1.Effects of acupuncture on the hypothalamic-pituitary-ovarian axis and FSH/cAMP signaling pathway in aged rats.
Yaoyao ZHU ; Yaqian YIN ; Huanfang XU ; Li YANG ; Weixin LI ; Chenchen SU ; Rong ZHANG ; Yigong FANG
Chinese Acupuncture & Moxibustion 2025;45(2):200-208
OBJECTIVE:
To explore the mechanism of acupuncture on improving ovarian hypofunction in aged rats from two perspectives: the overall regulation of the hypothalamic-pituitary-ovarian (HPO) axis and the local ovarian follicle stimulating hormone (FSH)/cyclic adenosine monophosphate (cAMP) signaling pathway.
METHODS:
Six 3-month-old female SPF-grade Sprague-Dawley (SD) rats were selected as the blank group. Another twelve 9-month-old female SD rats were randomly divided into a model group and an acupuncture group, with six rats in each. The acupuncture group received acupuncture at "Baihui" (GV20), "Guanyuan" (CV4), and bilateral "Ciliao" (BL32) for 20 min per session, once every other day, for a total of 10 sessions. Vaginal smear tests were performed daily to observe the estrous cycle of the rats. Ovarian morphology was observed using HE staining, and follicles at various stages were counted. ELISA was used to detect levels of serum FSH, luteinizing hormone (LH), estradiol (E2), anti-müllerian hormone (AMH), hypothalamic gonadotropin-releasing hormone (GnRH), pituitary FSH and LH, and ovarian cAMP. Immunohistochemistry and Western blot were used to detect the protein expression of ovarian cAMP protein kinase catalytic subunit, FSH receptor (FSHR), and P450. Real-time quantitative PCR was used to measure mRNA expression levels of FSHR and P450 in ovarian tissue.
RESULTS:
Compared with the blank group, the model group showed an increased rate of estrous cycle disorder (P<0.01), reduced granulosa cell layers with blurred boundaries and disordered arrangement, decreased numbers of developing follicles at all stages, and increased numbers of atretic follicles (P<0.01); the serum levels of FSH and LH were increased (P<0.01), while E2 and AMH levels were decreased (P<0.01); the hypothalamic GnRH and pituitary FSH and LH levels were elevated (P<0.01), and ovarian cAMP level was decreased (P<0.01); the positive expression and protein expression of ovarian P450, cAMP protein kinase catalytic subunit, and FSHR were reduced (P<0.01), and ovarian FSHR and P450 mRNA expression was decreased (P<0.01). Compared with the model group, the acupuncture group showed a reduced rate of estrous cycle disorder (P<0.01), clear granulosa cell margins, increased numbers of primordial and secondary follicles, and decreased numbers of atretic follicles (P<0.01); the serum FSH and LH levels were decreased (P<0.01, P<0.05), while E2 and AMH levels were increased (P<0.05, P<0.01); the hypothalamic GnRH and pituitary FSH and LH levels were decreased (P<0.01, P<0.05), and ovarian cAMP level was increased (P<0.01); the positive expression and protein expression of ovarian P450, cAMP protein kinase catalytic subunit, and FSHR were elevated (P<0.01), and ovarian FSHR and P450 mRNA expression was increased (P<0.01).
CONCLUSION
Acupuncture could delay ovarian hypofunction in aged rats, possibly through regulating the HPO axis and the FSH/cAMP signaling pathway.
