Objective To observe the onset of vascular smooth muscle cell (VSMC) senescence induced by tert-butyl hydroperoxide (t-BHP) and the intervention effect of dehydroepiandrosterone (DHEA). Methods The VSMCs were divided into four groups: blank control group, t-BHP group (incubated with 80 μmol/L t-BHP for 72 h), 10 nmol/L DHEA intervention group (pretreated with 10 nmol/L DHEA 30 min before t-BHP) and 100 nmol/L DHEA intervention group (pretreated with 100nmol/L DHEA 30 min before t-BHP). Two ageing markers of ageing associated β-galactosidase activity and cell proliferation activity were adopted as main indexes. β-galactosidase activity was measured with immunocytochemical method and cell proliferation activity was measured with flowcytometry. Results After continuous treatment with 80 mmol/L t-BHP for 72 h, the ratios of G0/G1 phase cells and SA-β-galactosidase staining positive cells increased as compared with blank controlgroup [(89.4±3.4)% vs. (49.5±5.5)%, (3.5±1.2)% vs. (75.3±4.3)%], which indicated that VSMCs senescence were successfully induced by t-BHP. While the above changes were smaller in 100 nmol/L DHEA intervention group than in t-BHP group. Conclusions With ageing,accumulation of damage produced by reactive oxygen species may be an important mechanism causing the onset of VSMCs senescence. DHEA may be able to retard the progression of VSMCs senescence through antioxidant effect.

Objective To analyze of the relationship between level of anxiety and depression before surgery and anus exhausting time for patients after gynecological abdominal surgery.Methods A descriptive study was conducted with a survey of 100 cases of patients with gynecological abdominal surgery where the level of anxiety and depression before surgery and post-anus exhausting time were analyzed to explore the correlation between them.Results In the 88 valid questionnaires,the incidences of stress and anxiety before surgery were respectively 38.64％(34/84) and 19.32％(17/88) and the average anus exhausting time after surgery was (44.55 ± 13.90) hours;the anus exhausting time of anxious patients was significantly longer than that of non-anxious patients [(48.35±12.84) hours vs.(42.50±14.13) hours,t=-2.12,P＜0.05] and bivariate Logistic regression analysis suggested that preoperative level of anxiety had a significant effect on anus exhausting time after surgery.Conclusions Stress,anxiety and other negative emotions can affect anus exhausting time after surgery and level of anxiety could be used as one predictor of the delay of anus exhausting time after surgery.

Objective To explore the effect of macrolide antibiotics(erythromycin) on tumor necrosis factor(TNF)-α and interleukin(IL)-8 in hyperoxia-induced lung tissue of premature newborn rats,and to study the intervention effect of erythromycin on hyperoxia-induced lung injury.Methods One-day old preterm Sprague Dawley rats were randomly divided into four groups by random number table method:air + sodium chloride group,air + erythromycin group,hyperoxia + sodium chloride group,hyperoxia + erythromycin group.Hyperoxia groups were continuously exposed to oxygen (oxygen ＞ 0.85) and air group in room air.After 1,7,14 days of exposure,the preterm rats of four groups were sacrificed,whole lung of these rats were isolated,the lung histological changes were observed by hematoxylin-eosin staining,TNF-α and IL-8 in pulmonary tissue homogenate were detected by ELISA.Results The results showed that:(1) Compared with air + sodium chloride group,TNF-α and IL-8 expression in hyperoxia + sodium chloride group were significantly increased(P ＜ 0.05) after 1,7 days of exposure [1 d:TNF-α:(16.163 ± 0.574) ng/ml vs.(21.923 ±2.066) ng/ml,IL-8:(18.214 ±3.649) ng/ml vs.(23.546 ± 5.240) ng/ml ;7 d:TNF-α:(15.940 ±0.821) ng/ml vs.(19.688 ±0.764) ng/ml,IL-8:(18.541 ± 4.114) ng/ml vs.(24.255 ±4.692) ng/ml],in particular,TNF-α expression appeared to increase earlier,their expression became significantly weak in 14 days (P ＜ 0.05).(2) Compared with hyperoxia + sodium chloride group,TNF-α and IL-8 expression in hyperoxia +erythromycin group became significantly weak after 1,7,14 days of exposure(P ＜0.05) after the intervention of erythromycin [1 d:TNF-α:(21.923 ± 2.066) ng/ml vs.(18.903 ± 1.851) ng/ml,7 d:IL-8:(24.255 ±4.692) ng/ml vs.(23.508 ±3.543) ng/ml,14 d:TNF-α:(16.443 ±5.466) ng/ml vs.(14.453 ±0.963)ng/ml],but their expression became weaker in 14 days than that in 1,7 days.Conclusion The release of inflammatory mediators TNF-α and IL-8 induced by oxidation outbreak participates in the development of hyperoxia induced lung injury,erythromycin may regulate immune function,inhibits the levels of oxidant-mediated TNF-α and IL-8 induced by oxidation outbreak,and alleviate hyperoxia lung injury in premature rats.

