1.Zhuluan Decoction Ameliorates Premature Ovarian Insufficiency by Inhibiting Excessive Autophagy of KGN Through Regulation of PI3K/Akt/mTOR Pathway
Yao CHEN ; Sainan TIAN ; Jing ZENG ; Xingxing YI ; Wen'e LIU ; Lei LEI ; Li TANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):89-98
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.
2.Zhuluan Decoction Ameliorates Premature Ovarian Insufficiency by Inhibiting Excessive Autophagy of KGN Through Regulation of PI3K/Akt/mTOR Pathway
Yao CHEN ; Sainan TIAN ; Jing ZENG ; Xingxing YI ; Wen'e LIU ; Lei LEI ; Li TANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):89-98
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.
3.Speech and language rehabilitation services for children with hearing impairment based on ICF:theoretical framework and service system
Jing ZHOU ; Ling YI ; Jianchao CHEN ; Xuefen CHEN ; Xingxing WEI ; Jingjing CHEN
Chinese Journal of Rehabilitation Theory and Practice 2024;30(7):745-751
Objective Based on the bio-psycho-social model of functioning and health of International Classification of Function-ing,Disability and Health(ICF)framework,this paper systematically analyzes rehabilitation-related policy docu-ments of the World Health Organization(WHO)to explore the theoretical and policy principles,service systems,priority development areas,and main policy and technical measures for speech and language rehabilitation for children with hearing impairment. Methods Under literature research and policy analysis methods,this paper systematically reviewed the functioning and health framework of ICF,relevant WHO policy reports and American Speech and Hearing Association technical documents,analyzed the current status and needs of rehabilitation services in China,and proposed a theoretical framework,priority areas,and main policy and technical measures for constructing an ICF-based speech and lan-guage rehabilitation service system for children with hearing impairment. Results There were eight major principles for the development of speech and language rehabilitation for children with hearing impairment:child-centered and respect for individual differences,life-cycle support and promoting con-tinuous development,evidence-based practice,multidisciplinary collaboration,building family participation sup-port networks,promoting integrated education and social participation,focusing on cultural sensitivity,and tech-nology-empowered rehabilitation innovation.Five service systems were elaborated,including comprehensive ear-ly screening,diagnosis,and intervention system;family and social support system;multidisciplinary team servic-es system;comprehensive,multi-level speech and language rehabilitation service system;and digitally empow-ered services to build new rehabilitation service models.The priority development areas,and main policy and technical measures for speech and language rehabilitation for children with hearing impairment were also dis-cussed. Conclusion Based on the bio-psycho-social model of functioning and health of ICF,comprehensive,personalized,con-tinuous,high-quality and universally accessible speech and language rehabilitation services can be developed by implementing measures,such as strengthening policy support,improving service systems,cultivating multidisci-plinary professionals,promoting digital empowerment technologies and enhancing support system,to promote the overall development,social participation and quality of life for children with hearing impairment.
4.Reliability on evaluation of quadriceps femoris muscle quality by ultrasonic echo intensity in patients with knee osteoarthritis
Junyi LI ; Zehua CHEN ; Zugui WU ; Yi WANG ; Congcong LI ; Shuai WANG ; Weijian CHEN ; Zixuan YE ; Xingxing SHEN ; Ruian XIANG ; Wengang LIU ; Xuemeng XU
Chinese Journal of Rehabilitation Theory and Practice 2023;29(6):738-744
ObjectiveTo evaluate the inter-rater and test-retest reliability of echo intensity of ultrasound image for quality of quadriceps femoris in patients with knee osteoarthritis (KOA). MethodsFrom March to December, 2021, 33 patients with unilateral KOA in Guangdong Provincial Second Hospital of Traditional Chinese Medicine were included. Ultrasound was used to observe the cross sections of rectus femoris (RF), vastus medialis (VM) and vastus lateralis (VL), and the images were saved. Two independent raters analyzed the average gray value of the picture with the Image J. One of the raters repeated the measurement. The intra-class correlation coefficient (ICC), standard error of measurement (SEM), minimum detectable change (MDC) and coefficient of variation (CV) were calculated and the Bland-Altman charts were drawn. ResultsThe gray value was higher in the RF, VM and VL in the affected side than in the healthy side (t > 2.262, P < 0.05). The inter-rater reliability was excellent, with ICC 0.982 to 0.995, SEM 1.60 to 3.82, CV 1.49% to 5.90%, and MDC value 4.43 to 10.59. The test-retest reliability of the rater was excellent, with ICC 0.969 to 0.990, SEM 2.37 to 5.41, CV 2.22% to 4.84%, and MDC value 6.57 to 15.00. Bland-Altman charts analysis showed that the consistency was good. ConclusionThe quadriceps femoris muscle quality is different between the affected and the healthy sides in patients with unilateral KOA. It is reliable to evaluate muscle echo intensity by Image J.
