1.Features of intestinal flora in patients with nonalcoholic fatty liver disease and Helicobacter pylori infection
Jingjing LIU ; Qike WANG ; Zhiqiang MA ; Yan LIANG ; Renping LI
Journal of Clinical Hepatology 2025;41(5):862-871
ObjectiveTo investigate the features and mechanism of action of intestinal flora in patients with nonalcoholic fatty liver disease (NAFLD) and Helicobacter pylori (HP) infection by comparing the changes in intestinal flora between the healthy population, the patients with HP infection, the patients with NAFLD, and the patients with NAFLD and HP infection. MethodsThis study was conducted among the 19 patients with NAFLD (NAFLD group), 19 patients with HP infection (HP group), and 19 patients with NAFLD and HP infection (NAFLD+HP group) who were admitted to The Second Affiliated Hospital of Henan University of Science and Technology from March 1, 2023 to April 30, 2024, and 20 individuals undergoing physical examination were enrolled as control group. Fecal samples were collected, total DNA was extracted for PCR amplification, and 16S rDNA sequencing was performed to compare the features of intestinal flora between the four groups. An analysis of variance was used for comparison of continuous data between multiple groups, and the chi-square test was used for comparison of categorical data between multiple groups. The Mann-Whitney U test or the Kruskal-Wallis H test was used for comparison of the species in intestinal flora. ResultsThe NAFLD+HP group showed a tendency of reduction in flora abundance compared with the other three groups. There was a significant difference in flora distribution between the NAFLD+HP group and the NAFLD group and between the NAFLD group and the control group (P<0.05). At the phylum level, the top three species in the NAFLD+HP group were Firmicutes (59.94%), Proteobacteria (17.00%), and Actinobacteria (14.75%), with an increase in the proportion of Proteobacteria and a reduction in the proportion of Actinobacteria compared with the other three groups. At the genus level, the top five dominant bacteria in the NAFLD+HP group were Bifidobacterium, Streptococcus, Escherichia-Shigella, Agathobacter, and Ruminococcus gnavus_group. Compared with the NAFLD group, the NAFLD+HP group had increases in the abundance of Streptococcus, Veillonella, and Rothia and reductions in the abundance of Dialister and Ruminococcus toraues_group. Compared with the HP group, the NAFLD+HP group had reductions in the abundance of Collinsella, Subdoligranulum, Catenibacterium, and Porphyromonas and increases in the abundance of Citrobacter and Olsenella (all P<0.05). ConclusionPatients with NAFLD and HP infection have changed in intestinal flora. These flora may be the intestinal microecological factors for HP infection in promoting the development and progression of NAFLD.
2.Clinical features of recompensation in autoimmune hepatitis-related decompensated cirrhosis and related predictive factors
Xiaolong LU ; Lin HAN ; Huan XIE ; Lilong YAN ; Xuemei MA ; Dongyan LIU ; Xun LI ; Qingsheng LIANG ; Zhengsheng ZOU ; Caizhe GU ; Ying SUN
Journal of Clinical Hepatology 2025;41(9):1808-1817
ObjectiveTo investigate the clinical features and outcomes of recompensation in patients with autoimmune hepatitis (AIH)-related decompensated cirrhosis, to identify independent predictive factors, and to construct a nomogram prediction model for the probability of recompensation. MethodsA retrospective cohort study was conducted among the adult patients with AIH-related decompensated cirrhosis who were admitted to The Fifth Medical Center of PLA General Hospital from January 2015 to August 2023 (n=211). The primary endpoint was achievement of recompensation, and the secondary endpoint was liver-related death or liver transplantation. According to the outcome of the patients at the end of the follow-up, the patients were divided into the recompensation group (n=16) and the persistent decompensation group(n=150).The independent-samples t test was used for comparison of normally distributed continuous data with homogeneity of variance, and the Mann-Whitney U rank sum test was used for comparison of non-normally distributed continuous data with heterogeneity of variance; the chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups; the Kaplan-Meier method was used for survival analysis; the Cox proportional-hazards regression model was used to identify independent predictive factors, and a nomogram model was constructed and validated. ResultsA total of 211 patients were enrolled, with a median age of 55.0 years and a median follow-up time of 44.0 months, and female patients accounted for 87.2%. Among the 211 patients, 