1.Shentong Zhuyutang Regulates SIRT1/Nrf2 Pathway to Ameliorate Intervertebral Disc Degeneration in Rats
Jiajun HUANG ; Diyou WU ; Guangyi TAO ; Yu ZHAO ; Junqing HUANG ; Bin YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):29-39
ObjectiveTo study the effect and mechanism of Shentong Zhuyutang in treating intervertebral disc degeneration (IDD) in rats. MethodsIn the cell experiment, male rats were administrated with normal saline or low-, medium-, and high-dose (3.38, 6.75,13.5 g·kg-1, respectively) Shentong Zhuyutang by gavage, respectively, and serum samples were collected after 7 days of continuous administration. Another 10 male rats were selected for the isolation of nucleus pulposus cells. The cell model of IDD was established by treatment with interleukin (IL)-1β. The modeled cells were then treated with Shentong Zhuyutang-containing serum and the ferroptosis inhibitor ferrostatin-1 (Fer-1), respectively, to investigate the effects of Shentong Zhuyutang-containing serum on the proliferation and ferroptosis of nucleus pulposus cells. To study the role of silent information regulator 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2) in the regulation of ferroptosis in nucleus pulposus cells by Shentong Zhuyutang-containing serum, this study treated the cells with the SIRT1 inhibitor Ex 527 and the Nrf2 inhibitor ML385, respectively, in addition to the treatment with IL-1β and high-dose Shentong Zhuyutang-containing serum. The cell-counting kit-8 (CCK-8) assay and EdU staining were employed to measure the cell viability and proliferation, respectively. The Fe2+, glutathione (GSH), and malondiadehyde (MDA) levels were measured by colorimetric assay. Western blot was employed to determine the protein levels of glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family 4 (ACSL4), Collagen Ⅱ, Aggrecan, SIRT1, and Nrf2. Immunofluorescence was used detect SIRT1 expression. In the animal experiment, male rats were treated with anulus puncture for the modeling of IDD. Rats were randomly assigned into sham operation, model, Shentong Zhuyutang-containing serum (13.5 g·kg-1), and positive control (nimesulide dispersible tablets, 0.18 mg·kg-1) groups. Rats in the drug intervention groups were administrated with corresponding agents at 1 mL·kg-1, and those in the sham operation and model groups were administrated with equal volumes of normal saline, once daily for 28 consecutive days. At the end of the last administration, the histopathological changes in the intervertebral discs of rats were observed by hematoxylin-eosin staining and scored by the Masuda method. Western blot was employed to determine the protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ in the nucleus pulposus tissue. ResultsCompared with the control group, the IL-1β group of nucleus pulposus cells showed elevated levels of Fe2+, MDA, and ACSL4 (P<0.05), decreased cell viability, lowered GSH level, and down-regulated protein levels of GPX4, Collagen Ⅱ, and Aggrecan (P<0.05). Shentong Zhuyutang-containing serum and Fer-1 reversed the effects of IL-1β on the viability and ferroptosis of nucleus pulposus cells and up-regulated the protein levels of Collagen Ⅱ and Aggrecan in nucleus pulposus cells (P<0.05). Compared with the control group, the IL-1β group showcased down-regulated expression of Sirt1 and Nrf2 in nucleus pulposus cells (P<0.05). Compared with the IL-1β group, the high-dose Shentong Zhuyutang-containing serum+IL-1β group showed up-regulated expression of SIRT1 and Nrf2 in nucleus pulposus cells (P<0.05). Compared with the high-dose Shentong Zhuyutang-containing serum+IL-1β group, the ML385 group showed down-regulated protein levels of Nrf2 and GPX4, lowered GSH level, and elevated Fe2+ and MDA levels (P<0.05). In addition, the Ex 527 group showed down-regulated protein levels of SIRT1, Nrf2, and GPX4 (P<0.05). The results of the animal experiment showed that compared with the sham operation group, the model group had severe degeneration of the intervertebral disc tissue with increased pathological score, up-regulated protein level of ACSL4 (P<0.05), and down-regulated protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ (P<0.05). Compared with the model group, the Shentong Zhuyutang group showed alleviated IDD with declined pathological score, down-regulated protein level of ACSL4 (P<0.05), and up-regulated protein levels of SIRT1, Nrf2, GPX4, and Collagen Ⅱ (P<0.05). ConclusionShentong Zhuyutang may activate the SIRT1/Nrf2 signaling pathway to inhibit the ferroptosis of nucleus pulposus cells, thereby delaying the process of IDD in rats.
