ObjectiveTo elucidate the underlying mechanism through which Zhuluan decoction suppresses excessive autophagy in human ovarian granulosa cells （KGN） and ameliorates premature ovarian insufficiency （POI） via the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. MethodsThe optimal concentration of cyclophosphamide for inducing a POI model in KGN cells was identified via the cell counting kit-8 （CCK-8） assay. Subsequently， the impacts of varying concentrations of Zhuluan decoction-containing serum on the viability of the KGN cell model were assessed. After the optimal drug concentration was determined， KGN cells were categorized into the following groups： blank control （20% blank serum）， model （20% blank serum + 5 μmol·L^-1 cyclophosphamide）， Zhuluan decoction-containing serum （20% Zhuluan decoction-containing serum + 5 μmol·L^-1 cyclophosphamide）， autophagy inhibitor （20% blank serum + 5 μmol·L^-1 cyclophosphamide + 20 μmol·L^-1 chloroquine phosphate）， autophagy inhibitor + Zhuluan decoction-containing serum （20% Zhuluan decoction-containing serum + 5 μmol·L^-1 cyclophosphamide + 20 μmol·L^-1 chloroquine phosphate）， and estradiol valerate （20% estradiol valerate-containing serum + 5 μmol·L^-1 cyclophosphamide）. Following 48 hours of incubation， flow cytometry was utilized to measure the apoptosis rate of KGN cells in each group. Western blotting was employed to quantify the protein levels of PI3K， phosphorylated （p）-Akt， Akt， p-mTOR， and mTOR， along with the expression levels of autophagy-related proteins such as Beclin1， autophagy-related 5 homolog （ATG5）， and microtubule-associated protein 1 light chain 3 （LC3）， in each group. Additionally， monodansylcadaverine （MDC） staining was performed to evaluate the extent of autophagy in each group. ResultsIncubation of KGN cells with 5 μmol·L^-1 cyclophosphamide for 48 h successfully established a POI model， marked by a significant inhibition of KGN cell proliferation. Notably， the inhibitory effect of cyclophosphamide on KGN cell proliferation exhibited a positive correlation with its concentration. Zhuluan decoction-containing serum at 20% and 30% promoted cell proliferation and mitigated the inhibitory effect of cyclophosphamide on KGN cell proliferation， with comparable therapeutic efficacy observed at both concentrations. Compared with the blank control group， the model group displayed an elevated apoptosis rate （P<0.01）， reduced protein levels of PI3K， p-Akt， and p-mTOR （P<0.01）， increased protein levels of Beclin1， LC3， and ATG5 （P<0.01）， no significant alterations in the protein levels of Akt and mTOR， and an enhanced MDC autophagy fluorescence intensity （P<0.01）. In comparison to that the model group， the apoptosis rates in the blank control group， model group， Zhuluan decoction-containing serum group， autophagy inhibitor group， autophagy inhibitor + Zhuluan decoction-containing serum group， and estradiol valerate group all reduced （P<0.05， P<0.01）， with the most pronounced reduction observed in the autophagy inhibitor + Zhuluan decoction-containing serum group. The protein levels of PI3K， p-Akt， and p-mTOR were higher in other groups than in the model group （P<0.05， P<0.01）， being the highest in the autophagy inhibitor + Zhuluan decoctio-containing serum group （P<0.01）. The protein levels of Beclin1 and ATG5 were lower in other groups than in the model group （P<0.05， P<0.01）. The expression level of LC3 declined in the Zhuluan decoction-containing serum group and the estradiol valerate group （P<0.05， P<0.01）， while it decreased without statistical significance in the autophagy inhibitor group and the autophagy inhibitor + Zhuluan decoction-containing serum group. ConclusionZhuluan decoction may activate the PI3K/Akt/mTOR pathway to inhibit excessive autophagy and counteract the detrimental effects of cyclophosphamide on the KGN cell model， thus managing POI.

