Objective: To investigate the effect of phosphatase and tensin homolog(PTEN) overexpression on autophagy in mouse primary cardiac fibroblasts. Methods: Neonatal mice(1-3 days old) were purchased and subjected to cardiac tissue harvesting. Cardiac fibroblasts were isolated through enzymatic digestion and cultured. After cellular adhesion, rapamycin(Rapa)-treated cardiac fibroblasts were used to establish an autophagy model. Following successful model construction, cells were transfected with eitherPTEN-overexpressing plasmid(PTENoverexpression group) or empty vector(control group), followed by 24-48 h incubation. The molecular expressions of PTEN, autophagy-related proteins [microtubule-associated protein light chain 3(LC3Ⅱ/Ⅰ), Beclin1], and fibrotic marker collagen Ⅰ were detected by Western blot and RT-qPCR. Autophagic flux was assessed using mCherry-GFP-LC3 fluorescence staining to evaluate changes in autophagic activity. Transmission electron microscopy(TEM) was employed to observe autophagosome formation in cardiac fibroblasts. Results : In the Rapa-induced cardiac fibroblast autophagy model, compared with the control group, the protein and mRNA levels of PTEN significantly decreased(P<0.05), while the expression of autophagy-related proteins(LC3Ⅱ/Ⅰ, Beclin1) and fibrotic marker Collagen Ⅰ was upregulated at both protein and mRNA levels(P<0.05). Additionally, in PTEN-overexpressing cardiac fibroblasts, the expression levels of LC3Ⅱ/Ⅰ, Beclin1, and Collagen Ⅰ were markedly reduced compared to the empty vector group(P<0.05). mCherry-GFP-LC3 fluorescence staining demonstrated that autophagic activity was significantly attenuated in thePTENoverexpression group versus the empty vector group(P<0.05). TEM further revealed a decreased number of autophagosomes in PTEN-overexpressing cardiac fibroblasts(P<0.05). Conclusion The overexpression ofPTENsignificantly inhibits autophagy in cardiac fibroblasts, suggesting thatPTENmay be a key gene involved in regulating autophagy in cardiac fibroblasts.

Debates persist regarding the efficacy and safety of azvudine, particularly its real-world outcomes. This study involved patients aged ≥60 years who were admitted to 25 hospitals in mainland China with confirmed SARS-CoV-2 infection between December 1, 2022, and February 28, 2023. Efficacy outcomes were all-cause mortality during hospitalization, the proportion of patients discharged with recovery, time to nucleic acid-negative conversion (T _NANC), time to symptom improvement (T _SI), and time of hospital stay (T _HS). Safety was also assessed. Among the 5884 participants identified, 1999 received azvudine, and 1999 matched controls were included after exclusion and propensity score matching. Azvudine recipients exhibited lower all-cause mortality compared with controls in the overall population (13.3% vs. 17.1%, RR, 0.78; 95% CI, 0.67-0.90; P = 0.001) and in the severe subgroup (25.7% vs. 33.7%; RR, 0.76; 95% CI, 0.66-0.88; P < 0.001). A higher proportion of patients discharged with recovery, and a shorter T _NANC were associated with azvudine recipients, especially in the severe subgroup. The incidence of adverse events in azvudine recipients was comparable to that in the control group (2.3% vs. 1.7%, P = 0.170). In conclusion, azvudine showed efficacy and safety in older patients hospitalized with COVID-19 during the SARS-CoV-2 omicron wave in China.

This paper systematically summarizes the practical experience of the 2025 Dingri earthquake emergency medical rescue in Tibet. It analyzes the requirements for earthquake medical rescue under conditions of high-altitude hypoxia, low temperature, and low air pressure. The paper provides a detailed discussion on the strategic layout of earthquake medical rescue at the national level, local government level, and through social participation. It covers the construction of rescue organizational systems, technical systems, material support systems, and information systems. The importance of building rescue teams is emphasized. In high-altitude and cold conditions, rapid response, scientific decision-making, and multi-party collaboration are identified as key elements to enhance rescue efficiency. By optimizing rescue organizational structures, strengthening the development of new equipment, and promoting telemedicine technologies, the precision and effectiveness of medical rescue can be significantly improved, providing important references for future similar disaster rescues.

[Objective] To analyze the serological characteristics and molecular biology results for a P^k phenotype. [Methods] One patient with P^k phenotype upon unexpected antibodies at Jining Blood Center in July 2022 was selected as the study subject. The blood groups and unexpected antibodies of the proband and his second son were identified using serological methods. The sequences of 3-β-N-acetylgalactosaminyltransferase gene (B3GALNT1) and the coding region of α-1,4-galactosyltransferase gene (A4GALT) were amplified and analyzed by PCR direct sequencing, and haploid sequence analysis was carried out on the variant sites of the B3GALNT1 gene. PROVEAN, SIFT, PolyPhen2 and Mutation Taster were used to analyze the effect of mutations on the protein. [Results] Serological test results suggested that the proband was a P2^k type, including anti-P antibody in his serum, and his son was the P2 phenotype. Sequencing analysis revealed that the proband had a compound heterozygous variants of the B3GALNT1 in c. 239T>A (p. Leu80Gln) and c. 433C>T (p. Arg145Ter). Further haploid analysis showed that the c. 239T>A had occurred in one haploid while c. 433C>T was present in the other haploid, and his son carried a heterozygous variant of c. 433C>T (p. Arg145Ter); the proband and his son all have homozygous mutation variants of the A4GALT in c. 903C>G. Bioinformatic analysis also predicted that the mutations were harmful to the protein function. [Conclusion] The compound heterozygous variants c. 239T>A (p. Leu80Gln) and c. 433C>T (p. Arg145Ter) in the B3GALNT1 gene may contribute to the P^k phenotype, of which the c. 239T>A variant has not been reported.

