF-MS was applied to non-targeted metabolomics study of mice infected with K. pneumoniae ATCC^® BAA 2146 to discover potential biomarkers and metabolic pathways that are associated with sepsis. Fifty-eight metabolites were identified by principal components analysis (PCA) and partial least-squares discriminant analysis (OPLS-DA), which was combined with variable projection importance (VIP) and nonparametric test. Eighteen of the 58 metabolites were further found to be involved in 8 metabolic pathways, including nicotinate and nicotinamide metabolism, pyrimidine metabolism, vitamin B6 metabolism, taurine and hypotaurine metabolism, arginine and proline metabolism, alanine, aspartate and glutamate metabolism, D-glutamine and D-glutamate metabolism and glycerophospholipid metabolism.

Objective To observe the choroidal blood flow and morphological changes in patients with severe stenosis of internal carotid artery stenosis (ICAS).Methods A retrospective case-control study. Forty-six patients (46 eyes) with ICAS were enrolled in this study. There was severe stenosis in one side (the eyes in this side were set as case group) and mild or no stenosis in other side (the eyes in this side were set as control group). Color doppler ultrasound (CDI) was used to observe the changes of hemodynamic parameters of the ophthalmic artery (OA) and posterior ciliary artery (PCA),the main parameters of ultrasound Doppler imaging are peak systolic velocity (PSV), end diastolic velocity (EDV), resistance indices (RI) and the calculation of the pulsation indices (PI) through the use of a formula. Enhanced binarization of deep imaging coherence tomography (EDI-OCT) was used to measure the subfoveal choroidal thickness (SFCT). The total subfoveal choroidal area (TCA), luminal (LA), stromal (SA) and choroidal vascularity index (CVI) were obtained by modified image binarization technique.Results In the case group, the PSV in the OA and PCA was significantly lower than that of the control group (t=?2.200, ?2.612;P=0.030, 0.011). There were no significant differences in EDV, RI, PI of OA (t=0.337, ?1.810, ?1.848;P=0.737, 0.074, 0.068) and PCA (t=?1.160, 1.400, 0.815;P=0.249, 0.165, 0.417). The SFCT (t=?3.711,P<0.001), TCA (t=?2.736,P=0.007), LA (t=?3.188, P=0.002) and CVI (t=?2.096,P=0.039) of the case group was significantly lower than that of the control group. There were no significant differences in SA (t=?1.262,P=0.210) and LA/SA (t=?1.696,P=0.093).Conclusion In severe stenosis ICAS eyes, the PSV in the PCA and SFCT, TCA, LA, CVI are decreased.