Objective To explore the effect of diammonium glycyrrhizinate(DG) and astragalus membranaceus (AM) injection on the clinical comprehensive score in patients with acute lung injury (ALI). Methods According to the random number table method,a prospective random controlled study was conducted in which 60 cases of patients with ALI were divided into a study group and a control group(each,30 cases). Both groups received a comprehensive treatment based on the new guidelines,and the study group was additionally given DG and AM injection(DG 150 mg+AM 20 ml)one time per day for 7 days. The scores of lung injury,acute physiology and chronic health evaluationⅡ(APACHEⅡ)and systemic inflammatory response syndrome(SIRS)were measured at baseline,3rd and 7th day after treatment,and ventilation support time and final disease mortality rate were also calculated in all the patients. Results There were no statistically significant differences between the two groups in the scores of lung injury,APACHEⅡand SIRS before treatment and after treatment for 3 days(all P>0.05),with prolonged treatment,the above indexes were significantly reduced compared with those before treatment in the two groups,and the decreases in scores of indexes in study group was more significant than those in control group after treatment(lung injury score:1.31±0.99 vs. 2.29±1.08,APACHEⅡscore:18.43±8.17 vs. 24.23±6.98,SIRS score:1.69±0.89 vs. 2.60±1.04,all P<0.01). The time(hour)for ventilator support in study group was shorter than that in the control group(176.10±57.81 vs. 286.07 ± 156.27,P<0.01),but there was no statistically significant difference in mortality rate between the two groups(13.33%vs.16.67%,P>0.05). Conclusion The results suggest that DG and AM injection improve the scores of lung injury,APACHEⅡand SIRS,and alleviate the lung injury,so that the injection is beneficial to the early weaning from the ventilator to support treatment in patients with acute lung injury,and has certain therapeutic effect on ALI.

OBJECTIVETo investigate the effect of anti-miR-145 on human airway smooth muscle cell (HASMC) proliferation and osteopontin systhesis in vitro and explore the mechanisms.

METHODSHASMCs were treated with 10-100 nmol/L anti-miR-145, and the cell proliferation and apoptosis were investigated using a CCK-8 assay and flow cytometry, respectively. The changes in osteopontin synthesis after the treatment was quantified with Western blotting.

RESULTSTreatment with 10 and 50 nmol/L anti-miR-145 significantly promoted the proliferation and osteopontin synthesis in HASMCs (P<0.05 or <0.01), and 50 nmol/L anti-miR-145 obviously inhibited the cell apoptosis (P<0.01).

CONCLUSIONAnti-miR-145 promotes HASMC proliferation and osteopontin synthesis and inhibits HASMC apoptosis in vitro, indicating the important role of anti-miR-145 in the pathogenesis of airway remodeling.

Airway Remodeling ; Apoptosis ; Cell Proliferation ; Cells, Cultured ; Humans ; MicroRNAs ; antagonists & inhibitors ; Myocytes, Smooth Muscle ; drug effects ; Osteopontin ; biosynthesis ; Respiratory System ; cytology

