Objective:To analyze the effects of SARS-CoV-2 infection and vaccination on virus mutation.Methods:The whole genome sequencing sequences of 2 659 local SARS-CoV-2 specimens from Jiangsu Province in 2023 were selected for analysis, and relevant information such as demographic and clinical characteristics were collected, and the effects of infection and vaccination on the genome-wide mutation rate and S gene′s selective pressure of the virus were analyzed by univariate and multivariate linear regression models.Results:The average age of these infected patients was 55.0 (31.0, 74.0) years, 1 150 cases (43.2%) in the age group of ≥60 years, 1 367 cases (51.4%) were males, 2 044 cases (76.9%) had a history of COVID-19 vaccination, and 1 629 cases (61.3%) had the first-time infection. The clinical symptoms of the infected patients were mainly mild, with a total of 2434 cases (91.5%), and 29 cases (1.1%) with severe symptoms or more. The average substitution rate of SARS-CoV-2 was 9.69 (9.38, 9.98)×10 -4 subs/site/year, and the dN/dS value of the S gene was 6.08 (5.56, 8.66), which was significantly greater than that of 1 ( P<0.001), indicating positive selection. The result of univariate and multivariate linear regression model analysis showed that the SARS-CoV-2 substitution rate was higher in those with vaccination history and reinfection, aged 20-30 years, ≥60 years, and the SARS-CoV-2 substitution rate was lower in males with moderate clinical symptoms and severe disease and above. Those with a history of vaccination and reinfection, aged 50-60 years old, ≥60 years old have smaller S gene dN/dS. Conclusions:Under the immune pressure exerted by vaccination and infection, the genome-wide mutation of SARS-COV-2 accelerated, but the non-synonymous mutation rate of the S gene decreased. The mechanism causing these phenomena needs further study.

Objective:To evaluate the detection ability of two kinds of high-throughput method to determine neutralizing antibodies based on a microneutralization test (MNT).Methods:Serum samples were collected from the early phase and follow-up period (117 samples in total) for neutralizing antibody testing. They were tested using MNT, pseudovirus neutralization assay (PBNA), competitive inhibition assay (including enzyme-linked immunosorbent assay (ELISA) and chemiluminescence immunoassay (CLIA)) to evaluate the correlation coefficients and threshold values for the effectiveness of these high-throughput neutralizing antibody assays.Results:The correlation coefficients for PBNA, ELISA, and CLIA relative to MNT were 0.760, 0.778, and 0.725, respectively, for individuals infected with 2019-nCoV. The area under the ROC curve was 0.901 for a cutoff value of 50 for the PBNA assay, 0.934 for a cutoff value of 1∶8 for the ELISA assay and 0.838 for a cutoff value of 1.28AU/ml for the CLIA assay when the threshold value for the microneutralization test was taken as 1: 10 (less than 1: 10 is considered negative).Conclusions:The high-throughput method for the detection of COVID-19 neutralizing antibodies are scientific and feasible, and provide an important technical tool for the regular prevention and control of the epidemic.

The high morbidity and fatality rates of cardiovascular and cerebrovascular diseases seriously harm human health，greatly affect the quality of life and increase the burden of disease. Thus， more and more attention has been paid to the relationship between heart and brain. "Simultaneous treatment of heart and brain" is the continuous deepening and development of the theory of "simultaneous treatment of different diseases" in traditional Chinese medicine （TCM）， and exploring the relationship between heart and brain as well as the current situation of Chinese medicine treatment has important clinical significance for the establishment of integrated traditional Chinese and western medicine diagnosis and treatment programs of cardiovascular and cerebrovascular diseases. Through a systematic theoretical tracing of the "simultaneous treatment of heart and brain"，the authors held that cardiovascular and cerebrovascular diseases had the same origin，and that heart and brain took blood as the material basis and governed mind jointly. They also expounded the modern biological relationship of the simultaneous treatment of heart and brain. In other words，the pathogenic factors of cardiovascular and cerebrovascular diseases were the same， with complicated comorbidities， and the disorders of neurohumoral reflex and endocrine system and immune system regulation could affect each other's function. Additionally，the present situation of clinical application of Chinese medicine for simultaneous treatment of heart and brain and its intervention effect on cardiovascular and cerebrovascular diseases were discussed. It was found that these Chinese medicines could protect cardiovascular and cerebrovascular diseases through multi-target，multi-pathway and multi-link regulation. Combined with the existing problems in the current research，the authors thought and looked forward to the practical strategies of treating cardiovascular and cerebrovascular diseases with Chinese medicine by paying attention to key groups，focusing on dominant diseases，giving full play to the value of Chinese medicine，and scientifically explaining the connotation of simultaneous treatment of heart and brain， in order to provide ideas and reference for the follow-up transformation application research and better guidance of clinical practice.

