ObjectiveTo investigate the effects of Maimendong Yinzi （MMDYZ） on cough with Yin deficiency and lung heat syndrome and explore its potential mechanism of action. MethodsForty-eight Institute of Cancer Research （ICR） mice were randomly divided into a control group， a model group， a Baihe Gujin Tablet （BHGJP） group （1.36 g·kg^-1）， and low-dose， medium-dose， and high-dose MMDYZ groups （5， 10， 20 g·kg^-1·d^-1， based on the weight of crude drug）， with eight mice in each group. The mouse model of cough with Yin deficiency and lung heat syndrome was prepared by a combination of smoke exposure， nasal drip of lipopolysaccharide （LPS）， intragastric gavage with thyroxine， and capsaicin atomization. After successful modeling， drug interventions were administered for seven days. During modeling， the mice were observed for changes in general status， anal temperature， fecal water content， and water intake. After medication， the above indicators were evaluated again， along with assessments of spontaneous activity， cough sensitivity， lung function， lung index， and tracheal phenol red secretion. Bronchoalveolar lavage fluid （BALF） was analyzed for cell differential counts， and the level of cyclic adenosine monophosphate （cAMP）， cyclic guanosine monophosphate （cGMP）， tumor necrosis factor-α （TNF-α）， and interleukin-6 （IL-6） in serum was measured via enzyme linked immunosorbent assay （ELISA）. Lung injury was assessed via hematoxylin-eosin （HE） staining. Network pharmacology was employed to predict the potential mechanism of MMDYZ in alleviation cough with Yin deficiency and lung heat syndrome. Western blot （WB） was used to measure protease-activated receptor1 （PAR1） and GTPhase α_i subunit （Gα_i） protein expressions in lung tissue. ELISA was used to determine lung cAMP content， and immunohistochemistry （IHC） was employed to evaluate transient receptor potential vanilloid subtype 1 （TRPV1） expression. ResultsCompared with the control group， the model group exhibited significantly increased water intake and anal temperature and significantly decreased fecal water content （P<0.05）. The total distance traveled in 5 min and the central zone duration were reduced， while standing frequency significantly increased （P<0.05）. Cough sensitivity and enhanced pause （PenH） were elevated. Peak expiratory flow （PEF） significantly declined （P<0.05）. BALF neutrophil （NEU） and white blood cell （WBC） counts rose. Serum cAMP and cAMP/cGMP ratio significantly increased， and cGMP significantly decreased （P<0.05）. Serum TNF-α and IL-6 levels were significantly elevated （P<0.05）. The lung injury was obvious， and the lung index was significantly elevated （P<0.05）. Compared with the model group， the medium-dose and high-dose MMDYZ groups and the BHGJP group showed significantly improved indicators mentioned above. Additionally， network pharmacology suggested that MMDYZ might alleviate cough with Yin deficiency and lung heat syndrome via cAMP， hypoxia inducible factor-1 （HIF-1）， and TNF signaling pathways. WB， ELISA， and IHC revealed that， compared with the control group， the model group exhibited significantly upregulated PAR1， Gα_i， and TRPV1 expressions and significantly downregulated cAMP in lung tissue （P<0.05）. Compared with the model group， MMDYZ reduced PAR1 （P<0.01）， Gα_i （P<0.05）， and TRPV1 （P<0.01） while increasing cAMP level （P<0.01）. ConclusionMMDYZ may alleviate cough with Yin deficiency and lung heat syndrome by modulating the PAR1/Gα_i/cAMP pathway and TRPV1 expression.

