To examine the molecular epidemiology of carbapenem-resistant Klebsiella pneumoniae (CRKP) and investigate the horizontal transmission of blaKPC and blaNDM genes for the prevention and treatment of CRKP. A total of 49 clinically isolated CRKP strains were retrospectively analyzed from January to December 2022 at The First Hospital of Hunan University of Chinese Medicine. Phenotypic screening was performed using modified carbapenem inactivation assay (mCIM) and EDTA-carbapenem inactivation assay (eCIM). Polymerase chain reaction (PCR) was utilized to identify carbapenem resistance genes, β-lactamase resistance genes, and virulence genes, while multi-locus sequence analysis (MLST) was employed to assess the homology of CRKP strains. Conjugation experiments were conducted to infer the horizontal transmission mechanism of blaKPC and blaNDM genes. The results showed that the study included 49 CRKP strains, with 44 carrying blaKPC and 8 carrying blaNDM, Three strains were identified as blaKPC+ blaNDM-CRKP. In this study, 28 out of 49 CRKP strains (57.2%) were found to carry virulence genes. Additionally, one CRKP strain tested positive in the string test and was found to carry both Aerobactin and rmpA virulence genes. MLST results revealed a total of 5 ST types, with ST11 being predominant (41/49, 83.7%). Successful conjugation was observed in all 3 blaKPC-CRKP strains, while only 1 out of 3 blaNDM-CRKP strains showed successful conjugation. The transconjugant exhibited significantly reduced susceptibility to imipenem and cephalosporin antibiotics. In conclusion, the resistance mechanism of CRKP in this study is primarily attributed to the production of KPC enzymes, along with the presence of multiple β-lactamase resistance genes. Additionally, there is a local prevalence of hv-CRKP and blaKPC+ blaNDM-CRKP. blaKPC and blaNDM can be horizontally transmitted through plasmids, with varying efficiency among different strains.

To examine the molecular epidemiology of carbapenem-resistant Klebsiella pneumoniae (CRKP) and investigate the horizontal transmission of blaKPC and blaNDM genes for the prevention and treatment of CRKP. A total of 49 clinically isolated CRKP strains were retrospectively analyzed from January to December 2022 at The First Hospital of Hunan University of Chinese Medicine. Phenotypic screening was performed using modified carbapenem inactivation assay (mCIM) and EDTA-carbapenem inactivation assay (eCIM). Polymerase chain reaction (PCR) was utilized to identify carbapenem resistance genes, β-lactamase resistance genes, and virulence genes, while multi-locus sequence analysis (MLST) was employed to assess the homology of CRKP strains. Conjugation experiments were conducted to infer the horizontal transmission mechanism of blaKPC and blaNDM genes. The results showed that the study included 49 CRKP strains, with 44 carrying blaKPC and 8 carrying blaNDM, Three strains were identified as blaKPC+ blaNDM-CRKP. In this study, 28 out of 49 CRKP strains (57.2%) were found to carry virulence genes. Additionally, one CRKP strain tested positive in the string test and was found to carry both Aerobactin and rmpA virulence genes. MLST results revealed a total of 5 ST types, with ST11 being predominant (41/49, 83.7%). Successful conjugation was observed in all 3 blaKPC-CRKP strains, while only 1 out of 3 blaNDM-CRKP strains showed successful conjugation. The transconjugant exhibited significantly reduced susceptibility to imipenem and cephalosporin antibiotics. In conclusion, the resistance mechanism of CRKP in this study is primarily attributed to the production of KPC enzymes, along with the presence of multiple β-lactamase resistance genes. Additionally, there is a local prevalence of hv-CRKP and blaKPC+ blaNDM-CRKP. blaKPC and blaNDM can be horizontally transmitted through plasmids, with varying efficiency among different strains.