As China is about to enter an era of deep aging， the coexistence of multiple diseases is gradually increasing. Coronary heart disease （CHD） and cognitive dysfunction also show increasing incidence year by year. The two diseases affect and cause each other， becoming the major chronic diseases harmful to the health of the elderly. It is of great clinical significance to explore the methods integrating traditional Chinese and western medicine for the prevention and treatment of the two diseases. The relationship between CHD and cognitive dysfunction in traditional Chinese medicine （TCM） was first recorded in Huangdi’s Internal Classic （Huang Di Nei Jing）. As the understanding of CHD and cognitive dysfunction is deepening， the influences of stasis and toxin on both diseases have attracted increasing attention. According to the theories of TCM， CHD and cognitive dysfunction have common points in the etiology and pathogenesis. Therefore， the theory of treating different diseases with same method provides a theoretical basis for the clinical treatment of different diseases with the same pathogenesis. Moreover， this theory conforms to the principle of integrated and individualized prevention and treatment of multi-disease coexistence in modern medicine. This paper systematically proposed that the coexistence of stasis and toxin is a major pathogenesis of CHD and cognitive dysfunction. We then explored the possible mechanisms of the blood-activating and toxin-removing method in the treatment of CHD and cognitive dysfunction based on the theory of treating different diseases with same method. The mechanisms include the regulation of ceramide metabolism， activation of silent mating-type information regulation 2 homolog 1 （SIRT1）， inhibition of pyroptosis， regulation of mitogen-activated protein kinase/nuclear factor-κB （MAPK/NF-κB） signaling pathway， inhibition of mitochondrial division， and regulation of DNA methylation. We hope this paper will provide an idea for the future research on the prevention and treatment of CHD and cognitive dysfunction with TCM.

Diabetic retinopathy（DR） and coronary heart disease（CHD） are both major chronic vascular complications that seriously jeopardize the health of the population and often occur together in clinical practice， it is of great clinical value to actively explore the association between the two in the process of disease development and methods of prevention and treatment of modern medicine and traditional Chinese medicine（TCM）. According to TCM， the heart and eyes physiologically communicate with each other by taking Qi， blood and veins as bridges， blood stasis obstructing collaterals is the common TCM etiology of DR and CHD， whose mechanism involves inflammation， oxidative stress and endothelial dysfunction. Promoting blood circulation and removing blood stasis plays an important role in the same treatment for different diseases and prevention and treatment of comorbidities， possibly by inhibiting the expression of interleukin-1β（IL-1β）， endothelin-1（ET-1） and hypoxia inducible factor-1α/vascular endothelial growth factor（HIF-1α/VEGF）， regulating phosphatidylinositol 3-kinases/protein kinase B/mammalian target of rapamycin（PI3K/Akt/mTOR） pathway， initiating adenosine monophosphate（AMP）-activated protein kinase/silent information regulator 1（AMPK/SIRT1） and nuclear transcription factor erythroid 2-related factor 2/heme oxygenase-1（Nrf2/HO-1） signaling pathways， inhibiting Hippo/Yes-associated protein（Hippo/YAP） signaling pathway， inhibiting mitochondrial permeability transition pore and anti-platelet agglutination for treating DR and CHD， which provides a multi-component， multi-pathway and multi-target selection strategies and ideas for the prevention and treatment of DR and CHD by TCM from a biological perspective. Based on this， subsequent studies should focus on constructing clinically relevant comorbidity models， conducting multicenter prospective studies， and fully utilizing artificial intelligence technology to gain a deeper understanding of the relationship between the two diseases， so as to elucidate the mechanism of promoting blood circulation and removing blood stasis in preventing and treating panvascular diseases.

