1.Treatment Principles and Paradigm of Diabetic Microvascular Complications Responding Specifically to Traditional Chinese Medicine
Anzhu WANG ; Xing HANG ; Lili ZHANG ; Xiaorong ZHU ; Dantao PENG ; Ying FAN ; Min ZHANG ; Wenliang LYU ; Guoliang ZHANG ; Xiai WU ; Jia MI ; Jiaxing TIAN ; Wei ZHANG ; Han WANG ; Yuan XU ; .LI PINGPING ; Zhenyu WANG ; Ying ZHANG ; Dongmei SUN ; Yi HE ; Mei MO ; Xiaoxiao ZHANG ; Linhua ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):272-279
To explore the advantages of traditional Chinese medicine (TCM) and integrative TCM-Western medicine approaches in the treatment of diabetic microvascular complications (DMC), refine key pathophysiological insights and treatment principles, and promote academic innovation and strategic research planning in the prevention and treatment of DMC. The 38th session of the Expert Salon on Diseases Responding Specifically to Traditional Chinese Medicine, hosted by the China Association of Chinese Medicine, was held in Beijing, 2024. Experts in TCM, Western medicine, and interdisciplinary fields convened to conduct a systematic discussion on the pathogenesis, diagnostic and treatment challenges, and mechanism research related to DMC, ultimately forming a consensus on key directions. Four major research recommendations were proposed. The first is addressing clinical bottlenecks in the prevention and control of DMC by optimizing TCM-based evidence evaluation systems. The second is refining TCM core pathogenesis across DMC stages and establishing corresponding "disease-pattern-time" framework. The third is innovating mechanism research strategies to facilitate a shift from holistic regulation to targeted intervention in TCM. The fourth is advancing interdisciplinary collaboration to enhance the role of TCM in new drug development, research prioritization, and guideline formulation. TCM and integrative approaches offer distinct advantages in managing DMC. With a focus on the diseases responding specifically to TCM, strengthening evidence-based support and mechanism interpretation and promoting the integration of clinical care and research innovation will provide strong momentum for the modernization of TCM and the advancement of national health strategies.
2.Mechanism of action of luteolin in treatment of liver injury
Ying JIANG ; Ning GAO ; Xiaxuan WANG ; Jie GONG ; Lili LIU
Journal of Clinical Hepatology 2026;42(2):472-478
Liver injury has become an increasingly serious global health problem, and existing chemical drugs face the limitations in efficacy and adverse reactions, resulting in the urgent need to develop safe and effective drugs. Recent studies have highlighted the potential of flavonoids from natural medicinal plants in the prevention and treatment of liver injury. As a typical natural flavonoid, luteolin shows a good protective effect against liver injury due to various etiologies, but there is still a lack of systematic elaboration on its mechanism of action. This article summarizes related research advances in China and globally and reviews the mechanism of action of luteolin in inhibiting oxidative stress, exerting an anti-inflammatory effect, regulating cell death, alleviating hepatic fibrosis, modulating lipid metabolism disorders, and regulating the gut-liver axis, as well as the application prospect of luteolin in the treatment of liver injury, in order to provide a scientific reference for further research on this compound.
3.Mechanism of action of luteolin in treatment of liver injury
Ying JIANG ; Ning GAO ; Xiaxuan WANG ; Jie GONG ; Lili LIU
Journal of Clinical Hepatology 2026;42(2):472-478
Liver injury has become an increasingly serious global health problem, and existing chemical drugs face the limitations in efficacy and adverse reactions, resulting in the urgent need to develop safe and effective drugs. Recent studies have highlighted the potential of flavonoids from natural medicinal plants in the prevention and treatment of liver injury. As a typical natural flavonoid, luteolin shows a good protective effect against liver injury due to various etiologies, but there is still a lack of systematic elaboration on its mechanism of action. This article summarizes related research advances in China and globally and reviews the mechanism of action of luteolin in inhibiting oxidative stress, exerting an anti-inflammatory effect, regulating cell death, alleviating hepatic fibrosis, modulating lipid metabolism disorders, and regulating the gut-liver axis, as well as the application prospect of luteolin in the treatment of liver injury, in order to provide a scientific reference for further research on this compound.
