ObjectiveTo explore the regulation of hypothalamic-pituitary-gonadal （HPG） axis by modified Erxian decoction in rats with perimenopausal syndrome （PMS） and to further analyze the expression of proteins related to the silent information regulator 1 （SIRT1）/hypothalamic kisspeptin （Kisspeptin）/gonadotropin-releasing hormone （GnRH） signaling pathway in the arcuate nucleus region （ARC） of the hypothalamus， so as to reveal the potential target of action and molecular biological mechanism of modified Erxian decoction for the treatment of perimenopausal syndrome. MethodsAn animal model was established via the incomplete castration method， with successful modeling confirmed by the exfoliated cervical cell smear method. The 48 rats were divided into six groups based on the randomization principle after successful modeling， including a sham operation group， a model group， an estradiol valerate group （0.09 mg∙kg^-1∙d^-1）， high-， medium-， and low-dose modified Erxian decoction groups （7.614， 3.807，1.903 5 g∙kg^-1∙d^-1）， with 8 rats in each group. The estradiol valerate group and the high-， medium- and low-dose modified Erxian decoction groups were continuously administered by gavage for 28 days， and the indicators were detected 24 hours after the last administration. Body weights and uterine indices were measured. The pathological changes of the uterus were observed by hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was performed to measure the levels of estradiol （E₂）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and gonadotropin-releasing hormone （GnRH）. Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to determine the expression levels of SIRT1， Kisspeptin， kisspeptin receptor （GPR54）， and GnRH in the ARC region of the hypothalamus and gonadotropin-releasing hormone receptor （GnRH-R） in pituitary. ResultsCompared with the sham operation group， rats in the model group had a significantly increased body weight （P0.01）， reduced wet weight and index of uterus （P0.01）， endometrial thinning or atrophy， glandular atrophy， and a decreasing number of glands. Additionally， serum levels of E₂ and the expression of SIRT1 in the ARC region of the hypothalamus significantly decreased （P0.01）. Serum levels of FSH， LH， and GnRH， the expression of Kisspeptin， GPR54， and GnRH in the ARC region of the hypothalamus， and GnRH-R in pituitary significantly increased （P0.01）. Compared with the model group， the estradiol valerate group and the high-， medium-dose modified Erxian decoction groups had significantly reduced body weight， serum levels of FSH， LH， and GnRH， and expression of Kisspeptin， GPR54， and GnRH in the ARC region of the hypothalamus and GnRH-R in pituitary （P0.05， P0.01） and significantly increased wet weight and index of uterus， serum level of E₂， and expression of SIRT1 in the ARC region of the hypothalamus （P0.05， P0.01）. In addition， they showed thickened endometrium， increased number of endometrial glands， and improved glandular atrophy. ConclusionModified Erxian decoction regulates the function of the HPG axis through multi-targets， and its mechanism of action may be related to the up-regulation of the expression of SIRT1 in the ARC region of the hypothalamus， the inhibition of the over-activation of the Kisspeptin/GnRH signaling pathway， the regulation of the expression of GnRH-R in the pituitary， the restoration of secretion balance of gonadotropins， and the elevation of the estrogen level. This study provides an experimental basis for the interpretation of the scientific connotation of modified Erxian decoction in the treatment of perimenopausal syndrome and a theoretical reference for the development of a novel therapeutic strategy based on the SIRT1/Kisspeptin/GnRH pathway.