Cognitive impairment secondary to cardiovascular disease is a common complication in the elderly population， which seriously threatens the health of patients and affects the long-term quality of life. Cardiovascular diseases can lead to vascular endothelial injury， cerebral hypoperfusion， and brain lesions， and then cause cognitive impairment. Cardiovascular risk factors also increase the risk of secondary cognitive impairment in patients. It is particularly important to seek for early diagnosis and traditional Chinese medicine （TCM） and western medicine prevention and treatment. Based on the "holistic concept"， TCM puts forward the theory of "the connection of heart and brain". The heart and brain are connected and both are in charge of mental activities. Heart injury affects the brain， resulting in abnormal consciousness， which is considered to be the pathogenesis of cognitive impairment secondary to cardiovascular disease in TCM. Later generations of doctors propose the theory of "simultaneous treatment of the heart and the brain" on the basis of "treating different diseases with the same treatment" and "the connection of heart and brain". The theory contains two meanings. One is that different diseases can be treated by the same method because of the same etiology and pathogenesis. The other is the coexistence of different diseases and the treatment should be targeted to the same risk factors. This paper systematically discussed the TCM pathogenesis of cognitive impairment secondary to cardiovascular disease and the prevention and treatment strategy of "simultaneous treatment of the heart and the brain". In this paper， the deficiency of qi and blood was the foundation， and the interaction of blood stasis and poison and the obstruction of the pulse and the loss of the body were the key factors. Activating blood， detoxifying arteries， and connecting the orifices were the main treatment method for the disease. This paper is expected to enrich the connotation of "simultaneous treatment of the heart and the brain"， and provide new ideas for the prevention and treatment of cognitive impairment secondary to the cardiovascular disease with TCM.

ObjectiveTo explore the correlation between the blood stasis score of coronary heart disease（CAD） and mild cognitive impairment（MCI）， as well as the changes in plasma metabolic profile of blood stasis in patients with CAD combined with MCI（CADMCI） through a cross-sectional study， and further explore the impact of different degrees of blood stasis on the plasma metabolite profile of CADMCI patients. MethodsAccording to the diagnostic criteria of CAD and CAD blood stasis， patients hospitalized in Xiyuan Hospital of China Academy of Chinese Medical Sciences from October 2022 to October 2023 were continuously included. According to the Montreal Cognitive Assessment（MoCA） scale score， the enrolled patients were divided into CADMCI blood stasis group and CAD blood stasis group. The association between blood stasis score and MCI was analyzed by multivariate Logistic regression model. The receiver operating characteristic（ROC） curve was drawn， and the area under the curve（AUC） was calculated to evaluate the sensitivity and specificity of the model. According to the blood stasis score， the first 30 patients in the CADMCI blood stasis group and CAD blood stasis group were divided into mild blood stasis and severe blood stasis. Ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） was used to detect plasma metabolites in each group of patients. The differential metabolites were screened according to variable importance in the projection（VIP） value≥1， fold change（FC）<0.67 or >1.5， and P<0.05. ROC curve analysis was further used to evaluate the discriminatory efficiency of the screened differential metabolites for each group of samples. ResultsA total of 266 CAD patients were included in this study. Multivariate Logistic regression analysis showed that the CAD blood stasis score was significantly correlated with MCI［odds ratio（OR）=1.619， 95% confidence interval（CI） 1.223-2.142， P<0.001， ROC curve AUC was 0.615（95% CI 0.547-0.683， P=0.001）］， indicating that the CAD blood stasis score has a certain predictive value for MCI. Plasma non-targeted metabolomics analysis showed that the main differential metabolites between CAD blood stasis and CADMCI blood stasis were lipid metabolites， among which phosphatidylcholine［20∶4（5Z， 8Z， 11Z， 14Z）/P-18∶1（11Z）］ had the best discriminatory efficiency（ROC curve AUC=0.867， 95% CI 0.754-0.942）. Further analysis of the differential metabolites between mild and severe blood stasis showed that lipid metabolites were also the main differential metabolites between mild and severe blood stasis. Among them， 1α，25-dihydroxy-2β-（2-hydroxyethoxy） vitamin D₃ had the best efficacy in distinguishing mild and severe CAD blood stasis（AUC=0.813， 95% CI 0.649-0.951）， and phosphatidylcholine 34∶2 had the best efficacy in distinguishing mild and severe CADMCI blood stasis（AUC=0.819， 95% CI 0.640-0.941）. ConclusionThere is a significant correlation between CAD blood stasis score and MCI. Phosphatidylcholine metabolites play an important role in the pathogenesis of CADMCI blood stasis and severe blood stasis. The CAD blood stasis score combined with the detection of phosphatidylcholine metabolites can provide a reference for the development of early and efficient identification strategies for CADMCI.