4.Exploration of ethical issues in the multiple disciplinary team model for children with 46, XY disorders of sex development
Lili PAN ; Zhe SU ; Wei SU ; Ying ZU ; Chenxi LI ; Huiping SU ; Jingyu YOU ; Kexin JIN ; Rongfei ZHENG
Chinese Medical Ethics 2026;39(6):718-723
ObjectiveTo explore the ethical issues and coping strategies in the diagnosis and treatment of 46, XY disorders of sex development (46, XY DSD) under a multidisciplinary team (MDT) model, and to deepen the understanding of relevant ethical issues while accumulating experience. MethodsA retrospective collection was conducted on the data of children with 46, XY disorders of sex development under a single-center MDT model from January 2017 to December 2024, and the ethical issues involved in the diagnosis and treatment process were analyzed. ResultsA total of 24 MDT discussion documents involving 20 children were collected, including 17 with female social gender and 3 with male social gender. The age ranged from 0.5 to 15.8 years, with children under 8 years accounting for 75%. The ethical issues primarily involved how to ensure adequate informed consent, whether surgery was necessary and its timing, and whether gender selection is reasonable and its timing. Among the 10 children raised as females who completed the gender role psychological scale assessment, 5 demonstrated masculine tendencies, all of whom had pathogenic genetic variants. Furthermore, only 1 patient changed the social gender to male. ConclusionThe diagnosis and treatment of 46, XY DSD is relatively complex, requiring multidisciplinary collaboration. This process involves numerous ethical issues and necessitates adhering to medical ethical principles. Ensuring adequate informed consent is crucial, especially for younger children. The surgery and its timing selection must adhere to the principles of beneficence, non-maleficence, and leaving room for the future. Gender selection constitutes a focus and challenge in diagnosis and treatment. Psychological scales and genetic etiologies are conducive to judgments regarding gender selection, yet multiple factors in the real world may interfere with the gender selection.
5.Clinical observation of coiling dragon needling plus chin tuck against resistance training for post-stroke deglutition disorders
Liying TANG ; Lili ZHU ; Ying ZHOU ; Mei YU ; Yuan ZHANG ; Fuqing ZHANG
Journal of Acupuncture and Tuina Science 2025;23(1):49-55
Objective:To explore the clinical efficacy of coiling dragon needling plus chin tuck against resistance(CTAR)training for post-stroke deglutition disorders and its impact on surface electromyography.Methods:A total of 100 patients with post-stroke deglutition disorders were randomly divided into two groups,with 50 cases in each group.Both groups received the same CTAR training,and the observation group was treated with additional coiling dragon needling.Both groups were treated for 2 weeks.The clinical efficacy,swallowing function,average value of the maximum amplitude of surface electromyography,quality of life,and the difference in adverse reactions were compared between the two groups.Results:The total effective rate of the observation group was higher than that of the control group(P<0.05).After treatment,the standardized swallowing assessment(SSA),aspiration score,and deglutition disorders score of the videofluoroscopic swallowing study(VFSS)in both groups were significantly lower than those before treatment(P<0.05).The average value of the maximum amplitude of surface electromyography in the relaxed state,dry swallowing state,and the state of swallowing water,and the swallowing quality of life questionnaire(SWAL-QOL)score were all significantly higher than those before treatment in the two groups(P<0.05).After treatment,the SSA score,VFSS aspiration score,and VFSS deglutition disorders score in the observation group were lower than those in the control group(P<0.05),and the average value of the maximum amplitude of surface electromyography in the relaxed state,dry swallowing state,and the state of swallowing water,and the SWAL-QOL score in the observation group were higher than those in the control group(P<0.05).There was no significant difference in the incidence of adverse reactions between the two groups(P>0.05).Conclusion:Compared to CTAR treatment alone,coiling dragon needling plus CTAR treatment can enhance the clinical efficacy in treating patients with post-stroke deglutition disorders,enhancing the contraction ability of swallowing muscles and improving their swallowing function and quality of life.