5.SOCS3 Attenuates GM-CSF/IFN-γ-Mediated Inflammation During Spontaneous Spinal Cord Regeneration.
Xuejie ZHANG ; Bingqiang HE ; Hui LI ; Yingjie WANG ; Yue ZHOU ; Wenjuan WANG ; Tiancheng SONG ; Nan DU ; Xingxing GU ; Yi LUO ; Yongjun WANG
Neuroscience Bulletin 2020;36(7):778-792
SOCS3, a feedback inhibitor of the JAK/STAT signal pathway, negatively regulates axonal regrowth and inflammation in the central nervous system (CNS). Here, we demonstrated a distinct role of SOCS3 in the injured spinal cord of the gecko following tail amputation. Severing the gecko spinal cord did not evoke an inflammatory cascade except for an injury-stimulated elevation of the granulocyte/macrophage colony-stimulating factor (GM-CSF) and interferon gamma (IFN-γ) cytokines. Simultaneously, the expression of SOCS3 was upregulated in microglia, and unexpectedly not in neurons. Enforced expression of SOCS3 was sufficient to suppress the GM-CSF/IFN-γ-driven inflammatory responses through its KIR domain by attenuating the activities of JAK1 and JAK2. SOCS3 was also linked to GM-CSF/IFN-γ-induced cross-tolerance. Transfection of adenovirus overexpressing SOCS3 in the injured cord resulted in a significant decrease of inflammatory cytokines. These results reveal a distinct role of SOCS3 in the regenerating spinal cord, and provide new hints for CNS repair in mammals.
6.Application of the moving epidemic method in the development of epidemic thresholds and tiered warning alert approachs for influenza prevention in Beijing
Yu WANG ; Li ZHANG ; Shuangsheng WU ; Wei DUAN ; Ying SUN ; Man ZHANG ; Xingxing ZHANG ; Yi ZHANG ; Chunna MA ; Quanyi WANG ; Peng YANG
Chinese Journal of Epidemiology 2020;41(2):201-206
Objective To calculate both the epidemic and intensity thresholds for different levels in Beijing and to establish a tiered alert system in the 2018-2019 influenza season as well as to evaluate the performance of calculated thresholds.Method Weekly count of influenza-like illness and percentage of influenza-like illness (ILI%) of the last five influenza seasons were modeled by 'moving epidemic method'(MEM) to calculate the influenza epidemic and intensity thresholds at different levels.A cross-validation procedure was used to evaluate the performance.Indicators of Matthew correlation coefficient,Youden's index,sensitivity and specificity were calculated.Results For weekly count of influenza-like illness,data showed that the epidemic threshold for 2018-2019influenza season was 12 984 and the medium,high and very high intensity thresholds were 22 503,37 589,47 157,respectively.Matthew correlation coefficient of the epidemic threshold was 62% and youden's index as 60%,sensitivity as 69%,specificity as 91%.Data on weekly ILI%,the epidemic threshold for 2018-2019 influenza season was 1.66%,with medium,high and very high intensity thresholds as 2.46%,3.84% and 4.66%,respectively.The overall Matthew correlation coefficient of the epidemic threshold was 59%,with 54% for the Youden's index,sensitivity as 60% and specificity as 94%.Conclusions MEM produced a good specific signal for detecting the influenza epidemics and the accuracy of the method was acceptable.The early warning performance regarding the application of weekly count on influenza-like illness was slightly better than ILI%.This method could be applied in the practical influenza epidemic alert "work in Beijing".