2.Duhuo Jisheng Decoction treats gouty arthritis by inhibiting NLRP3 inflammasome activation
Guangyi TAO ; Zhengzhen WANG ; Jiajun HUANG ; Diyou WU ; Xinwei WANG ; Shun YANG ; Bin YANG ; Junqing HUANG
Chinese Journal of Tissue Engineering Research 2024;28(32):5182-5189
BACKGROUND:The activation of NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome has been found to be an important factor in the pathogenesis of gouty arthritis,and the activation of NLPR3 inflammasome can be effectively inhibited by regulating the PTEN-induced kinas 1(PINK1)/Parkin signaling pathway. OBJECTIVE:To investigate the effect of Duhuo Jisheng Decoction on the PINK1/Parkin/NLRP3 signaling pathway. METHODS:(1)Animal experiment:Male Sprague-Dawley rats were randomly divided into control group,model group,positive control group(Qiushuixian tablets 0.3 mg/kg),and low-,medium-,and high-dose Duhuo Jisheng Decoction groups(6.9,13.8,and 27.6 g/kg,respectively).A rat gouty arthritis model was constructed by injecting monosodium urate crystal suspension into the right hind ankle joint cavity of rats in all the groups except for the control group.(2)Cell experiment:Human monocytes(THP-1)were divided into blank control group,model group,Duhuo Jisheng Decoction-containing serum group,and PINK1 siRNA+Duhuo Jisheng Decoction-containing serum group.The cells in each group except the blank control group were stimulated with sodium urate to establish an in vitro gouty arthritis model.(3)The swelling degree,joint circumference,gait score,joint inflammation index and degree of pathological grading in the ankle joints of rats were observed.Enzyme-linked immunosorbent assay was used to detect the levels of uric acid,C-reactive protein,NLRP3,and interleukin 1β.Immunoblotting assay was used to detect the levels of proteins related to the PINK1/Parkin/NLRP3 pathway.Tetramethyl azolidine blue assay was used to detect cell activity.Immunofluorescent double staining was performed to detect the level of mitophagy.Immunofluorescence was used to detect the expression of NLRP3 and interleukin 1β. RESULTS AND CONCLUSION:(1)Compared with the control group,rats in the model group showed elevated ankle swelling and ankle circumference at 6-48 hours(P<0.05),elevated gait scores and joint inflammation indexes at 24 and 48 hours(P<0.05),elevated serum uric acid and C-reactive protein levels,degree of pathological classification,expression of PINK1,Parkin,and NLRP3 proteins,and interleukin 1β level in synovial tissues(P<0.05).Compared with the model group,the protein expression levels of PINK1 and Parkin in all the Duhuo Jisheng Decoction groups were elevated,while the levels of the other indexes were decreased(P<0.05).(2)Compared with the model group,NLRP3 activity,interleukin 1β level and mitochondrial outer membrane translocator protein 20 protein level in the Duhuo Jisheng Decoction-containing serum group were decreased(P<0.05),while the proliferation inhibition rate,PINK1,Parkin and LC3B protein level were increased(P<0.05).Compared with the model group,the mitochondrial autophagy level of cells in the Duhuo Jisheng Decoction-containing serum group was elevated(P<0.05),while NLRP3 and interleukin 1β protein levels were decreased(P<0.05).(3)Compared with the Duhuo Jisheng Decoction-containing serum group,the mitochondrial autophagy level of cells in the PINK1 siRNA+Duhuo Jisheng Decoction-containing serum group was decreased(P<0.05),and the expression levels of NLRP3 and interleukin 1β were elevated(P<0.05).To conclude,Duhuo Jisheng Decoction inhibits the activation of NLRP3 inflammasome by regulating the PINK1/Parkin signaling pathway, thereby exerting a therapeutic effect on gouty arthritis.
