1.Effect of Serum Containing Zhenwutang on Apoptosis of Myocardial Mast Cells and Mitochondrial Autophagy
Wei TANG ; Meiqun ZHENG ; Xiaolin WANG ; Zhiyong CHEN ; Chi CHE ; Zongqiong LU ; Jiashuai GUO ; Xiaomei ZOU ; Lili XU ; Lin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):11-21
ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ (AngⅡ) and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment, AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h, and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 (CCK-8) to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of B-type natriuretic peptide (BNP) in cell culture supernatant, and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently, the experiment was divided into a blank group (20% blank serum), a model group (20% blank serum + 5×10-5 mol·L-1 AngⅡ), low-, medium-, and high-dose (5%, 10% and 20%) serum containing Zhenwutang groups, an autophagy inhibitor group (1×10-4 mol·L-1 3-MA), and autophagy inducer group (1×10-7 mol·L-1 rapamycin). The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe (Lyso Tracker) and mitochondrial probe (Mito Tracker) co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect Caspase-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), Bcl-2-related X protein (Bax), and cytochrome C (Cyt C) in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase (Parkin), phosphatase and tensin homolog (PTEN)-induced kinase 1 (PINK1), and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3, cleaved Caspase-9, Bax, Bcl-2, and Cyt C in apoptosis-related pathways, phosphorylated ubiquitin ligase (p-Parkin), phosphorylated PTEN-induced kinase 1 (p-PINK1), p62, and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways, and the change of microtubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10-5 mol·L-1, the cell activity was the lowest, and there was no cytotoxicity. At this concentration, the surface area of cardiomyocytes was significantly increased (P<0.01), and the content of BNP in the supernatant of culture medium was significantly increased (P<0.05). Therefore, AngⅡ with a concentration of 5×10-5 mol·L-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group, the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate (P<0.01) and significantly decreased mitochondrial membrane potential (P<0.01). The results of immunofluorescence co-localization showed that compared with the blank group, the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group, the relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 in the model group was significantly up-regulated (P<0.01), while the relative mRNA expression of Bcl-2, Parkin, and PINK1 was significantly down-regulated (P<0.01). In addition, the relative protein expression of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 was significantly up-regulated (P<0.01). The LC3Ⅱ/Ⅰ was significantly decreased, and the relative protein expression of Bcl-2, p-Parkin, p-PINK1, and Beclin1 was significantly down-regulated (P<0.01). Compared with the model group, the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate (P<0.01), and the decrease ratio of mitochondrial membrane potential is significantly lowered (P<0.01) in a dose-dependent manner. Additionally, both red and green fluorescence spots became more in these groups. In the 3-MA group, the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 was significantly down-regulated (P<0.05, P<0.01), while that of Bcl-2, Parkin, and PINK1 was significantly up-regulated (P<0.01). In the serum containing Zhenwutang groups, the relative protein expression levels of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 were significantly down-regulated (P<0.05,P<0.01). The LC3Ⅱ/Ⅰ was significantly increased, and the relative protein expression levels of Bcl-2, p-Parkin, p-PINK1, and Beclin1 were significantly up-regulated (P<0.01). ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.