61 (with a cumulative proportion of 35.5%) achieved recompensation. Compared with the persistent decompensation group, the recompensation group had significantly higher white blood cell count, platelet count (PLT), total bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bile acid, prothrombin time, international normalized ratio (INR), SMA positive rate, Model for End-Stage Liver Disease (MELD) score, Child-Pugh score, and rate of use of glucocorticoids (all P0.05), as well as significantly lower age at baseline, number of complications, and death/liver transplantation rate (all P0.05). At 3 and 12 months after treatment, the recompensation group had continuous improvements in AST, TBil, INR, IgG, MELD score, and Child-Pugh score, which were significantly lower than the values in the persistent decompensation group (all P0.05), alongside with continuous increases in PLT and albumin, which were significantly higher than the values in the persistent decompensation group (P0.05). The multivariate Cox regression analysis showed that baseline ALT (hazard ratio [HR]=1.067, 95% confidence interval [CI]: 1.010 — 1.127, P=0.021), IgG (HR=0.463,95%CI:0.258 — 0.833, P=0.010), SMA positivity (HR=3.122,95%CI:1.768 — 5.515, P0.001), and glucocorticoid therapy (HR=20.651,95%CI:8.744 — 48.770, P0.001) were independent predictive factors for recompensation, and the nomogram model based on these predictive factors showed excellent predictive performance (C-index=0.87,95%CI:0.84 — 0.90). ConclusionAchieving recompensation significantly improves clinical outcomes in patients with AIH-related decompensated cirrhosis. Baseline SMA positivity, a high level of ALT, a low level of IgG, and corticosteroid therapy are independent predictive factors for recompensation. The predictive model constructed based on these factors can provide a basis for decision-making in individualized clinical management.
3.Analysis of risk factors, pathogenic bacteria characteristics, and drug resistance of postoperative surgical site infection in adults with limb fractures.
Yan-Jun WANG ; Zi-Hou ZHAO ; Shuai-Kun LU ; Guo-Liang WANG ; Shan-Jin MA ; Lin-Hu WANG ; Hao GAO ; Jun REN ; Zhong-Wei AN ; Cong-Xiao FU ; Yong ZHANG ; Wen LUO ; Yun-Fei ZHANG
Chinese Journal of Traumatology 2025;28(4):241-251
PURPOSE:
We carried out the study aiming to explore and analyze the risk factors, the distribution of pathogenic bacteria, and their antibiotic-resistance characteristics influencing the occurrence of surgical site infection (SSI), to provide valuable assistance for reducing the incidence of SSI after traumatic fracture surgery.
METHODS:
A retrospective case-control study enrolling 3978 participants from January 2015 to December 2019 receiving surgical treatment for traumatic fractures was conducted at Tangdu Hospital of Air Force Medical University. Baseline data, demographic characteristics, lifestyles, variables related to surgical treatment, and pathogen culture were harvested and analyzed. Univariate analyses and multivariate logistic regression analyses were used to reveal the independent risk factors of SSI. A bacterial distribution histogram and drug-sensitive heat map were drawn to describe the pathogenic characteristics.
RESULTS:
Included 3978 patients 138 of them developed SSI with an incidence rate of 3.47% postoperatively. By logistic regression analysis, we found that variables such as gender (males) (odds ratio (OR) = 2.012, 95% confidence interval (CI): 1.235 - 3.278, p = 0.005), diabetes mellitus (OR = 5.848, 95% CI: 3.513 - 9.736, p < 0.001), hypoproteinemia (OR = 3.400, 95% CI: 1.280 - 9.031, p = 0.014), underlying disease (OR = 5.398, 95% CI: 2.343 - 12.438, p < 0.001), hormonotherapy (OR = 11.718, 95% CI: 6.269 - 21.903, p < 0.001), open fracture (OR = 29.377, 95% CI: 9.944 - 86.784, p < 0.001), and intraoperative transfusion (OR = 2.664, 95% CI: 1.572 - 4.515, p < 0.001) were independent risk factors for SSI, while, aged over 59 years (OR = 0.132, 95% CI: 0.059 - 0.296, p < 0.001), prophylactic antibiotics use (OR = 0.082, 95% CI: 0.042 - 0.164, p < 0.001) and vacuum sealing drainage use (OR = 0.036, 95% CI: 0.010 - 0.129, p < 0.001) were protective factors. Pathogens results showed that 301 strains of 38 species of bacteria were harvested, among which 178 (59.1%) strains were Gram-positive bacteria, and 123 (40.9%) strains were Gram-negative bacteria. Staphylococcus aureus (108, 60.7%) and Enterobacter cloacae (38, 30.9%) accounted for the largest proportion. The susceptibility of Gram-positive bacteria to Vancomycin and Linezolid was almost 100%. The susceptibility of Gram-negative bacteria to Imipenem, Amikacin, and Meropenem exceeded 73%.