ObjectiveBased on modern analytical techniques and a depressed mouse model established by chronic unpredictable mild stress（CUMS）， to evaluate the quality marker（Q-Marker） and pharmacodynamic difference of Baihe Dihuangtang prepared by different processes. MethodsHigh performance liquid chromatography（HPLC） was used to establish the characteristic profiles of Baihe Dihuangtang， and determine the content of Q-Marker in the samples prepared by ancient and modern processes. Seventy C57BL/6J mice were randomly divided into the normal group， model group， fluoxetine group（3 mg·kg^-1）， low and high dose groups of ancient process（6.5， 26 g·kg^-1）， and low and high dose groups of modern process（6.5， 26 g·kg^-1）， with 10 mice in each group. Except for the normal group， CUMS was used to induce depression in mice from the other groups for 28 d. After successful modeling， administration groups were given the corresponding drugs by gavage every day， and the normal and model groups were given an equal volume of pure water by gavage for 21 consecutive days. Change in body mass of mice was recorded， tail suspension test and open field test were used to evaluate the depressive behavior of mice， and enzyme-linked immunosorbent assay（ELISA） was employed to determine the contents of tumor necrosis factor-α（TNF-α）， interleukin（IL）-1β and IL-6 in serum. ResultsCharacteristic profiles of Baihe Dihuangtang prepared by the two processes were established， the similarity between the two was 0.951， and 8 characteristic peaks were recognized with the reference peak of regaloside A. The results of quantitative analysis showed that the Q-Marker content was similar in Baihe Dihuangtang prepared by ancient and modern processes. The results of pharmacodynamics showed that， compared with the normal group， the model group showed increased immobility time in the tail suspension test， reduced total movement distance in the open field test， and elevated IL-1β， IL-6 and TNF-α levels in the serum（P<0.01）. Compared with the model group， the behavioral indicators of mice in the Baihe Dihuangtang treatment group were significantly improved in terms of tail suspension time and open field exercise， and the levels of IL-1β， IL-6 and TNF-α in serum were significantly reduced（P<0.05， P<0.01）. Baihe Dihuangtang prepared by the two processes both had antidepressant effects， and the difference between the two was not statistically significant in improving depressive symptoms. ConclusionQ-Marker of Baihe Dihuangtang prepared by modern and ancient methods are equivalent in content， and the pharmacological effects are consistent， indicating that dried Lilii Bulbus can replace fresh products in the preparation of Baihe Dihuangtang. This study provides a scientific basis for the development of new drugs of Baihe Dihuangtang and a reference for its rational application and clinical use.

ObjectiveBased on modern analytical techniques and a depressed mouse model established by chronic unpredictable mild stress（CUMS）， to evaluate the quality marker（Q-Marker） and pharmacodynamic difference of Baihe Dihuangtang prepared by different processes. MethodsHigh performance liquid chromatography（HPLC） was used to establish the characteristic profiles of Baihe Dihuangtang， and determine the content of Q-Marker in the samples prepared by ancient and modern processes. Seventy C57BL/6J mice were randomly divided into the normal group， model group， fluoxetine group（3 mg·kg^-1）， low and high dose groups of ancient process（6.5， 26 g·kg^-1）， and low and high dose groups of modern process（6.5， 26 g·kg^-1）， with 10 mice in each group. Except for the normal group， CUMS was used to induce depression in mice from the other groups for 28 d. After successful modeling， administration groups were given the corresponding drugs by gavage every day， and the normal and model groups were given an equal volume of pure water by gavage for 21 consecutive days. Change in body mass of mice was recorded， tail suspension test and open field test were used to evaluate the depressive behavior of mice， and enzyme-linked immunosorbent assay（ELISA） was employed to determine the contents of tumor necrosis factor-α（TNF-α）， interleukin（IL）-1β and IL-6 in serum. ResultsCharacteristic profiles of Baihe Dihuangtang prepared by the two processes were established， the similarity between the two was 0.951， and 8 characteristic peaks were recognized with the reference peak of regaloside A. The results of quantitative analysis showed that the Q-Marker content was similar in Baihe Dihuangtang prepared by ancient and modern processes. The results of pharmacodynamics showed that， compared with the normal group， the model group showed increased immobility time in the tail suspension test， reduced total movement distance in the open field test， and elevated IL-1β， IL-6 and TNF-α levels in the serum（P<0.01）. Compared with the model group， the behavioral indicators of mice in the Baihe Dihuangtang treatment group were significantly improved in terms of tail suspension time and open field exercise， and the levels of IL-1β， IL-6 and TNF-α in serum were significantly reduced（P<0.05， P<0.01）. Baihe Dihuangtang prepared by the two processes both had antidepressant effects， and the difference between the two was not statistically significant in improving depressive symptoms. ConclusionQ-Marker of Baihe Dihuangtang prepared by modern and ancient methods are equivalent in content， and the pharmacological effects are consistent， indicating that dried Lilii Bulbus can replace fresh products in the preparation of Baihe Dihuangtang. This study provides a scientific basis for the development of new drugs of Baihe Dihuangtang and a reference for its rational application and clinical use.

