ObjectiveTo observe the effect of Yifei Jianpi prescription on the of signal transducer and activator of transcription protein 1 （STAT1）/interferon regulatory factor 3 （IRF3） signaling pathway in a pneumonia model induced by lipopolysaccharide （LPS） and to explore the mechanism of Yifei Jianpi prescription in improving lung immune and inflammatory responses. MethodsSixty male SPF SD rats were used in this study. Ten rats were randomly assigned to the normal control group， and the remaining 50 were instilled with LPS in the trachea to establish a pneumonia model. After successful modeling， the rats were randomly divided into the model group， dexamethasone group （0.5 mg·kg^-1）， and Yifei Jianpi prescription high-dose （12 mg·kg^-1）， medium-dose （6 mg·kg^-1）， and low-dose （3 mg·kg^-1） groups， with 10 rats in each group. Treatment was administered once daily， and the normal control and model groups received the same volume of normal saline. After 14 days， flow cytometry was used to detect the classification of whole blood lymphocytes. Enzyme-linked immunosorbent assay （ELISA） was used to measure serum levels of immunoglobulin G （IgG）， immunoglobulin A （IgA）， immunoglobulin M （IgM）， and the content of tumor necrosis factor-α （TNF-α）， interleukin-8 （IL-8）， interleukin-6 （IL-6）， and interleukin-10 （IL-10） in alveolar lavage fluid （BALF）. Hematoxylin-eosin （HE） staining was used to observe lung tissue pathology and score the damage. Thymus weight， spleen weight， and wet-to-dry weight ratio （W/D） were recorded. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of STAT1， IRF3， IL-6， and interferon-alpha （IFN-α） in lung tissues， while Western blot was performed to assess the protein expression of STAT1， IRF3， IL-6， and IFN-α. ResultsCompared with the normal control group， the model group showed significantly increased proportion of B lymphocytes in peripheral blood， decreased proportions of NK cells and CD4⁺/CD8⁺ （P<0.05， P<0.01）， decreased serum levels of IgG and IgA， significantly increased IgM levels （P<0.01）， significantly elevated content of TNF-α， IL-6， and IL-8 in BALF， and significantly decreased IL-10 levels （P<0.01）. Lung tissue damage was evident， with significant increases in thymus and spleen weights and a higher W/D ratio （P<0.01）. The mRNA and protein expression of STAT1， IRF3， IFN-α， and IL-6 in lung tissues was significantly upregulated （P<0.05，P<0.01）. Compared with the model group， the Yifei Jianpi prescription groups showed significantly reduced proportions of B lymphocytes in peripheral blood， increased proportions of NK cells and CD4⁺/CD8⁺ ratios （P<0.05， P<0.01）， significantly increased serum levels of IgG and IgA， significantly decreased IgM levels （P<0.05， P<0.01）， significantly reduced levels of TNF-α， IL-6， and IL-8 in BALF， and significantly increased IL-10 levels （P<0.01）. Lung tissue damage was alleviated， thymus and spleen weights were significantly reduced， and the W/D ratio was markedly decreased （P<0.01）. The mRNA and protein expression of STAT1， IRF3， IFN-α， and IL-6 in lung tissues was significantly downregulated （P<0.05， P<0.01）. ConclusionYifei Jianpi prescription can alleviate lung tissue damage and improve immune and inflammatory responses in LPS-induced pneumonia rats. The mechanism may be related to the inhibition of STAT1/IRF3 signaling pathway activation.