CONCLUSION
Orthopedic surgeons need to develop appropriate surgical plans based on the risk factors and protective factors associated with postoperative SSI to reduce its occurrence. Meanwhile, it is recommended to strengthen blood glucose control in the early stage of admission and for surgeons to be cautious and scientific when choosing antibiotic therapy in clinical practice.
Humans
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Surgical Wound Infection/epidemiology*
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Male
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Female
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Risk Factors
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Retrospective Studies
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Middle Aged
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Adult
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Case-Control Studies
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Fractures, Bone/surgery*
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Aged
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Drug Resistance, Bacterial
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Logistic Models
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Anti-Bacterial Agents/therapeutic use*
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Incidence
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Bacteria/drug effects*
4.The addition of 5-aminolevulinic acid to HBSS protects testis grafts during hypothermic transportation: a novel preservation strategy.
Meng-Hui MA ; Pei-Gen CHEN ; Jun-Xian HE ; Hai-Cheng CHEN ; Zhen-Han XU ; Lin-Yan LV ; Yan-Qing LI ; Xiao-Yan LIANG ; Gui-Hua LIU
Asian Journal of Andrology 2025;27(4):454-463
The aim of this investigation was to determine the optimal storage medium for testicular hypothermic transportation and identify the ideal concentration for the application of the protective agent 5-aminolevulinic acid (5-ALA). Furthermore, this study aimed to explore the underlying mechanism of the protective effects of 5-ALA. First, we collected and stored mouse testicular fragments in different media, including Hank's balanced salt solution (HBSS; n = 5), Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12; n = 5), and alpha-minimum essential medium (αMEM; n = 5). Storage of testicular tissue in HBSS preserved the integrity of testicular morphology better than that in the DMEM/F12 group ( P < 0.05) and the αMEM group ( P < 0.01). Testicular fragments were subsequently placed in HBSS with various concentrations of 5-ALA (0 [control], 1 mmol l -1 , 2 mmol l -1 , and 5 mmol l -1 ) to determine the most effective concentration of 5-ALA. The 2 mmol l -1 5-ALA group ( n = 3) presented the highest positive rate of spermatogonial stem cells compared with those in the control, 1 mmol l -1 , and 5 mmol l -1 5-ALA groups. Finally, the tissue fragments were preserved in HBSS with control ( n = 3) and 2 mmol l -1 5-ALA ( n = 3) under low-temperature conditions. A comparative analysis was performed against fresh testes ( n = 3) to elucidate the underlying mechanism of 5-ALA. Gene set enrichment analysis (GSEA) for WikiPathways revealed that the p38 mitogen-activated protein kinase (MAPK) signaling pathway was downregulated in the 2 mmol l -1 5-ALA group compared with that in the control group (normalized enrichment score [NES] = -1.57, false discovery rate [FDR] = 0.229, and P = 0.019). In conclusion, these data suggest that using 2 mmol l -1 5-ALA in HBSS effectively protected the viability of spermatogonial stem cells upon hypothermic transportation.
Male
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Animals
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Testis/cytology*
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Aminolevulinic Acid/pharmacology*
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Mice
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Organ Preservation/methods*
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Organ Preservation Solutions/pharmacology*
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Cryopreservation/methods*
5.Expert consensus on the diagnosis and treatment of cemental tear.