7.Investigation and analysis on the knowledge and attitude of nurses on common complications among bedridden patients
Zhengzheng DUAN ; Xinjuan WU ; Yilan LIU ; Deying HU ; Yi ZHU ; Hongmei LI ; Xiangkang ZENG ; Hongying JIA ; Lulu HUANG ; Xingxing LU ; Lin LAI
Chinese Journal of Modern Nursing 2018;24(14):1665-1667
Objective To investigate the knowledge and attitude of nurses on the prevention and nursing of common complications among bedridden patients in Wuhan City, and provide scientific basis for the implementation of targeted training in improving clinical nursing quality. Methods A total of 748 clinical nurses from 4 hospitals in Wuhan City were selected in November 2016 using convenient sampling method. A self-designed questionnaire was applied to investigate the common complications nursing knowledge and attitude of nurses on bedridden patients. Results The accuracy of nurses' knowledge on the prevention and nursing of common complications among bedridden patients were listed as follows: deep vein thrombosis (DVT) (77.44%), pneumonia (73.88%), urinary tract inflammation (70.63%), and pressure ulcers (61.44%). The scores of nurses' attitude were listed successively: pressure ulcers (88.21%), urinary tract inflammation (86.81%), pneumonia (86.66%), and DVT (85.92%). There were positive correlation between the scores of knowledge and attitude of nurses on four kinds of complications (P< 0.05). Conclusions Nursing administrators should improve the training and update of common complications knowledge on nurses, and promote their positive attitude to the prevention of common complication to increase the quality of nursing.
8. Infection characteristics of patients in acute gastroenteritis outbreaks caused by noroviruses
Zhiyong GAO ; Baiwei LIU ; Liyu HOU ; Hanqiu YAN ; Yi TIAN ; Yanwei CHEN ; Xingxing ZHANG ; Yi ZHANG ; Lei JIA ; Haikun QIAN ; Quanyi WANG
Chinese Journal of Experimental and Clinical Virology 2017;31(1):38-41
Objective:
To analyze the infection characteristics of patients in acute gastroenteritis outbreaks caused by noroviruses.
Methods:
Between April 2014 and March 2016, the clinical data and samples were collected from the patients in acute gastroenteritis outbreaks caused by noroviruses in Beijing. Noroviruses were detected and genotyped using real time RT-PCR, and the infection characteristics of norovirus gastroenteritis were analyzed using the descriptive epidemiological method.
Results:
A total of 1743 clinical diagnosed cases of norovirus gastroenteritis were collected, and children under 12 years old accounted for 77.68% (1354/1743). The detection rate of noroviruses was 73.98% (509/688). The detection rates of noroviruses in fecal, swab and vomitus samples were gradually decreased (
9.Prohibitin2 expression and its relation with astrocyte proliferation in rats after traumatic brain injury
Ting XU ; Hun WU ; Xingxing GU ; Long MA ; Yi WANG ; Yangyou YU
Chinese Journal of Neuromedicine 2017;16(7):671-675
Objective To research the prohibitin2 (PHB2) expression and cellular localization in the rat brain cortex after traumatic brain injury (TBI),and explore its relationship with astrocyte proliferation and neuron apoptosis.Methods TBI rat models were established by knife injurying the brain.Western blotting was used to detect the PHB2 expression variation trend in the brain cortex.Immunohistochemical method was used to detect the distribution of PHB2 in damaged cerebral cortex,and immunofluorescence was applied to research the PHB2 expression changes and cellular localization in the rat brain cortex after TBI,and explore its relationship with astrocyte proliferation and neuron apoptosis.Results (1) The TBI models were established successfully;12 h after TBI,the PHB2 expression started to increase obviously,and PHB2 expression reached to its peak level 5 d after TBI.(2)The PHB2 expression in the cortex of TBI rats was significantly increased as compared with that in the sham-operated group,control group and contralateral side of TBI rats.(3) PHB2 mainly located at the astrocytes and neurons of the cerebral cortex after TBI.(4) Co-localization was noted in astrocytes and cell proliferation marker PCNA,and in PHB2 and PCNA,which indicated that proliferation of astrocytes existed and PHB2 involved in the proliferation.(5) Co-localization was noted in astrocytes and cell apoptosis marker A-caspase-3,and in PHB2 and A-caspase-3,which indicated that TBI induced cell apoptosis,and PHB2 involved in the apoptosis.Conclusion PHB2 has a high expression in the cerebral cortex of rats after TBI,and these changes are related to astrocyte proliferation and neuronal apoptosis.
10.Research progress of long non-coding RNA in tumor epithelial-mesenchymal transition
Yi ZHAO ; Xingxing ZHANG ; Min XU
Journal of International Oncology 2015;(8):599-601
Long non-coding RNA (lncRNA)is a kind of RNA molecule which is longer than 200 nucleotides and has no capacity of coding proteins.Accumulating evidences have indicated that several lncRNAs,such as HOTAIR,MALAT-1 ,H1 9 and BANCR may promote tumor metastasis by inducing epitheli-al-mesenchymal transition.

Result Analysis
Print
Save
E-mail