ObjectiveTo investigate a suspected outbreak of healthcare-associated infection with carbapenem-resistant Klebsiella pneumoniae (CRKP) in a geriatric emergency ward, and to provide references for the prevention and control of multidrug-resistant bacteria in a hospital in Shanghai. MethodsOn-site epidemiological investigation, combined with environmental hygiene monitoring and pulsed field gel electrophoresis (PFGE) molecular typing method, were adopted to investigate a suspected outbreak of CRKP infection in the geriatric emergency ward of a hospital from October to November 2022, aiming at finding out factors caused the outbreak before taking corresponding control measures. ResultsA total of 3 cases of healthcare-associated CRKP infection were identified, of which 2 cases were homologous to a previous case of community-associated CRKP infection. What’s more, the 2 cases lived in the same ward with the latter and with adjacent beds, but the third case was non-homologous to the community-associated infection case. A total of 46 samples were collected from the environmental surfaces and the hands of healthcare workers, of which 7 samples tested positive for CRKP and were identical to the strains from the 2 healthcare-associated infection cases and the 1 community-associated infection case, originating from the bedrails, bedside tables, surface of non-invasive ventilator, bed curtains and panels of monitoring equipment, with a detection rate of 15.22%. But none of the 11 samples from the hands of healthcare workers tested positive for CRKP. The outbreak was effectively controlled after taking specific prevention and control measures such as strengthening personnel management, intensifying environmental cleaning and disinfection and strictly enforcing hand hygiene among healthcare workers. Subsequently, no similar new cases were reported during the 14-day follow-up period. ConclusionIncomplete environmental cleaning and disinfection, as well as inadequate enforcement of hand hygiene among heatheare workers may have contributed to the suspected outbreak of CRKP in the geriatric emergency ward. Early warning and timely investigation of suspected outbreaks of multidrug-resistant bacteria are crucial for preventing and controlling such outbreaks in hospitals.

Objective To explore the mechanism by which zinc-regulated transporters, iron-regulated transporter-likeprotein 4 (ZIP4) regulates glycolysis and its impact on tumor progression in cholangiocarcinoma (CCA), providing a theoretical basis for targeted therapy of CCA. Methods ZIP4 expression in CCA was analyzed using the GEPIA database. Immuno-histochemistry (IHC) was used to detect ZIP4 expression in 20 paired CCA and adjacent non-tumor tissues. Stable ZIP4-overexpressing CCA cell lines (ZIP4-OE) were established. Gene set enrichment analysis was used to screen differentially expressed genes and pathways in ZIP-OE CCA cells. ZIP4, N-myc proto-oncogene protein (MYCN), and histone-lysine N-methyltransferase 2E (KMT2E) were knocked down using small interfering RNAs (siRNAs). The expression of glycolysis-related gene (glucose transporter 1 [Glut1], hexokinase 2 [HK2], and lactate dehydrogenase A [LDHA]) was measured by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Glycolytic activity was assessed by measuring the extracellular acidification rate (ECAR). Cell proliferation was evaluated using colony formation assays, and cell migration was assessed using Transwell assays. A xenograft mouse model was constructed to examine CCA tumor growth. Protein levels of ZIP4, KMT2E, H3K4me3 (tri-methylation of lysine 4 on histone H3), and MYCN were detected by Western blotting. Results GEPIA database analysis and IHC results confirmed significantly higher ZIP4 expression levels in CCA tissues compared to adjacent non-tumor tissues (P＜0.05). Compared to the control group, the ZIP4-OE group exhibited a significantly increased ECAR, along with significantly enhanced proliferation and migration abilities (P＜0.01). Conversely, knockdown of ZIP4 suppressed CCA cells proliferation and migration. GEPIA analysis indicated that ZIP4 upregulates the transcription of oncogene MYCN, as well as glycolysis-related genes. Knockdown of MYCN abolished the ZIP4 overexpression-induced upregulation of Glut1, HK2, and LDHA gene transcription, reduced glycolysis, and significantly inhibited CCA cell proliferation and migration (P＜0.05). Mechanistic studies demonstrated that ZIP4 increases H3K4me3 level via KMT2E, leading to MYCN transcription. Knockdown of KMT2E in CCA cells suppressed the ZIP4 overexpression-induced enhancement in H3K4me3 modification, resulting in MYCN downregulation and significantly reduced CCA cells proliferation and migration (P＜0.05). Conclusions ZIP4 upregulates H3K4me3 modification through KMT2E, which recruits transcription factors to activate the transcription of MYCN. This subsequently enhances cellular glycolysis and promotes the proliferation and migration of CCA cells.