2.Effect and mechanism of BYL-719 on Mycobacterium tuberculosis-induced differentiation of abnormal osteoclasts
Jun ZHANG ; Jian GUO ; Qiyu JIA ; Lili TANG ; Xi WANG ; Abudusalamu·Alimujiang ; Tong WU ; Maihemuti·Yakufu ; Chuang MA
Chinese Journal of Tissue Engineering Research 2025;29(2):355-362
BACKGROUND:The phosphatidylinositol 3-kinase/protein kinase(PI3K/AKT)signaling pathway plays a pivotal role in regulating osteoclast activation,which is essential for maintaining bone homeostasis.Bone destruction in osteoarticular tuberculosis is caused by aberrant osteoclastogenesis induced by Mycobacterium tuberculosis infection.However,the role of the PI3K signaling pathway in Mycobacterium tuberculosis-induced aberrant osteoclastogenesis remains unclear. OBJECTIVE:To investigate the effects and mechanisms of the PI3K/AKT signaling pathway inhibitor BYL-719 on aberrant osteoclastogenesis induced by Mycobacterium tuberculosis. METHODS:RAW264.7 cells were infected with bovine Mycobacterium tuberculosis bacillus calmette-cuerin vaccine,and Ag85B was used for cellular immunofluorescence staining.The cell counting kit-8 assay was employed to determine the safe concentration of BYL-719.There were four groups in the experiment:blank control group,BYL-719 group,BCG group,and BCG+BYL-719 group.Under the induction of receptor activator of nuclear factor kappa-B ligand,the effects of BYL-719 on post-infection osteoclast differentiation and fusion were explored through tartrate-resistant acid phosphatase staining and phalloidin staining.RT-PCR and western blot were used to detect the expression of osteoclast-related genes and proteins,and further investigate the mechanism of action. RESULTS AND CONCLUSION:Immunofluorescence staining showed that RAW264.7 cells phagocytosed Mycobacterium tuberculosis.Cell counting kit-8 data indicated that 40 nmol/L BYL-719 was non-toxic to cells.Tartrate-resistant acid phosphatase staining and phalloidin staining showed that BYL-719 inhibited the generation and fusion ability of osteoclasts following infection.RT-PCR and western blot results also indicated that BYL-719 suppressed the upregulation of osteoclast-specific genes(including c-Fos,NFATc1,matrix metalloproteinase 9,and CtsK)induced by Mycobacterium tuberculosis infection(P<0.05).Western blot and immunofluorescence staining revealed that BYL-719 inhibited excessive osteoclast differentiation induced by Mycobacterium tuberculosis by downregulating the expression of IκBα-p65.To conclude,BYL-719 inhibits aberrant osteoclastogenesis induced by Mycobacterium tuberculosis through the downregulation of IκBα/p65.Therefore,the IκBα/p65 signaling pathway is a potential therapeutic target for osteoarticular tuberculosis,and BYL-719 holds potential value for the preventing and amelioration of bone destruction in osteoarticular tuberculosis.BYL-719 has the potential to prevent and ameliorate bone destruction in osteoarticular tuberculosis.
3.Exploration on the Formation,Evolution,and Alienation of Syndrome Differentiation and Treatment
Sicheng WANG ; Linhua ZHAO ; Rumeng TANG ; Lili ZHANG
Journal of Traditional Chinese Medicine 2025;66(12):1202-1206
Syndrome differentiation and treatment is an integral part of the traditional Chinese medicine (TCM) diagnostic and therapeutic system, whose development exhibits distinct stages. This paper systematically reviews the evolutionary trajectory of syndrome differentiation and treatment, from symptom-based treatment in Inner Canon of Yellow Emperor (《黄帝内经》), to ZHANG Zhongjing's establishment of the disease-pulse-syndrome-treatment framework, through its application and development in the Ming and Qing dynasties, and finally to its recognition as a fundamental characteristic of TCM in modern times. However, the overemphasis on syndrome differentiation and treatment, coupled with a diminished focus on disease concepts, has led to its gradual alienation as the primary diagnostic and therapeutic model. The alienation mainly manifests as a tendency to prioritize syndrome over disease, resulting in the overgeneralization and limited application of the concept, and causing a lack of specificity in practice. This paper emphasizes that a correct understanding of syndrome differentiation and treatment is a necessary premise for the deve-lopment and improvement of the TCM diagnostic and therapeutic system. By integrating modern medical diagnosis to clarify disease targets and applying TCM thinking to extract common patterns of diseases, precise alignment between syndrome differentiation and treatment and modern clinical demands can be achieved, providing a reference for addressing the contemporary challenges of syndrome differentiation and treatment.