ObjectiveTo elucidate the underlying mechanism through which Zhuluan decoction suppresses excessive autophagy in human ovarian granulosa cells （KGN） and ameliorates premature ovarian insufficiency （POI） via the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. MethodsThe optimal concentration of cyclophosphamide for inducing a POI model in KGN cells was identified via the cell counting kit-8 （CCK-8） assay. Subsequently， the impacts of varying concentrations of Zhuluan decoction-containing serum on the viability of the KGN cell model were assessed. After the optimal drug concentration was determined， KGN cells were categorized into the following groups： blank control （20% blank serum）， model （20% blank serum + 5 μmol·L^-1 cyclophosphamide）， Zhuluan decoction-containing serum （20% Zhuluan decoction-containing serum + 5 μmol·L^-1 cyclophosphamide）， autophagy inhibitor （20% blank serum + 5 μmol·L^-1 cyclophosphamide + 20 μmol·L^-1 chloroquine phosphate）， autophagy inhibitor + Zhuluan decoction-containing serum （20% Zhuluan decoction-containing serum + 5 μmol·L^-1 cyclophosphamide + 20 μmol·L^-1 chloroquine phosphate）， and estradiol valerate （20% estradiol valerate-containing serum + 5 μmol·L^-1 cyclophosphamide）. Following 48 hours of incubation， flow cytometry was utilized to measure the apoptosis rate of KGN cells in each group. Western blotting was employed to quantify the protein levels of PI3K， phosphorylated （p）-Akt， Akt， p-mTOR， and mTOR， along with the expression levels of autophagy-related proteins such as Beclin1， autophagy-related 5 homolog （ATG5）， and microtubule-associated protein 1 light chain 3 （LC3）， in each group. Additionally， monodansylcadaverine （MDC） staining was performed to evaluate the extent of autophagy in each group. ResultsIncubation of KGN cells with 5 μmol·L^-1 cyclophosphamide for 48 h successfully established a POI model， marked by a significant inhibition of KGN cell proliferation. Notably， the inhibitory effect of cyclophosphamide on KGN cell proliferation exhibited a positive correlation with its concentration. Zhuluan decoction-containing serum at 20% and 30% promoted cell proliferation and mitigated the inhibitory effect of cyclophosphamide on KGN cell proliferation， with comparable therapeutic efficacy observed at both concentrations. Compared with the blank control group， the model group displayed an elevated apoptosis rate （P<0.01）， reduced protein levels of PI3K， p-Akt， and p-mTOR （P<0.01）， increased protein levels of Beclin1， LC3， and ATG5 （P<0.01）， no significant alterations in the protein levels of Akt and mTOR， and an enhanced MDC autophagy fluorescence intensity （P<0.01）. In comparison to that the model group， the apoptosis rates in the blank control group， model group， Zhuluan decoction-containing serum group， autophagy inhibitor group， autophagy inhibitor + Zhuluan decoction-containing serum group， and estradiol valerate group all reduced （P<0.05， P<0.01）， with the most pronounced reduction observed in the autophagy inhibitor + Zhuluan decoction-containing serum group. The protein levels of PI3K， p-Akt， and p-mTOR were higher in other groups than in the model group （P<0.05， P<0.01）， being the highest in the autophagy inhibitor + Zhuluan decoctio-containing serum group （P<0.01）. The protein levels of Beclin1 and ATG5 were lower in other groups than in the model group （P<0.05， P<0.01）. The expression level of LC3 declined in the Zhuluan decoction-containing serum group and the estradiol valerate group （P<0.05， P<0.01）， while it decreased without statistical significance in the autophagy inhibitor group and the autophagy inhibitor + Zhuluan decoction-containing serum group. ConclusionZhuluan decoction may activate the PI3K/Akt/mTOR pathway to inhibit excessive autophagy and counteract the detrimental effects of cyclophosphamide on the KGN cell model， thus managing POI.