Objective To explore the drug selection, usage, dosage and effect of clinical pharmacists in the treatment of secondary Candida bloodstream infection in patients with digestive tract perforation, and provide reference for rational clinical use of such anti-infection therapy. Methods During the treatment of Candida infection, anti-infection specialist clinical pharmacists suggested replacing fluconazole for anti-infection treatment, and adjusted the usage and dosage of caspofungin to ensure the application of full treatment course. Results The patient’s Candida bloodstream infection was effectively controlled. Conclusion Echininocandins such as caspofungin are the first choice for Candida infection. The maintenance dose should be reduced to 35 mg qd in patient with moderate liver function injury, and the anti-infection treatment should be done at least 14 days after the blood culture being negative. Clinical pharmacists participate in clinical pharmaceutical care practices such as clinical ward rounds, and actively assist doctors to develop individualized anti-infection programs, which could improve the level of clinical drug use and the clinical outcome of diseases.

Objective To establish a cell bank of extrahepatic cholangiocarcinoma (ECC)-derived organoids and investigate the key factors influencing the organoids generation. Methods The tumor samples from patients with portal cholangiocarcinoma (pCCA) and distal cholangiocarcinoma (dCCA) were used to isolate cells, and these cells were cultured using three-dimensional (3D) technique to establish ECC organoids. Histological characteristics of the organoids were evaluated and identified through hematoxylin-eosin (HE) and immunohistochemistry stainings. The success rates of organoids generation from different tumor types were compared. And clinical characteristics of patients between successful and failure culture groups were compared. Results The success rates of organoids establishment from pCCA and dCCA were all low, with 42.4% (14/33), 51.9% (14/27), respectively. The tumor was larger in successful group than that in failure group (P＜0.001); there was no statistical difference in tumor differentiation status, microvascular invasion, and perineural invasion between the two groups. Conclusions The successful rate of ECC-derived organoids establishment is low, and larger tumor has higher successful culture rate.

Objective To explore the alterations in macrophage polarization and the expression of decorin (DCN) protein in the decidua of patients with missed abortion (MA), as well as to elucidate the regulatory effect of DCN on macrophage polarization. Methods Flow cytometry was employed to assess the polarization ratio of decidual macrophages in MA, recurrent spontaneous abortion (RSA) and normal pregnancy (NP); The expression and localization of DCN and hypoxia-inducible factor 1α (HIF-1α) in decidua and villi were assessed by immunohistochemistry staining, while their protein levels were measured by Western blot. Primary trophoblasts and peripheral blood mononuclear cell (PBMC)-derived macrophages were isolated and cultured. ELISA was conducted to quantify DCN levels in the culture supernatant of primary trophoblast and PBMC-derived macrophages. Additionally, flow cytometry was applied to evaluate the polarization ratio of PBMC-derived macrophages. Immunofluorescence cytochemical staining was conducted to examine HIF-1α expression in macrophages. Western blot was performed to detect the expression of proteins related to the gene associated with retinoid-IFN-induced mortality 19 (GRIM-19)/signal transducer and activator of transcription 3 (STAT3)/HIF-1α signaling pathway in macrophages. Results The polarization ratio of M1 macrophages in the decidua of abortion patients was significantly higher than that of NP, whereas the ratio of M2 macrophages was significantly lower. The expression of DCN and HIF-1α protein were significantly evaluated in abortion patients compared to NP. The supernatant DCN content and HIF-1α protein expression of primary trophoblast and PBMC-derived macrophages cultured under 10 mL/L O₂ for 24 hours were markedly increased compared to cells treated with 210 mL/L O₂. Compared with the PBS group, the proportion of M1 macrophage and GRIM-19 protein expression were significantly reduced in the DCN group, while phosphorylated STAT3 (p-STAT3) and HIF-1α protein expression were significantly increased. Conclusion The expression of DCN in decidua and villi of MA is higher than that of NP. DCN exhibits an inhibitory effect on the M1 polarization of PBMCs-derived macrophages, which is likely mediated through the GRIM-19/STAT3/HIF-1α signaling pathway.
Humans ; Female ; Decidua/metabolism* ; Macrophages/cytology* ; Decorin/genetics* ; Pregnancy ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Adult ; Leukocytes, Mononuclear/cytology* ; Abortion, Missed/genetics* ; STAT3 Transcription Factor/metabolism* ; Cells, Cultured ; Young Adult