Animals
;
Female
;
Rats
;
Rats, Sprague-Dawley
;
Follicle Stimulating Hormone/genetics*
;
Acupuncture Therapy
;
Ovary/metabolism*
;
Signal Transduction
;
Humans
;
Cyclic AMP/metabolism*
;
Hypothalamo-Hypophyseal System/metabolism*
;
Aging/metabolism*
;
Hypothalamus/metabolism*
;
Pituitary Gland/metabolism*
;
Gonadotropin-Releasing Hormone/metabolism*
2.Dosage of 4-Vinylcyclohexene Diepoxide in Induction of Premature Ovarian Insufficiency in Rats
Weixin LI ; Pengfei DU ; Yaoyao ZHU ; Chenchen SU ; Huanfang XU ; Li YANG ; Xiaojing SONG ; Yigong FANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):72-79
ObjectiveTo compare the effects of different doses and withdrawal time of 4-vinylcyclohexene diepoxide (VCD) on the reproductive endocrine levels of female rats, and to explore the effective, stable, and safe dosage of VCD for constructing a premature ovarian insufficiency (POI) rat model. MethodSD rats with regular estrous cycles were randomly divided into three groups: blank group, low-dose VCD group (80 mg·kg-1·d-1), and high-dose VCD group (160 mg·kg-1·d-1), with 24 rats in each group. After drug intervention, samples were collected on the 15th day (D15) and the 45th day (D45) after intervention. The general condition, rate of estrous cycle disturbance, serum hormone levels, ovarian histomorphology, follicle count, pregnancy outcome, and the protein and mRNA expression of transforming growth factor (TGF)-β and Smad2/3 were assessed. ResultCompared with the blank group, the low-dose VCD group showed no significant differences in the rate of estrous cycle disturbance or serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) levels. Ovarian tissue was damaged. Specifically, the number of primordial and primary follicles decreased on D15 (P<0.01), and the number of secondary follicles (P<0.01) and antral follicles (P<0.05) further decreased on D45. The litter number decreased on D15 (P<0.05), but there was no significant difference on D45. Furthermore, TGF-β protein levels increased on D15 (P<0.05) and D45 (P<0.01). The Smad2/3 levels increased on D45 (P<0.01), and TGF-β and Smad2/3 mRNA levels increased on D45 (P<0.05). Compared with the results in the blank group, the disturbance rate of the estrous cycle increased on D45 in the high-dose VCD group (P<0.01). The serum of FSH and LH increased (P<0.01), while E2 decreased (P<0.05). Ovarian tissue was damaged, and the downward trend of follicles at all levels was similar to that in the low-dose VCD group. The litter number significantly decreased on D15 and D45. TGF-β and Smad2/3 protein levels increased (P<0.05,P<0.01), and TGF-β mRNA increased on D45 (P<0.05). ConclusionHigh-dose VCD is an ideal method for constructing a POI rat model, being effective, stable, and safe.
3.Effect of Family Functioning on Adolescents'Internalizing and Externalizing Problem Behaviors:The Multiple Mediating Roles of Empathy and Emotional Competence
Yinhui PENG ; Li ZHAO ; Xiang LI ; Yu LONG ; Xinmao XU ; Huanfang ZHANG ; Lihua JIANG
Journal of Sichuan University (Medical Sciences) 2024;55(1):146-152
Objective To examine the mediating role of empathy and emotional competence in the association between family functioning and internalizing and externalizing problem behaviors among adolescents in China.Methods In this study,we used the data from the June-July 2022 survey of Chengdu Positive Child Development(CPCD)cohort.All respondents were 5th-9th graders from six primary or secondary schools in Chengdu.The Achenbach Child Behavior Checklist(CBCL),the Chinese Family Assessment Instrument(C-FAI),the empathy subscale of the Chinese version of the Interpersonal Reactivity Index(C-IRI),and the emotional competence(EC)subscale of the Chinese Positive Youth Development Scale(CPYD)were used to evaluate the respondents'internalizing and externalizing problem behaviors,family functioning,empathy,and emotional competence,respectively.The average score derived from the total score of a scale divided by the number of entries in each dimension was used as the final score of the scale.Independent samples t-tests or one-way analysis of variance(ANOVA)were performed to examine the differences in family functioning,empathy,emotional competence,and internalizing and externalizing problem behaviors across student groups with different demographic characteristics(sex,grade,and region).Pearson correlation analysis was conducted to examine the relationship between family functioning,empathy,emotional competence,and internalizing and externalizing problem behaviors.AMOS 24.0 was used to validate the hypothesized model and structural equation modeling was used to analyze the mediating effects of empathy and emotional competence between family functioning and internalizing and externalizing problem behaviors among adolescents.Results A total of 3026 eligible participants were included,including 1548(51.16%)male students and 1478(48.84%)female students.Among the respondents,798(26.37%)were 5th graders,738(24.39%)were 6th graders,567(18.74%)were 7th graders,614(20.29%)were 8th graders,and 