ObjectiveTo explore the mechanisms of Yangjing Tongluo prescription （YJTL） in the treatment of intrauterine adhesion （IUA） from the perspective of oxidative stress mediated by the Kelch-like ECH-associated protein 1/nuclear factor erythroid 2-related factor 2/heme oxygenase-1 （Keap1/Nrf2/HO-1） signaling pathway. MethodsA total of 48 rats with normal estrous cycles were selected and randomly divided into a normal group （n=8） and a modeling group （n=40）. An IUA rat model was established using a dual-injury method combining surgical curettage and infection. Eight rats were randomly selected from the modeling group for a pilot experiment to confirm successful model establishment. After successful modeling， the remaining 32 rats were randomly divided into a model group， a low-dose YJTL group （YJTL-L）， a high-dose YJTL group （YJTL-H）， and a Progynova group. Rats in the normal and model groups were administered purified water （15 mL·kg^-1） by gavage daily， while rats in the YJTL-L， YJTL-H， and Progynova groups received YJTL at doses of 6.43 and 12.86 g·kg^-1 and Progynova at 2.06 × 10^-4 g·kg^-1， respectively， for 14 consecutive days. The general condition， uterine morphology， and uterine index of the rats were monitored. Histopathological changes in uterine tissue were observed using hematoxylin-eosin （HE） staining. Serum levels of reactive oxygen species （ROS） and glutathione peroxidase （GSH-Px） were measured by enzyme-linked immunosorbent assay （ELISA）. Protein expression levels of Keap1， Nrf2， and HO-1 in endometrial tissue were detected by Western blot. Immunofluorescence （IF） was used to assess the distribution of Nrf2 and HO-1， as well as the expression of Nrf2 in the cytoplasm and nucleus. ResultsCompared with the normal group， rats in the model group exhibited poor mental status and reduced mobility， markedly edematous and tortuous uterine morphology， decreased gland number， and inflammatory reactions in the endometrium， along with an increased uterine organ index （P<0.05）. Serum ROS levels were significantly increased （P<0.05）， while serum GSH-Px levels were significantly decreased （P<0.05）. In endometrial tissue， Keap1 protein expression was increased （P<0.05）， whereas Nrf2 and HO-1 protein expression was decreased. Mild nuclear translocation of Nrf2 was observed， accompanied by increased relative fluorescence intensity of nuclear Nrf2 and decreased relative fluorescence intensity of cytoplasmic HO-1. Compared with the model group， all treatment groups showed varying degrees of improvement in the above symptoms and pathological changes. Serum ROS levels were reduced （P<0.05）， serum GSH-Px levels were increased （P<0.05）， Keap1 protein expression in endometrial tissue was decreased， and Nrf2 and HO-1 protein expression was increased in a dose-dependent manner （P<0.05）. Notably， significant nuclear translocation of Nrf2 was observed， with correspondingly increased relative fluorescence intensity of nuclear Nrf2 and enhanced relative fluorescence intensity of cytoplasmic HO-1. ConclusionYJTL may enhance antioxidant capacity and repair oxidative damage to the endometrial basal layer by regulating the Keap1/Nrf2/HO-1 signaling pathway.

