Objective To analyze the rates of drug resistance of carbapenem-resistant Klebsiella pneumoniae (CRKP) and extended-spectrum β-lactamase-producing Escherichia coli (ESBL-E.coli) in bronchoalveolar lavage fluid (BALF) of critically ill patients with severe pneumonia and artificial airway and risk factors of CRKP. Methods Patients with severe pneumonia and artificial airway treated in the Center of Emergency and Intensive Care Unit, Jinshan Hospital, Fudan University from May 2022 to May 2025 were selected. Klebsiella pneumoniae (K. pneumoniae) and Escherichia coli (E.coli) isolated and cultured from BALF of these patients were analyzed. The drug resistance rates of the two bacteria were compared. The multivariate logistic regression was used to analyze related risk factors for CRKP infection. Results A total of 384 patients were included, and 384 strains were cultured, including 330 strains (85.9%) of K. pneumoniae and 54 strains (14.1%) of E.coli. The infection rate of K. pneumoniae was higher. Compared with E.coli, the drug-resistant gene expression rate of K. pneumoniae was lower (37.3% vs 57.4%, P＜0.01). The resistance rates of CRKP to cefotaxime, cefepime, aztreonam, and imipenem were higher than those of ESBL-E.coli (P＜0.003). Both CRKP and ESBL-E.coli had high sensitivity to polymyxin and tigecycline. Multivariate logistic regression showed duration of mechanical ventilation ≥ 7 days, and combination of ≥ 2 antibiotics for over 7 days were independent risk factors for CRKP infection (P＜0.01). Conclusions For patients with severe pneumonia and artificial airway in ICU, K. pneumoniae is the main bacterial infection. CRKP and ESBL-E.coli in BALF exhibit different degrees of drug resistance to various antibacterial agents, both have the characteristic of multidrug resistance, and are sensitive to polymyxin and tigecycline. Prevention and control of risk factors related to CRKP should be formulated particularly.

Bacillus coagulans can utilize the hydrolyzed carbon source of agricultural waste to produce lactic acid via a homofermentative pathway. However, a significant carbon source metabolic repression effect was observed when the strain metabolized mixed sugars (glucose and xylose), reducing the productivity of lactic acid. In this study, we obtained the fermentation conditions for the simultaneous utilization of the mixed sugars by B. coagulans by changing the ratio of glucose to xylose in the medium. Through transcriptome sequencing, several key genes responsible for xylose utilization were identified. The critical role of xylose isomerase (XylA, EC 5.3.1.5) in the synchronous utilization of glucose/xylose in B. coagulans was investigated via qRT-PCR (quantitative real-time polymerase chain reaction). Subsequently, the heterologous expression and characterization of the XylA-encoding gene (XylA) were conducted. It was determined that the gene encoded a protein composed of 440 amino acid residues. The secondary structure of the encoded protein was predominantly composed of α-helixes and random coils, while the higher structure of the protein was identified as a homotetramer. Then, XylA was cloned and expressed in Escherichia coli BL21(DE3), and the recombinant protein Bc-XlyA was obtained with a molecular weight of approximately 50 kDa. The optimal pH and temperature of Bc-XylA were 8.0 and 60 ℃, respectively, and Mn²⁺, Mg²⁺, and Co²⁺ had positive effects on the activity of Bc-XlyA. The present study provides scientific data on the molecular modification of B. coagulans, offering theoretical support for the efficient utilization of xylose in the strain.
Xylose/metabolism* ; Cloning, Molecular ; Bacillus coagulans/enzymology* ; Aldose-Ketose Isomerases/metabolism* ; Fermentation ; Bacterial Proteins/metabolism* ; Glucose/metabolism*