Based on the androgen receptor(AR)/mammalian target of rapamycin(mTOR)signaling pathway, the effects of Xihuang Pills-medicated serum on the proliferation and apoptosis of prostate cancer LNCaP cells were investigated. The drug-containing serum of SD rats was prepared by intragastric administration of Xihuang Pills suspension. The effects of low-, medium-, and high-dose Xihuang Pills-containing serum on the in vitro proliferation of LNCaP cells were detected by cell counting kit-8(CCK-8). Flow cytometry was used to detect the apoptosis level of LNCaP cells after intervention with different concentrations of Xihuang Pills. Protein expression of cleaved cysteinyl aspartate-specific proteinase caspase-3(cleaved caspase-3), B-cell lymphoma-2(Bcl-2), and AR as well as the phosphorylation level of mTOR protein were detected by Western blot. The results showed that compared with the blank serum, the drug-medicated serum could blunt the activity of LNCaP cells. Low-, medium-, and high-dose Xihuang Pills-containing serum could significantly increase the cell apoptosis rate, increase the expression of cleaved caspase-3 protein, decrease the expression of Bcl-2 protein, reduce the expression of AR protein, and down-regulate the level of phosphorylated mTOR(p-mTOR). To study the effect of Xihuang Pills on the growth of LNCaP cells in vivo, different doses of Xihuang Pills were used to intervene in the subcutaneous graft model in nude mice inoculated with LNCaP cells. The expression levels of AR, mTOR, p-mTOR, Bcl-2, and cleaved caspase-3 were detected by Western blot. The results showed that the volumes of subcutaneous graft tumor in the low-dose, medium-dose, and high-dose Xihuang Pills groups significantly decreased compared with that in the model group. The weight of subcutaneous transplanted tumor in each group with drug intervention was significantly lower than that in the model group. Compared with the model group, the low-dose, medium-dose, and high-dose Xihuang Pills groups showed increased cleaved caspase-3 protein expression, decreased Bcl-2 and AR protein expression, and reduced p-mTOR protein expression. Further experiments showed that AR agonist R1881 could block the anti-proliferation and pro-apoptotic effects of Xihuang Pills. The mechanism of Xihuang Pills against prostate cancer is related to the inhibition of the AR/mTOR signaling pathway, inhibition of LNCaP cell proliferation, and induction of apoptosis in cancer cells.
Humans ; Male ; Mice ; Rats ; Animals ; Caspase 3/metabolism* ; Mice, Nude ; Cell Line, Tumor ; Rats, Sprague-Dawley ; Signal Transduction ; TOR Serine-Threonine Kinases/metabolism* ; Prostatic Neoplasms/pathology* ; Cell Proliferation ; Apoptosis ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; Mammals/metabolism*

This study aims to explore the effect of Buyang Huanwu Decoction glycosides on the inflammatory response of apolipoprotein E~(-/-)(ApoE~(-/-)) mice and RAW264.7 cells through nuclear factor kappa-B(NF-κB) signaling pathway. In the in vivo experiment, ApoE~(-/-) mice were fed with high-fat diets for 12 weeks to induce the animal model of atherosclerosis, and 75 μg·mL~(-1) oxidized low-density lipoprotein(Ox-LDL) incubated RAW264.7 cells for 24 h to establish the atherosclerosis cell model. Automatic biochemical analyzer, hematoxylin-eosin(HE) staining, enzyme-linked immunosorbent assay(ELISA), Western blot, and droplet digital polymerase chain reaction(PCR) were used to determine the blood lipid levels, aortic intimal thickness, inflammatory factor content, NF-κB pathway-related proteins, and mRNA expression levels, and evaluate arterial atherosclerotic lesions and anti-atherosclerotic mechanisms of the drug. The model of atherosclerosis was successfully established in ApoE~(-/-) mice after 12 weeks of feeding with high-fat diets. In the model group, the plasma levels of total cholesterol(TC), triglyceride(TG), and low-density lipoprotein cholesterol(LDL-C) were increased(P<0.01), the intima of the blood vessels was thickened, the levels of inflammatory factors tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) were increased, and the protein and mRNA expressions of NF-κB and inhibitor of NF-κB(IκBα) were significantly increased as compared with the control group. Compared with the model group, the high-dose Buyang Huanwu Decoction glycoside group decreased the plasma levels of TC, TG, and LDL-C, reduced the plaque area and thickness and the content of inflammatory factor TNF-α, and inhibited the protein and mRNA expressions of NF-κB and IκBα, with the effect same as Buyang Huanwu Decoction. In the in vivo experiment, 75 μg·mL~(-1) Ox-LDL stimulated RAW264.7 cells for 24 h to successfully establish a foam cell model. As compared with the control group, the nuclear amount of NF-κB and the protein and mRNA expressions of IκBα in the model group increased. Compared with the model group, the middle-dose and high-dose Buyang Huanwu Decoction glycoside groups decreased the nuclear amount of NF-κB and the protein and mRNA expressions of IκBα. The above results show that the glycosides are the main effective substances of Buyang Huanwu Decoction against atherosclerosis, which inhibit the NF-κB pathway and reduce the inflammatory response, thus playing the role against atherosclerotic inflammation same as Buyang Huanwu Decoction.