309(10.21%)were 9th graders.In addition,2064(68.21%)of all respondents were from urban areas and 962(31.79%)were from rural areas.The results of the difference analysis showed that the differences in adolescents'internalizing and externalizing problem behaviors were statistically significant between students of different grades(P=0.004),and that the differences in family functioning and empathy scores were also statistically significant between students of different grades(all P<0.001),whereas the differences in adolescents'internalizing and externalizing problem behaviors were not statistically significant between sexes and regions(P=0.919,0.959).The results of correlation analysis showed that family functioning scores(the higher the score,the worse the family functioning)were significantly negatively correlated with empathy and emotional competence(r=-0.482,-0.432,P<0.01),and significantly positively correlated with internalizing and externalizing problem behaviors(r=0.220,P<0.01).Empathy was significantly positively correlated with emotional competence(r=0.402,P<0.01).Empathy and emotional competence were significantly negatively correlated with all the dimensions of internalizing and externalizing problem behaviors(r=-0.115,-0.305,P<0.01).Emotional competence partially mediated the relationship between family functioning and adolescents'internalizing and externalizing problem behaviors,with a mediation effect value being 0.042(95%[confidence interval]CI:0.031-0.057).Empathy and emotional competence had chain mediation effect between family functioning and adolescents'internalizing and externalizing problem behaviors,with the value of the mediation effect being 0.010(95%CI:0.007-0.014).Conclusion Family functioning influences adolescents'internalizing and externalizing problem behaviors in a direct way and through the chain-mediating roles of empathy and emotional competence.
4.Relationship Between Empathy and Depression in Adolescents:The Mediating Role of Family Functioning
Yu LONG ; Xinmao XU ; Huanfang ZHANG ; Yinhui PENG ; Linlin FAN ; Lihua JIANG ; Geyang SONG ; Li ZHAO
Journal of Sichuan University (Medical Sciences) 2024;55(3):680-686
Objective To investigate the effect of empathy on depressive symptoms in adolescents and to explore the potential mediating role of family functioning in the effect of empathy on depressive symptoms.Methods The 2022 cross-sectional data from the Chengdu Positive Child Development(CPCD)cohort were analyzed in the study.A survey was conducted in Chengdu in June 2022,involving 3 020 students in grades 5-8 from three randomly selected stratified schools.The Interpersonal Reactivity Index(IRI-C),the Chinese Family Assessment Instrument(C-FAI),and the Center for Epidemiologic Studies Depression Scale for Children(CES-DC)were used in the survey.Chi-square test or one-way analysis of variance was performed to examine the differences in various demographic characteristics(sex,grade,region,and total monthly household income)between groups of respondents,as well as the differences in family functioning,empathy,and depression.Pearson correlation coefficient was used to examine the correlation between family functioning,empathy,and depressive symptoms.Structural equation modeling and SPSS PROCESS component Model 4 were used to analyze whether family functioning played a mediating role in the effect of empathy on depressive symptoms in adolescents.Results The detection rate of depressive symptoms among survey respondents was 25.40%.The results of the difference analysis revealed significant differences in the detection rates of depressive symptoms among respondents of different grades,regions,and monthly household incomes(P<0.05).There was no significant difference in the detection rates of depressive symptoms between male and female students.There was a significant difference in the detection rate of depressive symptoms between respondents with different scores for family dysfunction and empathy ability(P<0.001).Correlation analysis results showed that empathy scores were negatively correlated with depression(r=-0.11,P<0.001),that family dysfunction was positively correlated with depression(r=0.29,P<0.001),and that empathy scores were negatively correlated with family functioning(r=-0.37,P<0.001).The mediating role of family dysfunction in the relationship between empathy and depressive symptoms was established,with the direct effect being 0.039(95% confidence interval[CI]:0.010-0.069,P<0.001)and the indirect effect value being-0.096(95% CI:-0.115--0.079,P<0.001).The direct effect value accounted for 28.89% of the total effect value,while the mediation effect value accounted for 71.11% of the total effect value.Conclusion The empathy ability of adolescents is correlated to depressive symptoms,and family functioning plays a mediating role between empathy and depressive symptoms in adolescents.It is suggested that adolescents'empathy ability and family functioning should be enhanced through multiple channels to reduce the occurrence of depressive symptoms.
