1.Application Progress of Electrochemical Methods in Quality Control of Traditional Chinese Medicine
Yan-Bing PAN ; IHSAN AWAIS ; Min SHI ; Wen-Wen MA ; MURTAZA GHULAM ; Ke-Fei HU ; Jun LI ; Xian-Ju HUANG ; Han CHENG
Chinese Journal of Analytical Chemistry 2024;52(1):22-34
The quality control of traditional Chinese medicine(TCM)is the core issue to ensure the modernization,industrialization and internationalization of TCM.Compared with other detection methods,electrochemical analysis method has many advantages such as high sensitivity,fast detection speed and low cost,making it an important means of quality control for TCM and having broad development prospects.This article reviewed the research progress of electrochemical methods in quality control of TCM in recent years,discussed the application of electrochemical fingerprinting technique in identification of TCM,and comprehensively summarized the application of electrochemical technology in analyzing effective components and harmful substances in TCM,including flavonoids,alkaloids,quinones,glycosides,heavy metals and pesticide residues.Finally,the development prospects of electrochemical methods in the field of quality control of TCM were discussed.
2.Surgical strategies for atrial functional mitral regurgitation with atrial fibrillation
Nan MA ; Chunrong BAO ; Ke WEI ; Yunjiao ZHANG ; Li ZHANG ; Ju MEI
Chinese Journal of Thoracic and Cardiovascular Surgery 2024;40(10):607-611
Objective:Analysis of surgical strategies for atrial functional mitral regurgitation with atrial fibrillation.Methods:Retrospective analysis of 112 patients with mitral regurgitation and atrial fibrillation between June 2017 and January 2023. Among them, 56 cases were severe atrial functional mitral regurgitation with atrial fibrillation, and the other 56 cases were degenerative mitral regurgitation with atrial fibrillation. All patients underwent maze Ⅳ procedure and mitral valve surgery. Follow up will be conducted through outpatient follow-up and telephone calls. The condition of postoperative mitral valve is obtained through echo. The postoperative cardiac rhythm is based on the patient's conscious symptoms, electrocardiogram, 24 hour dynamic electrocardiogram.Results:The comparison of preoperative basic data shows that the age, duration of atrial fibrillation, and comorbidity of patients with atrial functional mitral regurgitation are significantly higher than those in the degenerative mitral regurgitation group. All patients successfully completed the surgery. Postoperative death occurred in 2 cases in the atrial mitral regurgitation group. The causes of death were ARDS and pulmonary infection, respectively. The main postoperative complications include bleeding, low cardiac output, pulmonary infection, and acute kidney injury. During follow-up, 43 patients (79.6%) in the atrial mitral regurgitation group maintained sinus rhythm, while 49 patients (87.5%) in the degenerative group. However, there was no statistically significant difference in the Kaplan- Meier curves. In the atrial mitral regurgitation group, there were 47 cases with no mitral regurgitation, 4 cases with mild regurgitation, and 1 case with moderate regurgitation. In the degenerative group, there were 42 cases with no mitral regurgitation, 6 cases with mild regurgitation, 1 case with moderate regurgitation, and 1 case with severe regurgitation. The risk for atrial fibrillation recurrence in the atrial mitral regurgitation is related to postoperative left atrial diameter greater than 50 mm, while in the degenerative group, atrial fibrillation recurrence is related to postoperative left atrial diameter greater than 50 mm and residual mitral regurgitation. Conclusion:Mitral valve repair combined with maze Ⅳ procedure is an effective treatment for patients with severe atrial functional mitral regurgitation and atrial fibrillation. Further improving the success rate of atrial fibrillation and reducing surgical trauma will benefit patients in the future.
3.Efficacy of surgical comprehensive therapy for 456 cases of hypopharyngeal carcinoma.