6.Diagnostic value of video-electroencephalography combined with CRP and PCT in infectious diseases of the central nervous system with convulsions
Haiyan ZHANG ; Lili WU ; Yuanyuan YIN ; Ying ZHANG
China Medical Equipment 2025;22(4):43-47
Objective:To analyze the diagnostic value of video-electroencephalography(VEEG)combined with C-reactive protein(CRP)and procalcitonin(PCT)in infectious diseases of the central nervous system with convulsions.Methods:This study included 100 pediatric patients with convulsions who were suspected as central nervous system infection at Bengbu First People's Hospital from May 2022 to May 2024.VEEG and laboratory tests were performed on them.These patients were divided into the infection group(36 cases)and the non-infection group(64 cases)according to whether they were judged as central nervous system infection by using testing cerebrospinal fluid.The abnormal situation of VEEG test,CRP and PCT levels of two groups were compared,and the consistency between the diagnostic result of VEEG combined with CRP and PCT,and the tested result of cerebrospinal fluid was further analyzed.The receiver operating characteristic(ROC)curve was adopted to analyze the diagnostic efficiency of VEEG combined with CRP and PCT.Results:In 36 pediatric patients of infection group,2 cases(5.56%)were within normal range of VEEG examination,and 11 cases(30.56%)were at cutoffvalue,and 23 cases(63.89%)were abnormal.In 64 pediatric patients of non-infection group,54 cases(84.38%)were normal,and 6 cases(9.38%)were within normal range,and 3 cases(4.68%)were at cutoffvalue,and 1cases(1.56%)were abnormal.The abnormal results of the infection group were significantly higher than those of the non-infection group(Z=8.81,P<0.05).The CRP and PCT levels of infection group were significantly higher than those of non-infection group(t=6.859,5.410,P<0.05).There was higher consistency between the diagnosis of VEEG combined with CRP and PCT,and the tested result of cerebrospinal fluid(Kappa=0.867).The ROC analysis showed that the AUC value,sensitivity and specificity of VEEG combined with CRP and PCT in diagnosing the central nervous system infection with convulsions were respectively 0.963,86.11%and 98.44%,which were higher than those of the single VEEG diagnosis.Conclusion:There are significant differences in VEEG abnormalities,CRP and PCT levels between pediatric patients with central nervous system infection with convulsions,and pediatric patients without central nervous system infection with convulsions.The diagnosis of applying VEEG combined with CRP and PCT has higher diagnostic efficiency in clinical work.
7.Effects of umbilical moxibustion combined with a multi-component exercise program on perioperative rehabilitation of frail elderly patients with intertrochanteric femoral fractures
Ying LI ; Lan WEI ; Ning ZHAO ; Lili CHEN ; Jing ZHANG
Chinese Journal of Trauma 2025;41(10):998-1007
Objective:To explore the effects of umbilical moxibustion combined with a multi-component exercise program on perioperative rehabilitation in frail elderly patients with intertrochanteric fractures so as to provide evidence-based basis for optimizing clinical rehabilitation nursing pathways.Methods:A quasi-experimental study design was used. Convenience sampling was applied to frail select elderly patients with intertrochanteric femoral fractures, who were admitted to Zhengzhou Orthopedic Hospital from January to October 2024. According to their admission time, the patients were divided into four groups: conventional nursing group, umbilical moxibustion group (conventional nursing+umbilical moxibustion), exercise group (conventional nursing+multicomponent exercise), and combination group (conventional nursing+umbilical moxibustion combined with multicomponent exercise), with 31 patients in each group. All the patients underwent internal fixation with proximal femoral nail antirotation under anesthesia. The umbilical moxibustion protocol focused on Shenque acupoint, combined with liver-kidney-nourishing Chinese herbal powder, with each session lasting 20 minutes. The multi-component exercise program was designed in phases, including resistance, balance, and aerobic training. The