Objective:To construct an ICU-acquired weakness training program for ICU nurses based on Gagne learning theory.Methods:From March to August 2021, the ICU-acquired weakness training program for ICU nurses was constructed through literature review, semi-structured interview, and Delphi expert consultation.Results:Among two rounds of expert consultation, 16 and 20 valid questionnaires were recovered respectively, and the expert authority coefficients were 0.897 and 0.903, and the Kendall harmony coefficients were 0.312 and 0.323, respectively, and the differences were all statistically significant ( P<0.01) . The ICU-acquired weakness training program for ICU nurses included 3 first-level indicators (training objectives, training process and content, training form) , 16 second-level indicators, and 41 third-level indicators. Conclusions:The ICU-acquired weakness training program for ICU nurses based on Gagne learning theory is scientific and reliable, and can provide a reference for ICU nurses' ICU-acquired weakness training.

After sequencing, we amplified and cloned foot-and-mouth disease virus (FMDV) O/QYYS/s/06 whole genome by three fragments. These three fragments were cloned into vector P43 one by one to construct recombinant plasmid P43C, which carried the full-length cDNA of FMDV O/QYYS/s/06. Then, plasmid P43C and plasmid T7 expressing T7 RNA polymerase were co-transfected into BHK-21 cells. After 48 h, we harvested the culture broth from transfected BHK-21 cells and inoculated into 2-3 day-old sucking mice. After four generation passage, the virus harvested from sucking mice was confirmed to be type O FMDV by the indirect hemagglutination test, sucking mice's neutralization test and sequencing. The results showed that we have successfully constructed the full-length cDNA clone of FMDV O/QYYS/s/06 strain.
Animals ; Animals, Newborn ; Cloning, Molecular ; DNA, Complementary ; genetics ; DNA, Viral ; biosynthesis ; genetics ; Foot-and-Mouth Disease ; virology ; Foot-and-Mouth Disease Virus ; classification ; genetics ; pathogenicity ; Mice ; Transcription, Genetic ; Transfection

Objective:To construct a stratified nursing prevention strategy for delirium in ICU patients to provide reference for the prevention of delirium in ICU patients.Methods:Preliminary items were screened based on evidence-based medicine method. After discussion by the research group, the first draft of stratified nursing prevention strategy for ICU patients with delirium was formed. Using the Delphi method, a total of two rounds of correspondence with 15 experts were conducted. Combined with expert opinions, a stratified nursing prevention strategy for delirium in ICU patients was constructed.Results:The effective recovery rates of the two rounds of questionnaires were 93.75% and 100.00%, respectively. The expert authority coefficients were respectively 0.893 and 0.897, and the Kendall's harmony coefficients of indicators at all levels were 0.203-0.213 ( P<0.01) . Finally, a delirium stratified nursing prevention strategy for ICU patients was formed, including 5 first-level indicators, 12 secondary indicators and 28 tertiary indicators. Conclusions:The stratified nursing prevention strategy constructed in this study is based on evidence-based and Delphi method, which is scientific, reliable, practical and targeted. It can provide reference for the prevention of delirium in ICU patients.

Objective:To evaluate the visibility of deep medullary veins (DMVs) in patients with recent subcortical small infarction (RSSI), and explore the influence of DMVs visibility in RSSI with cognitive impairment.Methods:One hundred and sixty-four first-onset RSSI patients admitted to Department of Neurology, Third Affiliated Hospital of Nanjing Medical University from February 2023 to May 2024 were selected. According to Montreal cognitive assessment (MoCA), they were divided into normal cognitive function group (MoCA scores≥26, n=78) and cognitive impairment group (MoCA scores<26, n=86); DMVs visibility on amplitude map or minimum intensity projection map in susceptibility weighted imaging was evaluated by visual scoring method. Univariate analysis was used to compare the differences in general clinical data and DMVs visibility scores between two groups; multivariate Logistic regression analysis was used to identify the independent influencing factors for RSSI combined with cognitive impairment. Spearman rank correlation was used to analyze the correlations of DMVs visibility score with MoCA score and cognitive scores in various cognitive domains, while mediation analysis was used to explore the correlations of MoCA score with total white matter hyperintensity (WMH) volume and DMVs visibility score. Results:The DMVs visibility score, age, hypertension proportion, periventricular white matter hyperintensity (PWMH) volume, and total WMH volume in RSSI patients with cognitive impairment were significantly higher, while body mass index (BMI) was statistically lower than those in patients with normal cognitive function ( P<0.05). Multivariate Logistic regression analysis showed that age ( OR=1.069, 95% CI: 1.017-1.123, P=0.008), total WMH volume ( OR=1.845, 95% CI: 1.050-3.241, P=0.033), and DMVs visibility score ( OR=1.239, 95% CI: 1.057-1.454, P=0.008) were independent influencing factors for cognitive impairment in RSSI patients. Spearman rank correlation showed negative correlations between DMVs visibility score and MoCA score ( rs=-0.472, P<0.001), between DMVs visibility score and score of executive function and visual-spatial skills ( rs=-0.329, P<0.001), between DMVs visibility score and attention score ( rs=-0.491, P<0.001), between DMVs visibility score and delayed recall score ( rs=-0.516, P<0.001), and between DMVs visibility score and directional ability score ( rs=-0.162, P=0.039) in RSSI patients. Mediation analysis results showed that DMVs visibility score not only had a direct effect of 87.5% on MoCA score, but also had an indirect effect of 12.5% on MoCA score through total WMH volume. Conclusion:Cognitive impairment trend to appear in RSSI patients with high DMVs visibility score, and WMH volume plays a mediating role in the effect of DMVs visibility on RSSI with cognitive impairment.