Zheng Hua LYU ; Wei XU ; Ju Ke MA ; Shou Hao FENG ; Pei Hang JING ; Xu Liang LIU ; Chao ZHOU
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2023;58(6):558-564
Objective: To retrospectively analyse the efficacy of surgerical comprehensive treatment for hypopharyngeal cancer. Methods: Four hundred and fifty-six cases of hypopharyngeal squamous cell carcinoma treated from Jan 2014 to Dec 2019 were analyzed retrospectively, including 432 males and 24 females, aged 37-82 years old. There were 328 cases of pyriform sinus carcinoma, 88 cases of posterior pharyngeal wall carcinoma, and 40 cases of postcricoid carcinoma. According to American Joint Committe on Cancer(AJCC) 2018 criteria, 420 cases were of stage Ⅲ or Ⅳ; 325 cases were of T3 or T4 stage. Treatment methods included surgery alone in 84 cases, preoperative planned radiotherapy plus surgery in 49 cases, surgery plus adjuvant radiotherapy or concurrent chemoradiotherapy in 314 cases, and inductive chemotherapy plus surgery and adjuvant radiotherapy in 9 cases. The primary tumor resection methods included transoral laser surgery in 5 cases, partial laryngopharyngectomy in 74 cases, of them 48 cases (64.9%) presented with supracricoid hemilaryngopharyngectomy, total laryngectomy with patial pharyngectomy in 90 cases, total laryngopharyngectomy or with cervical esophagectomy in 226 cases, and total laryngopharyngectomy with total esophagectomy in 61 cases. Among 456 cases, 226 cases received reconstruction surgery with free jejunum transplantation, 61 cases with gastric pull-up, and 32 cases with pectoralis myocutaneous flaps. All patients underwent retropharyngeal lymph node dissection, and high-definition gastroscopy was performed during admission and follow-up. SPSS 24.0 software was used to analyze the data. Results: The 3-year and 5-year overall survival rates were respectively 59.8%, and 49.5%. The 3-year and 5-year disease specific survival rates were respectively 69.0% and 58.8%. Total metastasis rate of retropharyngeal lymph nodes was 12.7%. A total of 132 patients (28.9%) suffered from simultaneous and metachronous multiple primary carcinoma of the hypopharynx. Multivariate Logistic regression analysis showed that T3-4 disease, cervical lymph node metastasis, retropharyngeal lymph node metastasis and postoperative adjuvant radiotherapy were independent factors affecting the prognosis of patients (all P<0.05). As of April 30, 2022, a total of 221 patients died during follow-up, of 109 (49.3%) with distant metastases, which were the main cause of death. Conclusions: The efficacy of comprehensive treatment for hypopharyngeal cancer can be improved by accurate preoperative evaluation, improved surgical resection, active retropharyngeal lymph node dissection and full process intervention of the second primary cancer.
Male
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Female
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Humans
;
Adult
;
Middle Aged
;
Aged
;
Aged, 80 and over
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Hypopharyngeal Neoplasms/pathology*
;
Carcinoma, Squamous Cell/pathology*
;
Lymphatic Metastasis
;
Retrospective Studies
;
Neck Dissection/methods*
;
Head and Neck Neoplasms/surgery*
4.Mei mini maze procedure for atrial fibrillation patients with atrial functional mitral regurgitation
Nan MA ; Chunrong BAO ; Ke WEI ; Yunjiao ZHANG ; Li ZHANG ; Ju MEI
Chinese Journal of Thoracic and Cardiovascular Surgery 2023;39(7):405-408
Objective:To study the influence of Mei mini maze procedure for atrial functional mitral regurgitation.Methods:The data of 33 patients with atrial fibrillation and atrial functional mitral regurgitation from January 2017 to June 2020 were retrospectively analyzed. All patients received Mei mini maze procedure for atrial fibrillation. The procedure is carried out thoracoscopically through the left thoracic approach. The ablation of atrial fibrillation includes bilateral circumferential pulmonary vein ablation, isolation of the left atrium posterior wall, left atrial appendage resection, ablation of Marshall's ligament and autonomic ganglion, etc. Follow-up was conducted by outpatient follow-up and telephone. Postoperative heart rhythm was recorded by the patient's symptoms, electrocardiogram, 24 h holter and other examinations. Postoperative mitral valve lesions were obtained by echocardiography.Results:33 patients successfully completed the operation. There was no conversion to thoracotomy and no perioperative death. Thirty patients(90.9%) maintained sinus rhythm at discharge. Before discharge, 16 patients had no mitral regurgitation in echocardiography, 8 patients had mild mitral regurgitation, and 9 patients had moderate mitral regurgitation. Follow-up was 1-4 years after discharge, with a mean of(2.6±1.1) years. Sinus rhythm was maintained in 23 patients(69.7%). 