5.Effects of 4-hydroxy-2(3H)-benzoxazolone on Inflammatory and Apoptosis Signaling Pathways in Non-alcoholic Fatty Liver Disease Model Rats
Wanpeng XU ; Jun LIN ; Yingqin LIANG ; Huanfang ZHOU ; Hua ZHANG ; Shizhen HUANG ; Xuemei SUN ; Xiugui WEI ; Hongyuan WANG ; Lin LIU
China Pharmacy 2021;32(11):1298-1303
OBJECTIVE:To inv estigate the effects of 4-hydroxy-2(3H)-benzoxazolone on inflammatory and apoptosis signaling pathways in non-alcoholic fatty liver disease (NAFLD)model rats. METHODS :SD rats were divided into normal control group(10 rats)and modeling group (50 rats). Normal control group was given basic diet ,and modeling group were given high-fat diet to induce NAFLD model. After modeling ,the rats were divided into normal control group ,model group ,silibinin group (26.25 mg/kg),and 4-hydroxy-2(3H)-benzoxazolone high-dose ,medium-dose and low-dose groups (100,50,25 mg/kg),with 8 rats in each group. Normal control group and modeling group were given 0.6% CMC-Na intragastrically ,and other groups were given relevant medicine 10 mL/kg intragastrically ,once a day ,for consecutive 4 weeks. After last medication ,the serum levels of albumin(ALB),total protein (TP),globulin(GLB),ALB/GLB and free fatty acid (FFA)were detected ;TUNEL staining was used to observe the apoptosis of rat hepatocytes. Western blot assay was used to detect the protein expression and phosphorylation level of inflammatory signaling pathway related proteins [Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88), nuclear factor-κB p65(NF-κB p65),NF-κB inhibitor protein(IκBα)] in liver tissue as well as the expression of apoptosis signaling pathway related proteins [B cell lymphoma 2(Bcl-2),Bax,caspase-3]. RESULTS :Compared with model group ,serum levels of TP (except for low-dose group ),GLB and FFA ,the protein expression of TLR 4(except for low-dose group ),MyD88 (except for medium-dose group )and caspase- 3,the phosphorylation levels of NF-κB p65 and IκBα protein were decreased significantly(P<0.05 or P<0.01). The ratio of A LB/GLB in serum and the ratio of Bcl- 2/Bax in liver tissue were significantly increased(P<0.05 or P<0.01),and the phenomenon of hepatocyte apoptosis was improved. CONCLUSIONS :4-hydroxy-2 (3H)-benzoxazolone can ameliorate NAFLD in rats ,and the mechanism may be associated with inhibiting the expression TLR 4/ MyD88/NF-κB signaling pathway-related proteins and apoptosis-related proteins in liver tissues.
6.Study on the Improvement Effect and Mechanism of Ilicifoliosids Alkaloid A on Non-alcoholic Fatty Liver Di- sease in Rats
Wanpeng XU ; Yingqin LIANG ; Xiugui WEI ; Hongyuan WANG ; Hua ZHANG ; Huanfang ZHOU ; Xing LIN ; Jun LIN
China Pharmacy 2020;31(16):1955-1960
OBJECTIVE:To s tudy improvement effect and mech anism of ilicifoliosids alkaloid A (HBOA)on non-alcoholic fatty liver disease in rats. METHODS :SD rats were randomly divided into blank control group ,model group ,Silybin capsule group(positive control ,26.25 mg/kg),HBOA high-dose ,medium-dose and low-dose groups (100,50,25 mg/kg),with 10 rats in each group. Except that blank control group fed normal feed ,the other groups were continuously fed with high-fat diet for 8 weeks to induce non-alcoholic fatty liver disease model. Form the 9th week ,blank control group and model group were given constant volume of 0.6% CMC-Na solution ,and administration groups were given corresponding drugs by intragastric admini- stration,once a day ,for consecutive 4 weeks. The general information of rats were observed and the body weight increase ,organ (liver,kidney and spleen )indexes were calculated ;the contents of AST ,ALT,TC,TG and NEFA in liver tissue were detected , and SOD,GSH-Px activities and MDA content in the serum were also determined. The protein expression of PPARα in liver tissue was detected by immunohistochemistry. RESULTS :Compared with blank control group ,the body mass increase and liver index of rats in model group were increased significantly (P<0.01);fat deposition could be observed in the liver ;the activities of SOD and GSH-Px in serum were reduced significantly ,and the contents of MDA ,the contents of AST ,ALT,TC,TG and NEFA in liver tissue were significantly increased ,and the protein expression of PPARα was decreased significantly(P<0.01). Compared with model group ,the body mass increase and liver index of the rats were decreased significantly in administration groups (P<0.05 or P<0.01),liver fat deposition was improved ,the activity of SOD and GSH-Px in serum (except for HBOA low-dose group )were increased significantly while MDA content ,the contents of AST ,ALT,TC(except for HBOA low-dose group ),TG(except for HBOA low-dose group ) and NEFA in liver tissue were decreased significantl y,while protein expression of PPAR α 15177460685@163.com was increased significantly (P<0.05 or P<0.01). Some of the above indexes of HBOA high-dose group were 电话:0771-5302433。