Mice ; Animals ; NF-kappa B/metabolism* ; NF-KappaB Inhibitor alpha/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Glycosides/pharmacology* ; Cholesterol, LDL ; Atherosclerosis/genetics* ; Signal Transduction ; Inflammation/drug therapy* ; Interleukin-6 ; Apolipoproteins E/pharmacology* ; RNA, Messenger/metabolism*

OBJECTIVES: Rheumatoid arthritis (RA) is a chronic autoimmune disease. MicroRNA has been shown to play an important role in RA. MicroRNA-124a (miR-124a) has anti-proliferative and anti-inflammatory effects in RA fibroblast synovial cells. This study aims to explore the effects of miR-124a overexpression on arthritis in collagen-induced arthritis (CIA) mice and the underlying mechanisms. METHODS: Bovine type II collagen and complete Ferris adjuvant were used to induce CIA model from DBA/1 mice. Twenty-eight days after initial immunization (D28), CIA mice were randomly divided into a model group, a miR-124a treatment group, and a negative control (NC) group. Physiological saline, miR-124a agomir, and miR-124a agomir NC were injected into the skin at the tail root of mice every 3 days for 4 times, respectively. The degree of joint swelling and arthritis index of mice were recorded accordingly. Sixty-three days after initial immunization (D63), the mice were sacrificed to obtain the synovial tissue of ankle joint. HE staining was used to observe the proliferation of synovial cell, infiltration of inflammatory cell, pannus, and bone erosion of synovial tissues; TUNEL staining was used to detect cell apoptosis; qRT-PCR was used to detect the mRNA expression of miR-124a, phosphatidylinositol-3-kinase catalytic subunit alpha (PIK3CA) and its downstream genes Bcl-2 and Bax. Immunohistochemistry was used to detect the protein expression of PIK3CA, Bcl-2, and Bax protein in synovial tissues of each group. RESULTS: Different degrees of swelling presented in the paws of DBA/1 mice at D28, which indicated the CIA model was constructed successfully. Forty-eight days after initial immunization (D48), the paws of mice in the miR-124a treatment group were only slightly red and swollen, while the paws of mice in the model group and the NC group were obviously red and swollen. The arthritis index of mice in the miR-124a treatment group were decreased significantly compared to the NC group at D51, D53, D59, and D62 (51, 53, 59, 62 days after initial immunization) (all P<0.05). Sixty-three days after initial immunization (D63), HE staining indicated that the scores of synovial cell proliferation, inflammatory cell infiltration, synovial pannus, and bone erosion were significantly reduced in the miR-124a treatment group (P<0.05 or P<0.01), while cell apoptosis was increased in the miR-124a treatment group compared with the model group and NC group (P<0.01 or P<0.001). Besides, the expression of miR-124a and Bax in the synovial tissue in miR-124a treatment group was significantly higher than those in the model group and NC group (P<0.01 or P<0.001), while the expressions of PIK3CA and Bcl-2 were decreased (P<0.05 or P<0.01 or P<0.001), and the ratio of Bcl-2 to Bax was significantly decreased (P<0.01 or P<0.001). CONCLUSIONS Overexpression of miR-124a can reduce arthritis in CIA mice bacause it could promote synovial cell apoptosis and inhibit synovial cell proliferation via targeting PIK3CA and regulating its downstream pathways.