ObjectiveTo investigate the mechanism by which Zhuluan decoction improves ovarian function in a cyclophosphamide （CTX）-induced premature ovarian insufficiency （POI） rat model through regulating the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/Unc-51-like kinase 1 （ULK1） signaling pathway. MethodsA POI rat model was established via intraperitoneal injection of CTX. Ten rats were randomly selected as the normal control group. The remaining 50 successfully modeled rats were randomly divided into the model group， the estradiol valerate group （0.1 mg·kg^-1）， low-dose （7 g·kg^-1）， medium-dose （14 g·kg^-1）， and high-dose （28 g·kg^-1） Zhuluan decoction groups， with 10 rats per group. All groups received intragastric administration for 21 consecutive days. Changes in body weight， uterine， and ovarian indices were observed across groups. Hematoxylin and eosin （HE） staining was used to examine ovarian pathological alterations and count follicles at various stages in cross-sections. Enzyme-linked immunosorbent assay （ELISA） measured serum estradiol （E₂）， follicle-stimulating hormone （FSH）， and anti-Müllerian hormone （AMH） levels. Western blot （WB） and immunohistochemistry （IHC） were used to detect the expression levels of AMPK， mTOR， ULK1 and their phosphorylated proteins， Beclin1， Bcl-2-associated X protein （Bax）， and Bcl-2 in ovarian tissues of each group. ResultsCompared with the normal group， model rats exhibited slower body weight gain （P<0.01）， significantly reduced uterine and ovarian organ indices （P<0.01）， reduced ovarian volume， irregular follicular morphology， unclear cumulus and oocyte structures， and a significant reduction in the number of primordial follicles （P<0.01）. Conversely，the numbers of secondary and atretic follicles significantly increased （P<0.01）. Serum E₂ and AMH levels significantly decreased， while FSH levels significantly elevated（P<0.01）. The expression of AMPK， ULK1， Bax， and Beclin1 was increased （P<0.01）， while that of mTOR and Bcl-2 was decreased （P<0.01）. Compared with the model group， the estradiol valerate group and the medium- and high-dose Zhuluan decoction groups showed significantly increased body weight， uterine organ index， and ovarian organ index （P<0.01）， while the low-dose Zhuluan decoction group showed no statistically significant difference. Ovarian volume increased across all treatment groups， with improved follicular morphology and structure. The number of primordial follicles significantly increased （P<0.01）， while the number of atretic follicles significantly decreased （P<0.01）. The number of secondary follicles decreased in the estradiol valerate group and the high-dose Zhuluan decoction group （P<0.01）， with no significant difference in the low- and medium-dose Zhuluan decoction groups. E₂ and AMH levels significantly increased， while FSH levels significantly decreased in all treatment groups （P<0.05， P<0.01）. The estradiol valerate group and medium- and high-dose Zhuluan decoction groups showed significantly reduced expression of AMPK， ULK1， Bax， and Beclin1 （P<0.05， P<0.01）， while mTOR and Bcl-2 expression levels were significantly elevated （P<0.01）. The low-dose Zhuluan decoction group exhibited significantly reduced expression of ULK1， Beclin1， and Bax （P<0.05）， while Bcl-2 expression was significantly increased （P<0.05）. ConclusionZhuluan decoction improves serum hormone levels in POI rats and mitigates chemotherapy （CTX）-induced damage to ovarian structure and function， exerting ovarian protective effect. This mechanism may involve regulating the AMPK/mTOR/ULK1 signaling pathway to inhibit excessive autophagy and apoptosis.

ObjectiveTo investigate the mechanism by which Zhuluan decoction improves ovarian function in a cyclophosphamide （CTX）-induced premature ovarian insufficiency （POI） rat model through regulating the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/Unc-51-like kinase 1 （ULK1） signaling pathway. MethodsA POI rat model was established via intraperitoneal injection of CTX. Ten rats were randomly selected as the normal control group. The remaining 50 successfully modeled rats were randomly divided into the model group， the estradiol valerate group （0.1 mg·kg^-1）， low-dose （7 g·kg^-1）， medium-dose （14 g·kg^-1）， and high-dose （28 g·kg^-1） Zhuluan decoction groups， with 10 rats per group. All groups received intragastric administration for 21 consecutive days. Changes in body weight， uterine， and ovarian indices were observed across groups. Hematoxylin and