Objective: To investigate the mechanism by which serum amyloid P component (SAP) alleviates periodontitis in mice, providing an experimental basis to establish SAP as a novel therapeutic agent for periodontitis. Methods: Ethical approval was obtained from the Institutional Animal Ethics Committee. Periodontitis models were established in wild-type (WT) mice and SAP-transgenic (SAP-Tg) mice, divided into four groups: WT control (WT group), WT periodontitis (WT+P group), SAP-Tg control (Tg group), and SAP-Tg periodontitis (Tg+P group). On day 7, the mice were euthanized, and periodontal tissues, teeth, and alveolar bone were collected. SAP protein expression was detected by enzyme-linked immunosorbent assay (ELISA). Micro-CT and HE staining were used to measure alveolar bone resorption (distance from the cementoenamel junction to the alveolar bone crest). Tartrate-resistant acid phosphatase (TRAP) staining was performed to assess osteoclast number, and immunohistochemistry (IHC) was employed to evaluate macrophage infiltration. The expression levels of inflammatory cytokines including interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were measured by qRT-PCR. Oral microorganism composition was analyzed using 16S ribosomal RNA (16S rRNA) gene sequencing. Additionally, macrophages from WT and SAP-Tg mice were isolated to establish an in vitro inflammation model, divided into WT+LPS and Tg+LPS groups. The expression of macrophage polarization-related genes including inducible nitric oxide synthase (iNOS), CD86, CD163, and CD206) were assessed by qRT-PCR. After the induction of osteoclast differentiation, TRAP staining was performed. Results: ELISA results demonstrated that periodontal tissues from Tg+P group mice exhibited higher levels of SAP expression compared to the WT+P group. Micro-CT and HE staining analyses revealed that the Tg+P group showed reduced alveolar bone resorption, indicated by a shorter distance between the cementoenamel junction and alveolar bone crest, compared to the WT+P group. Furthermore, TRAP staining results indicated a decrease in osteoclast numbers in the Tg+P group compared to the WT+P group. IHC and qRT-PCR results indicated reduced macrophage infiltration and decreased expression of IL-1β, IL-6, and TNF-α in the Tg+P group. Oral microorganism sequencing showed no significant difference in periodontitis-associated pathogenic bacteria between WT+P and Tg+P groups. In vitro experiments demonstrated that compared to the WT+LPS group, the Tg+LPS group exhibited downregulated M1 macrophage markers (iNOS and CD86) and upregulated M2 macrophage markers (CD163 and CD206). TRAP staining confirmed fewer osteoclasts in the Tg+LPS group. Conclusion SAP overexpression effectively alleviates periodontitis severity in mice by inhibiting M1 macrophage polarization, reducing pro-inflammatory cytokine expression, and suppressing osteoclast differentiation, thereby attenuating alveolar bone resorption.

OBJECTIVE: To observe the effect of moxibustion on small intestinal mucosal immune barrier in rats with diarrhea-predominant irritable bowel syndrome (IBS-D) and explore its underlying mechanisms. METHODS: Of 38 newborn rats from 4 healthy SPF pregnant rats, 12 neonatal rats were randomly selected in a normal group. IBS-D model was prepared by the combined measures for the rest rats, including neonatal maternal separation, acetic acid enema and chronic restraint stress. Twenty-four successfully-modeled rats were randomized into a model group and a moxibustion group, 12 rats in each one. In the moxibustion group, suspending moxibustion was delivered at bilateral "Tianshu" (ST25) and "Shangjuxu" (ST37), 20 min each time, once daily and for 7 consecutive days. Separately, before acetic acid enema (aged 35 days), after modeling (aged 45 days) and after intervention (aged 53 days), the body mass, loose stool rate (LSR) and and the minimum volume threshold when abdominal withdrawal reflex (AWR) scored 3 were observed in the rats of each group. After intervention (aged 53 days), using HE and PAS staining, the morphology of duodenum was observed, the length of villus and the depth of crypt were measured, the ratio of the length of villus to the depth of crypt was calculated; and the numbers of mucosal intraepithelial lymphocytes (IELs) and goblet cells were counted. With ELISA adopted, the contents of γ-interferon (IFN-γ), interleukin-4 (IL-4) and secretory immunoglobulin A (sIgA) in duodenal mucosa of rats were detected. The proportion of T cell subsets in duodenal mucosa was detected using flow cytometry. The microvilli and tight junctions of duodenal mucosal epithelial cells were observed by transmission electron microscopy, and the integrity of duodenal mucosa observed by scanning electron microscopy. RESULTS: Compared with the normal group, for the rats in the model group, the body mass, the minimum volume threshold when AWR scored 3, the length of duodenal villus and the the ratio of the length of villus to the depth of crypt, as well as the proportion of CD₈⁺ T subset were all reduced (P<0.01, P<0.05), the counts