Harris hip score (HHS), Tilburg frailty indicator (TFI), falls efficacy scale-international (FES-I), and Chinese version of the short form-36 health survey (SF-36) scores were compared among the four groups preoperatively, at 1 month, 3 months postoperatively, and at the last follow-up.Results:A total of 124 elderly patients were involved, including 50 males and 74 females, aged 60-80 years [(71.8±5.0)years]. All the patients were followed up for 6 months. The results of generalized estimating equation showed that the HHS, TFI, FES-I, and SF-36 scores differed significantly in terms of group effect, time effect, and interaction effect among the four groups across all the time points ( P<0.05). Simple effect analysis indicated no significant differences in preoperative HHS, TFI, FES-I, or SF-36 scores among the four groups ( P>0.05). The HHS scores at 1 and 3 months postoperatively were (54.3±5.1)points and (65.7±6.3)points in the umbilical moxibustion group, (61.3±5.3)points and (74.5±6.8)points in the exercise group, and (66.5±5.2)points and (86.3±6.7)points in the combination group, which were all significantly higher than (50.2±4.5)points and (60.7±5.1)points in the conventional nursing group ( P<0.05), with the combination group showing higher scores than the umbilical moxibustion group and exercise group ( P<0.05). At the last follow-up, the HHS score was (77.3±6.0)points in the conventional nursing group and (77.0±5.9)points in the umbilical moxibustion group ( P>0.05); the score was (83.8±4.7)points in the exercise group and (91.4±3.5)points in the combination group, which were significantly higher than that in the conventional nursing group ( P<0.05), with the combination group showing higher score than those in the umbilical moxibustion group and exercise group ( P<0.05). At 1 month, 3 months postoperatively, and at the last follow-up, the TFI total scores were 5.0(3.0, 7.0)points, 3.0(1.0, 5.0)points, and 3.0(0.0, 4.0)points in the umbilical moxibustion group, 4.0(1.0, 6.0)points, 2.0(0.0, 5.0)points, and 2.0(0.0, 3.0)points in the exercise group, and 2.0(0.0, 5.0)points, 1.0(0.0, 2.0)points, and 0.0(0.0, 1.0)points in the combination group, which were all significantly lower than those in the conventional nursing group [7.0(5.0, 10.0)points, 7.0(5.0, 9.0)points, and 6.0(5.0, 8.0)points, respectively] ( P<0.05), with the combination group showing lower scores than the umbilical moxibustion group and exercise group ( P<0.05). The physical and social dimension scores of TFI at 1 month, 3 months postoperatively, and at the last follow-up were significantly lower in the umbilical moxibustion group, exercise group, and combination group than those in the conventional nursing group ( P<0.05), with the combination group showing the lower scores than those in the umbilical moxibustion group and exercise group ( P<0.05). At 1 month postoperatively, no statistically significant difference was observed among the four groups in the psychological dimension scores of TFI ( P>0.05). At 3 months postoperatively and at the last follow-up, the psychological dimension scores of TFI in the umbilical moxibustion group, exercise group, and combination group were all lower than those in the conventional nursing group ( P<0.05), with the combination group showing the lower scores than those in the umbilical moxibustion group and exercise group ( P<0.05). At 1 month, 3 months postoperatively, and at the last follow-up, the FES-I scores were (45.3±4.3)points, (40.9±3.9)points, and (33.9±2.8)points, respectively in the umbilical moxibustion group, (38.7±3.8)points, (32.9±3.6)points, and (30.3±2.2)points, respectively in the exercise group, and (34.2±3.6)points, (30.2±2.6)points, and (27.9±1.3)points, respectively in the combination group, which were all significantly lower than those in the conventional nursing group [(50.9±4.9)points, (50.0±4.9)points, and (40.7±4.2)points] ( P<0.05), with the combination group showing lower scores than those in the umbilical moxibustion group and exercise group ( P<0.05). At 1 month, 3 months postoperatively, and at the last follow-up, the SF-36 physical health scores were (60.6±3.7)points, (76.1±3.5)points, and (78.4±2.6)points, respectively