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.Incidence characteristics of five major malignant tumors in Nanning City
ZHAO Lin ; JIANG Longyan ; XU Bin ; TANG Xianyan
Journal of Preventive Medicine 2025;37(2):135-138
Objective:
To investigate the incidence and trends of liver, lung, breast, colorectal and gastric cancers in Nanning City from 2018 to 2022.
Methods:
Data of new cases of liver, lung, breast, colorectal and gastric cancers in Nanning City from 2018 to 2022 were collected through the big data platform of the Nanning Health Propaganda and Information Center's Hospitalization System. The incidence rates were calculated and standardized using the data of the seventh national population census in 2020. The age, gender, and urban-rural distribution of the five malignant tumor cases were descriptively analyzed. The trends in incidence of the five malignant tumors were analyzed using annual percent change (APC).
Results:
From 2018 to 2022, the crude and standardized incidence rates of liver cancer in Nanning City were 24.46/105 and 26.39/105, respectively, showing upward trends (APC=15.122% and 13.111%, both P<0.05). The crude and standardized incidence rates of lung cancer were 23.42/105 and 25.83/105, respectively, showing upward trends (APC=13.714% and 10.056%, both P<0.05). The crude and standardized incidence rates of breast cancer were 19.13/105 and 20.29/105, respectively, with no significant trends (APC=-5.129% and -5.164%, both P>0.05). The crude incidence rate of colorectal cancer was 18.81/105, showing an upward trend (APC=8.164%, P<0.05), while the standardized incidence rate was 20.64/105, with no significant trend (APC=5.044%, P>0.05). The crude incidence rate of gastric cancer was 7.27/105, showing an upward trend (APC=5.984%, P<0.05), while the standardized incidence rate was 7.98/105, with no significant trend (APC=3.304%, P>0.05). The age of onset peak for liver cancer was 65 to <70 years, for lung cancer was 75 to <80 years, for breast cancer was 55 to <60 years, for colorectal cancer was 80 to <85 years, and for gastric cancer was 75 to <80 years. The standardized incidence rates of liver, lung, colorectal and gastric cancers were higher in males than in females (all P<0.05). The standardized incidence rates of liver, lung and gastric cancers were higher in rural areas than in urban areas, while the standardized incidence rates of breast and colorectal cancers were lower in rural areas (all P<0.05).
Conclusions
The incidence rates of liver, lung, colorectal and gastric cancers in Nanning City showed upward trends from 2018 to 2022, while the incidence rate of breast cancer remained stable. The standardized incidence rates of liver, lung, colorectal and gastric cancers were higher in males, and the standardized incidences of liver, lung and gastric cancers were higher in rural areas.
3.Application of CRISPR/Cas System in Precision Medicine for Triple-negative Breast Cancer
Hui-Ling LIN ; Yu-Xin OUYANG ; Wan-Ying TANG ; Mi HU ; Mao PENG ; Ping-Ping HE ; Xin-Ping OUYANG
Progress in Biochemistry and Biophysics 2025;52(2):279-289
Triple-negative breast cancer (TNBC) represents a distinctive subtype, characterized by the absence of estrogen receptors, progesterone receptors, and human epidermal growth factor receptor 2 (HER2). Due to its high inter-tumor and intra-tumor heterogeneity, TNBC poses significant chanllenges for personalized diagnosis and treatment. The advant of clustered regular interspaced short palindromic repeats (CRISPR) technology has profoundly enhanced our understanding of the structure and function of the TNBC genome, providing a powerful tool for investigating the occurrence and development of diseases. This review focuses on the application of CRISPR/Cas technology in the personalized diagnosis and treatment of TNBC. We begin by discussing the unique attributes of TNBC and the limitations of current diagnostic and treatment approaches: conventional diagnostic methods provide limited insights into TNBC, while traditional chemotherapy drugs are often associated with low efficacy and severe side effects. The CRISPR/Cas system, which activates Cas enzymes through complementary guide RNAs (gRNAs) to selectively degrade specific nucleic acids, has emerged as a robust tool for TNBC research. This technology enables precise gene editing, allowing for a deeper understanding of TNBC heterogeneity by marking and tracking diverse cell clones. Additionally, CRISPR facilitates high-throughput screening to promptly identify genes involved in TNBC growth, metastasis, and drug resistance, thus revealing new therapeutic targets and strategies. In TNBC diagnostics, CRISPR/Cas was applied to develop molecular diagnostic systems based on Cas9, Cas12, and Cas13, each employing distinct detection principles. These systems can sensitively and specifically detect a variety of TNBC biomarkers, including cell-specific DNA/RNA and circulating tumor DNA (ctDNA). In the realm of precision therapy, CRISPR/Cas has been utilized to identify key genes implicated in TNBC progression and treatment resistance. CRISPR-based screening has uncovered potential therapeutic targets, while its gene-editing capabilities have facilitated the development of combination