Metabolic dysfunction-associated steatotic liver disease (MASLD) comprises a spectrum of liver injuries, including steatosis to steatohepatitis (MASH), liver fibrosis, cirrhosis, and relevant complications. The liver mainly comprises hepatocytes, liver sinusoidal endothelial cells (LSECs), Kupffer cells (KCs), immune cells (T cells, B cells), and hepatic stellate cells (HSCs). Crosstalk among these different liver cells, endogenous aberrant glycolipid metabolism, and altered gut dysbiosis are involved in the pathophysiology of MASLD. This review systematically examines advances in understanding the molecular pathogenesis of MASLD, with a focus on emerging therapeutic targets and translational clinical trials. We first delineate the crucial regulatory mechanisms involving diverse liver cells and the gut-liver axis in MASLD development. These cell-specific pathogenic insights offer valuable perspectives for advancing precision medicine approaches in MASLD treatment. Furthermore, we evaluate potential therapeutic targets and summarize clinical trials currently underway. By comprehensively updating the MASLD pathophysiology and identifying promising strategies, this review aims to facilitate the development of novel pharmacotherapies for this increasingly prevalent condition.
Humans ; Fatty Liver/therapy* ; Animals ; Liver/pathology* ; Kupffer Cells/metabolism* ; Hepatocytes/metabolism* ; Hepatic Stellate Cells/metabolism*

Large bone defects in load-bearing bone can result from tumor resection, osteomyelitis, trauma, and other factors. Although bone has the intrinsic potential to self-repair and regenerate, the repair of large bone defects which exceed a certain critical size remains a substantial clinical challenge. Traditionally, repair methods involve using autologous or allogeneic bone tissue to replace the lost bone tissue at defect sites, and autogenous bone grafting remains the "gold standard" treatment. However, the application of traditional bone grafts is limited by drawbacks such as the quantity of extractable bone, donor-site morbidities, and the risk of rejection. In recent years, the clinical demand for alternatives to traditional bone grafts has promoted the development of novel bone-grafting substitutes. In addition to osteoconductivity and osteoinductivity, optimal mechanical properties have recently been the focus of efforts to improve the treatment success of novel bone-grafting alternatives in load-bearing bone defects, but most biomaterial synthetic scaffolds cannot provide sufficient mechanical strength. A fundamental challenge is to find an appropriate balance between mechanical and tissue-regeneration requirements. In this review, the use of traditional bone grafts in load-bearing bone defects, as well as their advantages and disadvantages, is summarized and reviewed. Furthermore, we highlight recent development strategies for novel bone grafts appropriate for load-bearing bone defects based on substance, structural, and functional bionics to provide ideas and directions for future research.
Humans ; Bone Transplantation/methods* ; Weight-Bearing ; Bone Regeneration ; Bone Substitutes ; Bone and Bones ; Animals ; Tissue Scaffolds

Repetitive transcranial magnetic stimulation (rTMS) is a rapid and effective therapy for major depressive disorder; however, there is significant variability in therapeutic outcomes both within and across individuals, with approximately 50% of patients showing no response to rTMS treatment. Many studies have personalized the stimulation parameters of rTMS (e.g., location and intensity of stimulation) according to the anatomical and functional structure of the brain. In addition to these parameters, the internal states of the individual, such as circadian rhythm, behavior/cognition, neural oscillation, and neuroplasticity, also contribute to the variation in rTMS effects. In this review, we summarize the current literature on the interaction between rTMS and internal states. We propose two possible methods, multimodal treatment, and adaptive closed-loop treatment, to integrate patients' internal states to achieve better rTMS treatment for depression.
Humans ; Transcranial Magnetic Stimulation/methods* ; Depressive Disorder, Major/physiopathology* ; Neuronal Plasticity/physiology* ; Brain/physiopathology*