Quantitative susceptibility mapping (QSM) is a noninvasive MR post-processing technique that can quantitatively analyze tissue magnetic susceptibility, which has important value in early diagnosis and assessment of cerebral small vessel disease (CSVD). In recent years, QSM has gradually been applied in clinical neuroimaging both domestically and internationally, becoming a new hotspot. This review focuses on the principle and classification of QSM, application of QSM in the gray matter microstructure, white matter microstructure, venous oxygen saturation assessment, cerebral microhemorrhage and cognitive function assessment of CSVD, so as to provide new perspective for its neuroimaging evaluation.

The field of gene therapy has been increasingly studied in the last four decades, and its clinical application has become a reality in the last 15 years. Traditional Chinese medicine (TCM), an important component of complementary and alternative medicine, has evolved over thousands of years with its own unique system of theories, diagnostics and therapies. TCM is well-known for its various roles in preventing and treating infectious and chronic diseases, and its usage in other modern clinical practice. However, whether TCM can be applied alongside gene therapy is a topic that has not been systematically examined. Here we provide an overview of TCM theories in relation to gene therapy. We believe that TCM theories are congruent with some principles of gene therapy. TCM-derived drugs may also act as gene therapy vehicles, therapeutic genes, synergistic therapeutic treatments, and as co-administrated drugs to reduce side effects. We also discuss in this review some possible approaches to combine TCM and gene therapy.
Gene Expression ; drug effects ; Genes ; drug effects ; Genetic Therapy ; methods ; Humans ; Medicine, Chinese Traditional ; methods

OBJECTIVEIn the present study, we systemically evaluated the ability of two bioactive compounds from traditional Chinese medicine, celastrol and pristimerin, to enhance recombinant adeno-associated virus (rAAV) serotype vector-mediated transgene expression both in human cell lines in vitro, and in murine hepatocytes in vivo.

METHODSHuman cell lines were infected with rAAV vectors with either mock treatment or treatment with celastrol or pristimerin. The transgene expression, percentage of nuclear translocated viral genomes and the ubiquitination of intracellular proteins were investigated post-treatment. In addition, nonobese diabetic/severe combined immunodeficient gamma (NSG) mice were tail vain-injected with rAAV vectors and co-administered with either dimethyl sulfoxide, celastrol, pristimerin or a positive control, bortezomib. The transgene expression in liver was detected and compared over time.

RESULTSWe observed that treatment with pristimerin, at as low as 1 μmol/L concentration, significantly enhanced rAAV2 vector-mediated transgene expression in vitro, and intraperitoneal co-administration with pristimerin at 4 mg/(kg·d) for 3 d dramatically facilitated viral transduction in murine hepatocytes in vivo. The transduction efficiency of the tyrosine-mutant rAAV2 vectors as well as that of rAAV8 vectors carrying oversized transgene cassette was also augmented significantly by pristimerin. The underlying molecular mechanisms by which pristimerin mediated the observed increase in the transduction efficiency of rAAV vectors include both inhibition of proteasomal degradation of the intracellular proteins and enhanced nuclear translocation of the vector genomes.

CONCLUSIONThese studies suggest the potential beneficial use of pristimerin and pristimerin-containing herb extract in future liver-targeted gene therapy with rAAV vectors.

Animals ; Cell Line ; Dependovirus ; genetics ; physiology ; Gene Expression ; drug effects ; Genetic Therapy ; Genetic Vectors ; genetics ; physiology ; Hepatocytes ; metabolism ; virology ; Humans ; Liver ; cytology ; metabolism ; virology ; Mice ; Transgenes ; drug effects ; Triterpenes ; pharmacology

OBJECTIVELittle effort has been made to study the protein-encoding genes isolated from traditional Chinese medicine (TCM) drugs, and the delivery of these genes into malignant cells through recombinant adeno-associated virus (rAAV) vectors has not been attempted.

METHODSWe synthesized the cDNAs of five known cytotoxic proteins isolated from TCM drugs and the FLAG epitope-tagged cDNAs were subcloned into a rAAV plasmid vector. The protein expression was confirmed by Western blot assay. Various cancer cell lines were transfected with the above plasmids and cell growth was monitored both in vitro and in vivo. The best cytotoxic gene was further packaged into rAAV vectors, under the control of a liver cancer-specific promoter. The liver tumor growth was then monitored following intratumor administration of the rAAV vectors.

RESULTSThe expression plasmids, encoding individual potential cytotoxic genes tagged with FLAG epitope, were successfully generated and sequenced. Among these genes, trichosanthin (TCS) gene yielded the most promising results for the inhibition of cancer cell growth in vitro. The over-expressed TCS functioned as a type I ribosome-inactivating protein, followed by inducing apoptosis that is associated with the Bcl-PARP signaling pathway. Furthermore, intratumor injection of rAAV vectors containing the TCS gene significantly inhibited the growth of human hepatocellular carcinoma tumors in a murine xenograft model.

CONCLUSIONOur studies suggest that the use of TCM cytotoxic genes is a useful therapeutic strategy for treating human cancers in general, and liver tumors in particular.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; physiology ; Cell Line, Tumor ; DNA, Complementary ; Dependovirus ; Genetic Vectors ; Humans ; Liver Neoplasms ; physiopathology ; Medicine, Chinese Traditional ; methods ; Mice ; Neoplasms ; physiopathology ; Trichosanthin ; genetics ; pharmacology