Objective: To investigate the regulatory effect of Jieduan Niwan Formula (JDNWF) drug-containing serum on AMPK-mediated mitochondrial quality control in D-GalN-induced HepG2 cells. Methods: Twenty male Wistar rats were randomly divided into blank control and JDNWF-containing serum groups, 10 rats per group. The JDNWF-containing serum group was gavaged with JDNWF (21.7 g/kg), whereas the blank control group was gavaged with saline. Blood was collected to prepare JDNWF-containing and blank control serum. Cell viability, mitochondrial damage indicators, and MQC pathway protein expression levels were evaluated to determine the optimal volume fraction of JDNWF. HepG2 cells were divided into control, D-GalN, DMSO, AMPK inhibitor, JDNWF drug-containing serum, and JDNWF drug-containing serum plus AMPK inhibitor groups, and corresponding drug interventions were administered to each group. Cells were collected after the interventions, and the CCK-8 assay was used to measure cell viability, the 2′-7′-dichlorodihydrofluorescein diacetate fluorescent probe was used to detect reactive oxygen species (ROS) levels, JC-1 was used to detect mitochondrial membrane potential, thiobarbituric acid was used to measure malondialdehyde (MDA) levels, WST-8 was used to measure superoxide dismutase (SOD) activity, and western blotting was used to detect the expression levels of mitochondrial quality control-related proteins, including p-AMPK, AMPK, PGC-1α, NRF1, TFAM, MFN2, and DRP1. Results: 5% JDNWF drug-containing serum most significantly restored cell viability, mitochondrial damage markers, and MQC pathway protein expression in the model group. Therefore, it was chosen for intervention in subsequent experiments. Compared to the control group, the cell viability of the D-GalN, DMSO, and AMPK inhibitor groups was significantly reduced (P<0.01). In contrast, the heterogeneity of mitochondrial membrane potential, ROS, and MDA levels was significantly increased (P<0.01), and SOD activity was significantly decreased (P<0.01). The p-AMPK, PGC-1α, NRF1, TFAM, MFN2, and DRP1 protein expression levels were significantly decreased (P<0.01). After JDNWF drug-containing serum intervention, compared to the DMSO group, cell viability significantly increased (P<0.01), mitochondrial membrane potential heterogeneity, ROS, and MDA levels significantly decreased (P<0.01), SOD activity significantly increased (P<0.01), and p-AMPK, PGC-1α, NRF1, TFAM, and MFN2 protein expression levels significantly increased (P<0.01), whereas DRP1 protein expression significantly decreased (P<0.01). Compared to the JDNWF drug-containing serum group, the cell viability in the JDNWF plus AMPK inhibitor group significantly decreased (P<0.01), mitochondrial membrane potential heterogeneity and ROS levels significantly increased (P<0.01), MDA levels significantly increased (P<0.05), SOD activity significantly decreased (P<0.05), p-AMPK, PGC-1α, NRF1, and TFAM protein expression levels significantly decreased (P<0.01), MFN2 protein expression significantly decreased (P<0.05), and DRP1 protein expression significantly increased (P<0.01). Conclusion JDNWF drug-containing serum may restore mitochondrial function and improve D-GalN-induced HepG2 cell injury by regulating AMPK-mediated mitochondrial quality control.

Heart and spleen deficiency syndrome is the most common syndrome type in patients with insomnia. Based on the theory of disease syndrome-combined animal model， this paper used multiple databases to search for the keywords "heart and spleen deficiency"， "insomnia"， "sleepless"， "disease syndrome-combined animal model"， "model evaluation"， etc. It selected the literature related to the animal model of insomnia with heart and spleen deficiency in the past 20 years to evaluate from the aspects of model establishment， modeling factors， syndrome model， disease model， macro characterization & macro characterization evaluation scale， micro indicators， etc. It is found that the existing animal model of insomnia with heart and spleen deficiency is not completely constructed by the method of disease syndrome combination of disease modeling factors and syndrome modeling factors. In the model using this method， the single establishment factor of heart and spleen deficiency does not conform to the clinical reality of disease， and the selection of the factors for the insomnia model is not closely related to or even separated from the syndrome performance. There is a problem of insufficient quantification of macro representation when the macro representation of the model replaces the symptoms related to heart and spleen deficiency syndrome and insomnia in an equivalent manner for macro representation evaluation， which can be improved according to the quantitative ideas and examples of the existing macro representation and macro representation evaluation scale. There are few specific indicators of heart and spleen deficiency syndrome in micro indicators. The micro research of heart and spleen deficiency syndrome and the essence of other traditional Chinese medicine （TCM） syndromes can be carried out by metabonomics and other technologies combined with the theory of corresponding prescription and syndrome， along the specific related ideas of "prescription and syndrome， treatment principle and selection of prescription， treatment principle and selection of acupoints， as well as therapeutic mechanism and syndrome essence". The future users and researchers of animal models of insomnia with heart and spleen deficiency can get improved methods and ideas through the shortcomings of animal models of heart and spleen deficiency listed in this paper and construct animal models of insomnia with heart and spleen deficiency that are more suitable for clinical practice， so as to establish a more perfect modeling method and evaluation system of disease syndrome-combined animal model.