17 patients had no mitral regurgitation, 9 had mild mitral regurgitation, 6 had moderate, and 1 had severe mitral regurgitation. The degree of regurgitation in 25 patients was reduced compared with pre-operation, 5 patients remained unchanged, and 3 patients mitral regurgitation aggravated. Unreduced atrial functional mitral regurgitation was associated with recurrence of atrial fibrillation by Cox multivariate analysis.Conclusion:This study found a close relationship between atrial fibrillation rhythm and atrial functional mitral regurgitation. Most moderate atrial functional mitral regurgitation can be alleviated by effective treatment for atrial fibrillation. It is not recommended that patients with severe atrial functional mitral regurgitation only receive treatment for atrial fibrillation.
5.Clinical analysis of retropharyngeal lymph node metastasis in patients with hypopharyngeal squamous cell carcinoma.
Shou Hao FENG ; Zheng Hua LYU ; Ju Ke MA ; Shan Feng LIU ; Xue Wen YU ; Yu Mei WEI ; Pei Hang JING ; Xu Liang LIU ; Chao ZHOU ; Na SA ; Wei XU
Chinese Journal of Oncology 2023;45(11):955-961
Objective: To analyze the incidence and the related risk factors of retropharyngeal lymph node metastasis in patients with hypopharyngeal squamous cell carcinoma, evaluate the accuracy of preoperative enhanced CT in judging retropharyngeal lymph node metastasis, and investigate the impact of retropharyngeal lymph node metastasis on the prognosis. Methods: Retrospective analyses were made on 398 patients with hypopharyngeal squamous cell carcinoma who underwent surgery as the primary therapy and accepted retropharyngeal lymph node exploration and clearance during surgery in Shandong Provincial ENT Hospital from January 2014 to December 2019. Multivariate logistic regression analysis was used to clarify the related risk factors of retropharyngeal lymph node metastasis. Multivariate Cox regression analysis was used to investigate the impact of retropharyngeal lymph node metastasis on prognosis. The retropharyngeal lymph nodes of 218 cases with available preoperative enhanced CT images were evaluated by two experienced radiologists and compared with postoperative pathological results. Results: Retropharyngeal lymph node metastasis were confirmed in 54 of 398 (13.6%) cases according to postoperative pathology. The sensitivity and specificity of preoperative enhanced CT in the diagnosis of retropharyngeal lymph node metastasis were 34.6% and 91.1%, respectively, and the overall accuracy was 84.4%. Multivariate logistic regression analysis showed that the site of the primary lesion and pathological N stage were independent risk factors for retropharyngeal lymph node metastasis in hypopharyngeal squamous cell carcinoma. Patients with primary lesion located in the posterior wall of hypopharynx (OR=4.83, 95% CI: 1.27-18.40), N2 stage (OR=6.30, 95% CI: 2.25-17.67), and N3 stage (OR=26.89, 95% CI: 5.76-125.58) were prone to retropharyngeal lymph node metastasis. The 5-year overall survival rate of the 398 patients was 50.4%, and the 5-year disease-free survival rate was 48.3%. Multivariate Cox regression analysis showed that T stage, N stage, retropharyngeal lymph node metastasis, and radiotherapy were independent influencing factors for overall survival (T stage: HR=1.28, 95% CI: 1.06-1.54; N stage: HR=1.26, 95% CI: 1.14-1.40; retropharyngeal lymph node metastasis: HR=2.13, 95% CI: 1.47-3.08; radiotherapy: HR=0.54, 95% CI: 0.38-0.76) and disease-free survival of patients with hypopharyngeal squamous cell carcinoma (T stage: HR=1.26, 95% CI: 1.06-1.51; N stage: HR=1.25, 95% CI: 1.13-1.37; retropharyngeal lymph node metastasis: HR=2.24, 95% CI: 1.56-3.21; radiotherapy: HR=0.55, 95% CI: 0.40-0.77). Conclusions: Metastasis of retropharyngeal lymph nodes in hypopharyngeal squamous cell carcinoma is not rare. Enhanced CT is of low accuracy and limited value in diagnosing retropharyngeal lymph node metastasis. Primary lesions located in the posterior wall of the hypopharyngx, N2 stage, and N3 stage are independent high-risk factors for retropharyngeal lymph node metastasis. The prognosis of hypopharyngeal cancer patients with retropharyngeal lymph node metastasis is worse, and active surgical exploration and clearance can effectively reduce the mortality caused by retropharyngeal lymph node metastasis.
