ObjectiveTo understand the infection characteristics of multidrug-resistant organisms (MDROs) in hospitalized patients in a tertiary sentinel hospital in Shanghai, so as to provide an evidence for the development of targeted prevention and control measures. MethodsData of MDROs strains and corresponding medical records of some hospitalized patients in a hospital in Shanghai from 2021 to 2023 were collected, together with an analysis of the basic information, clinical treatment, underlying diseases and sources of sample collection. ResultsA total of 134 strains of MDROs isolated from hospitalized patients in this hospital were collected from 2021 to 2023 , including 63 strains of methicillin-resistant Staphylococcus aureus (MRSA), 57 strains of carbapenem-resistant Acinetobacter baumannii (CRAB), and 14 strains of carbapenem-resistant Klebsiella pneumoniae (CRKP). Of the 134 strains, 30 strains were found in 2021, 47 strains in 2022 and 57 strains in 2023. The male-to-female ratio of patients was 2.05∶1, with the highest percentage (70.90%) in the age group of 60‒<90 years. The primary diagnosis was mainly respiratory disease, with lung and respiratory tract as the cheif infection sites. There was no statistically significant difference in the distribution of strains between different genders and infection sites (P>0.05). However, the differences in the distribution of strains between different ages and primary diagnosis were statistically significant (P<0.05). Patients who were admitted to the intensive care unit (ICU), had urinary tract intubation, were not artery or vein intubated, were not on a ventilator, were not using immunosuppresants or hormones, and were not applying radiotherapy or chemotherapy were in the majority. There was no statistically significant difference in the distribution of strains for whether received radiotherapy or chemotherapy or not (P>0.05), while the differences in the distribution of strains with ICU admission history, urinary tract intubation, artery or vein intubation, ventilator use, and immunosuppresants or hormones use or not were statistically significant (all P<0.05). The type of specimen was mainly sputum, the hospitalized ward was mainly comprehensive ICU, the sampling time was mainly in the first quarter throughout the year, the number of underlying diseases was mainly between 1 to 2 kinds, the application of antibiotics ≥4 kinds, and those who didn’t receive any surgery recently accounted for the most. There were statistically significant differences in the distribution of strains between different specimen types, wards occupied and history of ICU stay (P<0.05), but no statistically significant difference in the distribution of strains between different sampling times, number of underlying diseases and types of antibiotics applied (P>0.05). ConclusionThe situation of prevention and control on MDROs in this hospital is still serious. Focus should be placed on high-risk factors’ and infection monitoring and preventive measures should be strengthened to reduce the incidence rate of MDROs infection.