Heart and spleen deficiency syndrome is the most common syndrome type in patients with insomnia. Based on the theory of disease syndrome-combined animal model， this paper used multiple databases to search for the keywords "heart and spleen deficiency"， "insomnia"， "sleepless"， "disease syndrome-combined animal model"， "model evaluation"， etc. It selected the literature related to the animal model of insomnia with heart and spleen deficiency in the past 20 years to evaluate from the aspects of model establishment， modeling factors， syndrome model， disease model， macro characterization & macro characterization evaluation scale， micro indicators， etc. It is found that the existing animal model of insomnia with heart and spleen deficiency is not completely constructed by the method of disease syndrome combination of disease modeling factors and syndrome modeling factors. In the model using this method， the single establishment factor of heart and spleen deficiency does not conform to the clinical reality of disease， and the selection of the factors for the insomnia model is not closely related to or even separated from the syndrome performance. There is a problem of insufficient quantification of macro representation when the macro representation of the model replaces the symptoms related to heart and spleen deficiency syndrome and insomnia in an equivalent manner for macro representation evaluation， which can be improved according to the quantitative ideas and examples of the existing macro representation and macro representation evaluation scale. There are few specific indicators of heart and spleen deficiency syndrome in micro indicators. The micro research of heart and spleen deficiency syndrome and the essence of other traditional Chinese medicine （TCM） syndromes can be carried out by metabonomics and other technologies combined with the theory of corresponding prescription and syndrome， along the specific related ideas of "prescription and syndrome， treatment principle and selection of prescription， treatment principle and selection of acupoints， as well as therapeutic mechanism and syndrome essence". The future users and researchers of animal models of insomnia with heart and spleen deficiency can get improved methods and ideas through the shortcomings of animal models of heart and spleen deficiency listed in this paper and construct animal models of insomnia with heart and spleen deficiency that are more suitable for clinical practice， so as to establish a more perfect modeling method and evaluation system of disease syndrome-combined animal model.

OBJECTIVE To establish HPLC fingerprint for Jianpi hewei formula （JPHWF）， conduct chemical pattern recognition analysis， and determine the contents of seven components in the formula， aiming to provide a scientific basis for quality control and further research of JPHWF. METHODS Taking 15 batches of standard decoctions of JPHWF as samples， the HPLC fingerprint was established using the Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）. Subsequently， similarity evaluation， as well as identification and attribution analysis of chromatographic peaks， were conducted. Using the common peak areas from the 15 batches of samples as variables， chemical pattern recognition analyses were performed on the samples through hierarchical cluster analysis， principal component analysis， and orthogonal partial least squares-discriminant analysis. The contents of adenine， 5-hydroxymethylfurfural， tetrahydropalmatine， naringin， dehydrocorydaline， neohesperidin and glycyrrhizic acid in 15 batches of samples were determined by HPLC. RESULTS There were 19 common peaks in the characteristic chromatograms for 15 batches of samples with the similarities of more than 0.95. Results of chemical pattern recognition analysis showed that 15 batches of samples could be clustered into 3 categories， and 3 differential compounds were found [peak 7 （5- hydroxymethylfurfural）， peak 17 （neohesperidin）， and peak 15 （naringin）]. The 7 components were linearly good in the respective concentration ranges （R2≥0.999 4）； RSDs of precision， stability and repeatability tests were less than 2% （n＝6）； the average recovery rate of 98.95%-103.81%， RSD of 0.61%-2.75% （n＝6）； the contents of them were 0.031-0.106， 0.267-0.824， 0.089- 0.144， 1.344-2.091， 0.089-0.178， 1.328-2.028， 0.040-0.150 mg/g， respectively. CONCLUSIONS Established HPLC fingerprinting method coupled with multi-index content determination is validated to be accurate and reliable， and its combination with chemical pattern recognition analysis can be applied to the quality control of JPHWF.