4.Application of MRI compilation sequence for predicting lymphovascular space invasion status in early cervical cancer
Zebo HUANG ; Wenwei TANG ; Yao YAO ; Tong LIANG ; Zhongfu TIAN ; Lili WANG ; Hailei GU
Journal of Practical Radiology 2024;40(3):422-425,429
Objective To assess the value of magnetic resonance imaging compilation(MAGiC)sequence in predicting lympho-vascular space invasion(LVSI)in early cervical cancer.Methods The data of 48 patients with cervical cancer confirmed by pathology were collected retrospectively,and classified into LVSI-positive group(n=29)and LVSI-negative group(n=19)according to postop-erative pathological results.MAGiC sequence images of patients were obtained before injecting contrast agents,then the region of interest(ROI)was delineated along the largest dimension edge of the lesion,and T1,T2 and proton density(PD)values were automatically generated by the software.Predictors were screened by univariate analysis and receiver operating characteristic(ROC)curves were drawn to assess their diagnostic efficacy for predicting LVSI in cervical cancer.Results Significant differences were found in T1 and PD values between LVSI-positive and LVSI-negative groups(P=0.003,P=0.017).There were no significant differences in T2 values between the two groups(P=0.414).The area under the curve(AUC)for T1 and PD values to predict LVSI status were 0.73 and 0.721,respectively.Conclusion LVSI-positive group of cervical cancer has lower T1 and PD values than LVSI-negative group based on MAGiC sequence.The MAGiC sequence has a certain application value for predicting LVSI status in early cervical cancer.
5.Research progress on the relationship between m6A methylation modification and acute kidney injury
Lili TANG ; Xinyu WANG ; Jie ZHANG ; Yue ZHAO ; Xiaoyue LI
The Journal of Practical Medicine 2024;40(2):278-282
Acute kidney injury(AKI)is a global public health problem with high morbidity,high mortality and costly treatment cost.The pathogenesis of AKI is very complex,and the treatment strategies for AKI are lim-ited,then it is very matter to explore the pathophysiological mechanism and potential therapeutic targets of acute kidney injury.N6-methyladenosine(m6A)is the most abundant and extremely conservative epigenetic modification in eukaryotic,which is a dynamic and reversible process involving in splicing,nuclear export,translation,stabil-ity,and higher structure of RNA,and regulated by three regulatory factors:methyltransferase,demethylase and methylated reading protein.Current studies have found that m6A plays an important regulatory role in AKI and can be a potential therapeutic target for AKI.In this review,we provide a brief description of m6A and summarize the impact of m6A on AKI and possible future study directions for this research.
6.Research progress on the organ-protective effect of Maresin-1 in sepsis
Lili TANG ; Moxiang GONG ; Qiao CHEN ; Zhiyong SU ; Xiaoyue LI
Chinese Critical Care Medicine 2024;36(2):211-215
Sepsis is a fatal organ dysfunction caused by the uncontrolled inflammatory response of the host to infection. Excessive inflammatory reaction is the core factor in the occurrence and development of sepsis, the degree of organ dysfunction is directly related to the prognosis of sepsis. Timely intervention of excessive inflammatory response and alleviation of organ function damage are essential to improve the prognosis of sepsis. Maresin-1 (MaR-1) is a newly discovered endogenous specific pro-inflammatory resolution mediator, which plays a role of anti-inflammatory, pro-inflammatory regression and organ protection in sepsis, and may be a new target for the treatment of sepsis. This article reviews the research progress of the role of MaR-1 in the regulation of inflammation and organ protection in sepsis, in order to provide reference for the clinical development of new drugs for the treatment of sepsis.
7.Comparison of modified double-reverse traction and traditional open reduction in the treatment of tibial plateau fractures
Huankun LI ; Yanhong LI ; Dongjie HUANG ; Baijun HU ; Dawei GAO ; Yufeng WU ; Jianbang TANG ; Hongjun CHEN ; Lili SANG
Chinese Journal of Orthopaedic Trauma 2024;26(2):118-124
Objective:To compare the curative effects between modified double-reverse traction technique and traditional open reduction in the treatment of tibial plateau fractures.Methods:A retrospective study was conducted to analyze the data of 70 patients with Schatzker type Ⅳ-Ⅴ tibial plateau fracture who had undergone surgical treatment at The Third Department of Orthopaedics, Zhongshan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Traditional Chinese Medicine from January 2017 to December 2022. The patients were divided into an observation group treated with modified double-reverse traction and a control group treated with traditional open reduction. In the observation group of 37 cases, there were 20 males and 17 females with an age of (44.6±13.5) years, and 9 cases of type Ⅳ and 28 cases of type V by the Schatzker classification; in the control group of 33 cases, there were 18 males and 15 females with an age of (45.9±13.7) years, and 10 cases of type Ⅳ and 23 cases of type Ⅴ by the Schatzker classification. The 2 groups were compared in terms of operation time, length of main incision, intraoperative blood loss, Rasmussen imaging score before discharge, and knee function score of American Hospital for Special Surgery (HSS), Visual Analogue Scale (VAS), fracture healing and complications at 6 months postoperatively.Results:There was no significant difference in the preoperative general data between the 2 groups, showing group comparability ( P>0.05). All patients were followed up for (14.3±1.4) months. The observation group was significantly better than the control group in operation time [(113.9±11.4) min versus (151.82±10.37) min], length of main incision [4 (4, 5) cm versus 6 (6, 7) cm], intraoperative blood loss [30 (20, 35) mL versus 55 (50, 65) mL], VAS [0 (0, 0) point versus 0 (0, 1) points] and HSS score [(89.8±3.1) points versus (86.0±3.5) points] ( P<0.05). There were no significant differences between the 2 groups in Rasmussen imaging score before discharge, or fracture healing rate or complication rate at 6 months postoperatively ( P>0.05). Conclusion:In the treatment of Schatzker type Ⅳ-Ⅴ tibial plateau fractures, modified double-reverse traction technique is worthy of clinical application and promotion, because it is advantageous over traditional open reduction in terms of shorter operation time, smaller surgical incision, less intraoperative blood loss, less postoperative pain and better knee function.
