Objective:To investigate the role of mechanosensor Yes-associated protein 1 (YAP1) in urothelial cells in inducing bladder dysfunction in a partial bladder outlet obstruction (pBOO) model.Methods:Ten female C57BL/6 mice were included in this study and randomly divided into pBOO and sham groups based on body weight using a stratified pairing method, with 5 mice in each group. The pBOO group underwent proximal urethral ligation surgery, while the sham group underwent a sham operation. Two weeks after surgery, the urinary pattern was analyzed using the urine spot test. The significant increase in urine spot numbers indicated the successful establishment of the pBOO model. The mice were then sacrificed, and bladder tissues were weighed and stained with hematoxylin and eosin (HE) to observe morphological changes. The bladder urothelial layer was further isolated, and total cell proteins were extracted to detect the expression levels of YAP1 protein using Western blotting. Mouse immortalized bladder urothelial cells were divided into three experimental groups: the negative control (NC) group, which was treated with YAP1-NC lentivirus; the overexpression (OE) group, which was treated with YAP1-OE lentivirus to induce YAP1 protein overexpression; and the verteporfin treatment (VP) group, which was treated with verteporfin on the basis of the OE group. Real-time quantitative PCR and Western blotting were used to verify the transcription and expression levels of YAP1 protein, the co-transcriptional activator TEAD4 protein, and the phosphorylated protein DRP1-616 (at serine 616) of dynamin-related protein 1 (DRP1). An ATP detection kit was used to measure the ATP release concentration in the NC, OE, and VP groups. The interaction between YAP1 and TEAD4 was investigated using co-immunoprecipitation, and the expression of the mitochondrial marker translocase of the outer mitochondrial membrane 20 (Tom20) was observed using immunofluorescence staining.Results:The results of the urine spot test showed that the number of urine spots on the filter paper in the pBOO group was higher than that in the sham group within 6 hours [(283.0±9.1) spots vs. (3.7±0.3) spots, P<0.01], and the urine spots were scattered. The bladder wet weight in the pBOO group was significantly higher than that in the sham group [(105.70±6.84) mg vs. (22.33±1.20) mg, P<0.01]. Histological observations revealed reduced bladder mucosal folds and increased detrusor muscle thickness in the pBOO group. The expression of YAP1 protein in the bladder urothelial cells of the pBOO group was significantly upregulated compared to the sham group [(1.26±0.08) vs. (0.50±0.04), P<0.01]. In vitro experiments showed that compared to the NC group, the OE group had significantly increased expression of DRP1-616 [(0.94±0.05) vs. (0.33±0.01), P<0.01] and higher ATP release concentration [(24.45±0.16) μmol/mg vs. (19.67±0.42) μmol/mg, P<0.01]. In contrast, the VP group had significantly decreased expression of DRP1-616 [(0.29±0.04) vs. (0.94±0.05), P<0.01] and lower ATP release concentration [(10.55±0.01) μmol/mg vs. (24.45±0.16) μmol/mg, P<0.01] compared to the OE group. Co-immunoprecipitation experiments using YAP1 and TEAD4 antibodies showed that YAP1 and TEAD4 proteins could interact and form a transcriptional complex to regulate ATP release. Immunofluorescence staining revealed increased expression of Tom20 in the OE group compared to the NC group [(104.20±3.28) vs. (74.51±3.87), P<0.01]. Conclusions:In the pBOO-induced bladder dysfunction model, YAP1 is highly expressed in urothelial cells. YAP1 forms a transcriptional complex with TEAD4 to regulate ATP release by promoting mitochondrial fission via DRP1-616 expression, which is a key mechanism underlying pBOO-induced bladder dysfunction.

Objective:To investigate the role of mechanosensor Yes-associated protein 1 (YAP1) in urothelial cells in inducing bladder dysfunction in a partial bladder outlet obstruction (pBOO) model.Methods:Ten female C57BL/6 mice were included in this study and randomly divided into pBOO and sham groups based on body weight using a stratified pairing method, with 5 mice in each group. The pBOO group underwent proximal urethral ligation surgery, while the sham group underwent a sham operation. Two weeks after surgery, the urinary pattern was analyzed using the urine spot test. The significant increase in urine spot numbers indicated the successful establishment of the pBOO model. The mice were then sacrificed, and bladder tissues were weighed and stained with hematoxylin and eosin (HE) to observe morphological changes. The bladder urothelial layer was further isolated, and total cell proteins were extracted to detect the expression levels of YAP1 protein using Western blotting. Mouse immortalized bladder urothelial cells were divided into three experimental groups: the negative control (NC) group, which was treated with YAP1-NC lentivirus; the overexpression (OE) group, which was treated with YAP1-OE lentivirus to induce YAP1 protein overexpression; and the verteporfin treatment (VP) group, which was treated with verteporfin on the basis of the OE group. Real-time quantitative PCR and Western blotting were used to verify the transcription and expression levels of YAP1 protein, the co-transcriptional activator TEAD4 protein, and the phosphorylated protein DRP1-616 (at serine 616) of dynamin-related protein 1 (DRP1). An ATP detection kit was used to measure the ATP release concentration in the NC, OE, and VP groups. The interaction between YAP1 and TEAD4 was investigated using co-immunoprecipitation, and the expression of the mitochondrial marker translocase of the outer mitochondrial membrane 20 (Tom20) was observed using immunofluorescence staining.Results:The results of the urine spot test showed that the number of urine spots on the filter paper in the pBOO group was higher than that in the sham group within 6 hours [(283.0±9.1) spots vs. (3.7±0.3) spots, P<0.01], and the urine spots were scattered. The bladder wet weight in the pBOO group was significantly higher than that in the sham group [(105.70±6.84) mg vs. (22.33±1.20) mg, P<0.01]. Histological observations revealed reduced bladder mucosal folds and increased detrusor muscle thickness in the pBOO group. The expression of YAP1 protein in the bladder urothelial cells of the pBOO group was significantly upregulated compared to the sham group [(1.26±0.08) vs. (0.50±0.04), P<0.01]. In vitro experiments showed that compared to the NC group, the OE group had significantly increased expression of DRP1-616 [(0.94±0.05) vs. (0.33±0.01), P<0.01] and higher ATP release concentration [(24.45±0.16) μmol/mg vs. (19.67±0.42) μmol/mg, P<0.01]. In contrast, the VP group had significantly decreased expression of DRP1-616 [(0.29±0.04) vs. (0.94±0.05), P<0.01] and lower ATP release concentration [(10.55±0.01) μmol/mg vs. (24.45±0.16) μmol/mg, P<0.01] compared to the OE group. Co-immunoprecipitation experiments using YAP1 and TEAD4 antibodies showed that YAP1 and TEAD4 proteins could interact and form a transcriptional complex to regulate ATP release. Immunofluorescence staining revealed increased expression of Tom20 in the OE group compared to the NC group [(104.20±3.28) vs. (74.51±3.87), P<0.01]. Conclusions:In the pBOO-induced bladder dysfunction model, YAP1 is highly expressed in urothelial cells. YAP1 forms a transcriptional complex with TEAD4 to regulate ATP release by promoting mitochondrial fission via DRP1-616 expression, which is a key mechanism underlying pBOO-induced bladder dysfunction.