As a new manner of cell death,cuproptosis depends on the accumulation of copper ions in cells.Copper ion is an essential trace element in normal physiological state of organisms.The excess of free copper in cells not only has toxic effect on normal cells,but also plays its specific killing function on tumor cells.Copper transporter 1(CTR1)is a key transporter of transmembrane uptake of copper ions by cells.As a regulator of cuproptosis,its mutation and expression changes in tumors have an impact on the distribution of copper ions inside and outside the cells.It may participate in multiple biological processes such as proliferation,invasion and migration of tumor cells by regulating the pathway of cuproptosis.This article reviews the cuproptosis pathway mediated by CTR1 and the potential value of CTR1 in tumor treatment,elaborates the importance of copper ion homeostasis regulation for normal life activities and the mechanism of CTR1 in regulating cuproptosis,and discusses the potential value of CTR1 as a new target for tumor therapy,so as to provide a theoretical basis for the treatment of tumor patients.

Naval hospitals,the backbone of naval health service system,are the key to build a world-class naval health service system.On the basis of the requirements of military transformation and high-quality development of public hospitals,this paper summarized five aspects of high-quality development of naval hospitals,including directions and regulations,support and contribution of combats,efficiency,sustainable development momentum,and the expansion of service functions.It is very important to strengthen the top-level design and policy support for naval hospitals,to improve the level of governance and innovation,and to find the right target for the naval hospital construction in naval health service system reform.

ObjectiveTo explore the potential mechanisms of a baicalin-geniposide combination against cerebral ischemia using a network pharmacology strategy.MethodWe used network pharmacology integrating drug-target-disease interactions to identify key pathways which were validated in a rat middle cerebral artery occlusion model treated with baicalin (55 mg/kg), geniposide (5 mg/kg), or their 11:1 combination. Therapeutic efficacy and mechanistic insights were evaluated using triphenyltetrazolium chloride staining, Evans blue assay, enzyme-linked immunosorbent assay, and Western blot.ResultsThe results revealed that the nuclear factor-kappa B (NF-κB) signaling pathway is inhibited in combination treatment of cerebral ischemia. Ten targets were identified as key nodes in the protein–protein interaction network: interleukin 6 (IL-6), interleukin-1β, interleukin 18, C–C motif ligand 2, C–C motif ligand 4, interleukin 10, interferon-γ-inducible protein 10, C–C motif ligand 3, tumor necrosis factor-α (TNF-α), interleukin-1α. The baicalin-geniposide combination significantly reduced infarct volume, improved neurological deficits, and alleviated brain edema/blood–brain barrier leakage compared with monotherapy. Additionally, it significantly inhibited toll-like receptor 4 (TLR4)/NF-κB signaling and downregulated pro-inflammatory cytokines TNF-α and IL-6 levels.ConclusionThe baicalin-geniposide combination alleviated cerebral ischemia-reperfusion injury by synergistically suppressing the TLR4/NF-κB pathway and its downstream inflammatory factors.

ObjectiveTo evaluate the effectiveness and safety of neck seven-line method combined with periauricular acupuncture as salvage treatment for sudden hearing loss in non-acute stage. MethodsSixty-six patients with non-acute stage of sudden hearing loss with a disease duration of 15-90 days were randomly divided into 33 cases each in treatment group and control group. The treatment group was given neck seven-line method combined with acupuncture at periauricular points； the control group used sham acupuncture and sham electroacupuncture at the same points. Both groups were treated 3 times a week for 6 weeks. The pure tone average hearing threshold of impaired frequencies were examined before treatment， after treatment finish and at follow-up （week 10）， and the difference between pure tone average hearing threshold of impaired frequencies before and after treatment was calculated； calculate the proportion of patients with ≥10 dB improvement in pure tone average hearing threshold of impaired frequency after treatment and at follow-up； compare the patients' Tinnitus Evaluation Scale （TEQ） scores， Chinese Medicine Quality of Life Assessment Scale （CQ-11D） health utility values and the difference between before and after treatment， and record the occurrence of adverse events. ResultsThe pure tone average of impaired frequency in the treatment group were （50.57±18.07） dB and （47.70±17.42） dB at post-treatment and follow-up respectively， and （54.38±21.77） dB and （53.36±20.99） dB in the control group at post-treatment and follow-up. Compared with the pre-treatment period， the pure tone average hearing threshold of impaired frequency in the two groups significantly decreased （P＜0.05） at post-treatment and follow-up. The difference of pure tone average hearing threshold of impaired frequency in the treatment group after treatment and at the follow-up visit compared to that before treatment was lower than those in the control group （P＜0.05）. After treatment， 13 patients （39.39%） in the treatment group and 10 patients （30.30%） in the control group showed improvement of ≥10 dB in pure tone average hearing threshold of impaired frequency； at the follow-up visit， 18 patients （54.55%） in the treatment group and 10 patients （30.30%） in the control group showed improvement of ≥10 dB in the pure tone average hearing threshold of impaired frequency， and there was no statistical significance for comparison between groups at the time of post-treatment and follow-up （P＞0.05）. The TEQ score of the treatment group significantly lower than that before treatment （P＜0.05）； the TEQ score of the treatment group and the difference between before and after treatment significantly lower than that of the control group （P＜0.05）. The difference in CQ-11D health utility values and the difference between before and after treatment were not statistically significant between the two groups （P＞0.05）. Eight cases of acupuncture-related adverse events occurred among 66 patients， including subcutaneous haematomas after needling， severe pain during needling and needle fainting， which disappeared after symptomatic treatment and did not affect the following treatment. ConclusionNeck seven-line method combined with periauricular acupuncture could be used as salvage treatment for sudden hearing loss in non-acute stage to improve the pure tone average hearing threshold of impaired frequency， alleviate tinnitus， with safety.