8.A study of the correlation between oral insufficiency and mild cognitive impairment in community-dwelling older adults
Lili JIAO ; Yingxin TANG ; Si HONG ; Haiyan YIN ; Rumi WANG
Chinese Journal of Geriatrics 2024;43(3):378-383
Objective:To investigate the relationship between oral health and mild cognitive impairment(MCI)by using indicators that evaluate oral hypofunction.Methods:The study was conducted using a cross-sectional design.Participants were recruited from three communities(Shizi Ling, Wenyi Xincun, and Yaoling in Furong District)in Changsha in July 2021, using convenience sampling.Cognitive and oral functions were evaluated using the simple mental state examination, version 2(MMSE-2)and seven indicators of oral hypofunction, which included oral hygiene, oral dryness, occlusal force, tongue and lip movement, tongue pressure, mastication, and swallowing function.Results:A total of 144 subjects were included in this study, with 72 males.Except for education level, there were no statistically significant differences between the MCI group and the normal cognitive group in terms of demographic information and self-reported and measured oral functions(all P>0.05).There was a correlation between the subjects' self-reported oral function and the actual measured oral function.Multiple linear regression analysis revealed no significant correlation between the MMSE score and the seven indicators used to measure oral hypofunction(all P>0.05).The MMSE score of the female group showed a negative correlation with mastication( β=-0.003, P=0.043), while the MMSE score of subjects with elementary school education also showed a negative correlation with mastication( β=-0.022, P=0.016).Additionally, the MMSE score of subjects with middle school education showed a positive correlation with the number of residual teeth( β=0.090, P=0.030). Conclusions:The self-reported oral function can serve as an initial assessment of overall oral function.However, among elderly individuals who do not show significant decline in oral function, there was no significant correlation between oral function and MCI.To accurately identify individuals with MCI, a more detailed sub-analysis with a larger sample size is required.