E-mail:junlin898@126.com significantly better than HBOA medium- and low-dose group(P<0.05). CONCLUSIONS :HBOA has a certain improvement effect on non-alcoholic fatty liver disease in rats caused by high-fat diet ,and its mechanism may be related to improving lipid metabolism disorders ,anti-oxidative stress and up-regulating the expression of PPARα.
7.The relationship between compliance of analgesic drugs and self-management efficacy in patients with lung cancer
Hongxia HUA ; Huanfang ZHOU ; Yanwen XU
Chinese Journal of Modern Nursing 2019;25(1):85-88
Objective? To explore the relationship between compliance of analgesic drugs and self-management efficacy in patients with lung cancer, and to provide reference for formulating targeted interventions. Methods? A total of 287 cases of lung cancer patients in the Second Hospital of Wuxi Affiliated to Nanjing Medical University from January 2015 to December 2017 were recruited by convenience sampling method. General information questionnaire, Morisky Medication Adherece Scale and the Chinese version of Strategies Used by People to Promote Health (C-SUPPH) were applied to investigate the level of self-management efficacy and medication compliance status, so as to analyze the relationship between self-management efficacy and medication compliance. Results? The score of self-management efficacy of the 287 lung cancer patients was (91.58±28.06), and the score of medication compliance was (6.04±1.23). The correlation analysis showed that the positive attitude dimension, the stress reduction dimension and the making decisions dimension in C-SUPPH were all positively correlation with medication compliance (r=0.607, 0.497, 0.374; P< 0.01). Multivariate linear regression analysis showed that age, comorbidity and self-management efficacy were the influencing factors of medication compliance of analgesic drugs in patients with lung cancer (P<0.01). Conclusions? The compliance of lung cancer patients with analgesics needs to be improved. Patient's medication compliance increases with the self-management efficacy. Therefore, health education can strengthen patient's self-management efficacy, so as to improve patient's medication compliance.
8.Comments on " " from .
Huisheng YANG ; Xiaotong LI ; Jie SHANG ; Huanfang XU ; Yaqian YIN ; Mingzhao HAO ; Yigong FANG
Chinese Acupuncture & Moxibustion 2018;38(12):1341-1346
In recent years, acupuncture has been used widely as an adjuvant treatment for the in vitro fertilization (IVF). " " published in on May 15, 2018, the research findings do not support the use of acupuncture to improve the rate of live births among the women undergoing IVF. In order to avoid the misunderstanding of the scholars at home and abroad for the clinical effects of acupuncture on IVF assistance, the authors put forward the doubts after the analysis from the following 3 aspects, including the acupuncture scheme, outcomes and result explanation. Additionally, the thinking and suggestions are proposed for the future development of the clinical trials of acupuncture IVF assistance in terms of selecting the proper participants, being abided by the standards of the evidence-based medicine, designing multi-acupuncture schemes and setting up the rational control, as well as conducting the overall analysis of the trial data.
Acupuncture
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American Medical Association
;
Embryo Transfer
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Female
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Fertilization in Vitro
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Humans
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Live Birth
;
Pregnancy
;
Pregnancy Rate
;
United States
9.Acupuncture with regulating menstruation to promote pregnancy for diminished ovarian reverse: a prospective case series study.