in the umbilical moxibustion group, (57.6±3.7)points, (78.4±3.7)points, and (80.4±3.1)points, respectively in the exercise group, and (65.7±3.1)points, (85.9±2.9)points, and (87.4±2.2)points, respectively in the combination group, which were all significantly higher than those in the conventional nursing group [(47.5±4.6)points, (65.9±4.6)points, and (68.3±4.4)points] ( P<0.05), with the combination group showing higher scores than those in the umbilical moxibustion group and exercise group ( P<0.05). At 1 month, 3 months postoperatively, and at the last follow-up, the SF-36 mental health scores were (59.7±4.1)points, (74.5±3.6)points, and (76.2±3.0)points, respectively in the umbilical moxibustion group, (59.4±4.6)points, (74.1±3.8)points, and (74.4±3.9)points, respectively in the exercise group, and (66.9±4.1)points, (81.6±3.3)points, and (79.9±3.7)points, respectively in the combination group, which were all significantly higher than those in the conventional nursing group [(52.8±5.2)points, (64.0±4.9)points, and (65.3±5.1)points] ( P<0.05), with the combination group showing higher scores than those in the umbilical moxibustion group and exercise group ( P<0.05). Conclusion:For elderly patients with intertrochanteric femoral fracture combined with frailty, umbilical moxibustion combined with a multi-component exercise program can significantly improve hip function, reduce frailty and fall risk, and enhance quality of life.
8.Artificial light at night effects glucose metabolism in the developing jawbone by inhibiting melatonin secretion
Qingming TANG ; Ying YIN ; Lili CHEN
Chinese Journal of Stomatology 2025;60(2):140-150
Objective:To investigate the effects of artificial light at night on the growth of mandibles in mice and its regulatory mechanisms.Methods:A mouse model of artificial light at night (night light pollution group) and normal lighting (normal light group) was established by controlling light exposure time, with 4 mice in each group. Micro-CT was employed to analyze the differences in bone quantities of the mandibles between the two groups. Real-time fluorescence quantitative PCR (RT-qPCR) was used to examine the expression levels of osteogenic differentiation and metabolism-related genes in the cortical bone and condylar ossification center of the mandibles. Enzyme-linked immunosorbent assay was utilized to assess the diurnal variation of serum melatonin concentrations between the two groups. The artificial light at night experimental group received daily timed injections of a defined dose of melatonin to restore the diurnal variation of serum melatonin concentration in the mice, while the normal light group and the artificial light at night control group received the same volume of saline. Bone quantities, mandibular tissue morphologies, ossification differentiation in the condylar region and cortical bone, as well as glucose metabolism expression differences were assessed across the three groups.Results:The cortical bone thickness of the mandibles in the artificial light at night group [(0.196±0.017) mm] was significantly less than that in the control group [ (0.228±0.007) mm] ( P=0.029). The bone volume fraction of the condylar ossification center in the artificial light at night group [(36.90±1.09) %] was significantly lower than that in the normal light group [(54.24±1.49) %] ( P<0.001). The length of the mandible in the artificial light at night group [(10.86±0.17) mm] was significantly shorter than that in the normal light group [(11.41±0.32) mm] ( P=0.032). RT-qPCR results indicated that the expressions of osteogenic-related genes alkaline phosphatase, liver/bone/kidney (Alpl), osteocalcin (Ocn), Runt-related transcription factor 2 (Runx2), and osterix (Osx) in the cortical bone and condylar ossification center of mice in the artificial light at night group were significantly lower than those in the normal light group (all P<0.05). The expression of metabolism-related genes protein kinase, AMP activated alpha 1 (Prkaa1), V-type proton ATPase subunit d1 (Atp6v0d1), and cytochrome C oxidase subunit Ⅳ isoform 1 (Cox4i1) in the cortical bone and condylar ossification center of mice in the artificial light at night group were