Ye LIANG ; Hongrui LIU ; Chengjia XIE ; Yang YU ; Jinlong SHAO ; Chunxu LV ; Wenyan KANG ; Fuhua YAN ; Yaping PAN ; Faming CHEN ; Yan XU ; Zuomin WANG ; Yao SUN ; Ang LI ; Lili CHEN ; Qingxian LUAN ; Chuanjiang ZHAO ; Zhengguo CAO ; Yi LIU ; Jiang SUN ; Zhongchen SONG ; Lei ZHAO ; Li LIN ; Peihui DING ; Weilian SUN ; Jun WANG ; Jiang LIN ; Guangxun ZHU ; Qi ZHANG ; Lijun LUO ; Jiayin DENG ; Yihuai PAN ; Jin ZHAO ; Aimei SONG ; Hongmei GUO ; Jin ZHANG ; Pingping CUI ; Song GE ; Rui ZHANG ; Xiuyun REN ; Shengbin HUANG ; Xi WEI ; Lihong QIU ; Jing DENG ; Keqing PAN ; Dandan MA ; Hongyu ZHAO ; Dong CHEN ; Liangjun ZHONG ; Gang DING ; Wu CHEN ; Quanchen XU ; Xiaoyu SUN ; Lingqian DU ; Ling LI ; Yijia WANG ; Xiaoyuan LI ; Qiang CHEN ; Hui WANG ; Zheng ZHANG ; Mengmeng LIU ; Chengfei ZHANG ; Xuedong ZHOU ; Shaohua GE
International Journal of Oral Science 2025;17(1):61-61
Cemental tear is a rare and indetectable condition unless obvious clinical signs present with the involvement of surrounding periodontal and periapical tissues. Due to its clinical manifestations similar to common dental issues, such as vertical root fracture, primary endodontic diseases, and periodontal diseases, as well as the low awareness of cemental tear for clinicians, misdiagnosis often occurs. The critical principle for cemental tear treatment is to remove torn fragments, and overlooking fragments leads to futile therapy, which could deteriorate the conditions of the affected teeth. Therefore, accurate diagnosis and subsequent appropriate interventions are vital for managing cemental tear. Novel diagnostic tools, including cone-beam computed tomography (CBCT), microscopes, and enamel matrix derivatives, have improved early detection and management, enhancing tooth retention. The implementation of standardized diagnostic criteria and treatment protocols, combined with improved clinical awareness among dental professionals, serves to mitigate risks of diagnostic errors and suboptimal therapeutic interventions. This expert consensus reviewed the epidemiology, pathogenesis, potential predisposing factors, clinical manifestations, diagnosis, differential diagnosis, treatment, and prognosis of cemental tear, aiming to provide a clinical guideline and facilitate clinicians to have a better understanding of cemental tear.
Humans
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Dental Cementum/injuries*
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Consensus
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Diagnosis, Differential
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Cone-Beam Computed Tomography
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Tooth Fractures/therapy*
6.Lentivirus-modified hematopoietic stem cell gene therapy for advanced symptomatic juvenile metachromatic leukodystrophy: a long-term follow-up pilot study.
Zhao ZHANG ; Hua JIANG ; Li HUANG ; Sixi LIU ; Xiaoya ZHOU ; Yun CAI ; Ming LI ; Fei GAO ; Xiaoting LIANG ; Kam-Sze TSANG ; Guangfu CHEN ; Chui-Yan MA ; Yuet-Hung CHAI ; Hongsheng LIU ; Chen YANG ; Mo YANG ; Xiaoling ZHANG ; Shuo HAN ; Xin DU ; Ling CHEN ; Wuh-Liang HWU ; Jiacai ZHUO ; Qizhou LIAN
Protein & Cell 2025;16(1):16-27
Metachromatic leukodystrophy (MLD) is an inherited disease caused by a deficiency of the enzyme arylsulfatase A (ARSA). Lentivirus-modified autologous hematopoietic stem cell gene therapy (HSCGT) has recently been approved for clinical use in pre and early symptomatic children with MLD to increase ARSA activity. Unfortunately, this advanced therapy is not available for most patients with MLD who have progressed to more advanced symptomatic stages at diagnosis. Patients with late-onset juvenile MLD typically present with a slower neurological progression of symptoms and represent a significant burden to the economy and healthcare system, whereas those with early onset infantile MLD die within a few years of symptom onset. We conducted a pilot study to determine the safety and benefit of HSCGT in patients with postsymptomatic juvenile MLD and report preliminary results. The safety profile of HSCGT was favorable in this long-term follow-up over 9 years. The most common adverse events (AEs) within 2 months of HSCGT were related to busulfan conditioning, and all AEs resolved. No HSCGT-related AEs and no evidence of distorted hematopoietic differentiation during long-term follow-up for up to 9.6 years. Importantly, to date, patients have maintained remarkably improved ARSA activity with a stable disease state, including increased Functional Independence Measure (FIM) score and decreased magnetic resonance imaging (MRI) lesion score. This long-term follow-up pilot study suggests that HSCGT is safe and provides clinical benefit to patients with postsymptomatic juvenile MLD.