OBJECTIVE To determine the effect of Kai-Xin-San(KXS)combined with fluoxetine on the intestinal flora and the expression of inflammatory factors in chronic unpredictable mild stress(CUMS)depression mice and to elucidate the antidepressant mechanism of regulating"intestine-brain"axis.METHODS CUMS depression mice model was established and the effect of com-bined medication on improving depression-like behaviors of mice was evaluated by determination of sucrose preference rate,immobile time of tail suspension and forced swimming.Additionally,the levels of inflammatory factors IL-6,IL-1β,TNF-α and LPS were de-termined in cortex,serum and intestine using the ELISA method.The composition of intestinal flora in mouse feces was analyzed by 16S rRNA sequence sequencing.Furthermore,Western blot assay was utilized to determine the expression levels of intestinal barrier proteins such as ZO-1,Occludin and Claudin-5.RESULTS The combination of KXS and fluoxetine resulted in a significant in-crease in sucrose preference rate(P<0.01)and decreased immobile time of tail suspension and forced swimming(P<0.05,P<0.01)in CUMS mice.The antidepressant effect of KXS combined with middle dose of fluoxetine was equivalent to that of high dose of fluoxe-tine alone.Meanwhile,the combination could significantly inhibit the up-regulation of inflammatory factors in the cortex,serum and small intestine of model animals(P<0.05,P<0.01).Intestinal flora analysis showed that the combination could improve the ratio of Gram-positive bacteria to negative bacteria in the intestinal tract of model animals,and the improvement of the relative abundance of intestinal bacteria Lachnospiraceae,Bifidobacterium,Ruminococcus,Blautia,Eubacterium,Intestinimonas,Erysipelotrichaceae,Alis-tipes,Desulfovibrionia and Coriobacteriaceae UCG-002 and so on in the model animals was significantly related to the alleviation of de-pression-like behavior and the down-regulation of cortical inflammatory factors(P<0.05,P<0.01).Furthermore,the combination treatment could significantly up-regulate the expression of intestinal barrier protein(P<0.01).CONCLUSION KXS combined with fluoxetine can alleviate depression-like behavior and reduce fluoxetine dosage in CUMS model animals.The combination of the two drugs may regulate the composition of intestinal flora,inhibit the expression of intestinal inflammatory factors,up-regulate intestinal barrier proteins,and thus reduce the expression of serum and central inflammatory factors,which may be the mechanism of their regu-lation of the gut brain axis in antidepressant action.

Objective To investigate the beneficial effect of Kai-Xin-San combined with fluoxetine in improving depression-like behaviors on chronic unpredictable mild stress(CUMS)induced depression model mice.Methods The present study aimed to assess the potential of Kai-Xin-San in combination with fluoxetine to ameliorate depression-like behaviors in a CUMS induced mouse depression model.Behavioral tests,such as the sucrose preference test were employed to evaluate the efficacy of the treatment.Additionally,the levels of suppressed stress factors were measured using the ELISA method.The morphology of hippocampal tissue was evaluated using the HE staining method,Nissl Staining and TUNEL staining methods.Furthermore,western blotting analysis was utilized to determine the expression levels of proteins such as Caspase-3,and Caspase-9.Results The co-administration of Kai-Xin-San and fluoxetine resulted in a significant increase in sucrose preference rate in model mice.This effect was comparable to that of fluoxetine alone at the standard clinical dose.Furthermore,the combination treatment up-regulated the levels of suppressed stress factors,reduced the apoptosis of hippocampus induced by depression and regulated the apoptosis signaling pathway in hippocampus.Conclusion The combination of Kai-Xin-San and fluoxetine has been shown to be an effective treatment for depression-like behavior in animal models,resulting in a reduction in the required clinical dosage of fluoxetine.This effect may be attributed to the up-regulation of neurotransmitter expression,inhibition of stress axis activation,and central nervous inflammation.