also significantly reduced compared to normal mice (all P<0.01). The serum melatonin concentration peaked 8 hours after lights off in the normal light group, whereas the artificial light at night group exhibited a significantly reduced nocturnal serum melatonin concentration with no apparent peak compared to the normal light group. Micro-CT results demonstrated that after artificial light at night group mice received timed melatonin supplementation daily, the thickness of cortical bone, the bone volume fraction of the condylar ossification center, and the length of the mandible were all significantly higher than those in the artificial light at night group (all P<0.05). Histological staining results indicated that the cortical bone structure of the mandibles in the melatonin supplementation group was more organized than that of the artificial light at night group, with higher brain and muscle ARNT-like 1 (BMAL1) expression ( P=0.003). RT-qPCR results further showed that the expression levels of Prkaa1, Atp6v0d1, and Cox4i1 in the cortical bone and condylar ossification center significantly increased in the melatonin supplementation group compared to the artificial light at night group (all P<0.05), but still significantly lower than those in the normal light group (all P<0.05). Additionally, the RT-qPCR results further revealed that the expression levels of osteogenic differentiation-related genes Alpl, Ocn, and Runx2 in the cortical bone and condylar ossification center of the melatonin supplementation group were significantly higher than those in the artificial light at night group (all P<0.01). Western blotting analysis indicated that the expression levels of glucose metabolism and osteogenic-related proteins RUNX2, OSX, ATP6V0D1, and COX Ⅳ, along with the phosphorylation levels of AMPKα1/α2, were significantly higher in the melatonin supplementation group compared to the artificial light at night group (all P<0.01). Conclusions:Artificial light at night can inhibit melatonin secretion in mice, reduce glucose metabolism in mandibular tissues, and affect both intramembranous and chondrogenic ossification activities, ultimately leading to inadequate mandibular development.
9.Study on the mechanism of Cuscuta chinensis flavonoids promoting decidualization and improving recurrent spontaneous abortion
Fang FANG ; Ying CUI ; Jialü HUANG ; Lili CHEN ; Jia XU ; Yunhui WAN
China Pharmacy 2025;36(19):2379-2386
OBJECTIVE To explore the mechanism by which Cuscuta chinensis flavonoids (CCF) promote decidualization and improve recurrent spontaneous abortion (RSA). METHODS HTR-8/SVneo cells in logarithmic growth phase were randomly divided into blank group, lipopolysaccharide (LPS) group, CCF group, SGK2 inhibitor (GSK650394, abbreviated as “GSK”) group and CCF+GSK group. Each group was treated with the corresponding agents accordingly. HTR-8/SVneo cells with SGK2 knockdown were randomly divided into small interfering RNA of SGK2 (siSGK2) group and siSGK2+CCF group; additionally, blank group and LPS group were established; each group was treated with the corresponding agents accordingly. The cell survival rate, expression levels of WNK signaling pathway- and decidualization-related proteins and mRNAs, as well as mitochondrial membrane potential levels, were assessed in each group before and after SGK2 knockdown. RSA mice model was constructed and randomly divided into model group, CCF low-dose group, CCF high-dose group, GSK group, and combined dosing group, with 4 mice in each group. Other 4 normal pregnant female mice were selected as the control group. The number of implanted embryos, viable fetuses, and lost embryos in mice was recorded. The morphological changes of endometrium and decidualization were observed, and WNK signaling pathway- and decidualization-related proteins and mRNAs expressing levels as well as mitochondrial membrane potential levels were all detected. RESULTS Compared with the blank group, the cell survival rate, as well as the protein and mRNA expression levels of SGK2, WNK1, WNK4, prolactin, insulin-like growth factor- binding protein-1, oxidative stress responsive kinase 1, and Ste20-like proline-/alanine-rich kinase were significantly