Animals ; Arthritis, Experimental/metabolism* ; Arthritis, Rheumatoid/genetics* ; Cattle ; Cell Proliferation ; Class I Phosphatidylinositol 3-Kinases/metabolism* ; Mice ; Mice, Inbred DBA ; MicroRNAs/metabolism* ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; Synovial Membrane ; bcl-2-Associated X Protein/metabolism*

Objective:To explore the advantage of thyroid biopsy and evaluate detection in the application of thyroid cancer cervical lymph node dissection operation guidance, and provide evidence-based basis for guiding PTC patients whether to receive lateral dissection.Methods:The data of 258 patients with thyroid papillary carcinoma admitted to Department of Thyroid Surgery, Affiliated Hospital of Chengde Medical College from April 2018 to December 2019 were retrospectively analyzed. All patients were admitted to the hospital for ultrasonic examination of cervical lymph nodes. According to the examination results, thyroid biopsy and puncture eluent were performed for patients with suspicious signs of metastasis. Patients with positive test results and with metastatic signs in initial ultrasonic examination received lymph node dissection in the cervical region. All patients underwent pathological examination postoperatively to diagnose the lymph node metastasis, and to evaluate the sensitivity, specificity and accuracy of the detection of the patients with lateral clearance.The measurement data was expressed as mean±standard deviation ( Mean± SD), and the count data was expressed as n(%). The software of SPSS21.0 was used to conduct statistical analysis. Results:All 258 patients were successfully completed the diagnosis of various diagnostic methods. Combined with the postoperative pathological diagnosis results, the accuracy rate of ultrasonic diagnosis of lymph node metastasis was 74.42%, the accuracy rate of FNAC diagnosis was 82.95%, the accuracy rate of FNAC-Tg diagnosis was 87.98%, and the accuracy rate of FNAC-Tg diagnosis was 94.96%. The diagnostic sensitivity of ultrasound, FNAC, FNAC-Tg and FNAC-Tg was 73.60%, 81.72%, 91.01% and 95.83%, respectively, and the diagnostic specificity was 76.25%, 86.11%, 79.71% and 88.41%, respectively. In every four months for a time span since April 2018, the average length of hospital stay for patients with each span was (9.17±1.30), (8.39±1.21), (7.94±1.03), (7.46±0.94), (7.33±0.82) d, their neck area incidence of lymph node metastasis were 17.6%, 21.3%, 15.7%, 12.9%, 11.8%, side clear surgical patients accounted for 42.3%, 37.5%, 30.9%, 26.6%, 19.4%. The incidence of lymph node metastasis was 86.8%, 79.4%, 84.5%, 93.2% and 98.1%, respectively.Conclusion:FNAC-Tg method is used in the diagnosis of thyroid papillary carcinoma patients with high sensitivity and specificity of lymph node metastasis, which has certain value in guiding patients whether to perform lymph node dissection.

Diagnosis, Differential ; Humans ; Lung Neoplasms/genetics* ; Neoplasm Metastasis ; Neoplasms, Multiple Primary/diagnosis*

Diagnosis of Parkinson's disease (PD) based on speech data has been proved to be an effective way in recent years. However, current researches just care about the feature extraction and classifier design, and do not consider the instance selection. Former research by authors showed that the instance selection can lead to improvement on classification accuracy. However, no attention is paid on the relationship between speech sample and feature until now. Therefore, a new diagnosis algorithm of PD is proposed in this paper by simultaneously selecting speech sample and feature based on relevant feature weighting algorithm and multiple kernel method, so as to find their synergy effects, thereby improving classification accuracy. Experimental results showed that this proposed algorithm obtained apparent improvement on classification accuracy. It can obtain mean classification accuracy of 82.5%, which was 30.5% higher than the relevant algorithm. Besides, the proposed algorithm detected the synergy effects of speech sample and feature, which is valuable for speech marker extraction.

OBJECTIVE: : To study the feasibility and effect of PeriCam PSI system guiding the establishment of ischemia/reperfusion injury model in rats. METHODS: : A total of 70 adult male Sprague-Dawley rats were divided into the control group(=6), PSI monitoring group(=34) and traditional operation group(=30). Ischemia reperfusion model was established with reference to improve Zea-Longa line plug method. After the model established, the blood flow to the brain of control group, PSI monitoring group (ischemic 2 h, 24 h reperfusion) were observed and recorded respectively with PSI. The rats were then executed after 24 h, and the 2,3,5-triphenyltetrazolium chloride (TTC) staining and HE staining were used to observe the brain tissue. RESULTS: : The survival rate and modeling success rate of PSI monitoring group were higher than those of the traditional operation group(all <0.05). The blood perfusion in the brain and the distribution of blood vessels were clearly observed in the control group, and the data were normal. In 2 h ischemic group, the arterial flow was interrupted in the right cerebral artery, and the blood flow in the middle arterial blood supply was significantly decreased than that in the control group(<0.05). After the recovery of 24 h, the artery in the right side of the brain was restored to blood flow, but the blood flow in the partial supply area decreased, unable to recover to normal level. The TTC staining results indicated that there were obvious infarcts in the right brain tissue of PSI monitoring group,and the infarct area was more stable than that of the traditional operation group. The results of HE staining showed that the structure of brain tissue in the control group was normal, and the morphological rules of nerve cells were not change. While in brain tissue from PSI monitoring group, cortex and ischemia half dark stripe, nerve cell degeneration, necrosis and glial fiber disintegration, liquefaction, and light color, screen mesh in ischemic central area were observed. CONCLUSIONS : PSI system can guide ischemia reperfusion model building and improve the success rate of the model.