BACKGROUND:The formation of Lewy bodies due to abnormal α-synuclein aggregation is a characteristic pathological change in Parkinson's disease.In recent years,several studies have revealed that the formation of α-synuclein aggregates is closely related to its post-translational modifications.The modification of α-synuclein such as phosphorylation,nitration,acetylation,and ubiquitination has attracted extensive attention in the pathogenesis and progression of Parkinson's disease. OBJECTIVE:To review the research progress in the effect of modification types and sites of α-synuclein on the characteristic pathological formation and progression of Parkinson's disease. METHODS:PubMed and CNKI databases were searched by the first author with the key words of"α-synuclein,Parkinson's disease,phosphorylation,acetylation,ubiquitination,nitration"in English and Chinese respectively to collect and sort out the literature related to abnormal modification of α-synuclein in recent years.Finally,61 articles were included for further review. RESULTS AND CONCLUSION:Abnormal modification of α-synuclein is closely related to its protein structure and its positive and negative charges.Its amino terminus is positively charged and prone to ubiquitination and acetylation modifications.The central hydrophobic region is prone to forming β-pleated sheet due to its hydrophobic property.The carboxyl terminus is negatively charged,which is the main phosphorylation modification region.Phosphorylation modification sites promote phosphorylation modification and are closely related to α-synuclein aggregation,while protein kinases can target the activation of translational modifications,which may help to promote or inhibit aggregate formation.The degradation pathway of α-synuclein mainly plays a role in removing pathological proteins.Various kinase catalysts contribute to impaired protein ubiquitination modifications that lead to abnormal protein accumulation,thereby exacerbating neurodegeneration.The amino-terminal acetylation of α-synuclein improves the shuttle ability of the protein to the cell membrane and slows down the protein aggregation,which may be the protection target of nerve cells.However,the acetylation modification of the mutant protein produces the opposite effect.The protein nitration modification is mainly related to oxidative stress.The aggregation tendency of the protein modified by nitration is enhanced under the action of reactive oxygen species.Different post-translational modifications have different effects.Therefore,elucidating the main mechanisms of their post-translational modifications and inhibiting the post-translational modifications that contribute to protein aggregation may provide a reference for new targets for early diagnosis and treatment of Parkinson's disease.

BACKGROUND:C2 ceramide reduces the formation of Alpha-Synuclein(α-Syn)oligomers as the protein phosphatase 2A agonist,which has an important regulatory effect on cell aging in the central nervous system. OBJECTIVE:To investigate the protective mechanism of C2 ceramide on dopaminergic neurons. METHODS:Twenty-five C57BL/6 mice were randomly divided into control group,model group,C2 ceramide low-,medium-and high-dose groups(n=5 per group).Except for the control group,a mouse model of Parkinson's disease was established by injecting mutant A53T α-Syn oligomers into the left striatum in the other groups.On the 30th day after the striatal injection,three C2 ceramide groups were intragastrically administered with C2 ceramide(1,5,10 μg/g)dissolved in saline at one time,while the control and model groups were administered with the same amount of saline within 30-90 days after modeling,for a total of 60 days.Behavioral changes in each group of mice were observed during this period.On the 90th day after striatal injection,mouse brain tissue was extracted by perfusion under anesthesia,and the changes of dopaminergic neurons in the midbrain substantia nigra were analyzed by immunohistochemical staining.The levels of α-Syn oligomerization and phosphorylation in the midbrain of mice were detected by ELISA,and the changes of enzyme activities related to α-Syn phosphorylation were analyzed. RESULTS AND CONCLUSION:C2 ceramide had an ameliorating effect on Parkinson's disease-like dyskinesia in mice caused by the striatal injection of mutant A53T α-Syn oligomers.High-dose C2 ceramide showed better effects on dyskinesia in mice with Parkinson's disease(P<0.01).The mutant A53T α-Syn oligomers significantly reduced the number of dopaminergic neurons in the substantia nigra of mice(P<0.01),while the number of dopaminergic neurons in the substantia nigra increased significantly in the C2 ceramide high-dose group(P<0.01).The levels of α-Syn oligomers and phosphorylated α-Syn in the brain were significantly reduced in the C2 ceramide high-dose group compared with the model group(P<0.01),while the level of ceramide was increased(P<0.05)and the activity of protein phosphatase 2A was significantly upregulated(P<0.01).To conclude,C2 ceramide can attenuate the neurotoxic effects induced by oligomerized α-Syn by the phosphorylation modification environment of α-Syn in mouse midbrain tissue and protect against the reduction in the number of nigrostriatal dopaminergic neurons in mice,thereby reducing the degree of dyskinesia in Parkinson's disease.