3.Fulminant type 1 diabetes mellitus with acute pancreatitis:A case report and litera-ture review
Peiheng ZHANG ; Ying GAO ; Honghua WU ; Jian ZHANG ; Junqing ZHANG
Journal of Peking University(Health Sciences) 2024;56(5):923-927
The objective was to report a relatively rare case of fulminant type 1 diabetes(FT1 DM)complicated with acute pancreatitis(AP),to summarize the characteristics as well as experience of diag-nosis and treatment,and to explore its pathogenesis.Clinical data of a case of FT1DM complicated with AP in the Department of Endocrinology of our hospital were analyzed retrospectively.A 66-year-old male presented with acute fever and abdominal pain,accompanying with the significantly elevated pancreatic enzymes,and his abdominal CT scan showed exudation around the pancreas.The clinical manifestations mentioned above were consistent with the diagnosis of AP.Five days after onset,the patient developed clinical symptoms,such as obvious thirst,polyuria,polyasthenia and fatigue.Meanwhile,his plasma glucose increased significantly and the diabetic ketoacidosis(DKA)occurred.The patient's fasting and postprandial 2 hours C peptide decreased significantly(all 0.02 μg/L),glycated hemoglobin level was not high(6%),and his islet-related autoantibodies were undetectable.Thus,the patient could be diag-nosed with FT1DM.After the treatment of fasting,fluid replacement,anti-infection,somatostatin,anti-coagulation and intravenous insulin sequential subcutaneous insulin pump,the patient gained the allevia-tion of pancreatitis,restoration of oral intake,and relatively stable blood glucose levels.Summarizing the characte-ristics of this case and reviewing the literature,FT1DM complicated with AP was relatively rare in FT1DM.Its common characteristics were described below:(1)Most cases started with AP and the blood glucose elevated within 1 week,or some cases had the simultaneously onset of AP and FT1DM.(2)The clinical course of AP was short and relieved no more than 1 week;Pancreatic imaging could completely return to normal within 1 to 4 weeks after onset.(3)The etiology of AP most was idiopathic;The elevation of pancreatic enzyme level was slight and the recovery was rapidly compared with AP of other etiologies.FT1DM could be complicated with AP,which was different from the physiological mani-festations of pancreatic disease in general FT1DM patients.Virus infection mignt be the common cause of AP and FT1DM,and AP might be the early clinical manifestation of some FT1DM.The FT1DM patients developed with abdominal pain was easy to be missed,misdiagnosed and delayed,which should receive more attention in clinic.
4.Clinical investigation of enamel demineralization during clear aligner treatment
Danyan HU ; Huifen CHEN ; Junqing WU ; Hu LI ; Bin YAN ; Qinghua SHAO
STOMATOLOGY 2024;44(10):742-746
Objective To investigate the incidence and severity of enamel demineralization during clear aligner treatment and provide reference for clinical medical nursing work.Methods A total of 151 patients(3 507 teeth)undergoing clear aligner treatment in our hospital from 2015 to 2022 were collected.The enamel demineralization before and after treatment was compared,and the enamel demi-neralization index(EDI)was calculated to evaluate the severity of demineralization.The incidence and severity of enamel demineraliza-tion were analyzed statistically,and the differences among different genders,ages and dental positions were compared.Results Enam-el demineralization occurred in 84 patients during treatment,with an incidence rate of 55.63%.Gender and treatment time had no effect on the incidence,and the incidence of adolescent patients was higher than that of adult patients.Among the investigated 3 507 teeth,601 teeth had enamel demineralization,and the incidence rate was 17.14%.The incidence of maxillary anterior teeth and mandibular posterior teeth was higher;the incidence of right teeth was significantly higher than that of left teeth,and the incidence of teeth with ad-hesive attachments was also significantly increased.88%of the enamel demineralization lesions were 1-2 degrees,and the degree of enamel demineralization was similar in male and female patients.But the degree in adolescents was significantly higher than that in adult patients.The average EDI value of maxillary anterior teeth was significantly higher than that of mandible,but that of maxillary posterior teeth was significantly lower than that of mandible.The average EDI value of right teeth was significantly higher than that of left.Conclusion The incidence of enamel demineralization is high in the process of clear aligner treatment,and teenagers are suscep-tible to it.The demineralization of teeth is significantly different among different tooth positions,and adhesive attachments will increase the risk of enamel demineralization.Therefore,medical staff should take active and effective measures in the prevention and treatment of enamel demineralization.