of goblet cells in duodenal mucosa decreased (P<0.01); LRS, the proportion of CD₄⁺ T subset and CD₄⁺/CD₈⁺, as well as the contents of IFN-γ, IL-4 and sIgA in duodenal mucosa and IFN-γ/IL-4 were all elevated (P<0.01); and the numbers of IELs rose (P<0.01). The morphology of duodenal mucosa was irregular, the villi got shorter, sparse and scattered, with uneven density. The morphology of epithelial cells was destroyed and the tight junctions damaged, with larger spaces. When compared with the model group, in the moxibustion group, the body mass, the minimum volume threshold when AWR scored 3, the length of duodenal villus and the ratio of the length of villus to the depth of crypt, as well as the counts of goblet cells in duodenal mucosa increased (P<0.01); LRS, the proportion of CD₄⁺ T subset, and CD₄⁺/CD₈⁺, as well as the contents of IFN-γ, IL-4 and sIgA in duodenal mucosa and IFN-γ/IL-4 were reduced (P<0.01); and the numbers of IELs was dropped (P<0.01). The morphology of duodenal mucosa was more regular, the villi were grew, got longer and arranged regularly, with even density. The morphology of epithelial cells was slightly destroyed, and the tight junctions partially damaged. CONCLUSION Moxibustion at "Tianshu" (ST25) and "Shangjuxu" (ST37) can reduce visceral hypersensitivity in IBS-D rats and relieve abdominal pain, diarrhea and other symptoms. Its effect mechanism may be related to the repair of small intestinal mucosal immune barrier and the improvement in the immune function in IBS-D.
Animals ; Irritable Bowel Syndrome/immunology* ; Rats ; Moxibustion ; Intestinal Mucosa/immunology* ; Female ; Diarrhea/therapy* ; Intestine, Small/immunology* ; Male ; Humans ; Rats, Sprague-Dawley ; Disease Models, Animal

Objective: To explore the therapeutic mechanism of puerarin in treating rheumatoid arthritis (RA) rats based on the serine/tyrosine protein kinase B (AKT)-phosphorylated forkhead box protein O1 (FOXO1)-interleukin-9 (AKT-FOXO1-IL-9) signaling pathway. Methods : 36 rats were randomly divided into a blank group , a model group , a positive control group , and low , medium , and high dose groups of puerarin. Except for the blank group , the other groups were induced with type Ⅱ collagen to establish a RA rat model. After successful modeling , different doses of puerarin and methotrexate were given to treat the rats. The body mass and toe thickness of the rats were measured , and biochemical indicators of rat blood rheology were detected. X-ray was used to observe changes in rat joint morphology. Safranin green staining were used to observe the pathology of rat joint tissue. ELISA was used to detect the levels of IL-9 and rheumatoid factors in rat serum , and Western blot was used to detect changes in levels of AKT and FOXO1 . 36 rats were randomly divided into a blank group , a model group , a positive control group , and low , medium , and high dose groups of puerarin. Except for the blank group , the other groups were induced with type Ⅱ collagen to establish a RA rat model. After successful modeling , different doses of puerarin and methotrexate were given to treat the rats. The body mass and toe thickness of the rats were measured , and biochemical indicators of rat blood rheology were detected. X-ray was used to observe changes in rat joint morphology. Safranin green staining were used to observe the pathology of rat joint tissue. ELISA was used to detect the levels of IL-9 and rheumatoid factors in rat serum , and Western blot was used to detect changes in levels of AKT and FOXO1 . Results: Compared with the blank group , the model group had the lowest toe thickness , and X-ray images showed more obvious segmental stenosis and more severe marginal bone invasion ; scaly like changes appeared at the edges of joints stained with safranin green , accompanied by the exudation of inflammatory cells and increased proliferation and secretion of chondrocytes ; the expression levels of inflammatory factors IL-9 and rheumatoid factors were the highest , and the expression levels of AKT and FOXO1 proteins were the highest (P < 0. 05) . Compared with the model group , the toe thickness of rats treated with different doses of puerarin decreased ; X-ray images showed that the puerarin treatment group of rats showed improvement in plantar joint stenosis and marginal bone invasion ; the results of safranin green staining showed that after treatment with different doses of puerarin , the infiltration of inflammatory cells decreased , and the expression levels of inflammatory factor IL-9 , rheumatoid factors , AKT , and FOXO1 proteins decreased significantly ( P < 0. 05 ) , with the high-dose puerarin group showing the most significant difference. Compared with the high-dose puerarin group , the positive control group showed a significant decrease in the above results and statistical differences (P < 0. 05) . Conclusion Puerarin has a good therapeutic effect on rats with RA by inhibiting the AKT-FOXO1-IL-9 pathway. The high-dose puerarin group (60 mg/kg) has the best therapeutic effect and the results show a dose-response relationship.