OBJECTIVES: To synthesize a temperature-responsive multimodal motion microrobot (MMMR) using temperature and magnetic field-assisted microfluidic droplet technology to achieve targeted drug delivery and controlled drug release. METHODS: Microfluidic droplet technology was utilized to synthesize the MMMR by mixing gelatin with magnetic microparticles. The microrobot possessed a magnetic anisotropy structure to allow its navigation and targeted drug release by controlling the temperature field and magnetic field. In the experiment, the MMMR was controlled to move in a wide range along a preset path by rotating a uniform magnetic field, and the local circular motion was driven by a planar rotating gradient magnetic field of different frequencies. The MMMR was loaded with simulated drugs, which were released in response to laser heating. RESULTS: Driven by a rotating magnetic field, the MMMR achieved linear motion following a predefined path. The planar gradient rotating magnetic field controlled circular motion of the MMMR with an adjustable radius, utilizing the centrifugal force generated by rotation. The drug-loaded MMMR successfully reached the target location under magnetic guidance, where the gelatin matrix was melted using laser heating for accurate drug release, after which the remaining magnetic particles were removed using magnetic field. CONCLUSIONS The MMMR possesses multimodal motion capabilities to enable precise navigation along a predefined path and dynamic regulation of drug release within the target area, thus having great potential for a wide range of biomedical applications.
Drug Delivery Systems/methods* ; Temperature ; Drug Liberation ; Magnetic Fields ; Robotics ; Gelatin/chemistry* ; Delayed-Action Preparations ; Microfluidics ; Motion

Background Obsessive-compulsive personality disorder and obsessive-compulsive disorder(OCD)are common psychological disorders with similar clinical symptoms,but the differences between the two need further clarification.Objective To explore the clinical features of and influencing factors of obsessive-compulsive personality disorder in patients with OCD,so as to provide references for further relevant clinical diagnosis and treatment.Methods A total of 195 patients with OCD were selected as the research subjects,who received treatment at the outpatient and inpatient departments of the Affiliated Brain Hospital of Nanjing Medical University from July 2022 to December 2023 and met the diagnostic criteria for OCD in the International Classification of Diseases,tenth edition(ICD-10).Evaluation was conducted by using the Yale-Brown Obsessive Compulsive Scale(Y-BOCS),Personality Diagnostic Questionnaire-4+(PDQ-4+),Obsessive-Compulsive Inventory-Revised(OCI-R),Beck Depression Inventory(BDI),Beck Anxiety Inventory(BAI)and Sheehan Disability Scale(SDS).In accordance with the score of Obsessive-Compulsive Personality Disorder Scale in PDQ-4+,patients were divided into the OCD group with obsessive-compulsive personality disorder(n=58)and the OCD group without obsessive-compulsive personality disorder(n=137).Pearson correlation analysis and Spearman correlation analysis were adopted to examine the correlation between clinical features and the score of the Obsessive-Compulsive Personality Disorder Scale.Multiple linear regression analysis was used to explore the influencing factors of OCD patients with obsessive-compulsive personality disorder.Results Statistically significant differences were observed between OCD patients with and without obsessive-compulsive personality disorder in the age,family history of mental illness,time without treatment,hoarding and ranking dimension scores in OCI-R,OCI-R total score,score of Obsessive-Compulsive Personality Disorder Scale in PDQ-4,and BDI score(P≤0.05).OCD patients with obsessive-compulsive personality disorder in the time without treatment,OCI-R total score,hoarding and ranking dimension scores in OCI-R and BDI score are all positively correlated with the score of the Obsessive-Compulsive Personality Disorder Scale(r=0.120,0.526,0.364,0.492,0.414,P<0.05).The results of multiple linear regression analysis showed that time without treatment(β=0.132,P<0.05),hoarding dimension score(β=0.283,P<0.05)and ranking dimension score in OCI-R(β=0.418,P<0.05)were the influencing factors of OCD patients with obsessive-compulsive personality disorder.Conclusion OCD patients with obsessive-compulsive personality disorder may have longer untreated periods,more pronounced functional impairments in hoarding and sorting and more severe depressive symptoms.Untreated time,hoarding symptoms and sorting symptoms may be influencing factors for OCD patients with obsessive-compulsive personality disorder.