Humans
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Squamous Cell Carcinoma of Head and Neck/pathology*
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Lymphatic Metastasis/pathology*
;
Retrospective Studies
;
Carcinoma, Squamous Cell/surgery*
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Lymph Nodes/pathology*
;
Hypopharyngeal Neoplasms/surgery*
;
Prognosis
;
Head and Neck Neoplasms/pathology*
;
Neoplasm Staging
6.Clinical analysis of retropharyngeal lymph node metastasis in patients with hypopharyngeal squamous cell carcinoma.
Shou Hao FENG ; Zheng Hua LYU ; Ju Ke MA ; Shan Feng LIU ; Xue Wen YU ; Yu Mei WEI ; Pei Hang JING ; Xu Liang LIU ; Chao ZHOU ; Na SA ; Wei XU
Chinese Journal of Oncology 2023;45(11):955-961
Objective: To analyze the incidence and the related risk factors of retropharyngeal lymph node metastasis in patients with hypopharyngeal squamous cell carcinoma, evaluate the accuracy of preoperative enhanced CT in judging retropharyngeal lymph node metastasis, and investigate the impact of retropharyngeal lymph node metastasis on the prognosis. Methods: Retrospective analyses were made on 398 patients with hypopharyngeal squamous cell carcinoma who underwent surgery as the primary therapy and accepted retropharyngeal lymph node exploration and clearance during surgery in Shandong Provincial ENT Hospital from January 2014 to December 2019. Multivariate logistic regression analysis was used to clarify the related risk factors of retropharyngeal lymph node metastasis. Multivariate Cox regression analysis was used to investigate the impact of retropharyngeal lymph node metastasis on prognosis. The retropharyngeal lymph nodes of 218 cases with available preoperative enhanced CT images were evaluated by two experienced radiologists and compared with postoperative pathological results. Results: Retropharyngeal lymph node metastasis were confirmed in 54 of 398 (13.6%) cases according to postoperative pathology. The sensitivity and specificity of preoperative enhanced CT in the diagnosis of retropharyngeal lymph node metastasis were 34.6% and 91.1%, respectively, and the overall accuracy was 84.4%. Multivariate logistic regression analysis showed that the site of the primary lesion and pathological N stage were independent risk factors for retropharyngeal lymph node metastasis in hypopharyngeal squamous cell carcinoma. Patients with primary lesion located in the posterior wall of hypopharynx (OR=4.83, 95% CI: 1.27-18.40), N2 stage (OR=6.30, 95% CI: 2.25-17.67), and N3 stage (OR=26.89, 95% CI: 5.76-125.58) were prone to retropharyngeal lymph node metastasis. The 5-year overall survival rate of the 398 patients was 50.4%, and the 5-year disease-free survival rate was 48.3%. Multivariate Cox regression analysis showed that T stage, N stage, retropharyngeal lymph node metastasis, and radiotherapy were independent influencing factors for overall survival (T stage: HR=1.28, 95% CI: 1.06-1.54; N stage: HR=1.26, 95% CI: 1.14-1.40; retropharyngeal lymph node metastasis: HR=2.13, 95% CI: 1.47-3.08; radiotherapy: HR=0.54, 95% CI: 0.38-0.76) and disease-free survival of patients with hypopharyngeal squamous cell carcinoma (T stage: HR=1.26, 95% CI: 1.06-1.51; N stage: HR=1.25, 95% CI: 1.13-1.37; retropharyngeal lymph node metastasis: HR=2.24, 95% CI: 1.56-3.21; radiotherapy: HR=0.55, 95% CI: 0.40-0.77). Conclusions: Metastasis of retropharyngeal lymph nodes in hypopharyngeal squamous cell carcinoma is not rare. Enhanced CT is of low accuracy and limited value in diagnosing retropharyngeal lymph node metastasis. Primary lesions located in the posterior wall of the hypopharyngx, N2 stage, and N3 stage are independent high-risk factors for retropharyngeal lymph node metastasis. The prognosis of hypopharyngeal cancer patients with retropharyngeal lymph node metastasis is worse, and active surgical exploration and clearance can effectively reduce the mortality caused by retropharyngeal lymph node metastasis.
Humans
;
Squamous Cell Carcinoma of Head and Neck/pathology*
;
Lymphatic Metastasis/pathology*
;
Retrospective Studies
;
Carcinoma, Squamous Cell/surgery*
;
Lymph Nodes/pathology*
;
Hypopharyngeal Neoplasms/surgery*
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Prognosis
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Head and Neck Neoplasms/pathology*
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Neoplasm Staging
7.The effect of cervical lymph node metastasis density on prognosis of hypopharyngeal carcinoma.
Zhi Chao LIU ; Zheng Hua LYU ; Ju Ke MA ; Shou Hao FENG ; Xu Liang LIU ; Yu Mei WEI ; Wei XU
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2022;57(8):957-962
Objective: To investigate the relationship between the cervical lymph node density (LND) and the prognosis of hypopharyngeal carcinoma. Methods: The clinical and pathological data of 241 patients with hypopharyngeal carcinoma who underwent surgery in Shandong Provincial ENT Hospital from January 2014 to December 2017 were retrospectively analyzed, including 229 males and 12 females, aged 37-81 years. The LND was calculated, i.e. the ratio of the number of lymph nodes with metastasis to the total number of lymph nodes removed. The patients were divided into low LND group and high LND group based on the cutoff value of LND determined by receiver operating characteristic curve (ROC curve). The univariate and multivariate analyses of the disease-free survival (DFS) and the overall survival (OS) were performed in two groups. Results: With the cutoff value of 0.068, 165 patients were in the low LND group (<0.068) and 76 patients in the high LND group (≥0.068). T stage, N stage, maximum lymph node diameter, extracellular invasion of lymph node, and postpharyngeal lymph node metastasis were associated with LND (statistical values were -3.15, -6.82, 23.37, 20.44, and 30.18, respectively, all P values were<0.05). The univariate analysis showed that age, T stage, N stage, maximum diameter of cervical lymph nodes, extracapsular invasion, retropharyngeal lymph node metastasis and LND were the main factors affecting the patients' DFS (χ2=9.31, 7.30, 20.09, 15.30, 9.04, 19.44, 50.27, all P values<0.05) and OS (χ2 were 5.02, 12.94, 18.28, 15.91, 7.95, 16.88, 49.45, all P values<0.05). Multivariate analysis showed that patients with age≤60 years old and LND≥0.068 had reduced DFS [HR values were 0.61 (95%CI 0.43-0.88) and 2.23 (95%CI 1.44-3.45), both P values<0.05]; patients with advanced T stage and LND≥0.068 had reduced OS [HR values were 1.73 (95%CI 1.02-2.93) and 2.39 (95%CI 1.51-3.80), both P values<0.05]. Conclusion: LND is a prognostic factor for patients with hypopharyngeal carcinoma after surgery, with worse prognosis in patients with LND≥0.068.