therapies with traditional chemotherapy drugs, enhancing their efficacy. Despite its promise, the clinical translation of CRISPR/Cas technology remains in its early stages. Several clinical trials are underway to assess its safety and efficacy in the treatment of various genetic diseases and cancers. Challenges such as off-target effects, editing efficiency, and delivery methods remain to be addressed. The integration of CRISPR/Cas with other technologies, such as 3D cell culture systems, human induced pluripotent stem cells (hiPSCs), and artificial intelligence (AI), is expected to further advance precision medicine for TNBC. These technological convergences can offer deeper insights into disease mechanisms and facilitate the development of personalized treatment strategies. In conclusion, the CRISPR/Cas system holds immense potential in the precise diagnosis and treatment of TNBC. As the technology progresses and becomes more costs-effective, its clinical relevance will grow, and the translation of CRISPR/Cas system data into clinical applications will pave the way for optimal diagnosis and treatment strategies for TNBC patients. However, technical hurdles and ethical considerations require ongoing research and regulation to ensure safety and efficacy.
4.Strategies and advances in hepatitis B vaccination in China
Lin TANG ; Xia XU ; Zemei ZHOU ; Xiaoqi WANG ; Jizhen LI ; Zundong YIN ; Fuzhen WANG
Journal of Clinical Hepatology 2025;41(2):210-215
Hepatitis B is a major global public health issue. Through the implementation of comprehensive prevention and control strategies centered on hepatitis B vaccination, China has achieved remarkable progress in hepatitis B prevention and control, while there are still many issues and challenges. This article reviews the development of hepatitis B vaccination strategies in China, analyzes the goal and advances in vaccination in different populations, and problems and challenges, in order to provide a reference for further optimizing vaccination strategies and improving the levels of prevention and control.
5.Progress on antisense oligonucleotide in the field of antibacterial therapy
Jia LI ; Xiao-lu HAN ; Shi-yu SONG ; Jin-tao LIN ; Zhi-qiang TANG ; Zeng-ming WANG ; Liang XU ; Ai-ping ZHENG
Acta Pharmaceutica Sinica 2025;60(2):337-347
With the widespread use of antibiotics, drug-resistant bacterial infections have become a significant threat to human health. Finding new antibacterial strategies that can effectively control drug-resistant bacterial infections has become an urgent task. Unlike small molecule drugs that target bacterial proteins, antisense oligonucleotide (ASO) can target genes related to bacterial resistance, pathogenesis, growth, reproduction and biofilm formation. By regulating the expression of these genes, ASO can inhibit or kill bacteria, providing a novel approach for the development of antibacterial drugs. To overcome the challenge of delivering antisense oligonucleotide into bacterial cells, various drug delivery systems have been applied in this field, including cell-penetrating peptides, lipid nanoparticles and inorganic nanoparticles, which have injected new momentum into the development of antisense oligonucleotide in the antibacterial realm. This review summarizes the current development of small nucleic acid drugs, the antibacterial mechanisms, targets, sequences and delivery vectors of antisense oligonucleotide, providing a reference for the research and development of antisense oligonucleotide in the treatment of bacterial infections.
6.The effect of rutaecarpine on improving fatty liver and osteoporosis in MAFLD mice
Yu-hao ZHANG ; Yi-ning LI ; Xin-hai JIANG ; Wei-zhi WANG ; Shun-wang LI ; Ren SHENG ; Li-juan LEI ; Yu-yan ZHANG ; Jing-rui WANG ; Xin-wei WEI ; Yan-ni XU ; Yan LIN ; Lin TANG ; Shu-yi SI
Acta Pharmaceutica Sinica 2025;60(1):141-149
Metabolic-associated fatty liver disease (MAFLD) and osteoporosis (OP) are two very common metabolic diseases. A growing body of experimental evidence supports a pathophysiological link between MAFLD and OP. MAFLD is often associated with the development of OP. Rutaecarpine (RUT) is one of the main active components of Chinese medicine Euodiae Fructus. Our previous studies have demonstrated that RUT has lipid-lowering, anti-inflammatory and anti-atherosclerotic effects, and can improve the OP of rats. However, whether RUT can improve both fatty liver and OP symptoms of MAFLD mice at the same time remains to be investigated. In this study, we used C57BL/6 mice fed a high-fat diet (HFD) for 4 months to construct a MAFLD model, and gave the mice a low dose (5 mg·kg-1) and a high dose (15 mg·kg-1) of RUT by gavage for 4 weeks. The effects of RUT on liver steatosis and bone metabolism were then evaluated at the end of the experiment [this experiment was approved by the Experimental Animal Ethics Committee of Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences (approval number: IMB-20190124D303)]. The results showed that RUT treatment significantly reduced hepatic steatosis and lipid accumulation, and significantly reduced bone loss and promoted bone formation. In summary, this study shows that RUT has an effect of improving fatty liver and OP in MAFLD mice.