Periodontitis is a widespread oral disease characterized by continuous inflammation of the periodontal tissue and an irreversible alveolar bone loss, which eventually leads to tooth loss. Four-octyl itaconate (4-OI) is a cell-permeable itaconate derivative and has been recognized as a promising therapeutic target for the treatment of inflammatory diseases. Here, we explored, for the first time, the protective effect of 4-OI on inhibiting periodontal destruction, ameliorating local inflammation, and the underlying mechanism in periodontitis. Here we showed that 4-OI treatment ameliorates inflammation induced by lipopolysaccharide in the periodontal microenvironment. 4-OI can also significantly alleviate inflammation and alveolar bone loss via Nrf2 activation as observed on samples from experimental periodontitis in the C57BL/6 mice. This was further confirmed as silencing Nrf2 blocked the antioxidant effect of 4-OI by downregulating the expression of downstream antioxidant enzymes. Additionally, molecular docking simulation indicated the possible mechanism under Nrf2 activation. Also, in Nrf2^-/- mice, 4-OI treatment did not protect against alveolar bone dysfunction due to induced periodontitis, which underlined the importance of the Nrf2 in 4-OI mediated periodontitis treatment. Our results indicated that 4-OI attenuates inflammation and oxidative stress via disassociation of KEAP1-Nrf2 and activation of Nrf2 signaling cascade. Taken together, local administration of 4-OI offers clinical potential to inhibit periodontal destruction, ameliorate local inflammation for more predictable periodontitis.
Alveolar Bone Loss/prevention & control* ; Animals ; Antioxidants/pharmacology* ; Inflammation ; Kelch-Like ECH-Associated Protein 1/metabolism* ; Mice ; Mice, Inbred C57BL ; Molecular Docking Simulation ; NF-E2-Related Factor 2/metabolism* ; Periodontitis/prevention & control* ; Succinates

Periodontitis is the most widespread oral disease and is closely related to the oral microbiota. The oral microbiota is adversely affected by some pharmacologic treatments. Systemic antibiotics are widely used for infectious diseases but can lead to gut dysbiosis, causing negative effects on the human body. Whether systemic antibiotic-induced gut dysbiosis can affect the oral microbiota or even periodontitis has not yet been addressed. In this research, mice were exposed to drinking water containing a cocktail of four antibiotics to explore how systemic antibiotics affect microbiota pathogenicity and oral bone loss. The results demonstrated, for the first time, that gut dysbiosis caused by long-term use of antibiotics can disturb the oral microbiota and aggravate periodontitis. Moreover, the expression of cytokines related to Th17 was increased while transcription factors and cytokines related to Treg were decreased in the periodontal tissue. Fecal microbiota transplantation with normal mice feces restored the gut microbiota and barrier, decreased the pathogenicity of the oral microbiota, reversed the Th17/Treg imbalance in periodontal tissue, and alleviated alveolar bone loss. This study highlights the potential adverse effects of long-term systemic antibiotics-induced gut dysbiosis on the oral microbiota and periodontitis. A Th17/Treg imbalance might be related to this relationship. Importantly, these results reveal that the periodontal condition of patients should be assessed regularly when using systemic antibiotics in clinical practice.
Humans ; Mice ; Animals ; Dysbiosis ; Anti-Bacterial Agents/pharmacology* ; Virulence ; Microbiota ; Periodontitis/chemically induced* ; Cytokines