Objective:To observe the efficacy and safety of vitrectomy combined with subretinal injection of alteplase (tPA) and intravitreal injection of Conbercept in the treatment of large area submacular hemorrhage (SMH) secondary to polypoidal choroidal vasculopathy (PCV).Methods:A retrospective clinical study. From January to September 2021, 32 eyes of 32 patients with massive SMH secondary to PCV diagnosed in the Affiliated Eye Hospital of Nanchang University were included in the study. Large SMH was defined as hemorrhage diameter ≥4 optic disc diameter (DD). There were 32 patients (32 eyes), 20 males and 12 females. The mean age was (72.36±8.62) years. All patients had unilateral disease.The duration from onset of symptoms to treatment was (7.21±3.36) days. All patients underwent best corrected visual acuity (BCVA) and optical coherence tomography (OCT) examination. BCVA examination was performed using the international standard visual acuity chart, which was converted to the logarithm of the minimum angle of resolution (logMAR) visual acuity during statistics. The central macular thickness (CMT) was measured by spectral domain-OCT. The average size of SMH was (6.82±1.53) DD. The logMAR BCVA 1.73±0.44; CMT was (727.96±236.40) μm. All patients were treated with 23G pars plana vitrectomy combined with subretinal injection of tPA and intravitreal injection of Conbercept. At 1, 3, 6 and 12 months after treatment, the same equipment and methods were used for relevant examinations before treatment. The changes of BCVA and CMT, the clearance rate of macular hemorrhage, and the complications during and after surgery were observed. BCVA and CMT before and after treatment were compared by repeated measures analysis of variance.Results:Compared with before treatment, BCVA gradually increased at 1, 3, 6 and 12 months after treatment, and the differences were statistically significant ( F=77.402, P<0.001). There was no significant difference in BCVA between any two groups at different time points after treatment ( P>0.05). Correlation analysis showed that BCVA at 12 months after treatment was negatively correlated with the course of disease ( r=-0.053, P=0.774). One week after treatment, macular hemorrhage was completely cleared in 30 eyes (93.75%, 30/32). The CMT was (458.56±246.21), (356.18±261.46), (345.82±212.38) and (334.64±165.54) μm at 1, 3, 6 and 12 months after treatment, respectively. Compared with before treatment, CMT decreased gradually after treatment, and the difference was statistically significant ( F=112.480, P<0.001). There were statistically significant differences in different follow-up time before and after treatment ( P<0.001). The number of treatments combined with Conbercept during and after surgery was (4.2±1.8) times. At the last follow-up, there was no recurrence of SMH, retinal interlamellar effusion and other complications. Conclusion:Subretinal injection of tPA combined with intravitreal injection of Conbercept is safe and effective in the treatment of large SMH secondary to PCV, and it can significantly improve the visual acuity of patients.

Objective To identify and verify the interacting protein of α-11 giardin, so as provide the experimental evidence for studies on the α-11 giardin function. Methods The yeast two-hybrid cDNA library of the Giardia lambia C2 strain and the bait plasmid of α-11 giardin were constructed. All proteins interacting with α-11 giardin were screened using the yeast two-hybrid system. α-11 giardin and all screened potential interacting protein genes were constructed into pBiFc-Vc-155 and pBiFc-Vn-173 plasmids, and co-transfected into the breast cancer cell line MDA-MB-231. The interactions between α-11 giardin and interacting proteins were verified using bimolecular fluorescence complementation (BiFC). Results The yeast two-hybrid G. lambia cDNA library which was quantified at 2.715 × 10⁷ colony-forming units (CFU) and the bait plasmid containing α-11 giardin gene without an autoactivation activity were constructed. Following two-round positive screening with the yeast two-hybrid system, two potential proteins interacting with α-11 giardin were screened, including eukaryotic translation initiation factor 5A (EIF5A), calmodulin-dependent protein kinase (CAMKL) and nicotinamide adenine dinucleotide phosphate-specific glutamate dehydrogenase (NADP-GDH), hypothetical protein 1 (GL50803_95880), hypothetical protein 2 (GL50803_87261) and a protein from Giardia canis virus. The α-11 giardin and EIF5A genes were transfected into the pBiFc-Vc-155 and pBiFc-Vn-173 plasmids using BiFC, and the recombinant plasmids pBiFc-Vc-155-α-11 and pBiFc-Vn-173-EIF5A were co-tranfected into MDA-MB-231 cells, which displayed green fluorescence under a microscope, indicating the interaction between α-11 giardin and EIF5A protein in cells. Conclusion The yeast two-hybrid cDNA library of the G. lambia C2 strain has been successfully constructed, and six potential protein interacting with α-11 giardin have been identified, including EIF5A that interacts with α-11 giardin in cells.