Objective:To investigate the value of nomogram based on radiomics features of CT enterography (CTE) combined with clinical characteristics to predict secondary loss of response (SLOR) after infliximab (IFX) treatment in patients with Crohn′s disease (CD).Methods:This study was a case-control study. Clinical and imaging data of 155 patients with CD diagnosed at the First Affiliated Hospital of Anhui Medical University from March 2015 to July 2022 were retrospectively collected. The patients were divided into a training set ( n=108) and a testing set ( n=47) in the ratio of 7∶3 by stratified sampling method. All patients were treated according to the standardized protocol and were classified as SLOR (43 in the training set and 18 in the testing set) and non-SLOR (65 in the training set and 29 in the testing set) according to treatment outcome. Based on the data from the training group, independent clinical predictors of SLOR after IFX treatment were screened in the clinical data using univariate and multivariate logistic regression analysis to establish a clinical model. Intestinal phase images were selected to be outlined layer by layer along the margin of the lesion to obtain the volume of the region of interest to extract the radiomics features. The radiomics features were screened using univariate analysis and the minimum absolute shrinkage and selection operator to establish the radiomics model. Multivariate logistic regression analysis was used to build a combined clinical-radiomics model based on the screened clinical independent predictors and radiomics characters, then a nomogram was drawn. The predictive efficacy of the 3 models for SLOR after IFX treatment was assessed by receiver operating characteristic curves, and the area under the curve (AUC) was calculated. The decision curve analysis was applied to evaluate the clinical utility of the models. Results:Disease duration ( OR=1.983, 95% CI 1.966-2.000, P=0.046) and intestinal stenosis ( OR=1.246, 95% CI 1.079-1.764, P=0.015) were identified as the independent predictors of SLOR in the clinical data, and a clinical model was established. Totally 9 radiomics features were included in the radiomics model. The AUCs of clinical, radiomics, and combined models for predicting SLOR after IFX treatment in CD patients were 0.691 (95% CI 0.591-0.792), 0.896 (95% CI 0.836-0.955), and 0.910 (95% CI 0.855-0.965) in the training set, and 0.722 (95% CI 0.574-0.871), 0.866 (95% CI 0.764-0.968), and 0.889 (95% CI 0.796-0.982) in the testing set. Decision curve analysis in the testing set showed higher net clinical benefits for both the radiomics model and combined model than the clinical model, and combined model had higher net clinical benefits than the radiomics model over most threshold probability intervals. Conclusions:CTE-based radiomics model can effectively predict SLOR after IFX treatment in patients with CD, and a combined model by incorporating clinical characteristics of disease duration and intestinal stenosis can further improve the predictive efficacy.

Objectives To evaluate the effectiveness and safety of peramivir trihydrate in patients with influenza.Methods This was a randomized,double-blind,double-dummy,placebo and positive control,multicenter clinical trial,comparing peramivir trihydrate with oseltamivir and placebo.The inclusive criteria were 15-70 years old,onset within 48 h,positive rapid influenza antigen test,and febrile(＞38℃) accompanied with at least two associated symptoms.The severe cases complicated with chronic pulmonary and cardiac diseases,malignancies,organ transplantation,hemodialysis,uncontrolled diabetes,immunocompromised status,pregnancy and coexistence of bacterium infections were excluded.All patients were randomized 2:2:1 to receive peramivir,oseltamivir and placebo respectively.The primary endpoint was the disease duration,the secondary endpoints included time to normal axillary temperature and normal living activities,viral response,and adverse effects.Results Following informed consent,133 patients were included in this study.Four patients were exclude due to missing medical records,not fitting inclusion or exclusion criteria and poor compliance.A total of 129 patients were finally analyzed,including 49 cases,54 cases and 26 cases in peramivir group,oseltamivir group and placebo group.The median disease duration were 96 (76,120)hours,105(90,124) hours,and 124 (104,172)hours in three groups respectively(P＞0.05).The time to normal axillary temperature,normal living activities and viral response were not significantly different in three groups(P＞0.05).Conclusion The value of antiviral therapy in patients with mild influenza needs to be further determined.

Noninvasive high-frequency ventilation (NIHFV) is a new form of noninvasive ventilation which distends positive pressure continuously with oscillations superimposed on spontaneous tidal breathing, through a noninvasive interface.NIHFV, which provides the advantages of nasal continuous positive airway pressure and high frequency ventilation, can rapidly improve oxygenation, effectively remove carbon dioxide, and does not require synchronous techno-logy.It is considered as a new and effective noninvasive ventilation mode.In this paper, the application of NIHFV to neonates were reviewed, including mechanics of gas exchange, research advances and approach to the use of NIHFV.