Humans
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Leukodystrophy, Metachromatic/genetics*
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Pilot Projects
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Genetic Therapy/methods*
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Hematopoietic Stem Cell Transplantation
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Male
;
Follow-Up Studies
;
Female
;
Lentivirus/genetics*
;
Child
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Child, Preschool
;
Hematopoietic Stem Cells/metabolism*
;
Cerebroside-Sulfatase/metabolism*
;
Adolescent
7.Development of a High-throughput Sequencing Platform for Detection of Viral Encephalitis Pathogens Based on Amplicon Sequencing
Li Ya ZHANG ; Zhe Wen SU ; Chen Rui WANG ; Yan LI ; Feng Jun ZHANG ; Hui Sheng LIU ; He Dan HU ; Xiao Chong XU ; Yu Jia YIN ; Kai Qi YIN ; Ying HE ; Fan LI ; Hong Shi FU ; Kai NIE ; Dong Guo LIANG ; Yong TAO ; Tao Song XU ; Feng Chao MA ; Yu Huan WANG
Biomedical and Environmental Sciences 2024;37(3):294-302
Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.
8.Nanozyme and Abiogenesis
Long MA ; Zi-Mo LIANG ; Yin-Yin HOU ; Jing FENG ; Ke-Long FAN ; Xi-Yun YAN
Progress in Biochemistry and Biophysics 2024;51(10):2274-2291
Nanozymes, a groundbreaking discovery by Chinese scientists, represent a novel and remarkable property of nanomaterials. They not only exhibit catalytic activity comparable to natural enzymes, but also boast exceptional stability, tunable reactivity, and the ability to catalyze reactions under mild conditions. The identification of nanozymes has unveiled the biocatalytic potential of inorganic nanomaterials. In parallel, inorganic minerals have long been regarded as pivotal catalysts in the origin of life, driving the synthesis of early biomolecules. These minerals not only facilitate redox reactions that convert simple inorganic compounds into organic molecules but also enable chiral selection, the synthesis of biomacromolecules, and radioprotective functions via their surface structures. Recent advances suggest that inorganic nanomaterials can delicately catalyze the formation of biomolecules, aid in macromolecular assembly, and provide radiation shielding. Furthermore, nanominerals are found in abundance across Earth and extraterrestrial environments. This paper seeks to explore the potential of nanozymes as catalytic agents in the processes that gave rise to life, integrating the catalytic roles of inorganic minerals with the unique attributes of nanozymes, which will provide a new perspective for research of origin of life.
9.Effect of Modified Dahuang Huanglian Xiexintang on Oxidative Stress Injury of Liver in Type 2 Diabetes Mellitus Rats Based on Nrf2/HO-1 Axis
Chengjun MA ; Fengzhe YAN ; Lixia YANG ; Yonglin LIANG ; Xiangdong ZHU ; Dong AN ; Yankui GAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):121-130
ObjectiveTo explore the effects and mechanisms of modified Dahuang Huanglian Xiexintang on hepatic oxidative stress injury in type 2 diabetes mellitus (T2DM) rats based on the nuclear factor erythroid 2 related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) axis. MethodSix ZDF (fa/+) rats were as assigned to the blank group, and 30 ZDF (fa/fa) rats were used to induce the T2DM model by feeding a high-fat diet. After successful modeling, the rats were randomly divided into the model group, metformin group (0.18 g·kg-1), and low, medium, and high dose groups of modified Dahuang Huanglian Xiexintang (0.54, 1.08, 2.16 g·kg-1), with six rats in each group. After 12 weeks of drug intervention, the body mass, liver mass, fasting blood glucose (FBG), and oral glucose tolerance test (OGTT) levels were measured. Serum total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL), low-density lipoprotein cholesterol (LDL), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels were detected using an automatic biochemical analyzer. The pathological changes of liver tissue were observed by hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay (ELISA) was used to detect the activity of superoxide dismutase (SOD), reactive oxygen species (ROS), glutathione peroxidase (GSH-Px), and the level of malondialdehyde (MDA) in liver tissues. Immunohistochemistry was used to detect the expression of