9.Effect of Qihuang Jianpi Zishen Granules on STAT3/mTOR Signaling Pathway in Patients with Systemic Lupus Erythematosus
Zhongfu TANG ; Chuanbing HUANG ; Lili CHENG ; Junjie CHEN ; Shuangshuang SHANG ; Ming LI ; Sidi LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):118-125
ObjectiveTo investigate the clinical efficacy of Qihuang Jianpi Zishen Granules in the treatment of systemic lupus erythematosus (SLE) and its effect on the signal transducer and activator of tranSCription 3/mammalian target of rapamycin (STAT3/mTOR) signaling pathway, and to decipher the possible mechanism. MethodSixty female SLE patients who met the criteria in the First Affiliated Hospital of Anhui University of Chinese Medicine from May 2022 to May 2023 were selected and randomized into a control group and an observation group (30 cases in each group). The control group was treated with prednisone acetate + hydroxychloroquine sulfate orally, and the observation group was additionally treated with Qihuang Jianpi Zishen granules. The treatment lasted for 8 weeks. The SLE disease activity (SLEDAI), TCM syndrome score, erythrocyte sedimentation rate (ESR), hypersensitive C-reactive protein (hs-CRP), immune indexes [immunoglobulin G (IgG), C3, C4, CD4+, and CD8+], interleukin (IL)-17, IL-23, interferon (IFN)-γ, 24 h urinary protein (24 h PRO), serum creatinine (SCr), and expression of proteins [STAT3, phosphorylated (p)-STAT3, mTOR protein and STAT3,mTOR mRNA] in the STAT3/mTOR signaling pathway were determined before and after treatment. In addition, the adverse reactions were recorded. ResultAfter 8 weeks of treatment, the total response rate in the observation group was 93.33% (28/30), which was higher than that (70.00%, 21/30) in the control group (χ2=4.007, P<0.05). After treatment, both groups showed declined SLEDAI, TCM syndrome score, ESR, hs-CRP, IgG, CD8+, IL-17, IL-23, IFN-γ, 24 h PRO, SCr, and expression of proteins in the STAT3/mTOR pathway (P<0.01) and elevated levels of C3, C4, and CD4+ (P<0.01). Moreover, the observation group had lower SLEDAI, TCM syndrome score, ESR, hs-CRP, IgG, CD8+, IL-17, IL-23, IFN-γ, 24 h PRO, SCr, and expression of proteins in the STAT3/mTOR pathway (P<0.05, P<0.01) and higher levels of C3, C4, and CD4+ (P<0.05, P<0.01) than the control group after treatment. Neither group showed serious adverse reactions during the treatment period. ConclusionQihuang Jianpi Zishen Granules can ameliorate the inflammatory response, reduce the disease activity, and mitigate the kidney injury in SLE by inhibiting the STAT3/mTOR signaling pathway to regulate the immune function.
10.Research on reliability management of medical equipment supply in cardiology ward
Xi TANG ; Ting ZENG ; Yanhua DU ; Lili WU
China Medical Equipment 2024;21(3):123-128
Objective:To construct a reliability management index system of cardiology medical equipment supply,to form a reliability evaluation strategy,and to improve the effectiveness of reliability management.Methods:13 supply management items were decomposed from the three links of spare parts before use,tracking during use and post-use processing of 126 medical equipment,and the probability of work error,probability of quality problems and risk expectation of diagnosis and treatment were evaluated in the whole process management,and the whole-process management countermeasures of grading and stratification,quality standardization and department cooperation was formed.2,000 pieces(sets)of medical equipment used in The People's Hospital of Longhua from 2021 to 2022 were selected by random sampling method,and the department cooperative management[referred to as collaborative management mode,1,000 pieces(sets)times]and reliability evaluation management[referred to as evaluation management mode,1,000 pieces(sets)times]were adopted respectively.The reliability and satisfaction of medical equipment under different management modes were compared.Results:The work error probabilities of pre-use spare parts,in-use tracking,post-use processing and supply of medical equipment adopting evaluation management mode were(4.74±2.19)%,(4.39±1.85)%,(5.75±1.88)%and(8.29±1.30)%,respectively,which were lower than the collaborative management mode,the difference was statistically significant(t=5.367,5.663,5.432,6.847;P<0.05);the probabilities of quality problems were(4.13±1.67)%,(3.89±1.25)%,(5.28±1.84)%and(7.64±1.18)%,respectively,which were lower than the collaborative management mode,the difference was statistically significant(t=6.504,6.229,5.123,6.166,P<0.05);the risk expectations of diagnosis and treatment were(2.74±0.89)%,(2.47±0.96)%,(3.42±0.95)%and(4.02±1.09)%,respectively,which were lower than the collaborative management mode,the difference was statistically significant(t=7.027,6.509,8.915,9.266,P<0.05).The satisfaction of the relevant staff on the performance quality,sterilization specifications,supply timeliness,level of collaboration and safety of the medical equipment adopting the evaluation management mode were(93.54±4.65)%,(91.67±4.11)%,(94.58±4.34)%,(92.39±3.82)%and(90.97±3.76)%,respectively,which were higher than the collaborative management mode,the difference was statistically significant(t=2.809,3.030,2.843,4.939,3.739,P<0.05).Conclusion:The reliability management model can reduce the work errors and security risks in medical equipment supply,improve the reliability of supply and improve the quality of supply service.

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