eosin （HE） staining was used to examine ovarian pathological alterations and count follicles at various stages in cross-sections. Enzyme-linked immunosorbent assay （ELISA） measured serum estradiol （E₂）， follicle-stimulating hormone （FSH）， and anti-Müllerian hormone （AMH） levels. Western blot （WB） and immunohistochemistry （IHC） were used to detect the expression levels of AMPK， mTOR， ULK1 and their phosphorylated proteins， Beclin1， Bcl-2-associated X protein （Bax）， and Bcl-2 in ovarian tissues of each group. ResultsCompared with the normal group， model rats exhibited slower body weight gain （P<0.01）， significantly reduced uterine and ovarian organ indices （P<0.01）， reduced ovarian volume， irregular follicular morphology， unclear cumulus and oocyte structures， and a significant reduction in the number of primordial follicles （P<0.01）. Conversely，the numbers of secondary and atretic follicles significantly increased （P<0.01）. Serum E₂ and AMH levels significantly decreased， while FSH levels significantly elevated（P<0.01）. The expression of AMPK， ULK1， Bax， and Beclin1 was increased （P<0.01）， while that of mTOR and Bcl-2 was decreased （P<0.01）. Compared with the model group， the estradiol valerate group and the medium- and high-dose Zhuluan decoction groups showed significantly increased body weight， uterine organ index， and ovarian organ index （P<0.01）， while the low-dose Zhuluan decoction group showed no statistically significant difference. Ovarian volume increased across all treatment groups， with improved follicular morphology and structure. The number of primordial follicles significantly increased （P<0.01）， while the number of atretic follicles significantly decreased （P<0.01）. The number of secondary follicles decreased in the estradiol valerate group and the high-dose Zhuluan decoction group （P<0.01）， with no significant difference in the low- and medium-dose Zhuluan decoction groups. E₂ and AMH levels significantly increased， while FSH levels significantly decreased in all treatment groups （P<0.05， P<0.01）. The estradiol valerate group and medium- and high-dose Zhuluan decoction groups showed significantly reduced expression of AMPK， ULK1， Bax， and Beclin1 （P<0.05， P<0.01）， while mTOR and Bcl-2 expression levels were significantly elevated （P<0.01）. The low-dose Zhuluan decoction group exhibited significantly reduced expression of ULK1， Beclin1， and Bax （P<0.05）， while Bcl-2 expression was significantly increased （P<0.05）. ConclusionZhuluan decoction improves serum hormone levels in POI rats and mitigates chemotherapy （CTX）-induced damage to ovarian structure and function， exerting ovarian protective effect. This mechanism may involve regulating the AMPK/mTOR/ULK1 signaling pathway to inhibit excessive autophagy and apoptosis.

Objective To explore the differences in autophagic expression levels between hypertrophic scar (HS) tissue and normal skin tissue,and further investigate the relationship between hypertrophic scar formation and autophagy protein expression through the rabbit ear hypertrophic scar model.Methods 30 patients with hypertrophic scar were collected.One hypertrophic scar tissue and one normal skin tissue were harvested.The relative expressions of LC3,P62 and Beclin-1 in each tissue specimen were detected by immunohistochemistry and Western blot.Western blot was used to detect the autophagic-associated protein LC3 (MAPLC3),P62 and Beclin-1 in the hypertrophic scar tissue of rabbit ear and the corresponding normal tissue of rabbit ears at 4 weeks,8 weeks,12 weeks,and 24 weeks,and further explore their clinical significance.Results In vivo,the expression of hypertrophic scar tissue protein LC3 and Beclin-1 was significantly stronger than that in normal skin tissue (P ＜ 0.05).The expression of P62 was significantly weaker than that in normal skin tissue (P ＜ 0.05).In animal experiments,during the process of HS formation,the protein expression of LC3 gradually increased,while the protein expression of P62 gradually decreased;the protein expression of Beclin-1 was higher than that of normal rabbit ears tissue,with statistically significant differences (P ＜ 0.05).Conclusions The expression of LC3 and Beclin-1 in human hypertrophic scar tissues is higher than that in normal tissues.While the expression of P62 is lower than that in normal tissues.That is,the expression of autophagy in human hypertrophic scar tissue showed an upward trend in a certain period of time,and was significantly higher than that in normal tissue.