7.A Case Report of Hypothyrotropin Hypothyroidism Caused by Roxadustat
Xuelian YAN ; Bingying TANG ; Xuan QU ; Ning ZHANG ; Lin KANG
Medical Journal of Peking Union Medical College Hospital 2025;16(2):519-522
Roxadustat is the world's first small molecule hypoxia-inducible factor prolyl hydroxylase inhibitor. Its adverse effect of causing hypothyroidism with low thyroid-stimulating hormone (TSH) is relatively rare and manifests subtly in elderly patients with multiple coexisting diseases. This article reports a case of an elderly patient with renal anemia who developed reversible low-TSH hypothyroidism after taking roxadustat for 12 days, with a significant decrease in thyroid hormone levels. After discontinuing roxadustat for 15 days, the thyroid hormone levels gradually returned to normal. Due to the worsening of renal anemia, the patient took roxadustat again, and 9 days later, the thyroid function-related indicators decreased upon re-examination, leading to the initiation of levothyroxine replacement therapy. In conjunction with relevant literature, this article analyzes the adverse reactions that occur during the oral administration of roxadustat in elderly patients with chronic kidney disease, aiming to provide reference for drug treatment of such patients.
8.Characteristic ion Identification of Different Original Haliotidis Concha and Its Counterfeits
Xiaojie LIANG ; Guowei LI ; Lin ZHOU ; Qiping HU ; Muxiang LUO ; Jiehao TANG ; Xiangdong CHEN ; Liye PAN ; Dongmei SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):263-269
ObjectiveTo establish a method for the identification of Haliotidis Concha and its counterfeits, and to improve its quality evaluation method. MethodsA total of 17 batches of Haliotis discus hannai, 4 batches of H. ruber, 3 batches of H. laevigata, 3 batches of H. ovina, 3 batches of H. diversicolor, 3 batches of H. asinina, 3 batches of H. iris were collected. Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive-Orbitrap-MS/MS) was used to analyze the hydrolysates of different original Haliotidis Concha and its counterfeits, and the potential characteristic ions of each species were screened by Venn diagram. UPLC-triple quadrupole tandem mass spectrometry(UPLC-QqQ-MS/MS) was used to validate the characteristic ions, and the specific detection method of the characteristic ions was established. ResultsA total of 1 182, 167, 47, 89, 104, 203, 424 potential characteristic ions were screened from H. discus hannai, H. ruber, H. laevigata, H. ovina, H. diversicolor, H. asinina and H. iris, respectively. And 9 characteristic ions were selected. The precision, stability and repeatability of the 9 characteristic ions in the established identification method met the requirements. Different original Haliotidis Concha and its counterfeits could detect their own characteristic ions, including m/z 631.83-886.48(double charge) and m/z 631.83-443.74(double charge) of H. discus hannai, m/z 699.28-232.11(double charge) and m/z 699.28-544.27(double charge) of H. ruber, m/z 535.76-752.37(double charge) and m/z 535.76-548.28(double charge) of H. laevigata, m/z 708.35-442.28(double charge) and m/z 708.35-215.14(double charge) of H. ovina, m/z 561.33-614.86(triple charge), m/z 561.33-468.28(triple charge), m/z 608.29-618.32(double charge) and m/z 608.29-390.21(double charge) of H. diversicolor, m/z 769.85-274.10(double charge), m/z 769.85-532.75(double charge), m/z 827.43-646.36(single charge), m/z 827.43-257.12(single charge) of H. asinina, and m/z 468.24-576.29(double charge) and m/z 468.24-505.26(double charge) of H. iris. ConclusionIn this study, a total of 9 characteristic ions are screened from 6 kinds of original Haliotidis Concha and its counterfeits, and a specific identification method is established, which is helpful to solve the limitations of the existing quality evaluation methods of Haliotidis Concha, and provide a basis for the production, circulation and medication quality.