Objective To investigate the effect of polymyxin B(PMB)combined with tigecycline(TGC)on delaying Klebsiella pneumoniae(KP)resistance to PMB,and to analyze the possible mechanisms involved in the induction and delay of resistance.Methods Six clinical isolates of KP strains from the Intensive Care Unit of First Affiliated Hospital of Army Medical University were subjected and then induced with PMB at 1/2 minimal inhibitory concentration(MIC)alone or combined with TGC at 1/2 and 1/4 MIC,respectively.The MIC changes of PMB in these strains were monitored over 14 consecutive passages.The strain 686K,which showed the most significant delay in resistance,was selected for further analysis.Differences in gene and protein expression were examined among the wild-type strain 686K,PMB-induced resistant strain(686K·R),and PMB combined with TGC delayed resistant strain(686K·DR)using transcriptome sequencing,qRT-PCR,and proteomics.Relevant target genes during the delay of resistance were analyzed through literature and bioinformatics analyses.Additionally,cpxR gene knockout strain 686K/ΔcpxR∷Apr and its complementation strain 686K/ΔcpxR∷Apr/pRK415-cpxR were constructed using homologous recombination technology to assess the expression levels of resistance-related genes and changes in MIC after induction in vitro.Results Under sub-MIC(1/2)PMB alone,resistance developed in all 6 KP strains within 2 d,while,the combination with TGC significantly delayed the development of resistance.Transcriptomic and proteomic analyses indicated that in strain 686K·R,the expression levels of the PhoP/Q two-component system,lipopolysaccharide(LPS)modification enzymes,and efflux pump systems were significantly up-regulated(|Log2FC|≥2,P<0.0001),while TGC co-administration markedly inhibited these expression changes.The cpxR deletion and complementation strains were successfully constructed.The expression levels of resistance-related genes phoP,pmrD,and acrA were decreased in the cpxR deletion strain(P<0.001),and the resistance was delayed until day 6 under PMB monotherapy,whereas the complementation strain restored the resistance phenotype by day 2.In the absence of cpxR,the effect of PMB when combined with TGC on delaying resistance did not differ from that observed with PMB monotherapy.Conclusion The combination of PMB and TGC can delay KP resistance to PMB.cpxR,as a critical regulatory factor,can impact PMB resistance by modulating LPS modifications and the expression of the AcrAB-TolC efflux pump,and plays an important regulatory role in the process of resistance induction.

Objective To construct an evidence-based discharge preparation service plan for patients after receiving percutaneous transhepatic biliary drainage(PTBD)so as to provide a theoretical reference basis for improving the quality of discharge nursing service.Methods A computerized retrieval of academic papers concerning the discharge preparation service plan for patients after receiving PTBD was conducted.The quality of the included literature was evaluated and the evidences were summarized.According to the clinical actual requirements,the first draft of discharge preparation service for patients after receiving PTBD was formed.Using Delphi method,two rounds of letter inquiries were conducted in 17 experts to determine the final version.Results In the first round of expert consultation,17 questionnaires were distributed and 15 questionnaires were recovered;and in the second round of expert consultation,15 questionnaires were distributed and 15 questionnaires were recovered.In the first round of expert correspondence,11 experts made suggestions for modification,and in the second round of expert correspondence,4 experts made suggestions,indicating that the experts were more motivated to participate in the research.The coefficient of expert consultation judgment(Ca)was 0.90,the degree of familiarity(Cs)was 0.91,the coefficient of authority(Cr)was 0.91,and the Kendall's w for round 1 and round 2 were 0.363 and 0.368 respectively.The final discharge preparation service scheme consisted of 13 items at six different time points from patient admission to after discharge.Conclusion The established discharge preparation service plan for patients after receiving PTBD is scientific and reliable,which can provide theoretical basis for patients'discharge service.