reduced in the LPS group (P<0.05); compared with the LPS group, the cell survival rate and the expression levels of the above- mentioned proteins and mRNAs were significantly increased in the CCF group, while the cell survival rate and the expression levels of the above-mentioned proteins and mRNAs were significantly decreased in the GSK group (P<0.05); compared with the CCF group, the cell survival rate and the expression levels of the above-mentioned proteins and mRNAs were significantly reduced in the CCF+GSK group (P<0.05). After knocking down SGK2, compared with the LPS group, the cell survival rate, red/green fluorescence intensity ratio, and the expression levels of the above-mentioned proteins and mRNAs were significantly reduced in the siSGK2 group (P<0.05); compared with the siSGK2 group, the cell survival rate, red/green fluorescence intensity ratio, and the expression levels of the above-mentioned proteins and mRNAs were significantly increased in the siSGK2+CCF group (P<0.05). The in vivo experimental results showed that CCF treatment can significantly improve the number of implanted embryos and viable fetuses in RSA model mice and reduce lost embryos, the expression levels of the above-mentioned proteins and mRNAs in endometrial tissue were significantly increased, and the red/green fluorescence intensity ratio was significantly increased (P< 0.05); the combined dosing group could reverse the effect of CCF (P<0.05). CONCLUSIONS CCF can activate SGK2, up- regulate the WNK signaling pathway, promote endometrial decidualization, and improve RSA.
10.Effect of SOX4 on the behavior of small cell lung cancer cells by regulating Beclin1 mediated autophagy
Zhongcheng Han ; Lili Ma ; Ying Su ; Jiang Liu
Acta Universitatis Medicinalis Anhui 2025;60(4):684-691
Objective :
To investigate the effect and mechanism of sex-determining region Y-box transcription factor 4(SOX4) on autophagy in small cell lung cancer(SCLC) cells.
Methods :
Human SCLC cell line NCI-H446 was transfected with small interfering RNA(siRNA) to knockdown SOX4. RT-qPCR and Western blot were used to verify the transfection efficiency. NCI-H446 cells were divided into control group, si-SOX4 group, si-SOX4+oe-Beclin1 group and oe-Beclin1 group. Western blot analysis was performed to detect the ratio of microtubule-associated protein 1 light chain 3(LC3)-Ⅱ/LC3-Ⅰ expression and the expression of Beclin1 and p62 in different groups of cells. The transcriptional regulation of Beclin1 by SOX4 was detected by dual luciferase reporter assay and chromatin immunoprecipitation PCR(ChIP-PCR) assay. CCK-8 assay was used to detect the proliferation ability of NCI-H446 cells in different groups. Flow cytometry was used to detect the apoptosis rate of NCI-H446 cells in different groups. Transwell assay was performed to determine the cell migration and invasion ability in different groups.
Results:
Compared with the control group or the si-NC group, the relative mRNA and protein expression level of SOX4 in si-SOX4 group were down-regulated(P<0.05), and the ratio of LC3-Ⅱ/LC3-Ⅰ protein expression and the relative protein expression level of Beclin1 in si-SOX4 group decreased(P<0.05), the relative protein expression of p62 increased(P<0.05). The relative luciferase activity of Beclin1 WT in the si-SOX4 group was lower than that in the si-NC group(P<0.05); the relative enrichment of Beclin1 promoter in the Anti-SOX4 group was higher than that in the Anti-Ig G group( P<0. 05). Compared with control group,cell proliferation activity decreased,cell apoptosis rate increased,migration number and invasion number decreased in si-SOX4 group( P<0. 05). Compared with the si-SOX4 group,the ratio of LC3-Ⅱ/LC3-Ⅰ protein expression and the relative protein expression of Beclin1 increased in si-SOX4 + oe-Beclin1 group,while the relative protein expression of p62 decreased,cell proliferation activity increased,apoptosis rate decreased,migration number and invasion number increased( P<0. 05).
Conclusion
Down-regulation of SOX4 can inhibit autophagy,decrease proliferation activity of NCI-H446 cells,and inhibit cell migration and invasion by inhibiting Beclin1 expression.


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