Objective: To establish a method for determining the molecular weight and molecular weight distribution of Poly(Lactide-co-Glycolide Acid) (PLGA) using Size Exclusion Chromatography-Refractive Index-Multiangle Laser Light Scattering (SEC-RI-MALLS) and Size Exclusion Chromatography-Refractive Index (SEC-RID), and to compare the results obtained from these two methods.
Methods: For SEC-RI-MALLS, tetrahydrofuran was used as the mobile phase, Shodex GPC KF-803L was employed as the chromatographic column with a flow rate of 1 mL·min^-1, column temperature at 30 ℃, and an injection volume of 100 μL. For SEC-RID, tetrahydrofuran was also used as the mobile phase, Agilent PLgel 5 μm MIXD-D was used as the chromatographic column with a flow rate of 1 mL·min^-1, column temperature at 30 ℃, differential detector temperature at 35 ℃, and an injection volume of 20 μL. The molecular weight and molecular weight distribution were calculated using Agilent’s GPC software. The newly established methods were validated methodologically, and the molecular weight and molecular weight distribution of 13 batches of samples were determined.
Results: The precision, accuracy, stability, and repeatability tests for SEC-RI-MALLS showed RSD values of 1.35%, 1.58%, 1.53%, and 1.26%, respectively. The SEC-RID method exhibited good linearity (r=0.999 9), with RSD values for precision, accuracy, stability, and repeatability tests (n=6) of 2.05%, 1.62%, 1.30%, and 2.97%, respectively. The results obtained from SEC-RI-MALLS were lower than those from SEC-RID, and the molecular weight distribution coefficient was smaller, but the results from the paired T-test performed with the value measured by SEC-RID method and the value measured by SEC-RI-MALLS method multiplied a conversion coefficient of 1.5 showed no significant difference between the two methods.
Conclusion: Both methods are stable and reliable, and can be used for the determination of PLGA molecular weight and molecular weight distribution based on the specific situations.

BACKGROUND:Lingual movable wing is a new type of lingual orthodontic technique and the different stretching lengths of the wring affect the torque control effect of anterior teeth.However,there is yet no related biomechanical research. OBJECTIVE:To investigate the displacement trend of dentition during adduction of mandibular anterior teeth and the effect of different wing stretching lengths on the biomechanical effect of mandibular anterior teeth. METHODS:The data of the mandible and lower dentition were collected by cone-beam CT and reconstructed using Mimics software to establish a three-dimensional finite element model of mandibular anterior teeth adducted by the lingual movable wing.The ANSYS software was used to analyze the initial displacement of the mandibular anterior teeth under the following conditions:A,2 mm stretching length;B,2.5 mm stretching length;C,3 mm stretching length;and D,3.5 mm stretching length. RESULTS AND CONCLUSION:The trend of initial displacement of lower dentition:The central incisors moved lingually with depression,the lateral incisors and canines moved mildly lingually with mesial lingual torsion,the second premolar was tilted distally with a marked lingual inclination and the first molar showed an overall mesial inclination with mesial crown eversion.Therefore,in the adduction cases of mandibular tooth extraction,attention should be paid to the lingual movement of the second premolar,which could be offset by corresponding techniques in clinic.The trend of anterior tooth displacement in all directions:from condition A to condition D,in the sagittal direction,the difference value in crown-root displacement of central incisors changed from-11.891 μm to-5.757 4 μm,indicating that the central incisor changes from oblique movement to overall movement.The difference value in crown-root displacement of lateral incisors changed from-11.828 1 μm to-6.711 45 μm,and that of canines changed from-7.572 3 μm to-4.695 5 μm,indicating that the oblique movement of the lateral incisors and canines is also changing to an overall movement.In the vertical direction,from condition A to condition D,the reduction of incisors was gradually increased,while that of canines was gradually decreased.These findings indicate that the stretching length of the wing can affect the oblique movement trend of the anterior teeth.As the wing continues to stretch,the torque control of the lower anterior teeth will become better.