Animals ; Brain Ischemia ; diagnostic imaging ; surgery ; Disease Models, Animal ; Hemodynamics ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion ; instrumentation ; Reperfusion Injury ; diagnostic imaging

Objective To investigate the clinical efficacy of neoadjuvant chemotherapy (oxaliplatin +capecitabine,XELOX) for the resectable locally advanced adenocarcinoma of esophageal-gastric junction (AEG).Methods The prospective study was conducted.The clinicopathological data of 106 locally advanced AEG patients who were admitted to the Cancer Hospital of Shantou University Medical College from January 2011 to December 2014 were collected.All the patients were divided into the treatment group and control group by single blind,randomized,controlled random number table.Patients underwent preoperative neoadjuvant chemotherapy (XELOX) + surgery + postoperative adjuvant chemotherapy (XELOX) in the treatment group and surgery + postoperative adjuvant chemotherapy (XELOX) in the control group.Total gastrectomy + Roux-en-Y esophagojejunostomy + D2 lymphadenectomy were applied to patients by the same team of doctors.Observation indicators:(1) treatment situations;(2) results of postoperative pathological examination;(3) follow-up and survival situations.Follow-up using outpatient examination was performed to detect the postoperative tumor recurrence or metastasis and patients' survival up to February 2017.Measurement data with normal distribution were represented as (-x)±s,and comparisons between groups were evaluated with the t test.Measurement data with skewed distribution were described as M (range) and analyzed by the nonparametric test.Comparisons of count data were analyzed using the chi-square test.The ordinal data were compared using the nonparametric test.Survival rate and curve were respectively calculated and drawn by the Kaplan-Meier method and survivals were compared using the Long-rank method.Results One hundred and six patients were screened for eligibility,including 54 in the treatment group and 52 in the control group.(1) Treatment situations:① preoperative neoadjuvant chemotherapy:54 in the treatment group received 2-4 cycle neoadjuvant chemotherapy.During the chemotherapy,gastrointestinal reaction,grade 1-2 granulocytopenia,elevated alanine transaminase (ALT) and grade 3 granulocytopenia were detected in 21,17,8,1 patients,and 7 patients had no adverse reaction.The complete response(CR),partial response (PR),stable disease (SD) and progressive disease (PD) of neoadjuvant chemotherapy in the treatment group were detected in 4,27,20 and 3 patients,respectively.Of 54 patients in the treatment group,4,13,25 and 12 were in grade 0,1,2 and 3 of response to preoperative chemotherapy,respectively.② Surgical situations:preoperative carcinoembryonic antigen (CEA) in the treatment and control groups were respectively 4.71 μg/L (range,0.20-36.19 μg/L) and 14.09 μg/L (range,0.71-178.20 μg/L),with a statistically significant difference between groups (Z =-1.92,P＜ 0.05).All patients underwent total gastrectomy + Roux-en-Y esophagojejunostomy + D2 lymphadenectomy.Operation time in the treatment and control groups were respectively (210± 31) minutes and (195 ±26) minutes,with a statistically significant difference between groups (t =-2.45,P ＜ 0.05).Volume of intraoperative blood loss,cases with intraoperative blood transfusion,time to postoperative anal exsufflation,time to defecation,time for initial diet intake,cases with postoperative complications and duration of hospital stay were respectively (216± 172) mL,6,(4.3± 1.0) days,(4.5±0.8)days,(3.1±0.5)days,11,(15.0±5.0)days in the treatment group and (174±108)mL,4,(4.2± 1.0) days,(4.4± 0.8) days,(3.1 ± 0.5) days,15,(15.0± 5.0) days,with no statistically significant difference between groups (t=-1.01,x2 =0.36,t=-0.31,-0.88,-0.36,x2 =1.03,t=-0.38,P＞0.05).