Carcinoma, Squamous Cell/pathology*
;
Female
;
Humans
;
Hypopharyngeal Neoplasms/surgery*
;
Lymph Nodes/pathology*
;
Lymphatic Metastasis/pathology*
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Male
;
Middle Aged
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Neoplasm Staging
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Prognosis
;
Retrospective Studies
8. Effect of Astragaloside Ⅳ on Regulation of PI3K/Akt/FoxO1 Signal in Kidney of Type 2 Diabetic Nephropathy Rats
Ke-ke MA ; Ying-hui JU ; Qing-qing CHEN ; Wei-zu LI ; Wei-ping LI
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(2):74-81
Objective:To study the protective effect of astragaloside (AS) Ⅳ on kidney in type 2 diabetic nephropathy rats and its regulation effect on phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/forkhead box O1(FoxO1) signaling, and investigate the mechanism of glycosides against type 2 diabetic nephropathy. Method:After 6 weeks of high-glucose and high-fat diet, rat models of type 2 diabetes were established by intraperitoneal injection of streptozotocin (STZ) (35 mg·kg-1) and randomly divided into model group, AS-Ⅳ (20, 40, 80 mg·kg-1) groups and metformin hydrochloride (positive) group. After 8 weeks of continuous administration, changes in body weight, kidney index, blood glucose, 24-hour urinary protein, urinary microalbumin, urinary creatinine, serum creatinine, and blood urea nitrogen were measured in each group; hematoxylin-eosin (HE) staining was used to observe the pathological changes of renal tissues; Masson trichrome staining was used to observe the collagen expression level. Periodontium hexaammine silver (PASM) staining was used to observe the changes of basement membrane. Immunohistochemistry assay was used to detect phospho-FoxO1(p-FoxO1) protein expression, and Western blot was used to analyze the expression levels of autophagy marker protein PI3K, microtubule-associated protein 1 light chain 3 (LC3)/Ⅱ, B-cell lymphoma-2 (Bcl-2)/adenovirus E1B19 kDa interacting protein 3 (BNIP3), Beclin1, p-Akt, and Akt. Result:As compared with the normal group, the glomerular basement membrane of the model group was thicker; the extracellular matrix was increased; the mesangial was expanded; the collagen was significantly increased; the PI3K/Akt/FoxO1 signal was increased(P<0.01); and the autophagic activity was weakened (P<0.01). As compared with the model group, the phosphorylation of PI3K, Akt, and FoxO1 in renal tissue was significantly inhibited in AS-Ⅳ high and medium dose groups (P<0.05,P<0.01); autophagy was increased; and the expression of BNIP3, LC3 Ⅱ/LC3 I, and Beclin1 was up-regulated (P<0.05,P<0.01). Conclusion:AS-Ⅳ may increase the autophagic activity of renal cells by inhibiting PI3K/Akt/FoxO1 signal, slowing down the development of type 2 diabetic nephropathy.
9.Study on superposition law of drug bitterness based on tongue taste evaluation and electronic tongue evaluation.