Nrf2 in the liver. Real time quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the mRNA and protein expression levels of Nrf2 and HO-1 in liver tissues. ResultCompared with the normal group, the model group showed a significant increase in body mass, liver mass, and liver index (P<0.01). Compared with the model group, the metformin group and the medium and high dose groups of modified Dahuang Huanglian Xiexintang showed a significant decrease in body weight, liver mass, and liver index (P<0.01). Compared with the normal group, the model group showed significantly increased TC, TG, and LDL levels (P<0.01), and significantly decreased HDL levels (P<0.01). Compared with the model group, the metformin group and all doses of modified Dahuang Huanglian Xiexintang showed significantly reduced TC levels (P<0.01), and significantly reduced TG levels (P<0.05). The medium and high dose groups of modified Dahuang Huanglian Xiexintang showed significantly reduced LDL levels (P<0.05). The metformin group and all doses of modified Dahuang Huanglian Xiexintang showed significantly increased HDL levels (P<0.05). Compared with the normal group, the model group showed significantly increased ALT and AST activities (P<0.01). Compared with the model group, all doses of modified Dahuang Huanglian Xiexintang and the metformin group showed significantly reduced ALT activities (P<0.05) and significantly reduced AST activities (P<0.01). Compared with normal group, the model group showed significantly increased FBG at all time points (P<0.01). Compared with the model group, the metformin group and all doses of modified Dahuang Huanglian Xiexintang showed significantly reduced FBG at 8, 10, 12 weeks. The OGTT results showed that compared with the normal group, the model group had significantly increased blood glucose at all time points (P<0.01). Compared with the model group, the metformin group showed significantly reduced blood glucose at all time points (P<0.01), and the medium and high dose groups of modified Dahuang Huanglian Xiexintang showed significantly reduced blood glucose at 90, 120 min (P<0.01). HE pathology showed clear and regular liver cell structure in the normal group, while the model group showed disordered liver cell structure with visible fat vacuoles and a large number of deformed necrotic cells. The liver tissue structure improved in the metformin group and all doses of modified Dahuang Huanglian Xiexintang, with fewer necrotic cells. Compared with the normal group, the model group showed significantly reduced SOD and GSH-Px levels (P<0.01), and significantly increased ROS and MDA levels (P<0.01). Compared with the model group, the metformin group and all doses of modified Dahuang Huanglian Xiexintang showed significantly increased SOD and GSH-Px levels (P<0.01), and significantly reduced MDA levels (P<0.01). The medium and high dose groups of modified Dahuang Huanglian Xiexintang showed significantly reduced ROS levels (P<0.05). Compared with the normal group, the model group showed significantly reduced Nrf2 and HO-1 mRNA expression levels (P<0.01). Compared with the model group, the metformin group and the medium and high dose groups of modified Dahuang Huanglian Xiexintang showed significantly increased Nrf2 and HO-1 mRNA expression levels (P<0.05). Immunohistochemistry showed that compared with the normal group, the model group had significantly reduced positive expression of Nrf2 and HO-1 (P<0.05). Compared with the model group, the metformin group and all doses of modified Dahuang Huanglian Xiexintang showed increased positive expression of Nrf2 and HO-1, with a significant increase in brown-yellow granules around the cell nucleus (P<0.05). Western blot results showed that compared with the normal group, the model group had significantly reduced protein expression of Nrf2 and HO-1 (P<0.01). Compared with the model group, the metformin group and all doses of modified Dahuang Huanglian Xiexintang showed significantly increased protein expression of Nrf2 and HO-1 (P<0.01). ConclusionModified Dahuang Huanglian Xiexintang can significantly improve the general condition and pathological changes of liver tissues in T2DM model rats. This improvement is likely achieved through ameliorating hepatic oxidative stress injury via regulating the Nrf2/HO-1 axis.