Objective:To construct a biospecimen bank of patient derived organoids (PDOs) from pancreatic cancer tissues and to explore the feasibility of PDOs drug sensitivity assay technology to guide chemotherapy drug selection for pancreatic cancer.Methods:Pancreatic cancer tissue specimens obtained after surgical resection and puncture biopsy from Mar 2020 to Dec 2022 at Drum Tower Hospital, Nanjing University School of Medicine were collected. Pancreatic cancer PDOs were cultured in vitro and histologically identified; PDOs were treated with gemcitabine, Nab-paclitaxel, fluorouracil, Oxaliplatin, and Irinotecan and cell viability was measured to analyze the correlation between PDOs drug sensitivity and the actual clinical treatment response.Results:The PDOs can reproduce the pathological features of corresponding tumor tissues; the sensitivity of different PDOs to the same chemotherapeutic drug is significantly different; The sensitivity of PDOs was highly consistent with the actual treatment effect of the corresponding patients 75.76% (25/33); organoid organ-based susceptibility testing had predictive value for the treatment response of patients (AUC=0.733, 95% CI: 0.546-0.919, P<0.05). Conclusion:A biobank of pancreatic cancer PDOs was successfully constructed, and the drug susceptibility test results were significantly correlated with the actual medication response of patients, suggesting that the drug susceptibility test technology based on PDOs has the potential to guide individualized chemotherapy for pancreatic cancer.

Objective : To explore the safety and effectiveness of transvaginal ischia spinous fascia fixation for pelvic organ prolapse. Methods : The retrospective analysis of 124 patients who underwent surgical treatment for stage III - IV pelvic organ prolapse was conducted. Among them , 53 cases of transvaginal ischia spinous fascia fixation (IS⁃ FF) were performed as a study group (ISFF group) while 71 cases of transvaginal sacrospinous ligament fixation (SSLF) were performed as a control group (SSLF group) . The operation time , postoperative hospitalization days , preoperative and postoperative hemoglobin values , indwelling urinary catheter time , postoperative pain scores , and the occurrence of complications were compared between the two groups , and the efficacy of the operation was objectively evaluated by using the staging method of pelvic organ prolapse (POP⁃Q) . Also the scores of the pelvic floor impact questionnaire⁃7 (PFIQ⁃7) , the pelvic floor dysfunction questionnaire⁃20 (PFDI⁃20) , and the questionnaire of quality of life 12 (PISQ⁃12) were used to evaluate the patients ′ postoperative quality of life. Results : The operation time and postoperative hospitalization days of patients in the ISFF group were less than those in the SSLF group , and the differences were statistically significant (P < 0. 05) . The preoperative and postoperative hemoglobin values , retention time of urinary catheter, postoperative pain scores , and hospitalization costs of patients in the two groups were compared , and the differences were not statistically significant. At the 3 ⁃month postoperative outpatient follow⁃up , the objective success rate was 100% in two groups. The median follow⁃up time of patients in both groups was 24 months ( 12 - 41 months) , and there were 2 cases of recurrence in the ISFF group , with a recurrence rate of 3. 77% and a subjective success rate of 96. 23% . While there were 3 cases of recurrence in the SSLF group and 2 cases of loss of visit , with a recurrence rate of 4. 34% and a subjective success rate of 95. 65% . 1 patient in the SSLF group presented with a pelvic hematoma with a diameter of about 5 cm after surgery. The hematoma disappeared after hemostasis and other symptomatic treatment. There was no organ injury or blood transfusion in both groups. Conclusion Transvaginal ischia spinous fascia fixation is a safe and effective treatment for pelvic organ prolapse , and it has the advantages of short operation time , fast postoperative recovery , fewer complications , and improvement of patients ′ quality of life.

Radiotherapy is widely used in the management of advanced colorectal cancer (CRC). However, the clinical efficacy is limited by the safe irradiated dose. Sensitizing tumor cells to radiotherapy via interrupting DNA repair is a promising approach to conquering the limitation. The BRCA1-BARD1 complex has been demonstrated to play a critical role in homologous recombination (HR) DSB repair, and its functions may be affected by HERC2 or BAP1. Accumulated evidence illustrates that the ubiquitination-deubiquitination balance is involved in these processes; however, the precise mechanism for the cross-talk among these proteins in HR repair following radiation hasn't been defined. Through activity-based profiling, we identified PT33 as an active entity for HR repair suppression. Subsequently, we revealed that BAP1 serves as a novel molecular target of PT33 via a CRISPR-based deubiquitinase screen. Mechanistically, pharmacological covalent inhibition of BAP1 with PT33 recruits HERC2 to compete with BARD1 for BRCA1 interaction, interrupting HR repair. Consequently, PT33 treatment can substantially enhance the sensitivity of CRC cells to radiotherapy in vitro and in vivo. Overall, these findings provide a mechanistic basis for PT33-induced HR suppression and may guide an effective strategy to improve therapeutic gain.