Objective Based on the best evidences to establish the practice plan of discharge preparation service for patients after receiving percutaneous transhepatic biliary drainage(PTBD),and to assess its clinical application value.Methods According to the PIPOST principle the clinical questions were proposed,the best evidences of discharge preparation service for patients after receiving PTBD were retrieved and summarized.The review indicators and review methods were formulated.The baseline review was carried out,the facilitators and barrier factors were analyzed,the change strategies were developed,the clinical transforms were implemented,and the patient outcomes were evaluated.Results After application of the evidences,the implementation of the review indicators of discharge preparation services after PTBD was improved.After discharge,the incidence of catheter complications(including catheter falling-off and puncture site skin infection)was decreased,and the difference was statistically significant(P＜0.05).There were no statistically significant differences in the incidences of tube obstruction and fluid extravasation.Conclusion The evidence-based practice of discharge preparation service for patients after receiving PTBD is helpful for improving the self-care ability of patients after discharge,reducing the incidence of tubular complications and improving the clinical outcome of patients.

Objective To investigate the role of reactive oxygen species(ROS)and the extracellular signal-regulated kinase(ERK)/cAMP response element binding protein(CREB)signaling pathway in arsenic-induced apoptosis in HT-22 cells.Methods HT-22 cells were cultured in vitro and exposed to NaAsO2.The cells were divided into the following groups:control group,4,6,8,and 10 μmol/L NaAsO2 groups,NAC(5 mmol/L)group,and NAC(5 mmol/L)+NaAsO2(10 μmol/L)group.ROS levels were measured using a DCFH-DA assay.The expression of ERK/CREB signaling pathway-related and apoptosis-related proteins were analyzed by Western blotting.Apop-tosis rates were evaluated using flow cytometry.Results Compared with the control group,ROS levels in HT-22 cells significantly increased,while p-ERK protein level in the nucleus and p-CREB and Bcl-2 protein levels in the cells significantly decreased in the NaAsO2 only exposed groups(P＜0.05).Additionally,the apoptosis rate significantly increased in the 8 and 10 μmol/L NaAsO2 groups compared with the control group(P＜0.05).In the NAC+NaAsO2 group,ROS levels and the apoptosis rate significantly decreased while p-ERK protein level in the nucleus and p-CREB and Bcl-2 protein levels in the cells significantly increased compared with the 10 μmol/L NaAsO2 group(P＜0.05).Conclusion Arsenic exposure induces oxidative stress,inhibits the nuclear translocation of p-ERK,and dis-rupts the CREB signaling pathway,leading to apoptosis.

Objective:To investigate the impact and potential mechanisms of Sorbin and SH3 domain-containing protein 2 (Sorbs2) on ventricular arrhythmias in mice.Methods:In the animal experiments, mating was performed using six 8-week-old Sorbs2 +/- mice (3 males and 3 females) weighing 20-22 g. Wild-type (Sorbs2 +/+, n=8) and homozygous (Sorbs2 -/-, n=6) offspring were selected as experimental subjects through genotyping. Echocardiography was performed at 16 weeks of age to record cardiac function parameters in both groups. Resting-state and caffeine-dobutamine-induced electrocardiograms were also conducted. Real-time quantitative reverse transcription polymerase chain reaction was used to detect Sorbs2 messenger RNA expression in the heart, liver, spleen, lung, kidney, brain, small intestine, and skeletal muscle tissues of wild-type mice. Western blotting was employed to measure the protein expression levels of Sorbs2 and voltage-dependent sodium channel alpha subunit 1.5 (Na v1.5) in myocardial tissues from both groups. In the cell experiments, H9C2 cells were transfected with Sorbs2 small interfering RNA as the si-Sorbs2 group, with a corresponding si-negative control group established. Western blot was performed to detect the protein expression levels of Sorbs2 and Na v1.5 in both groups. Results:Sorbs2 was abundantly expressed in cardiac tissue. Compared with wild-type mice, homozygous mice exhibited larger left ventricular end-systolic diameter, along with lower left ventricular ejection fraction and fractional shortening ( P all<0.05). Resting-state electrocardiograms revealed no spontaneous arrhythmias in either group; however, homozygous mice showed shorter RR intervals but longer QRS and QTc intervals versus wild-type mice ( P all<0.05). Following caffeine and dobutamine induction, homozygous mice demonstrated a higher incidence of ventricular arrhythmias, longer arrhythmia duration, and higher ventricular arrhythmia scores than wild-type mice ( P all<0.05). Western blot analysis revealed that Na v1.5 protein expression was markedly lower in myocardial tissues of homozygous mice compared to wild-type mice. Similarly, si-Sorbs2-transfected H9C2 cells exhibited lower Na v1.5 protein levels compared to the si-negative control group ( P<0.05). Conclusion:Sorbs2 plays a critical role in maintaining normal cardiac electrophysiological function. Deficiency of Sorbs2 may lead to impaired cardiac function and increased susceptibility to ventricular arrhythmias in mice, which could be associated with reduced expression of Na v1.5 protein.

Serotonin syndrome (SS), also known as serotonin toxicity, is a rare but life-threatening drug reaction syndrome. This case involves a 17-year-old female patient who developed tremors, fatigue, and tachycardia after taking fluoxetine combined with bupropion and tandospirone for five days. SS was highly suspected, and her symptoms improved following treatment targeted at serotonin syndrome. This case is reported to raise awareness among clinicians about the potential adverse reactions of drug combinations, the importance of early identification of SS symptoms, and precautions when prescribing combined medications to avoid serious consequences.