Objective To set up and apply a standardized communication system of adolescents and young adults (AYAs) cancer patients,in order to improve AYAs cancer patients' psychological distress and other negative emotions,as well as promote social support and quality of life of patients.Methods A AYAs cancer patients standardized communication system,suitable for China's national conditions,was preliminarily built.Using randomized controlled trials,a total of 171 subjects,selected from 486 cases of AYAs cancer patients,in the Third Xiangya Hospital of Central South University and Hunan Cancer Hospital from August to September in 2016,were intervened with a set of standardized communication system.The Mental Distress Thermometer (DT),Hospital Anxiety and Depression Scale (HADS),Social Support Rating Scale (SSRS) and Concise Health Status Questionnaire (SF-36) were used as evaluation indexes to observe the psychological distress,emotion,social support and quality of life of the three groups of subjects before intervention,immediately after intervention,1 month after intervention and 3 months after intervention.Results At the follow-up of 3 months after intervention,38 cases were lost,14 cases in communication group (final n =43),11 cases in music therapy group (final n =46),and 13 cases in routine group (final n =44)..There was no significant difference in the evaluation indexes between the communication group,music treatment group and the routine group before intervention (P ＞ 0.05).There were statistically significant differences in scores of psychological pain,anxiety and depression,and social support in the 3 groups,before and immediately after intervention,1 month and 3 months after intervention (P ＜ 0.05).The scores of psychological pain,anxiety and depression,social support and quality of life in the communication group 1 month after intervention were statistically significant compared with those in the music group and the routine group (P ＜ 0.05).Conclusions Compared to music therapy and regular care,the standardized communication system has significant effect on improving the psychological distress of AYAs with cancer,and can also improve their social support level and quality of life.The clinical validation of the standardized communication system can provide reference for psychological rehabilitation of cancer survivors.

Objective: To analyze the clinical characteristics of patients with adult onset Still's disease (AOSD) with interstitial lung diseases (ILD), and review the literature. Methods: The medical records of inpatients with AOSD and ILD from January 2000 to October 2017 were retrospectively analyzed, and pa-pers were searched and summarized with the key words "adult onset Still's disease" and "interstitial lung diseases". Kolmogorov-Smirnov test was used to test if variables met normal distribution. Measurement data which was normally distributed was described as Mean±SD. Measurement data which was not normally distributed was described as median and interquartile range. Results: Among the 15 patients included in the study, six were male, and nine were female, and the mean age was (50±12) years. All of the 15 patients had fever, and the average temperature was (39.4±0.4) ℃. Eleven patients had rash, and 12 patients had arthralgia, seven patients presented with cough, and eight patients presented with short of breath. The high resolution computed tomography of the chest presented as ground glass opacity in nine patients, grid shadow in three patients and consolidation in three patients. All the 15 patients received glucocorticoids, and 10 patients received immunosuppressants at the same time. One patient was lost to follow up, four patients died (three patients died of respiratory failure and one patient died of myocardial infarction), 10 patients improved. Conclusion Patients with AOSD can also have ILD, which should be alerted by clinicians. Early treatment with glucocorticoids and immunosuppressants if infection is excluded may bring good prognosis, and it is easy to relapse when glucocorticoids is tapered off.

Cancer is the most cause death of among adolescents and young adults (AYAs).Psychological distress caused by cancer affects AYAs' effective coping abilities of disease,physical symptoms and treatment.This paper mainly introduces the related concepts,screening tools and intervention progress of psychological distress of AYAs cancer patients to deepen the understanding of these among clinical professionals and provide reference for implement effective interventions to patients.

OBJECTIVE: To explore the application and the effect of early recognition of deteriorating patient program in department of cardiac surgery. METHODS: We used the early recognition of deteriorating patient program in the cardiac surgery groups, including cardiac surgeons, nurses in ward, ICU and operation rooms of the cardiac surgery department, and compared the satisfaction of nurses and doctors, handover time, handover score of critical patients, and rate of unplanned ICU admission before and after the intervention. RESULTS: After using the early recognition of deteriorating patient program, the satisfaction of doctors and nurses was increased, the handover time was lowered 0.56 min/time (t=2.22, P<0.05), the handover score of critical patients enhanced by 19.59 points (t=30.57, P<0.001), the rate of unplanned ICU readmission after the operation reduced by 4.8% (χ2=4.14, P<0.05). CONCLUSION Early recognition of deteriorating patient program can improve the safety of cardiac patients, enhance the self-confidence of nurses and work efficiency.
Cardiology Service, Hospital ; organization & administration ; Critical Illness ; Humans ; Intensive Care Units ; Outcome and Process Assessment, Health Care ; Patient Handoff ; Surgery Department, Hospital ; organization & administration