5.Construction of a prediction model for difficult tracheal intubation in patients with obstructive sleep apnea-hypopnea syndrome
Junqing LI ; Ning CAI ; Dongfeng LIANG ; Di WU ; Guanghong CHEN ; Hao YUAN
Chinese Journal of Anesthesiology 2023;43(6):692-696
Objective:To construct a prediction model for difficult tracheal intubation in the patients with obstructive sleep apnea-hypopnea syndrome (OSAHS).Methods:A total of 324 patients with OSAHS undergoing surgery with general anesthesia, admitted to our hospital from June 2019 to June 2021, were included in model group, and 175 patients with OSAHS undergoing surgery with general anesthesia, admitted from July 2021 to July 2022, were selected and served as validation group. The patients in model group were divided into occurrence group and non-occurrence group according to whether difficult tracheal intubation occurred. Logistic regression was used to construct the prediction model, and R4.2.1 software was used to draw the risk nomogram and calibration curve. The predictive accuracy of the model was evaluated by the area under the receiver operating characteristic curve.Results:Body mass index (BMI), sagittal minimum cross-sectional area, horizontal minimum cross-sectional area and mandibular distance were risk factors for difficult tracheal intubation in OSAHS patients ( P<0.05). A prediction model was developed using the above factors: Logit P=33.726+ 1.411×BMI score-0.014×sagittal airway minimum cross-sectional area-0.013×airway horizontal minimum cross-sectional area-0.312× mandibular distance. The area under the receiver operating characteristic curve was 0.846, Youden index 0.585, sensitivity 0.831, specificity 0.755, and the accuracy 0.889 (Hosmer-Lemeshow test χ2=9.24, P=0.322) in model group. The area under the external validation curve was 0.802, Youden index 0.545, sensitivity 0.636, specificity 0.908, and the accuracy 0.893 (Hosmer-Lemeshow test χ2=10.24, P=0.287) in validation group. Conclusions:The prediction model based on BMI, sagittal minimum cross-sectional area of airway, horizontal minimum cross-sectional area of airway and mandibular distance has a high value in predicting the risk of difficult tracheal intubation in patients with OSAHS.
6.Cost-effectiveness analysis of different screening modes for thalassemia in Hunan Province
Hui XI ; Qin LIU ; Donghua XIE ; Xu ZHOU ; Wanglan TANG ; Deguo TANG ; Chunyan ZENG ; Qiong WANG ; Xinghui NIE ; Jinping PENG ; Xiaoya GAO ; Hongliang WU ; Haoqing ZHANG ; Li QIU ; Zonghui FENG ; Shuyuan WANG ; Shuxiang ZHOU ; Jun HE ; Shihao ZHOU ; Faqun ZHOU ; Junqing ZHENG ; Hua WANG ; Junqun FANG ; Changbiao LIANG
Chinese Journal of Perinatal Medicine 2023;26(6):468-475
Objective:To analyze the costs and effectiveness of five common screening modes and genetic screening for thalassemia in China in order to find the optimal way and provide evidence for the implementation of thalassemia prevention and control projects in Hunan Province.Methods:From June 2020 to April 2021, 12 971 couples from 14 cities and autonomous prefectures in Hunan Province were selected as the study population. The diagnosis of thalassemia was based on the results of genetic testing. Results of routine blood test and hemoglobin electrophoresis were collected and analyzed. The efficacy of five screening modes, at the cut-off value of <80 fl or 82 fl for the mean corpuscular volume (MCV), was analyzed by positive predictive value, negative predictive value, Jorden index and cost-effectiveness ratio. Sensitivity analysis was used to assess the feasibility of genetic screening at different costs after fixing the costs of routine blood and hemoglobin electrophoresis. The five thalassemia screening models are as follows: Mode 1: The woman had a blood routine test first. If the result was positive, the spouse required a blood routine test. If both results were positive, a thalassemia gene test should be offered to the couple. Mode 2: Both husband and wife were screened by blood routine and hemoglobin electrophoresis. If one or both of them were positive, both would be tested for thalassemia gene. Mode 3: The couple received blood routine tests initially. If either was positive, both should receive hemoglobin electrophoresis testing. If either was positive, both parties will conduct thalassemia gene testing. Mode 4: The woman was screened by blood routine and hemoglobin electrophoresis. If any one of them was positive, the woman would be tested for thalassemia gene. If the gene test result was positive, the spouse should receive thalassemia gene. Mode 5: Both spouses conducted a blood routine test. If either was positive, both would conduct hemoglobin electrophoresis test. If both were positive, both spouses should receive thalassemia gene testing. Gene testing mode: The woman would be tested for thalassemia, and her spouse would have thalassemia test too if her result was positive.Results:When using MCV<80 fl as the cut-off for diagnosing thalassemia, the Youden indices of the five prenatal screening modes in Hunan Province were 0.551, 0.639, 0.898, 0.555 and 0.356, while when using MCV<82 fl as the cut-off, the Youden indices were 0.549, 0.629, 0.851, 0.548 and 0.356. When the MCV cut-off value was <80 fl, the missed diagnosis rates of the five screening modes were 44.44%, 0.00, 0.00, 18.52% and 62.96%, and the cost-effectiveness ratios were 21 709, 250 939, 76 870, 138 463 and 92 860 yuan (RMB)/couple, respectively. When the price of genetic testing was lower than 55 yuan (RMB), the cost-effectiveness ratio of genetic screening was lower than that of Mode 3.Conclusions:MCV<80 fl can be considered as the positive criteria in blood routine screening for thalassemia in Hunan Province, and the cost-effectiveness ratio of Mode 3 (the couple received blood routine tests initially. If either was positive, both should receive hemoglobin electrophoresis testing. If either was positive, both parties will conduct thalassemia gene testing) is the best. Genetic screening has certain advantages with the decreasing price.