Xiaotong LI ; Huanfang XU ; Yigong FANG ; Jie SHANG ; Huisheng YANG ; Xuelin ZHOU ; Minglong ZHANG
Chinese Acupuncture & Moxibustion 2017;37(10):1061-1065
OBJECTIVETo investigate the effect and safety of acupuncture with regulating menstruation to promote pregnancy for diminished ovarian reverse (DOR).
METHODSAccording to prospective case series, 46 patients were observed and finally 40 cases were included. The acupoints were ① Baihui (GV 20), Shenting (GV 24), Guanyuan (CV 4) and bilateral Benshen (GB 13), Huangshu (KI 16), Dahe (KI 12), Luanchao (Extra), Zusanli (ST 36), Sanyinjiao (SP 6), Taixi (KI 3), Taichong (LR 3) and ② bilateral Shenshu (BL 23) and Ciliao (BL 32). The points in the two groups were used alternately. Acupuncture was given for 3 courses, 12 times as a course and 3 times a week. Before and after treatment, and 3 months after treatment, follicle-stimulating hormone (FSH), follicle-stimulating hormone/luteinizing hormone (FSH/LH), estradiol (E), antral follicle count (AFC) and TCM symptom score were observed. The safety was evaluated.
RESULTSCompared with before treatment, the levels of FSH, FSH/LH decreased, and the levels of Eand AFC increased after treatment and at follow-up (all<0.05). And the TCM symptom scores were significantly lower than those before treatment (both<0.05). The rate of pregnancy after treatment was 15% (6/40). There was no infection and organ injury.
CONCLUSIONAcupuncture with regulating menstruation to promote pregnancy can safely improve the ovarian reserve of patients with DOR.
10.Influence of EGFR in B[a]P-induced SIRT1 activation in human bronchial epithelial cells
Min ZHANG ; Yunqin CUI ; Huanfang SONG ; Jianyi LYU ; Xiaohong XU ; Jimin GAO
Journal of Jilin University(Medicine Edition) 2016;42(4):731-736
Objective:To investigate the role of epidermal growth factor receptor (EGFR)in the regulation of B[a]P-induced silent information regulator 1 (SIRT1 ) activation in the human bronchial epithelial cells (BEAS-2B),and to clarify the relationship between EGFR/SIRT1 signal transduction pathway and the occurrence and development lung cancer.Methods:The prediction analysis of transcriptional factor binding sites of SIRT1 was performed by MOTIF SearchTM software.The primary cultivated BEAS-2B cells were divided into control group (without B[a]P exposure)and B[a]P groups (the cells were exposed to B[a]P for 6,12,24,48 h).RT-PCR and Western blotting method were carried out to detect the EGFR mRNA and protein expression levels.The BEAS-2B cells transfected with SIRT1 promotor luciferase reporter gene plasmid were treated with human epidermal growth factor (hEGF)and Genistein (tyrosine protein kinase inhibitor)for 24 h,the morphology of BEAS-2B cells was observed by inverted microscope, and luciferase reporter assay was used to test the SIRT1 transcriptional activity.The human lung tissue biopsies were acquired and the immunohistochemical analysis was used to determine the EGFR protein expression level.Results:The transcriptional factor binding sites of SIRT1 contained EGF, 2Fe-2S and vWF.Compared with control group,the expression levels of EGFR mRNA in B [a]P groups were increased in a time-dependent manner,and reached the peak at 12 h (P < 0.05);the EGFR protein expression levels were also increased (P <0.05).The SIRT1 luciferase activity in hEGF group was increased compared with control group (P <0.05);when hEGF and B[a]P worked together,the SIRT1 luciferase activity was increased even further (P <0.001). The cells showed arrangement and morphologic changes gradually when the B[a]P concentration was above 30 μmol · L-1. Genistein (30 μmol · L-1 )inhibited the increase of SIRT1 luciferase activity induced by B [a]P ,and there was significant difference compared with control group (P <0.05).The immunohistochemistry results showed that EGFR expression level in lung cancer tissue was higher than that in normal lung tissue (P < 0.001).Conclusion:EGFR can regulate the B [a]P-induced SIRT1 expression in BEAS-2B cells,and to cause lung chronic inflammation;EGFR/SIRT1 signal transduction pathway may play a role in the occurrence and development of lung cancer.

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