③Postoperative adjuvant chemotherapy:all the patients completed the postoperative adjuvant chemotherapy.The granulocytopenia,elevated ALT and gastrointestinal reaction occurred in 25,5,28 patients in the treatment group and 21,7,30 patients in the control group,respectively,with no statistically significant difference between groups (x2 =0.38,0.47,0.36,P＞0.05).Some of the patients were merged with multiple adverse reactions.(2) Results of postoperative pathological examination:maximum tumor dimension,cases with lymphovascular invasion,perineural invasion,T0,T2,T3,T4 (T stage),stage 0,Ⅰ,Ⅱ and Ⅲ1 (TNM stage) were respectively (3.6±1.4)cm,5,10,4,10,20,20,4,7,15,28 in the treatment group and (4.5±1.7)cm,24,30,0,2,13,37,0,1,12,39 in the control group,with statistically significant differences between groups (t=-2.88,x2 =18.14,17.30,Z=14.74,8.13,P＜0.05).(3) Follow-up and survival situations:of 54 patients in the treatment group,52 were followed up for 4-72 months,with a median time of 32 months;of 52 patients in the control group,49 were followed up for 5-71 months,with a median time of 36 months.The postoperative diseasefree survival time,1-,3-and 5-year tumor-free survival rates,postoperative overall survival time and 1-,3-and 5-year overall survival rates were respectively 26 months (range,3-72 months),79.5％,64.7％,61.3％,27 months (range,5-72 months),88.3％,69.2％ and 62.1％ in the treatment group.Seventeen patients had tumor recurrence,including 2 with intraperitoneal local recurrence and 15 with distant metastasis.The postoperative disease-free survival time,1-,3-and 5-year tumor-free survival rates,postoperative overall survival time and 1-,3-and 5-year overall survival rates were respectively 33 months (range,2-71 months),89.7％,55.4％,55.4％,33 months (range,5-71 months),91.8％,72.1％ and 59.7％ in the control group.Nineteen patients had tumor recurrence,including 8 with intraperitoneal local recurrence and 11 with distant metastasis.There was no statistically significant difference in disease-free survival and overall survival between groups (x2 =0.018,0.596,P＞0.05).There was a statistically significant difference in cases with local recurrence between groups (x2=4.41,P＜ 0.05) The tumor-free survival time and overall survival time in the treatment group were respectively 29 months (range,8-72 months),38 months (range,10-72 months) in 31 patients with CR and PR and 11 months (range,3-68 months),18 months (range,4-68 months) in 23 patients with SD and PD,showing statistically significant differences in tumor-free and overall survival times (x2=5.27,7.72,P＜0.05).Concluslon Neoadjuvant chemotherapy with oxaliplatin and capecitabine is safe and effective for patients with the resectable locally advanced AEG,it can also decrease tumor stage and reduce local recurrence,but fail to demonstrate a survival benefit.

Objective:To study the effects and safety of triamcinolone acetonide oral ointment in the treatment of erosive oral lichen planus(EOLP).Methods:62 cases with EOLP were randomly divided into 2 groups.The patients in test group(n =29) were treated by local application of triamcinolone acetonide oral ointment (TAOO),oral administration of hydmxychloroquine(HCQ) and gargle with cetylpyridinium chloride buccal tablets(CCBT),in control group (n =33) by HCQ and CCBT.The patients were followed up for 4 weeks.The changes of the erosion area and the VAS pain degree of the patients were analysed at the first visit,the second week and the fourth week follow-up respectively.The safety indexes were examined at the first visit and the fourth week follow-up respectively.Results:TAOO accelerated the erosion surface healing of EOLP in 2 weeks(P ＜ 0.05),and reduced the pain index of the patients (P＜0.05),but after 4 week treatment,no significant difference was observed between 2 groups (P ＞ 0.05).Fungal infection was found in 4 cases in test group.Conclusion:Local application of TAOO combined with oral administration of HCQ and gargle with CCBT is more effective than HCQ and CCBT in the treatment of EOLP,attention should be paid to prevent fungal infection.