7.Diagnostic value of a combined serology-based model for minimal hepatic encephalopathy in patients with compensated cirrhosis
Shanghao LIU ; Hongmei ZU ; Yan HUANG ; Xiaoqing GUO ; Huiling XIANG ; Tong DANG ; Xiaoyan LI ; Zhaolan YAN ; Yajing LI ; Fei LIU ; Jia SUN ; Ruixin SONG ; Junqing YAN ; Qing YE ; Jing WANG ; Xianmei MENG ; Haiying WANG ; Zhenyu JIANG ; Lei HUANG ; Fanping MENG ; Guo ZHANG ; Wenjuan WANG ; Shaoqi YANG ; Shengjuan HU ; Jigang RUAN ; Chuang LEI ; Qinghai WANG ; Hongling TIAN ; Qi ZHENG ; Yiling LI ; Ningning WANG ; Huipeng CUI ; Yanmeng WANG ; Zhangshu QU ; Min YUAN ; Yijun LIU ; Ying CHEN ; Yuxiang XIA ; Yayuan LIU ; Ying LIU ; Suxuan QU ; Hong TAO ; Ruichun SHI ; Xiaoting YANG ; Dan JIN ; Dan SU ; Yongfeng YANG ; Wei YE ; Na LIU ; Rongyu TANG ; Quan ZHANG ; Qin LIU ; Gaoliang ZOU ; Ziyue LI ; Caiyan ZHAO ; Qian ZHAO ; Qingge ZHANG ; Huafang GAO ; Tao MENG ; Jie LI ; Weihua WU ; Jian WANG ; Chuanlong YANG ; Hui LYU ; Chuan LIU ; Fusheng WANG ; Junliang FU ; Xiaolong QI
Chinese Journal of Laboratory Medicine 2023;46(1):52-61
Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.
8.Impact of different diagnostic criteria for assessing mild micro-hepatic encephalopathy in liver cirrhosis: an analysis based on a prospective, multicenter, real-world study
Xiaoyan LI ; Shanghao LIU ; Chuan LIU ; Hongmei ZU ; Xiaoqing GUO ; Huiling XIANG ; Yan HUANG ; Zhaolan YAN ; Yajing LI ; Jia SUN ; Ruixin SONG ; Junqing YAN ; Qing YE ; Fei LIU ; Lei HUANG ; Fanping MENG ; Xiaoning ZHANG ; Shaoqi YANG ; Shengjuan HU ; Jigang RUAN ; Yiling LI ; Ningning WANG ; Huipeng CUI ; Yanmeng WANG ; Chuang LEI ; Qinghai WANG ; Hongling TIAN ; Zhangshu QU ; Min YUAN ; Ruichun SHI ; Xiaoting YANG ; Dan JIN ; Dan SU ; Yijun LIU ; Ying CHEN ; Yuxiang XIA ; Yongzhong LI ; Qiaohua YANG ; Huai LI ; Xuelan ZHAO ; Zemin TIAN ; Hongji YU ; Xiaojuan ZHANG ; Chenxi WU ; Zhijian WU ; Shengqiang LI ; Qian SHEN ; Xuemei LIU ; Jianping HU ; Manqun WU ; Tong DANG ; Jing WANG ; Xianmei MENG ; Haiying WANG ; Zhenyu JIANG ; Yayuan LIU ; Ying LIU ; Suxuan QU ; Hong TAO ; Dongmei YAN ; Jun LIU ; Wei FU ; Jie YU ; Fusheng WANG ; Xiaolong QI ; Junliang FU
Chinese Journal of Hepatology 2023;31(9):961-968
Objective:To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test.Methods:This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ2 test. A kappa test was used to compare the consistency between groups. Results:After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea ( Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences ( P < 0.001). Conclusion:The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.
