OBJECTIVE: To observe the effect of Huayu Tongluo moxibustion on the NOD-like receptor protein 3 (NLRP3)/cysteine-aspartic acid protease-1 (Caspase-1)/gasdermin D (GSDMD) signaling pathway in rats with vascular dementia (VD), and to explore its mechanism in improving learning and memory ability and alleviating neuronal injury in the hippocampal CA1 region. METHODS: A total of 80 SPF-grade male Wistar rats were included. Three rats were excluded based on the Morris water maze test. From the remaining rats, 12 were randomly selected as the sham operation group. The rest were used to establish VD models via modified bilateral common carotid artery ligation. Thirty-six successfully modeled rats were randomly divided into a model group, a medication group, and a moxibustion group, with 12 rats in each group. The medication group was treated with nimodipine solution (12 mg/kg) via gavage. The moxibustion group was treated with Huayu Tongluo moxibustion. The suspended moxibustion was applied at Shenting (GV24) and Dazhui (GV14), and aconite cake-separated moxibustion was applied at Baihui (GV20), with each acupoint treated for 20 min. All treatments were administered once daily for 21 consecutive days. Before and after modeling, and after intervention, the Morris water maze test was used to assess cognitive function. After intervention, the activation and morphology of microglia in the hippocampal CA1 region were observed by immunofluorescence. Ultrastructure of hippocampal CA1 neurons was examined by transmission electron microscopy. Western blot was used to detect protein expression of NLRP3, apoptosis-associated speck-like protein (ASC), Caspase-1, GSDMD, and interleukin-1β (IL-1β) in the hippocampal CA1 region. ELISA was used to detect the content of IL-6, IL-8, and tumor necrosis factor-α (TNF-α) in the hippocampal CA1 region. RESULTS: Compared with the sham operation group, the model group showed longer mean escape latency (P<0.01) and fewer platform crossings (P<0.01); the microglial processes in the hippocampal CA1 region were thickened, cytoplasm was hypertrophic, and relative fluorescence intensity of ionized calcium-binding adapter molecule 1 (IBA-1) was increased (P<0.05); the neuronal ultrastructure in the CA1 region was severely damaged, rough endoplasmic reticulum was swollen, mitochondria were deformed and swollen, some cristae were ruptured or dissolved, showing vacuolar changes; the protein expression of NLRP3, ASC, Caspase-1, GSDMD, and IL-1β, as well as levels of IL-6, IL-8, and TNF-α were significantly elevated (P<0.001). Compared with the model group, both the medication group and the moxibustion group showed shortened mean escape latency (P<0.01) and increased platform crossings (P<0.01); the microglial processes were thinner, and IBA-1 fluorescence intensity was decreased (P<0.05); the neuronal ultrastructure in the CA1 region was partially improved; the protein expression of NLRP3, ASC, Caspase-1, GSDMD, and IL-1β, and levels of IL-6, IL-8, and TNF-α were significantly reduced (P<0.001). Compared with the medication group, the moxibustion group showed shortened mean escape latency (P<0.05) and more platform crossings (P<0.05); the IBA-1 fluorescence intensity was decreased (P<0.05); the neuronal ultrastructure in the CA1 region was improved; the protein expression of NLRP3, ASC, Caspase-1, GSDMD, and IL-1β, as well as levels of IL-6, IL-8, and TNF-α, were significantly lower (P<0.001). CONCLUSION The Huayu Tongluo moxibustion could enhance learning and memory abilities in VD rats, inhibit excessive activation of microglia, and alleviate neuronal injury in the hippocampal CA1 region. Its mechanism may involve modulation of the NLRP3/Caspase-1/GSDMD signaling pathway, reduction of inflammatory responses.