Objective:To understand the current status of core measures of enhanced recovery after surgery (ERAS) nursing in Anhui Province and analyze its promotion strategies, so we can provide reference for further implementation and promotion of clinical practice guidelines of ERAS.Methods:Design a Development Questionnaire of Enhanced Recovery After Surgery in Anhui Province based on the Clinical Practice Guidelines for Enhanced Recovery After Surgery in China (2021 edition) (I) and, a cross-sectional survey was conducted on surgical nurses in 132 secondary and above hospitals in Anhui Province from July to September 2023 by the convenience sampling method.Results:A total of 1 375 nurses from 13 cities in Anhui province participated in the survey. The nurses′ awareness rate of ERAS concept was 100.00%, but only 52.51% (722/1 375) of the nurses had received standardized ERAS training, and only 26.11% (359/1 375) of the nurses indicated that they had a multidisciplinary team such as medical care, anesthesia and nutrition were involved in the implementation of the ERAS. Nearly 38.33% (527/1 375) of the nurses said that the department lacked the relevant nursing quality control system, and the overall implementation rate of quality control training, development, feedback and monitoring was not high. Only 55.71% (766/1 375) of nurses reported that a quality control team has been established within their department, and only 40.44% (556/1 375) of departments conduct monthly checks on the implementation and progress of ERAS. In terms of the implementation of core measures, preoperative thromboprevention therapy, intraoperative temperature monitoring and management, postoperative multimodal analgesia were better; however, the implementation of preoperative fasting time ≤ 6 h, water ban time≤ 2 h, oral carbohydrate drinks or clean streaming drinks, oral intake of enteral nutritional supplementation when less than 60% of the normal amount was poor.Conclusions:There are still many deficiencies in the overall development of ERAS nursing in Anhui Province, and the implementation level of core measures also needs to be improved. It is necessary to combine the concept of precision nursing, strengthen the whole process management and quality control, so as to promote the clinical practice and promotion of ERAS.

Objective:To understand the current status of core measures of enhanced recovery after surgery (ERAS) nursing in Anhui Province and analyze its promotion strategies, so we can provide reference for further implementation and promotion of clinical practice guidelines of ERAS.Methods:Design a Development Questionnaire of Enhanced Recovery After Surgery in Anhui Province based on the Clinical Practice Guidelines for Enhanced Recovery After Surgery in China (2021 edition) (I) and, a cross-sectional survey was conducted on surgical nurses in 132 secondary and above hospitals in Anhui Province from July to September 2023 by the convenience sampling method.Results:A total of 1 375 nurses from 13 cities in Anhui province participated in the survey. The nurses′ awareness rate of ERAS concept was 100.00%, but only 52.51% (722/1 375) of the nurses had received standardized ERAS training, and only 26.11% (359/1 375) of the nurses indicated that they had a multidisciplinary team such as medical care, anesthesia and nutrition were involved in the implementation of the ERAS. Nearly 38.33% (527/1 375) of the nurses said that the department lacked the relevant nursing quality control system, and the overall implementation rate of quality control training, development, feedback and monitoring was not high. Only 55.71% (766/1 375) of nurses reported that a quality control team has been established within their department, and only 40.44% (556/1 375) of departments conduct monthly checks on the implementation and progress of ERAS. In terms of the implementation of core measures, preoperative thromboprevention therapy, intraoperative temperature monitoring and management, postoperative multimodal analgesia were better; however, the implementation of preoperative fasting time ≤ 6 h, water ban time≤ 2 h, oral carbohydrate drinks or clean streaming drinks, oral intake of enteral nutritional supplementation when less than 60% of the normal amount was poor.Conclusions:There are still many deficiencies in the overall development of ERAS nursing in Anhui Province, and the implementation level of core measures also needs to be improved. It is necessary to combine the concept of precision nursing, strengthen the whole process management and quality control, so as to promote the clinical practice and promotion of ERAS.

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.

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.

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 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.

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

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