OBJECTIVE: To compare the histopathological features of the synovium between rheumatoid arthritis (RA) and osteoarthritis (OA). METHODS: We retrospectively analyzed the synovial specimens obtained after synovial surgery in 72 cases of RA and 24 cases of OA. Two independent pathologists reviewed the sections of the synovial tissues with HE staining, quantitatively scored the degree of fibroblast-like synoviocyte (FLS) hyperplasia, vascular hyperplasia, fibroplasia, and lymphocyte infiltration, and examined the presence plasma cell infiltration. The pathological morphology of the synovial tissues was evaluated in relation with the clinical data of the patients. RESULTS: Pannus formation was also detected in the synovium of OA patients, which showed a lesser degree of OA-FLS hyperplasia, fibrosis and lymphocyte infiltration and a significantly lower rate of plasma cell infiltration compared with the pannus in RA patients. Vascular proliferation was also milder in the pannus of OA patients than in RA pannus, but the difference was not statistically significant. In OA patients, the pannus could be observed under a microscope and was difficult to distinguish from that in RA patients. CONCLUSIONS Pannus formation occurs also in the synovium of OA patients but with milder FLS hyperplasia, fibrosis and lymphocyte infiltration and a lower rate of plasma cell infiltration compared with the pannus in RA patients. These differences in the pannus between OA and RA can be of potential value in the diagnosis and treatment of the patients.
Arthritis, Rheumatoid ; Cells, Cultured ; Humans ; Osteoarthritis, Knee ; Retrospective Studies ; Synovial Membrane

Objective To investigate the clinical significance of procalcitonin(PCT)and serum amyloid A (SAA)in early bacterial infection in preschool children.Methods 67 children with bacterial infection(bacte-rial infection group),62 children with viral infection(viral infection group)and 60 healthy children(healthy control group)were enrolled in this study,latex enhanced turbidimetric immunoassay was used to detect SAA and turbidimetric immunoassay was used to detect the level of PCT,the levels of SAA and PCT and the posi-tive rate were compared among all groups,and the sensitivity,specificity and positive predictive value,negative predictive value and Youden index of SAA and PCT levels.Results The levels of SAA and PCT in the bacte-rial infection group before treatment were significantly higher than those in the viral infection group and the healthy control group(P<0.05),7 days after treatment,the levels of SAA and PCT decreased significantly (P<0.05);the level of PCT in viral infection group was not significantly different from that in healthy control group(P>0.05),but the positive rate of SAA was significantly different from that of the healthy control group(P<0.05).The sensitivity,specificity,positive predictive value and negative predictive value of PCT for early bacterial infection in preschool children were 92.5%,93.5%,93.9%,92.1%,respectively,of the SAA values were 97.0%,59.7%,72.2% and 94.9%,respectively,there were significant differences between the two groups in specificity and positive predictive value(P<0.05).Conclusion Detection of SAA and PCT lev-els was helpful for early diagnosis,differential diagnosis and prognosis of bacterial infection in preschool chil-dren.

Objective To investigate the effect of inhibition of Rabll expression on the proliferation and invasion of human bladder cancer cell line T24. Methods T24 cells were transfected with Rabll siRNA, and RNA interference efficiency was determined by performing Western blotting. The effect of inhibition of Rabll expression on cell proliferation, cell cycle progression, and cell invasion was analyzed by performing CCK8, cell cycle detection, and Matrigel invasion assays, respectively. Expression of cell cycle-related proteins cyclin D1 and cyclin E and invasion-related protein matrix metalloproteinase 9 (MMP9) was determined by performing Western blotting and RT-PCR. Results Inhibition of Rabl 1 expression inhibited the proliferation and invasion of bladder cancer cells and downregulated the expression of cell cycle-related proteins cyclin D1 and cyclin E and invasion-related protein MMP9. Conclusion Our results suggest that Rabl 1 functions as a tumor protein and is involved in the progression of bladder cancer.