Animals ; Male ; Dementia, Vascular/physiopathology* ; Rats ; Signal Transduction ; Moxibustion ; Rats, Wistar ; CA1 Region, Hippocampal/injuries* ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Caspase 1/genetics* ; Memory ; Humans ; Neurons/metabolism* ; Learning

ObjectiveTo observe the clinical efficacy of Chinese medicine combined with indirect moxibustion plaster on corona virus disease 2019 （COVID-19） patients during recovery period. MethodNinety patients of COVID-19 during the recovery period were randomly divided into a Chinese medicine group， an indirect moxibustion plaster group， and a combination group，with 30 cases in each group. According to the 10th edition of COVID-19 Diagnosis and Treatment Protocol，patients in the Chinese medicine group received oral Chinese medicine based on syndrome differentiation，one dose per day， twice a day. Patients in the indirect moxibustion plaster group were treated with indirect moxibustion plaster at Zusanli （ST 36）， Pishu （BL 20）， Dazhui （GV 14）， Feishu （BL 13）， Kongzui （LU 6）， and Tiantu （CV 22），once a day，40 min each time. Patients in the combination group were treated with Chinese medicine combined with indirect moxibustion plaster. Treatment lasted two weeks. Before and after treatment，the traditional Chinese medicine （TCM） symptom score，pulmonary computed tomography （CT） score，St. George's Respiratory Questionnaire （SGRQ） score，blood routine indexes ［white blood cell count （WBC），neutrophil count （NEUT），and lymphocyte count （LYM）］， and inflammatory indexes ［C-reactive protein （CRP），serum ferritin， and interleukin-6 （IL-6）］ were observed in the three groups. The clinical efficacy was evaluated. ResultAfter treatment，the scores of TCM symptoms，pulmonary CT， and SGRQ，CRP，IL-6，and ferritin in the three groups decreased（P<0.05），while WBC and LYM increased（P<0.05）， but there was no significant difference in NEUT. The above indexes in the combination group were better than those in the other two groups（P<0.05）. After treatment， the cured and markedly effective rate was 76.7% （23/30） in the combination group， 50.0% （15/30） in the Chinese medicine group， and 46.7% （14/30） in the indirect moxibustion plaster group. The cured and markedly effective rate of the combination group was significantly higher than that of the Chinese medicine group （χ²=4.593， P<0.05） and the indirect moxibustion plaster group （χ²=5.711， P<0.05）. The total effective rate was 96.7 % （29/30） in the combination group， 93.3% （28/30） in the Chinese medicine group， and 86.7% （26/30） in the indirect moxibustion plaster group. The total effective rate of the combination group was higher than that of the Chinese medicine group and the indirect moxibustion plaster group， but the differences were not statistically significant. ConclusionChinese medicine combined with indirect moxibustion plaster can effectively improve the clinical symptoms，promote pulmonary inflammation，blood routine indexes， and inflammatory indexes， and improve the quality of life of COVID-19 patients during the recovery period，which is more advantageous than Chinese medicine alone or indirect moxibustion plaster.

Batch and continuous fermentation were adopted to investigate the effect of specific growth rate and amino acid components on RNA accumulation in Candida tropicalis ATCC 20408 in fermentation medium ( FM), yeast peptone dextrose medium (YPD), molasses fermentation medium ( MFM) and FM without corn steep liquor. The data showed that obvious differences in intracellular RNA accumulation were observed at different cell growth phases in bath fermentation prosess, and RNA level reached 11. 8% (g-RNA /g-DCW) during exponential phase, and only 6.9% during stationary phases. It was also found that intracellular RNA accumulation increased with the increase of specific growth rate in continue fermentation prosess, and the highest RNA level reached 15. 6% with the glucose conversion rate of 42. 8% at the dilution rate of 0. 5 h-1. Furthermore, the data showed that RNA lever was notably increased in batch fermentation process when amino acids or peptone was added into the fermentation medium containing no corn steep liquor. Taken together, it was reported for the first time that specific growth rate and amino acid components plays a leading role on the intracellular RNA accumulation in C. tropica lis, and specific growth rate is more important.