Pure-tone audiometry can be performed to evaluate the type and degree of hearing loss， whose results can be divided into four types including low-frequency descending， high-frequency descending， flat descending and total deafness. The low-frequency descending type of sudden hearing loss （SHL） is more likely to be spleen deficiency and dampness exuberance， the high-frequency descending type is often due to yin deficiency of liver and kidney， the flat descending type is commonly associated with qi and blood depletion， and the type of total deafness is often linked to blood stasis. Our team has further developed a comprehensive diagnostic and therapeutic approach for SHL， emphasizing "the integration of disease and syndrome， the combination of acupuncture and herbal medicine， and dynamically administering treatment". Firstly， it advocates integrating disease diagnosis with syndrome differentiation. Secondly， it recommends combining acupuncture and herbal medicine， with local acupoints such as Ermen （TE 21）， Tinggong （SI 19）， Tinghui （GB 2）， and Yifeng （TE 17） used to unblock the auditory orifice， and herbal prescriptions tailored to the hearing curve patterns. For the low-frequency descending type， it is recommended to fortify the spleen and percolate dampness by taking distal points of spleen channel and stomach channel and using Shenling Baizhu Powder （参苓白术散）. For the high-frequency descending type， the method of nourishing kidney and calming liver is recommended， using distal points of kidney and liver channels and taking Erlong Zuoci Pills （耳聋左慈丸）. Regarding the flat descending type， tonifying qi and nourishing blood is advised， for which acupoints of Conception Vessel， spleen， stomach and large intestine channels can be needled， and Yiqi Congming Decoction （益气聪明汤） can be administered. For the total deafness type， it is recommended to activate blood and dissipate stasis， often with Xuehai （SP 10） and Geshu （BL 17） needled and Tongqiao Huoxue Decoction （通窍活血汤） administered. All these are conducted to treat the root and branch simultaneously. At the same time， it emphasizes the need to consider complex syndrome presentations and disease progression， dynamically analyze the disease causes and mechanisms， and adjust treatment according to the changing syndromes. In conclusion， this article is expected to inspire clinical diagnosis and treatment of SHL.

Rabies is a zoonotic disease that poses a global public health threat.Rabies virus(RABV)is neurotropic and can cause severe neurological disorders and behavioral abnormalities in host,with a fatality rate nearly 100％.In order to identify the key genes for RABV affecting neuronal cell function,we established and analyzed the mRNA expression profile of Neuro-2a(N2a)cell in-fected with challenge virus standard(CVS)-11 by RNA sequencing(RNA-seq).Biological func-tions of differentially expressed genes(DEGs)were determined by GO and KEGG enrichment a-nalysis.The results showed that there were 415 differentially expressed genes in N2a infected with CVS-11 strain,of which 89 were up-regulated and 326 were down-regulated.These genes were re-lated to a variety of biological processes,such as axon guidance pathway,cholesterol metabolism pathway,nitrogen metabolism pathway,and glycosphingolipid biosynthesis pathway,many of them have been shown to be closely associated with RABV infection.A total of 12 DEGs related to axon conduction,antigen processing and presentation pathways were selected and detected by real-time PCR,and their expression trends were consistent with the RNA-seq results.The genomic tran-scriptomic data on N2a cell under RABV infection will provide new clues for probing the mecha-nisms of RABV infection and transmission in the nervous system in the future.