ObjectiveTo investigate the effects of Anmeidan （AMD） on neuroinflammation in the hippocampus of sleep-deprived rats. MethodsSD rats were randomly divided into four groups （n = 10 per group）： control group， model group， AMD group， and melatonin group. A sleep deprivation model was established using the modified multiple platform water environment method. The AMD group received AMD at a dose of 18.18 g·kg^-1·d^-1， the melatonin group received melatonin at 100 mg·kg^-1·d^-1， and the control and model groups were given an equal volume of pure water. All treatments were administered by gavage for four weeks. Spontaneous activity was assessed using an animal behavior video system. Serum levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay （ELISA）. Hippocampal pyramidal neuron morphology was examined using hematoxylin-eosin （HE） staining， and ultrastructural changes of hippocampal neurons were observed via transmission electron microscopy. Immunofluorescence was used to detect the expression of brain-derived neurotrophic factor （BDNF） and nerve growth factor （NGF） in the hippocampus. Western blot analysis was performed to measure the expression of nuclear factor-κB （NF-κB）， phosphorylated NF-κB （p-NF-κB）， NOD-like receptor protein 3 （NLRP3）， and Caspase-1 proteins. ResultsCompared with the control group， the model group showed a significant increase in activity duration and frequency （P<0.01）， increased hippocampal pyramidal cell structural damage and decreased cell count， aggravated hippocampal ultrastructural damage， mitochondrial cristae disruption， and exacerbated vacuolization. The expression of p-NF-κB p65， NLRP3， and Caspase-1 proteins was upregulated， serum IL-1β， IL-6， and TNF-α levels were significantly elevated （P<0.01）， and the fluorescence intensity of BDNF and NGF proteins was significantly reduced （P<0.01）. Compared with the model group， the AMD group showed a significant reduction in activity duration and frequency （P<0.01）， increased hippocampal pyramidal cell count with reduced structural damage， alleviated hippocampal ultrastructural damage， significantly downregulated p-NF-κB p65， NLRP3， and Caspase-1 protein expression （P<0.01）， decreased serum IL-1β， IL-6， and TNF-α levels （P<0.01）， and significantly increased the fluorescence intensity of BDNF and NGF proteins （P<0.01）. ConclusionAnmeidan alleviates hippocampal neuronal damage in sleep-deprived rats， potentially by downregulating the NLRP3 signaling pathway， reducing inflammatory cytokine release， and increasing neurotrophic factor levels.

ObjectiveTo investigate the effects of Anmeidan （AMD） on cognitive function， cytochrome C （Cyt C） signaling pathway protein expression， and apoptosis in a geriatric sleep deprivation model. MethodSixty aged C57 mice were randomly divided into a blank group， a model group， AMD high， medium， and low dose （26.26， 13.13， 6.565 g·kg^-1·d^-1） groups， and a melatonin group （1.3 mg·kg^-1·d^-1）， with 10 mice in each group. Continuous sleep deprivation was performed for 4 weeks using a homemade sleep deprivation box. Cognitive function was assessed using the Morris water maze， and morphological changes in pyramidal cells in the CA1 area of the hippocampus were observed by hematoxylin-eosin （HE） staining and Nissl staining. Transmission electron microscopy was used to observe the mitochondrial morphology and structure of hippocampal neurons. Western blot was used to detect Cyt C， cysteine-aspartate protease-3 （Caspase-3）， cysteine-aspartate protease-9 （Caspase-9）， brain-derived neurotrophic factor （BDNF）， mitochondrial transcription factor A （TFAM）， and voltage-dependent anion channel 1 （VDAC1） protein expression. Immunohistochemistry was used to detect protein expression levels of Cyt C， Caspase-3， and Caspase-9， and immunofluorescence was used to detect the protein expression of B-cell lymphoma 2 （Bcl-2） and Bcl-2-associated X protein （Bax）. ResultCompared with the blank group， the model group showed prolonged platform latency （P<0.01）， reduced number of platform crossings， and reduced time and distance in the target quadrant （P<0.01）. The mitochondrial structure was damaged， with disappearance or breakage of cristae， and increased swelling and deformation. The protein expression levels of Cyt C， Caspase-3， Caspase-9， Bax， and VDAC1 were significantly increased （P<0.01）， while BDNF， TFAM， and Bcl-2 protein expression levels were decreased （P<0.01）. Compared with the model group， the AMD high， medium， and low dose groups improved spatial exploration and navigation abilities in geriatric sleep-deprived mice （P<0.05， P<0.01）， alleviated mitochondrial damage， and increased the number of Nissl bodies. Additionally， the expression levels of Cyt C， Caspase-3， Caspase-9， Bax， and VDAC1 proteins were significantly reduced （P<0.05， P<0.01）， while the expression levels of BDNF， TFAM， and Bcl-2 proteins were significantly increased （P<0.05， P<0.01）. ConclusionAMD improved the cognitive function of geriatric sleep-deprived mice， and its effect may be related to the reduction of apoptosis mediated by the Cyt C signaling pathway.