Background With the change of the national energy development layout, Qingyang has seen a situation where oil exploitation and agriculture go hand in hand, which may lead to local soil pollution if not taken seriously. Objective To evaluate the distribution characteristics, possible sources, and ecological risks of heavy metals in farmland soils around the main production areas of Changqing Oilfield. Methods A total of 60 farmland soil samples were collected from Zhengning County, Zhenyuan County, and Qingcheng County of Qingyang City, and the contents of heavy metals such lead (Pb), mercury (Hg), chromium (Cr), cadmium (Cd), and arsenic (As) in farmland soil were detected according to GB 15618-2018 Soil environmental quality—Risk control standard for soil contamination of agricultural land (on trial). The soil background value of Gansu Province was used as the denominator in the calculation of pollution index, and the pollution characteristics and ecological characteristics of selected five heavy metals in farmland soil were evaluated by single-factor pollution index (P_i), Nemerow comprehensive pollution index (P_N), and potential ecological risk index. Results The levels of Pb, As, and Hg in farmland soils around Changqing Oilfield, the levels of Cr and Cd in Qingcheng County, and the level of Cd in Zhengning County were higher than the corresponding soil background values of Gansu Province, but lower than the national soil environmental quality standard. The single-factor pollution indexes (P_i) were: Hg, 2.14; Pb, 1.24; As, 1.13; Cr, 0.78; Cd, 0.67, which indicated that Hg were graded as moderate pollution, Pb and As were slight pollution, and Cd and Cr were not polluted. The Nemerow comprehensive pollution indexes (P_N) were: Cr, 0.92; Cd, 1.08; As, 1.20; Pb, 1.68; Hg, 3.85, which indicated that Cr was graded as no pollution, Cd, Pb and As were mild pollution, and Hg was severe pollution. The variation coefficients of Hg and Cd in Zhenyuan County and that of Hg in Qingcheng County were 60.00%, 50.00%, and 50.00%, respectively, which were all greater than 50%, indicating that the pollution of above heavy metals in the location was subject to human activities. The potential ecological risk indexes (E_r) of Pb, Cr, Cd, As, and Hg were 6.20, 1.55, 20.05, 11.28, and 81.64, respectively, indicating that Hg was graded as strong ecological risk, and the other four heavy metals were mild ecological risk. The comprehensive potential ecological risk index (R_RI) was 124.48. Combined with the potential ecological risk index of Hg, the comprehensive potential ecological risk of the five heavy metals in local farmland soils was considered to be at a strong ecological risk level. Conclusion Although the average values of selected five heavy metals in farmland soils surrounding the main production areas of Changqing Oilfield are qualified with the national soil environmental quality standards, they exceed corresponding soil background values of Gansu Province, and there are signs of human influence and potential ecological risks of different degrees.

Objective:To identify 49 Aeromonas caviae strains isolated form foodborne and environmental samples and analyze their virulence and antibiotic resistance. Methods:All strains were identified by VITEK and API 20NE. Two housekeeping genes, gryB and rpoD, were amplified by PCR for phylogenetic analysis. Virulence genes including act, alt, ast, lip, ahp, aerA, hlyA, ompA1 and fla were detected by PCR. In vitro antimicrobial susceptibility of these strains was tested with AST-GN16 kit. Results:Fifty-four Aeromonas caviae/ Aeromonas hydrophila strains were identified by biochemical tests. Phylogenetic analysis revealed that there were 49 Aeromonas caviae strains, four Aeromonas hydrophila strains and one Aeromonas taiwanensis strain. The positive rates of alt, lip, ompA1, fla, act, aerA and hlyA genes were 100.00%, 100.00%, 79.59%, 14.29%, 2.04%, 2.04% and 2.04%, respectively. None of the isolates carried ast or ahp gene. A total of four virulence gene combination patterns were detected, and the predominant pattern was alt/ lip/ ompA1 (32/49). Antibiotic resistance rates of the Aeromonas caviae strains to ampicillin, cefazolin, cefoxitin, amoxicillin/clavulanic acid, ceftriaxone, trimethoprim/sulfamet hoxoazole and aztreonam were 79.59%, 14.29%, 10.20%, 6.12%, 4.08%, 4.08% and 2.04%, respectively. All strains were susceptible to piparacillin/tazobactam, imipenem, amikacin, gentamicin, levofloxacin, tigacycline and nitrofurantoin. Two multidrug-resistant strains were detected. Conclusions:Aeromonas caviae/ Aeromonas hydrophila can be effectively identified by the housekeeping genes gyrB and rpoD, and the closely related Aeromonas caviae and Aeromonas taiwanensis strains can be identified by rpoD. All Aeromonas caviae strains carried two or more virulence genes and one strain isolated from environment was positive for seven virulence genes. Aeromonas caviae strains were generally resistant to penicillin, which was mainly relate to the production of β-lactamase. No strain was resistant to carbapenems, aminoglycosides, fluoroquinolones, tetracyclines or furans. Multidrug-resistant strains were observed in food and drinking water.