9.Effect of Gegen Qinliantang-medicated Serum on Nonalcoholic Steatohepatitis Based on Nrf2/TXNIP Signaling Pathway
Yue WEI ; Junqing SHENG ; Ziwen CHENG ; Xiaoquan LUO ; Zhenzhen SONG ; Ailan WU ; Lan CAO ; Changhua ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(20):8-16
ObjectiveTo investigate the effect of Gegen Qinliantang (GGQLT)-medicated serum on free fatty acid (FFA)-induced nonalcoholic steatohepatitis (NASH) in vitro model of human hepatoma cells HepG2. MethodNASH model of HepG2 cells was established in vitro, and the cells were intervened with different volume fractions of GGQLT-medicated serum and resveratrol. Intracellular lipid deposition in each group was detected by oil red O staining, the level of reactive oxygen species (ROS) in each group were detected by flow cytometry, the levels of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), triglyceride (TG) and malondialdehyde (MDA) in each group were detected by kits. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to measure the mRNA expression levels of nuclear transcription factor (NF)E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), quinone oxidoreductase 1 (NQO1), Kelch-like epichlorohydrin-associated protein-1 (Keap1), NF-κB, thioredoxin interacting protein (TXNIP), interleukin-1β (IL-1β) in HepG2 cells of each group. The protein expression of Nrf2, TXNIP in cells of each group was detected by Western blot. ResultFFA induced large accumulation of intracellular lipids. Compared with the normal group, the activities of GSH-Px and SOD were significantly decreased (P<0.01) and the contents of TG, ROS and MDA were significantly increased (P<0.05, P<0.01) in the model group. Compared with the model group, all GGQLT groups and resveratrol group could elevate intracellular SOD activity to different degrees (P<0.05, P<0.01) and significantly reduce the levels of intracellular ROS and MDA (P<0.05, P<0.01), GGQLD high- and medium-dose groups and resveratrol group significantly elevated GSH-Px activity (P<0.01), GGQLD medium- and low-dose groups and resveratrol group significantly decreased TG content (P<0.05, P<0.01). Compared with the model group, GGQLT high- and medium-dose groups and resveratrol group could significantly upregulate the mRNA expression levels of Nrf2, HO-1 and NQO1 (P<0.01), all GGQLT groups and resveratrol group could significantly downregulate the TXNIP protein expression level, as well as significantly downregulate the mRNA expression levels of Keap1, NF-κB (P<0.05, P<0.01). Nrf2-siRNA transfection of cells revealed that Nrf2 expression was significantly downregulated (P<0.01) in the Nrf2-siRNA group of cells by comparing with NC-siRNA group at the corresponding dose of drugs, and the inhibitory effects of GGQLT and resveratrol on TXNIP, IL-1β were attenuated. ConclusionFFA induces the production of ROS and inflammatory factors in HepG2 cells, and GGQLT can improve the anti-inflammatory and antioxidant capacities of cells, and its mechanism may be related to the regulation of Nrf2/TXNIP signaling pathway, so as to improve NASH.
10.Modified endoscopic retrograde appendicitis therapy for appendicitis with intussusception in children: a primary study (with video)
Jianqin KANG ; Wei ZHANG ; Chongkang HU ; Yan LIN ; Junqing WU ; Yupin LI ; Ye LI ; Xiangzeng LIU ; Baoxi WANG ; Lingchao ZENG ; Xun JIANG
Chinese Journal of Digestive Endoscopy 2022;39(3):231-234
To investigate the efficacy and safety of modified endoscopic retrograde appendicitis therapy (ERAT) in children with acute uncomplicated appendicitis complicated with intussusception. Data of 6 patients with acute uncomplicated appendicitis complicated with intussusception who received modified ERAT were collected from October 2018 to February 2020 in Pediatrics Department of Tangdu Hospital. The reduction rate of intussusception, the success rate of ERAT appendiceal intubation and endoscopic treatment, the remission time of clinical symptoms, the time of white blood cells return to normal, the length of hospital stay, complications and recurrence were summarized. All the 6 patients were confirmed to be acute uncomplicated appendicitis combined with ileocecal intussusceptions under colonoscopy. Endoscopic intussusception reduction and ERAT were performed successfully. The mean time of disappearance of abdominal tenderness was 1.3 days after the therapy, and the time of white blood cells return to normal was 1 day after surgery. The endoscopic treatment were all successful and the length of hospital stay was 3-6 days. No complications or recurrence of intussusception and appendicitis occurred during the follow-up period of 6-12 months. Modified ERAT can effectively, safely and minimally invitingly treat acute uncomplicated appendicitis with intussusception in children on the basis of definite diagnosis.

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