Xue-Lin LI ; Yao ZHANG ; Peng-Ju CHEN ; Jun-Han SHI ; Jing YAO ; Qing-Xiao WANG ; Xin-Jing GUI ; Lu ZHANG ; Ke-Han WANG ; Jing MA ; Yan-Li WANG ; Rui-Xin LIU
China Journal of Chinese Materia Medica 2019;44(23):5134-5142
Traditional Chinese medicine( TCM) decoction contains complex bitterness. In this paper,the simple mixing of TCM monomer bitter substances is used as the entry point to study the law of bitterness superposition. With berberine hydrochloride( alkaloids),geniposide( terpenoids),and arbutin( glycosides) as mother liquor,sophoridine( alkaloids),gentiopicroside( terpenoids),and puerarin( glycosides) as additive substances,these different additive substances were mixed with different mother liquor according to concentration gradients to form different liquid mixtures. The bitterness of the additive solution and the mixtures was evaluated by traditional human taste panel method( THTPM) and electronic tongue; the bitterness-concentration fitting model of the additive solution and the liquid mixtures was established by Weibull and logarithmic curves. By comparing and analyzing the bitterness-concentration model and the bitterness difference( ΔI_0/ΔI_e) of the additive solution and the mixture,the influence of mother liquor on the bitterness of the mixture was investigated. The results showed that both the additive solution bitterness model and the liquid mixture bitterness model were consistent with the Weibull model and the logarithmic model( THTPM: R~2≥0. 887 0,P<0. 01; electronic tongue test:R~2≥0. 753 2,P<0. 05). The fitting degree of the Weibull model was generally higher than that of the logarithmic model; the bitterness difference( ΔI_0) was monotonously decreasing; the fitting equation of tongue bitterness and electronic tongue bitterness: R~2≥0. 874 2,P<0. 01. In this article,through the superposition of different kinds of TCM bitter substances,THTPM and electronic tongue test was combined. It was found that the bitterness after superposition was still in Weibull or logarithmic relationship with the concentration of additive substances; THTPM showed that the effect of bitter mother liquor on the bitterness of the mixture decreased with the increase of the concentration of the additive; the bitterness of the electronic tongue was obviously related to the bitterness of THTPM. However,further verification is needed later by optimizing the concentration gradient and expanding the sample size.
Alkaloids/analysis*
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Electronic Nose
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Glycosides/analysis*
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Humans
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Medicine, Chinese Traditional
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Taste
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Terpenes/analysis*
10.miRNA-99b inhibits the invasion ability of glioma cells by targeting mTOR
Peng-Ju MA ; Qian-Kun JI ; Xiang-Sheng LI ; Hai-Gang CHANG ; Wen-Ke ZHOU ; Bao-Zhe JIN
Chinese Pharmacological Bulletin 2018;34(4):473-479
Aim To detect the expression of miRNA-99b and mTOR in glioma tissues and to investigate the effect of miRNA-99b on the cell invasion ability of hu-man glioma cell line U251. Methods The expres-sions of miRNA-99b and mTOR mRNA in glioma tis-sues and normal brain tissues were detected by real-time PCR. After co-transfection with miRNA-99b mim-ics and wild or mutation type mTOR 3′UTR recombina-tion vector,the specific binding ability of miRNA-99b to 3′-UTR in mTOR gene was examined by luciferase gene reporter system. The expression levels of miRNA-99b,mTOR mRNA and mTOR protein in glioma cell line U251 after transfected with miRNA-99b mimics were measured by real-time PCR and Western blot,re-spectively. The cell invasion was measured by Tran-swell assay. The changes of mTOR and miRNA-99b expression levels in U251 cells after transfection with mTOR PsiCHECK were detected by real-time PCR. The correlation between the expression of miRNA-99b and prognosis was analyzed statistically. Results The expressed level of miRNA-99b was lower, and the ex-pression level of mTOR was higher in the glioma tissues than that in the normal brain tissues. The expression of miRNA-99b was up-regulated, and the expressions of mTOR mRNA and protein were down-regulated in U251 cells after transfection with miRNA-99b mimics. However, the abilities of invasion of U251 cells after transfection with miRNA-99b mimics were inhibited. The relative protein expression levels of mTOR in mTOR PsiCHECK group were significantly different from those in negative control group, but the relative expression levels of miRNA-99b had no signifi-cant difference compared with those in negative control group. Over-expression of mTOR restored the abilities of cell invasion in U251 cells, which was reduced by miRNA-99b. The Kaplan—Meier analysis and Log-Rank Test showed that there were significant differ-ences in overall survival (OS) between the miRNA-99b high-expression and low-expression group. Con-clusions The expression level of miRNA-99b is low in human glioma tissue. miRNA-99b may inhibit the cell invasion activity of glioma cell line U251 in vitro via inhibiting mTOR expression.

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