Objective:To study the correlation between spontaneous cerebrospinal fluid rhinorrhea and increased intracranial pressure.Methods:Clinical data of patients with spontaneous cerebrospinal fluid rhinorrhea treated in the Department of Otorhinolaryngology of the Provincial Hospital of Shandong First Medical University from January 2019 to December 2023 and their epidemiology, clinical symptoms and signs, preoperative cerebrospinal fluid pressure, imaging data, leakage site, repair method and the presence or absebce of increased intracranial pressure were analysed. SPSS 23.0 software was used for statistical analysis.Results:Of the 57 patients with cerebrospinal fluid nasolacrimal leakage, 84.2%(48/57) were females and 15.8%(9/57) were males; 80.7%(46/57) were between 40 and 60 years old; and overweight and obese patients accounted for 75.4%(43/57). Of 57 patients, 35 patients received cranial MRI+magnetic resonance venography, and among them, 12(34.3%) patients were clinically with headache symptoms; the incidences of empty pterygoid and venous sinus stenosis were 40.0%(14/35) and 51.4%(18/35), respectively; and 33 patients underwent preoperative lumbar puncture examination, of whom 25(75.8%) patients had the increased cerebrospinal fluid pressure and 1 patient had idiopathic intracranial hypertension. All 57 patients underwent dural repair of the skull base, the postoperative follow-up period ranged from 3 to 60 months, and the success rate of surgery was 94.7%. The success rate was 100% in female patients and 33.3%(3/9) in male patients, including one with surgical failure receiving reoperation and two with postoperative recurrence in other areas of the skull base, with significant difference in the success rate of surgery between males and females (χ 2=16.890, P<0.001). Conclusions:Most patients with spontaneous cerebrospinal fluid rhinorrhea have the increased cerebrospinal fluid pressures, but very few fulfil the diagnosis of idiopathic intracranial hypertension. The success rate of surgical repair alone is high, but some recurrences still exist, especially in male patients.

Objective To investigate the clinical features in children with invasive pneumococcal disease (IPD) and early warning indicators of severe case.Methods The case data of 101 children inpatients with IPD isolated Streptococcus pneumoniae (Sp) at least once blood culture in this hospital from January 2007 to De-cember 2021 were collected retrospectively.The children patients were divided into the pediatric intensive care unit (PICU) group (35 cases) and general ward group (66 cases) according to whether or not entering PICU during hospitalization.The general clinical data,infection types and results of drug sensitivity test and bacteri-al strain serotype identification were compared between the two groups and analyzed.Results Compared with the general ward group,the male proportion,proportions of complicating nervous system disease and iron-defi-ciency anemia,winter incidence rate,disturbance of consciousness,convulsion,dyspnea,heart rate increase,oc-currence rate of vomiting and diarrhea,proportion of peripheral blood neutrophilic granulocytes,C reactive protein (CRP),lactic dehydrogenase (LDH) level,Hb<90 g/L,LDH>700 U/L,creatine kinase isoenzyme-MB (CK-MB) abnomality,proportions of bacterial meningitis,sepsis and severe pneumonia in the PICU group were higher,the proportion of 3-97 percentile of body weight,bronchopneumonia and no-focus blood stream infection were lower,WBC,Hb and albumin level were lower,and the differences were statistically significant (P<0.05).Eleven kinds of serotypes were identified in this study,ranking the top five in order were 6B,14,19F,23F and 19A.Pneumococcal 13-valent conjugate vaccine (PCV13) could cover 96.9％ of the IPD sero-types.The multivariate logistic regression analysis showed that consciousness disturbance,convulsion,per-centage of neutrophils increase,Hb<90 g/L,LDH>700 U/L and CK-MB abnormality indicated the condition of IPD children patients was critical,which was the early-warning indicator of entering PICU (P<0.05).Con-clusion Severe IPD lacks the early specific early-warning indicator,and the cover rate of PCV13 is high.