Objective To investigate the changes of component and morphology in internal carotid vulnerable plaque,for helping to make clinical intervention strategy individually. Methods A total of 47 patients with internal carotid vulnerable plaques and primary hypertension underwent 2 high-resolution and multi-contrast MRI scans, from March 2008 to April 2014 were retrospectively reviewed. At baseline, the plaque was mainly located at the proximal internal carotid artery,and maximum plaque thickness ≥1.5 mm with intraplaque hemorrhage(IPH)and(or)thin or ruptured fibrous cap.Interscan interval was 0.5 years and above. Patients with carotid occlusion or surgery were excluded. Morphological measurements included maximum plaque thickness, maximum plaque area and cross-sectional vessel area (CSVA) on the level of plaque with maximum thickness. The paired-samples t test was performed to compare the difference of plaque morphology between baseline and follow-up carotid MRI.Results The interscan interval was 1.83 (1.59,1.99)years for 47 internal carotid vulnerable plaques.One case(interscan interval 2.16 years)showed IPH within those 11 plaques without IPH at baseline,and one case(interscan interval 1.42 years)had new incident IPH within those 36 plaques with IPH at baseline. Maximum plaque thickness increased significantly from(3.94±1.44)mm to(4.24±1.68)mm(t=2.30,P<0.05)by 5.14%(-3.83,11.34)% per year. Maximum plaque area increased significantly from(49.19±21.15)mm2to(56.03±24.91)mm2(t=3.87,P<0.01)by 6.67%(-2.26,19.60)% per year.CSVA increased significantly from(66.22±27.51)mm2to(73.68±31.47)mm2(t=4.08,P<0.01)by 5.18%(-1.63,12.34)% per year.Conclusion The progression of component,burden and outer remodeling in the internal carotid vulnerable plaque may be faster in hypertension, therefore reasonable intervention strategy and regular follow-up carotid MRI should be performed.

Objective: To produce latent membrane protein 2A (LMP2A) chimeric antigen receptor (CAR)-T cells and detect the lethal effect of LMP2A CAR-T cells on nasopharyngeal carcinoma (NPC) cells. Methods: The study was conducted from September 2016 to December 2017.Genetic engineering technology was used to construct anti-LMP2A CAR lentiviral expression vector and sequencing was identified. The expression of anti-LMP2A CAR in the 293T cells was confirmed by western blot. CCK8 assay was used to evaluate the cytotoxicity of LMP2A CAR-T cells to NPC cells. ELISA assay was performed to test IL-2 and IFN-γ releasing of activated LMP2A CAR-T cells. The inhibition effect of LMP2A CAR-T cells on NPC xenograft tumor was observed in vivo. Statistical analysis was performed by statistical software SPSS 21.0. Results: The results of PCR and sequencing showed that anti-LMP2A CAR lentiviral expression vector was constructed successfully. The result of western blot indicated the expression of anti-LMP2A CAR in the 293T cells effectively. The results of CCK-8 assay showed that the killing activities of LMP2A CAR-T cells to LV-LMP2A-CNE1 cells were (72.11±9.75)%, (54.65 ±5.42)% and (36.68±3.80)% at 20∶1, 10∶1 and 5∶1 ratio of effective cells to target cells, and had a statistical difference compared to CD19 CAR-T cells and T cells (P<0.05). There was no significant difference in the killing activities of LMP2A CAR-T cells to CNE1 cells compared with CD19 CAR-T cells and T cells. The results of ELISA showed that the content of IL-2 and IFN-γ in the co-culture supernatant of LMP2A CAR-T cells and LV-LMP2A-CNE1 cells was significantly higher than that of LMP2A CAR-T cells and CNE1 cells which had statistical difference (P<0.05); In vivo experiment, the volume of LMP2A CAR-T cell group was (80.3±10.0) mm³ which was significantly lower than that of the control groups, and the difference was statistically significant (P<0.05). Conclusion LMP2A CAR-T cells are successfully prepared and have an obvious targeting cytotoxicity on LMP2A-positive NPC cells.