Cystic echinococcosis (CE) in sheep is a hazardous zoonotic parasitic disease that is caused by Echinococcus granulosus (Eg). At present, serological test is an important diagnostic method for Eg infection in domestic animals. Here, a fusion protein Eg mefAg-1 harboring 8 dominant B-cell epitopes of Eg such as antigen B, tetraspanin 1, tetraspanin 6, reticulon and Eg95 was produced in E. coli and evaluated for CE in sheep by indirect ELISA. Eg mefAg-1 showed in ELISA a high sensitivity (93.41%) and specificity (99.31%), with a coincidence rate of 97.02%. Overall, it is suggested that the Eg mefAg-1 could be a potential antigen candidate for CE serodiagnosis in sheep.
Animals, Domestic ; Echinococcosis ; Echinococcus granulosus ; Enzyme-Linked Immunosorbent Assay ; Epitopes, B-Lymphocyte ; Methods ; Parasitic Diseases ; Sensitivity and Specificity ; Serologic Tests ; Sheep

Steady improvement in mass spectrometers technology has transformed the targeted proteome analysis into a new stage. Parallel reaction monitoring (PRM) technology has evolved from the basic multiple reaction monitoring (MRM) targeted proteomics methods in recent years. PRM performs with a higher sensitivity, throughput and reproducibility in targeted quantification, however its limitations in effectiveness and accurate quantification of samples with higher complexity still remain unsolved. In this study through improving the chromatographic conditions of PRM we established a simple and robust platform for targeted proteomic quantification. The newly established PRM system is equipped with columns with increased inner diameter (150 μm) and decreased total length (8 cm); faster liquid phase elution rate (800 nL/min) and shortened elution gradient (35 min). These modifications enable PRM platform to combine with dual reverse phase chromatography, to quantify up to 400 low abundance peptides in human 293T cells whole cell extract. Our findings would benefit the promotion of PRM technology, especially providing a technical option for accurate quantification of low abundance proteins.

Objective To investigate the effect of coupled plasma filtration adsorption (CPFA) on plasma cytokines:TNF-α,IL-1β,IL-6,cellular immunity,blood lactate acid concentration,heart rate,respiration rate,oxygenation index,hemodynamics,blood cells counts,and prognosis in patients with multiple organ dysfunction syndromes (MODS).Methods This was a prospective,randomized clinical trial in 45 patients diagnosed as MODS.Patients were randomly assigned to hemoperfution with resin adsorption (HP) + continuous venous-venous hemofiltration (CVVH) group,CPFA group and CVVH group.The general clinical data,APACHE Ⅱ score,number of failure organ and previous mentioned biomarkers were documented.Blood samples were collected before and after blood filtration with any one of these procedures.The plasma samples were isolated and stored with frozen at-60 ℃.Data were statistically analyzed with SPSS 13.0 version software.Results In CPFA group,plasma cytokines,TNF-α、IL-1β、IL-6,decreased markedly after plasma adsorption for two hours (P ＜ 0.01);and plasma concentrations of IL-6 were further descended after subsequent CVVH for 10 hours (P ＜ 0.05).In HP + CVVH group,plasma cytokines,TNF-α、IL-1β、IL-6,decreased markedly after HP (P ＜ 0.01),and plasma concentrations of IL-6 were further descended after subsequent CVVH for 10 hours (P ＜ 0.05).In CVVH group,plasma cytokines,TNF-α、IL-1β、IL-6,decreased after CVVH for 12 hours (P ＜ O.05).Blood lactate acid concentration,heart rate,respiration rate,oxygenation index,T-lymphocytes subgroups (CD3 +,CD4 +,CD8 +,CD4 +/CD8 + ratio),clinical symptoms were improved and dose of vasoactive agent was reduced in the patients of three groups without differences among them.The counts of red blood cells,white blood cells and platelets after CPFA and CVVH showed no significant changes.There was no significant difference in blood cell counts between CPFA and CVVH groups.After HP + CVVH,there was a trend of decrease in platelet count (P ＜ 0.05).Platelet counts were significanfly higher in patients treated with CPFA and CVVH group than those in patients treated with HP + CVVH group (P ＜ 0.05).There were 6 patients died in HP + CVVH group,6 patients died in CPFA group and 5 patients died in CVVH group within 28days.Conclusions The comparison of efficacy of blood filtration among 3 modalities of HP + CVVH,CPFA and CVVH showed CPFA had higher capacity of Inflammatory medium scavenging than CVVH,and had less damage effect on blood visible component,especially on platelet compared with HP + CVVH.CPFA was an effective and safety modality in the treatment of the patients with multiple organ dysfunction syndrome.