Objective:To juxtapose laparoscopic cholecystectomy combined with common bile duct exploration and stone extraction (LC+ LCBDE) against endoscopic retrograde cholangiopancreatography/sphincterotomy with laparoscopic cholecystectomy (LC+ ERCP/EST) in the therapeutic context of acute biliary pancreatitis.Methods:The clinical data of patients with acute biliary pancreatitis in Department of Hepatobiliary Surgery, Datong Third People's Hospital from January 2017 to January 2021 were retrospectively analyzed. A total of 44 patients were inrolled, including 23 males and 21 females, with the age of (60.6±11.7) years. Based on different treatment approaches, the patients were divided into the LC+ LCBDE group ( n=33) and the LC+ ERCP group ( n=11, LC+ ERCP/EST). Total bilirubin, direct bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood amylase, operation time, postoperative hospitalization stays, total hospitalization cost, postoperative anal exhaust time, and postoperative complications (bile leakage, fever, bleeding) were compared between the two groups. Results:There were no significant differences in preoperative total bilirubin, direct bilirubin, ALT, AST, and blood amylase between LC+ ERCP group and LC+ LCBDE groups (all P>0.05). In LC+ LCBDE group, operation time was 110.0 (96.3, 147.5) min, postoperative hospitalization time was 9.0 (7.5, 11.0) d, postoperative exhaust time was 2.0 (1.0, 2.0) d, and in LC+ LCBDE group, operation time was 60.0 (32.0, 65.0) min, postoperative hospitalization time was 7.0 (4.0, 8.0) d, postoperative exhaust time was 1.0 (1.0, 1.0) d. Comparisons with LC+ LCBDE group, LC+ ERCP group had shorter postoperative hospitalization stay and earlier postoperative exhaust time, the total hospitalization cost of LC+ LCBDE group was 23 829.3 (21 779.6, 27 221.9) yuan, which was higher than 36 894.8 (31 963.5, 41 172.2) yuan in LC+ ERCP group, and the differences were statistically significant (all P<0.05). Comparison of postoperative total bilirubin, direct bilirubin, ALT and AST between LC+ ERCP group and LC+ LCBDE group, with no significant difference(all P>0.05). No postoperative complications such as bile leakage, residual stones, fever and bleeding occurred in both groups. Conclusion:Compared with LC+ ERCP/EST, LC+ LCBDE in the treatment of acute biliary pancreatitis, although the operation time and hospital stay are longer, but the total hospitalization cost is less, there is no need for multiple operations, and it can be used as the first choice for acute biliary pancreatitis.

Objective To investigate the effects of monocyte chemotactic protein-4(MCP-4)on pyroptosis and p38 mitogen-activated protein kinase(p38 MAPK)activation in human nasal epithelial cells(HNEpCs).Methods Forty patients diagnosed with chronic rhinosinusitis(CRS)were selected as the case group,and another 40 healthy individuals were selected as the normal group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of MCP-4 in serum samples from both groups.Receiver operator characteristic(ROC)curve analysis was performed to assess the diagnostic efficacy of serum MCP-4 levels for CRS.HNEpCs were divided into six groups:control group,5 mg/L lipopolysaccharide(LPS)model group,LPS+siRNA negative control(si-Con)group,LPS+siRNA MCP-4(si-MCP-4)group,LPS+pcDNA3.1-vector group,and LPS+pcDNA3.1-MCP-4 group.Cell viability was assessed using the cell counting kit-8(CCK-8),and cell proliferation activity was detected by 5-Ethynyl-2'-deoxyuridine(EdU)staining.Western blot was used to detect the expression levels of phosphorylated p38 MAPK(p-p38 MAPK),Gasdermin D(GSDMD),NOD-like receptor family,pyrin domain containing 3(NLRP3),apoptosis-associated speck-like protein(ASC),and caspase-1.ELISA was performed to measure the levels of tumor necrosis factor-α(TNF-α),interleukin-18(IL-18),and IL-1β in cell supernatants.The Transwell chamber was utilized to assess the effects of MCP-4 on the migratory capacity of HNEpCs.HNEpCs were divided into four groups:control group,10 ng/ml MCP-4 group,and 20 ng/ml MCP-4 group.Results Compared with the normal group,the level of MCP-4 in the serum of patients in the case group was significantly upregulated.ROC curve analysis showed that the area under the curve(AUC)for serum MCP-4 levels in the diagnosis of CRS was 0.921.In vitro experiments demonstrated that LPS stimulation reduced the survival rate and proliferation ability of HNEpCs,increased the expression of pyroptosis proteins,and upregulated p-p38 MAPK levels,and increased the release of TNF-α,IL-1β,and IL-18(P<0.01).Compared with the LPS model group,the LPS+si-MCP-4 group showed increased survival rate and proliferation ability of HNEpCs,decreased expression levels of p-p38 MAPK,pyroptosis proteins and reduced levels of TNF-α,IL-1β,and IL-18(P<0.01).In contrast,compared with the LPS model group,the LPS+pcDNA3.1-MCP-4 group exhibited further decreased survival rate and proliferation ability of HNEpCs,increased expression levels of p-p38 MAPK,pyroptosis proteins,and elevated levels of inflammatory cytokines TNF-α,IL-1β,and IL-18(P<0.01).Compared with the control group,there was no significant increase in the migration ability of HNEpCs in the 10 ng/mL MCP-4 and 20 ng/ml MCP-4 groups.Conclusion Inhibiting the expression of MCP-4 can block the activation of p38 MAPK,reducing pyroptosis and the release of inflammatory cytokines in HNEpCs.