9.Characteristic ion Identification of Different Original Haliotidis Concha and Its Counterfeits
Xiaojie LIANG ; Guowei LI ; Lin ZHOU ; Qiping HU ; Muxiang LUO ; Jiehao TANG ; Xiangdong CHEN ; Liye PAN ; Dongmei SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):263-269
ObjectiveTo establish a method for the identification of Haliotidis Concha and its counterfeits, and to improve its quality evaluation method. MethodsA total of 17 batches of Haliotis discus hannai, 4 batches of H. ruber, 3 batches of H. laevigata, 3 batches of H. ovina, 3 batches of H. diversicolor, 3 batches of H. asinina, 3 batches of H. iris were collected. Ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive-Orbitrap-MS/MS) was used to analyze the hydrolysates of different original Haliotidis Concha and its counterfeits, and the potential characteristic ions of each species were screened by Venn diagram. UPLC-triple quadrupole tandem mass spectrometry(UPLC-QqQ-MS/MS) was used to validate the characteristic ions, and the specific detection method of the characteristic ions was established. ResultsA total of 1 182, 167, 47, 89, 104, 203, 424 potential characteristic ions were screened from H. discus hannai, H. ruber, H. laevigata, H. ovina, H. diversicolor, H. asinina and H. iris, respectively. And 9 characteristic ions were selected. The precision, stability and repeatability of the 9 characteristic ions in the established identification method met the requirements. Different original Haliotidis Concha and its counterfeits could detect their own characteristic ions, including m/z 631.83-886.48(double charge) and m/z 631.83-443.74(double charge) of H. discus hannai, m/z 699.28-232.11(double charge) and m/z 699.28-544.27(double charge) of H. ruber, m/z 535.76-752.37(double charge) and m/z 535.76-548.28(double charge) of H. laevigata, m/z 708.35-442.28(double charge) and m/z 708.35-215.14(double charge) of H. ovina, m/z 561.33-614.86(triple charge), m/z 561.33-468.28(triple charge), m/z 608.29-618.32(double charge) and m/z 608.29-390.21(double charge) of H. diversicolor, m/z 769.85-274.10(double charge), m/z 769.85-532.75(double charge), m/z 827.43-646.36(single charge), m/z 827.43-257.12(single charge) of H. asinina, and m/z 468.24-576.29(double charge) and m/z 468.24-505.26(double charge) of H. iris. ConclusionIn this study, a total of 9 characteristic ions are screened from 6 kinds of original Haliotidis Concha and its counterfeits, and a specific identification method is established, which is helpful to solve the limitations of the existing quality evaluation methods of Haliotidis Concha, and provide a basis for the production, circulation and medication quality.
10.Analysis of distortion product otoacoustic emissions results of noise-exposed workers at a metal shipbuilding enterprise
Jieting ZHOU ; Jianyu GUO ; Hairu YANG ; Linyan SHU ; Zhixing FAN ; Jia TANG ; Xinqiang NIE ; Guoyong XU ; Hansheng LIN ; Bin XIAO
China Occupational Medicine 2025;52(1):99-105
Objective To evaluate the role of distortion product otoacoustic emissions (DPOAE) testing in evaluating early hearing loss among noise-exposed workers. Methods A total of 174 noise-exposed workers in a metal shipbuilding enterprise were selected as the research subjects by the convenience sampling method. Pure tone audiometry (PTA), DPOAE and the level of noise exposure were conducted on the workers. The rank correlation analysis was used to analyze the correlation between DPOAE amplitude and PTA threshold. The multilevel model was used to analyze the effects of gender, age, noise exposure intensity, cumulative noise exposure (CNE), hearing loss classification and PTA threshold on DPOAE results. Results At the frequencies of 0.50, 1.00, 2.00, 3.00, 4.00, 6.00 and 8.00 kHz, the DPOAE amplitude was negatively correlated with the PTA threshold (rank correlation coefficients were -0.12, -0.48, -0.47, -0.18, -0.23, -0.44, -0.19, respectively, all P<0.01). At the most frequencies, DPOAE amplitude was negatively correlated with age and CNE (all P<0.05). The results of multilevel model analysis showed that there were significant differences in DPOAE amplitudes at certain frequencies across gender, age, noise intensity, CNE, and hearing loss classification (all P<0.05). Significant differences in DPOAE responses were found among different CNE and hearing loss groups (all P<0.01). Conclusion DPOAE testing can objectively reflect the hearing status of noise-exposed workers and could be considered for inclusion in routine hearing monitoring to facilitate early detection of noise-induced hearing loss.


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