Objective To investigate the expression of scaffold protein palladin in pancreatic ductal adenocarcinoma (PDAC) tissues and to discuss its clinicopathological significance.Methods 56 samples of PDAC and corresponding adjacent normal pancreas (NP) tissues were collected.Another 10 samples of chronic pancreatitis (CP) tissues were collected.Western blot analysis and immunohistochemistry assay were performed to detect the expression of protein palladin.The correlation of palladin expression with clinicopathological factors of PDAC was analyzed.Results Western blot analysis revealed that the expression of palladin in PDAC,CP and NP tissues were respectively 0.93±0.07,0.41±0.07 and 0.20±0.06,and the expression of palladin was significantly increased in PDAC tissues compared with NP and CP tissues (P ＜ 0.05),and its expression was significantly increased in CP tissues compared with NP tissues (P ＜ 0.05).Immunohistochemical staining showed that palladin was mainly expressed in activated myofibroblasts in PDAC tissues.The rate of palladin expression was 79 ％ (44/56),which was higher than that in NP tissues (2/10) and CP tissues (4/10),and its expression was found to be correlated with the degree of tumor differentiation,lymph node metastasis and clinical TNM classification (P ＜ 0.05),and it had no correlation with patient' s sex,age,tumor location and distant metastasis (P ＞ 0.05).Conclusions Scaffold protein palladin is highly expressed in PDAC tissues,and it is expressed in the activated myofibroblasts within tumor microenvironment.Scaffold protein palladin may be involved in the invasion and metastasis of PDAC.

SUMOylation is a post‐translational modification involved in various cellular processes .SUMO‐specific protease (SENP) regulates SUMOylation by removing SUMO from conjugated substrates (deSUMOylation) and promoting maturation of SUMO precursor .In order to express Giardia lambia (C2 strain) SENP catalytic domain in E .coli ,the full‐length open reading frame of SENP was amplified by PCR from Giardia lamblia genome DNA .The PCR product about 1 620 bp was cloned into cloning vector pGM‐T .Sequencing result showed the sequence of SENP in C2 strain was identical with that in Gi‐ardia WB strain .Bioinformatics analysis showed that SENP protein possessed a 372 aa discontinuous ULP catalytic domain at C‐terminal .The catalytic domain of SENP was cloned into prokaryotic expression vector pET‐28a(+ ) .The recombinant vector pET‐28a(+ )‐SENPc was transformed into E .coli Rosetta(DE3) ,then the recombinant SENPc protein was expressed by IPTG induction .SDS‐PAGE and Western blot using anti‐His Tag antibody showed that the expression product of SENPc was a fusion protein with a molecular weight of 43 kD .The successful prokaryotic expression and bioinformatics analysis of Giardia lamblia SENP protein provide basis for further functional study of SENP .