Objective To investigate the effects of monocyte chemotactic protein-4(MCP-4)on pyroptosis and p38 mitogen-activated protein kinase(p38 MAPK)activation in human nasal epithelial cells(HNEpCs).Methods Forty patients diagnosed with chronic rhinosinusitis(CRS)were selected as the case group,and another 40 healthy individuals were selected as the normal group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of MCP-4 in serum samples from both groups.Receiver operator characteristic(ROC)curve analysis was performed to assess the diagnostic efficacy of serum MCP-4 levels for CRS.HNEpCs were divided into six groups:control group,5 mg/L lipopolysaccharide(LPS)model group,LPS+siRNA negative control(si-Con)group,LPS+siRNA MCP-4(si-MCP-4)group,LPS+pcDNA3.1-vector group,and LPS+pcDNA3.1-MCP-4 group.Cell viability was assessed using the cell counting kit-8(CCK-8),and cell proliferation activity was detected by 5-Ethynyl-2'-deoxyuridine(EdU)staining.Western blot was used to detect the expression levels of phosphorylated p38 MAPK(p-p38 MAPK),Gasdermin D(GSDMD),NOD-like receptor family,pyrin domain containing 3(NLRP3),apoptosis-associated speck-like protein(ASC),and caspase-1.ELISA was performed to measure the levels of tumor necrosis factor-α(TNF-α),interleukin-18(IL-18),and IL-1β in cell supernatants.The Transwell chamber was utilized to assess the effects of MCP-4 on the migratory capacity of HNEpCs.HNEpCs were divided into four groups:control group,10 ng/ml MCP-4 group,and 20 ng/ml MCP-4 group.Results Compared with the normal group,the level of MCP-4 in the serum of patients in the case group was significantly upregulated.ROC curve analysis showed that the area under the curve(AUC)for serum MCP-4 levels in the diagnosis of CRS was 0.921.In vitro experiments demonstrated that LPS stimulation reduced the survival rate and proliferation ability of HNEpCs,increased the expression of pyroptosis proteins,and upregulated p-p38 MAPK levels,and increased the release of TNF-α,IL-1β,and IL-18(P<0.01).Compared with the LPS model group,the LPS+si-MCP-4 group showed increased survival rate and proliferation ability of HNEpCs,decreased expression levels of p-p38 MAPK,pyroptosis proteins and reduced levels of TNF-α,IL-1β,and IL-18(P<0.01).In contrast,compared with the LPS model group,the LPS+pcDNA3.1-MCP-4 group exhibited further decreased survival rate and proliferation ability of HNEpCs,increased expression levels of p-p38 MAPK,pyroptosis proteins,and elevated levels of inflammatory cytokines TNF-α,IL-1β,and IL-18(P<0.01).Compared with the control group,there was no significant increase in the migration ability of HNEpCs in the 10 ng/mL MCP-4 and 20 ng/ml MCP-4 groups.Conclusion Inhibiting the expression of MCP-4 can block the activation of p38 MAPK,reducing pyroptosis and the release of inflammatory cytokines in HNEpCs.

Objective To investigate the infection status and drug resistance of urogenital tract mycoplasma in Lianyungang district,Jiangsu.Methods Based on retrospective collection of 22 889 cases of suspected mycoplasma genital infection in Lianyungang Mater-nity and Child Health Hospital from 2015 to 2022,the infection and drug resistance rate of genital mycoplasma were analyzed.Results A total of 8 943 genital mycoplasma positive specimens were detected.Among them,the infection rate of Ureaplasma urealyticum(Uu)was the highest,accounting for 33.64％(7 700/22 889),which was significantly higher than that of Mycoplasma hominis(Mh),3.91％(894/22 889),and that of mixed(Uu+Mh)type,1.52％(349/22 889).The difference was statistically significant(P＜0.05).The infection rates of genital mycoplasma and Uu showed increasing trend from 2015 to 2022.The overall infection rates of genital mycoplasma in females were 42.42％,which was significantly higher than that in males of 22.47％.The highest infection rate of 40.84％(6 108/14 955)was found in the group of 20 to 40 years old as compared to those younger than 20 years old as well as those over 40 years old.The genital mycoplasma was highly sensitive to doxycycline,minocycline and josamycin,and the drug resistance rate was lower than 5.00％.From 2015 to 2022,there was an increasing trend of resistance rates to erythromycin,ofloxacin and ciprofloxa-cin in Uu-infected patients(P＜0.05)and erythromycin,roxithromycin,azithromycin and ofloxacin in Mh-infected patients(P＜0.05).Conclusion The incidence of urogenital tract mycoplasma infection in Lianyungang district shows a gradual increase for recent years.Doxycycline,minocycline and josamycin are suitable as the first choice for the treatment of genital mycoplasma infection.Most quinolo-ne drugs have shown high resistance rate,indicating that this type of drugs are possibly overused.The strengthened relevant monitoring of antibacterial drugs should be necessary.