As a basic source for the theoretical system of traditonal Chinese medicine （TCM）， the Inner Canon of Yellow Emperor （《黄帝内经》） made the point of “treatment with combination of diverse methods and prescriptions”，which is one of the characteristics of TCM in treating diseases. This paper discussed the therapeutic idea of “diverse methods and prescriptions” from five dimensions， including time， population， regions， diseases， and treatments， and proposed that “combined treatment” is an effective measure to embody the idea of “diverse methods and prescriptions”. That is to say， by taking the combination of the concept of holism and syndrome differentiation and treatment as the starting point， combining pharmacological and non-pharmacological therapies， and treating both internal and external diseases through multiple ways and methods， the goal to treat the disease at the root， and keep yin at peace and yang compact can be achieved. Finally， it is suggested to guide the clinical practice with “treatment with combination of diverse methods and prescriptions”， providing new ideas for clinical diagnosis and treatment.

Objective: To observe the effect of traditional Chinese medicine foment package in spontaneous heat position pad to prevent lower extremity deep venous thrombosis (LEDVT) during general anesthesia. Methods: From April 2018 to April 2019, 160 cases during general anesthesia were selected and divided into the control group(common hot pad for leg and knee) and the experimental group(self-control position pad for leg and knee) with 80 cases each by the random number table method. The coagulation function and deep venous thrombosis were observed and compared between the two groups. Results: There was no significant difference in coagulation function between the two groups on the 1 day before operation (P > 0.05). The level of D-dimer, prothrombin time, fibrinogen, platelet count on the 5 days after operation was (1.28 ± 0.39) g/L, (11.75 ± 0.81) s, (4.37 ± 0.71) g/L, (203.39 ± 32.12)×10⁹/L in the control group, and (0.96 ± 0.17) g/L, (11.45 ± 0.67) s, (3.93 ± 0.65) g/L, (186.10 ± 34.69)×10⁹/L in the experimental group, and there was significant difference between the two groups (t=2.553-6.728, P <0.01). The incidence of LEDVT on the 5 day after operation was 4%(3/80) in the experimental group and 18% (77/80) in the control group, and there was significant difference between the two groups (χ²=5.005, P<0.05). Conclusions The spontaneous fever knee pad with traditional Chinese medicine foment package can improve the blood coagulation function of general anesthesia patients, and the incidence of LEDVT is lower than that of the control group 5 days after operation.

ObjectiveTo explore the effect and mechanism of the classic famous prescription Anmeidan （AMD） developed in the Qing Dynasty in regulating the hepatic neurotransmitters and circadian rhythm in the rat model of insomnia via the orexin-1 receptor （OX1R）/phosphatidylinositol-specific phospholipase Cβ-1 （PLCβ-1）/protein kinase Cα （PKCα）/extracellular signal-regulated kinase 1/2 （ERK1/2） signaling pathway. MethodSixty SPF-grade SD rats were randomized into blank， model， suvorexant （30 mg·kg^-1·d^-1）， and low-， medium-， and high-dose （4.55， 9.09， 18.09 g·kg^-1·d^-1， respectively） AMD groups， with 10 rats in each group. The rats in other groups except the blank group were modeled by intraperitoneal injection of p-chlorophenylalanine （PCPA） and administrated with corresponding drugs by gavage， and the blank group received an equal volume of normal saline. The general condition， body mass， and 24 h autonomic activity of each group were observed. The pathological changes of the liver tissue were observed by hematoxylin-eosin（HE）staining and Masson staining. The expression of gamma-aminobutyric acid （GABA）， 5-hydroxytryptamine （5-HT）， epinephrine （EPI）， norepinephrine （NE）， and acetylcholine （ACh） in the liver tissue was detected by enzyme-linked immunosorbent assay. The glutamate （Glu） expression in the liver tissue was detected by the biochemical method. The mRNA levels of biological clock genes Per1， Per2， Cry1， Cry2， Bmal1， and Bmal2 in the liver were determined by Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. The protein and mRNA levels of factors in the OX1R/PLCβ-1/PKCα/ERK1/2 signaling pathway in the liver were determined by Western blot and Real-time PCR， respectively. ResultCompared with the blank group， the modeling decreased the body mass （P<0.05， P<0.01） and caused mania and disturbed resting rhythms （P<0.01）， hepatic muscle fiber fracture， and edema with inflammatory cell infiltration. In addition， the modeling decreased the GABA， 5-HT， EPI， NE， and ACh content， increased Glu content （P<0.01）， down-regulated the mRNA levels of Per1， Per2， Cry1， and Cry2 （P<0.01）， up-regulated the mRNA levels of Bmal1 and Bmal2 （P<0.01）， and promoted the expression of OX1R， PLCβ-1， PKCα， and ERK1/2 at both protein and mRNA levels （P<0.01）. Compared with the model group， suvorexant and AMD increased the body mass （P<0.05， P<0.01）， alleviated the mania， and increased the resting time and frequency （P<0.05， P<0.01）. Moreover， the medications elevated the levels of GABA， 5-HT， EPI， NE， and ACh， lowered the Glu level， up-regulated the mRNA levels of Per1， Per2， Cry1， and Cry2 （P<0.05， P<0.01）， down-regulated the mRNA levels of Bmal1 and Bmal2， and inhibited the expression of OX1R， PLCβ-1， PKCα， and ERK1/2 at both mRNA and protein levels （P<0.05， P<0.01）. ConclusionAMD can regulate hepatic neurotransmitters and improve circadian rhythm in insomniac rats by inhibiting the OX1R/PLCβ-1/PKCα/ERK1/2 signaling pathway， and high-dose AMD demonstrated the strongest effect.