10.Effect of Modified Dahuang Huanglian Xiexintang on Mitochondrial Autophagy and Browning of Visceral Fat in Obese Type 2 Diabetes Mellitus Rats
Dong AN ; Yonglin LIANG ; Yankui GAO ; Fengzhe YAN ; Sichen ZHAO ; Zhongtang LIU ; Chengjun MA ; Xiangdong ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):131-140
ObjectiveTo observe the effect of modified Dahuang Huanglian Xiexintang on mitochondrial autophagy and browning of visceral adipose tissue in obese type 2 diabetes mellitus (T2DM) model ZDF rats. MethodForty ZDF rats were induced with a high-fat diet to establish an obese T2DM model. The rats were randomly divided into five groups: Model group, metformin group (0.18 g·kg-1), and high, medium, and low dose groups of modified Dahuang Huanglian Xiexintang (2.16, 1.08, 0.54 g·kg-1), with eight rats in each group. Additionally, eight ZDF (fa/+) rats were assigned to the normal group. All groups received an intragastric volume of 10 mL·kg-1, with the model and normal groups receiving the same volume of purified water once daily for 12 weeks. Fasting blood glucose (FBG) was regularly measured. After 12 weeks of intervention, the body weight, epididymal fat weight, and serum levels of glucose (GLU), glycated serum protein (GSP), triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were measured. Hematoxylin-eosin (HE) staining was used to observe pathological changes in epididymal fat tissue. Transmission electron microscopy (TEM) was employed to observe mitochondrial autophagy in adipocytes. Real-time PCR was used to detect the mRNA expression of hypoxia-inducible factor-1α (HIF-1α), Bcl-2/adenovirus E1B 19 kDa interacting protein 3 (BNIP3), microtubule-associated protein 1 light chain 3B (LC3B), p62/SQSTM1, uncoupling protein 1 (UCP1), iodothyronine deiodinase 2 (Dio2), and PR domain containing 16 (Prdm16) in epididymal fat. Western blot was used to detect the protein expression of HIF-1α, BNIP3, LC3B, p62, and UCP1 in epididymal fat. ResultCompared with the normal group, the model group showed pathological changes in epididymal fat, with adipocyte mitochondrial condensation and numerous autophagosomes indicating mitochondrial autophagy. The model group also exhibited significantly increased body weight, epididymal fat weight, FBG, GLU, GSP, TC, TG, and LDL-C levels (P<0.01), significantly decreased HDL-C levels (P<0.01), significantly elevated mRNA and protein expression of HIF-1α, BNIP3, and LC3B (P<0.01), significantly reduced mRNA and protein expression of p62 and UCP1 (P<0.01), and significantly reduced mRNA expression of Dio2 and Prdm16 (P<0.01). Compared with the model group, all intervention groups showed varying degrees of improvement in epididymal fat pathology. The metformin group and high-dose modified Dahuang Huanglian Xiexintang group displayed intact mitochondrial morphology, clear cristae, uniform matrix, and few autophagosomes and autophagosomes in the adipocyte cytoplasm. The metformin group and high- and medium-dose groups of modified Dahuang Huanglian Xiexintang showed significantly reduced body weight and epididymal fat weight (P<0.01). The epididymal fat index was reduced in all intervention groups (P<0.05), and FBG was lowered in all intervention groups (P<0.01).Serum GSP, GLU, TG, and LDL-C levels were reduced in the metformin group and the high- and medium-dose groups of modified Dahuang Huanglian Xiexintang (P<0.05, P<0.01). The serum TC level was significantly reduced in the metformin group and high-dose group of modified Dahuang Huanglian Xiexintang (P<0.01), and HDL-C levels were significantly increased in all intervention groups (P<0.05, P<0.01). The mRNA and protein expression of HIF-1α, BNIP3, and LC3B were significantly reduced, and UCP1 protein expression was significantly increased in the metformin group and high- and medium-dose groups of modified Dahuang Huanglian Xiexintang (P<0.05, P<0.01). The mRNA and protein expression of p62, Dio2, and Prdm16 were significantly increased in the metformin group and high-dose group of modified Dahuang Huanglian Xiexintang (P<0.05, P<0.01). ConclusionModified Dahuang Huanglian Xiexintang may inhibit mitochondrial autophagy and promote the browning of visceral adipose tissue through the HIF-1α/BNIP3/LC3B pathway, thereby improving glucose and lipid metabolism in obese T2DM rats.

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