Lingguizhugan Decoction (LGZG) has been investigated in basic studies, with satisfactory effects on insulin resistance in non-alcoholic fatty liver disease (NAFLD). This translational approach aimed to explore the effect and underlying mechanism of LGZG in clinical setting. A randomized, double-blinded, placebo-controlled trial was performed. A total of 243 eligible participants with NAFLD were equally allocated to receive LGZG (two groups: standard dose and low dose) or placebo for 12 weeks on the basis of lifestyle modifications. The primary efficacy variable was homeostasis model assessment of insulin resistance (HOMA-IR). Analyses were performed in two populations in accordance with body mass index (BMI; overweight/obese, BMI ⩾ 24 kg/m²; lean, BMI < 24 kg/m²). For overweight/obese participants, low-dose LGZG significantly decreased their HOMA-IR level compared with placebo (-0.19 (1.47) versus 0.08 (1.99), P = 0.038). For lean subjects, neither dose of LGZG showed a superior effect compared with placebo. Methylated DNA immunoprecipitation sequencing and real-time qPCR found that the DNA N6-methyladenine modification levels of protein phosphatase 1 regulatory subunit 3A (PPP1R3A) and autophagy related 3 (ATG3) significantly increased after LGZG intervention in overweight/obese population. Low-dose LGZG effectively improved insulin resistance in overweight/obese subjects with NAFLD. The underlying mechanism may be related to the regulation of DNA N6-methyladenine modification of PPP1R3A and ATG3. Lean subjects may not be a targeted population for LGZG.
Humans ; Non-alcoholic Fatty Liver Disease/drug therapy* ; Overweight/drug therapy* ; Insulin Resistance ; Obesity/drug therapy* ; China ; DNA/therapeutic use*

Objective: To investigate the inhibitory effect of polydopamine (PDA) on enamel demineralization in isolated teeth and the induction of hydroxyapatite (HA) production on the surface of demineralized enamel to provide a novel protocol for the prevention and treatment of enamel demineralization. Methods: Twenty isolated bovine teeth were cut into 20 enamel slices and randomly divided into an experimental group and a control group, with 10 slices in each group. The enamel slices in the experimental group were immersed in 2 mg/mL freshly prepared dopamine solution and incubated for 24 hours at room temperature in the dark to prepare the PDA coating, while the control group was left untreated. Then, the isolated bovine teeth, with and without PDA coating, were immersed in artificial demineralization solution at 37 °C for 3 days, followed by 7 days in simulated body fluid (SBF), and the immersion solution was changed daily. The surface morphology of enamel was observed by scanning electron microscopy (SEM), the calcium/phosphorus ratio of the enamel surface was analyzed by energy dispersive spectroscopy (EDS), and the characteristic functional groups in enamel deposits were analyzed by Fourier transform infrared spectroscopy (FTIR). Results: Compared with the control group, the number of demineralized pores produced after 3 d of enamel demineralization with polydopamine coating was less, and the diameter was smaller. EDS elemental analysis showed that the Ca/P ratio after enamel demineralization was 2.37 in the experimental group, which was smaller than the 2.53 ratio in the control group. In the remineralization experiment, after 7 days of remineralization of PDA coated enamel in the experimental group, lamellar grains were produced on the enamel surface, and the growth showed obvious directionality, growth regularity and uniform arrangement. In the control group, the surface of enamel was flocculent mineral deposit, and the crystallinity was poor. The FTIR results proved that the enamel surface deposit of PDA-coated enamel was HA after 7 d of remineralization. Conclusion PDA can affect the nucleation process of HA and promote the production of HA on the surface of demineralized enamel.