Objective:To visualize the relationship between different combinations of mechanical power exposure intensity-duration and death risk in mechanical ventilation patients using a visualization method.Methods:Critically ill patients receiving mechanical ventilation were selected from the Medical Information Mart for Intensive Care-Ⅳ v1.0 (MIMIC-Ⅳ v1.0) database. The patients were divided into four subgroups according to oxygenation index (PaO 2/FiO 2) including > 300 mmHg (1 mmHg≈0.133 kPa) group, 201-300 mmHg group, 101-200 mmHg group and ≤100 mmHg group. The baseline characteristics, ventilator parameters, and prognostic indicators for different patient populations were collected. For each patient, the mechanical power thresholds from low to high (5-30 J/min, increasing at intervals of 1 J/min) were used to evaluate the different exposures of mechanical power (above the set threshold was recorded as one exposure), and the number of events with different exposure intensity-duration combinations was counted based on their corresponding durations. Based on the 28-day survival/non-survival status, the number of exposures for survivors and non-survivors in each exposure intensity-duration combination was calculated, and the survival odds ratio ( OR) for different mechanical power exposure intensity-duration combinations was subsequently computed. Two-dimensional tables were generated with mechanical power exposure duration on the x-axis and exposure intensity on the y-axis, and the heatmap and its corresponding equipotential line view were used to visualize the OR value to assess the risk of death. Results:A total of 5 378 patients receiving mechanical ventilation were enrolled in the study, of whom 2 069 patients in the PaO 2/FiO 2 > 300 mmHg group, 813 patients in the 201-300 mmHg group, 1 493 patients in the 101-200 mmHg group, and 1 003 patients in the ≤100 mmHg group. The severity scores of patients, including sequential organ failure assessment (SOFA) score and simplified acute physiology score Ⅱ (SAPSⅡ), gradually increased following the decrease in PaO 2/FiO 2, and the incidence of co-morbidities also gradually increased. In terms of ventilator parameters, mechanical power was increased gradually with decrease in PaO 2/FiO 2, measuring 10.4 (7.8, 13.9), 11.3 (8.5, 14.7), 13.6 (10.0, 18.2), and 16.7 (12.5, 22.0) J/min ( P < 0.01). In terms of prognosis, 28-day mortality of patients was gradually increased with decrease in PaO 2/FiO 2 [29.1% (601/2 069), 26.9% (219/813), 28.1% (420/1 493), and 33.3% (334/1 003), respectively, P < 0.05]. In the heatmap, it could be observed that the 28-day death risk of mechanical ventilation patients was gradually increased with increase in mechanical power exposure intensity and long duration, showing two distinct areas: a region near the bottom left corner (representing low mechanical power exposure intensity and short duration) was blue, indicating a greater chance of survival. In contrast, another region near the top right corner (representing high mechanical power exposure intensity and long duration) was red, indicating a higher risk of death. According to the fitted lines of death risk, for the same risk of death, a shorter mechanical power exposure duration was required for higher exposure intensity, while lower mechanical power exposure intensity required a longer exposure duration. The above trend of change was similarly reflected in the overall population and different oxygenation populations. Conclusions:Cumulative mechanical power exposure to higher intensity and/or longer duration is associated with worse outcomes in mechanical ventilation patients. Considering both the mechanical power exposure intensity and duration may help to evaluate the effectiveness of lung protection in mechanical ventilation patients and guide adjustments in mechanical ventilation strategy to reduce the risk of ventilator-induced lung injury.

Background The incidence rate of missed abortion is increasing year by year, but the etiology has not been fully elucidated. Adverse pregnancy history and exposure to polycyclic aromatic hydrocarbons (PAHs) may increase the risk of missed abortion. Objective To investigate the interaction between adverse pregnancy history and PAHs exposure on missed abortion in early pregnancy, and to provide evidence for the etiologic research of missed abortion. Methods A total of 114 pregnant women diagnosed with missed abortion in the Department of Obstetrics of the First Hospital of Shanxi Medical University from March to December 2019 were selected as the case group, and 139 pregnant women who visited the same hospital for voluntary induced abortion in the same period as the control group, to collect basic information and medical information of abortion, stillbirth, intrauterine growth retardation, and other adverse pregnancy history. Abortion villus tissues were collected to detect PAH-DNA adducts levels, stratified by pregnancy and adverse pregnancy history and grouped by quartile method: Q₁ (< 404.61 ng·L⁻¹), Q₂ (404.61−453.75 ng·L⁻¹), Q₃ (453.76−506.72 ng·L⁻¹), and Q₄ (≥506.73 ng·L⁻¹). SPSS 25.0 statistical software was used for χ² test and multiple logistic regression, and additive and multiplicative models were used to investigate the interaction between adverse pregnancy history and PAH-DNA adducts level on missed abortion. The PAH-DNA adducts were grouped by tertiles and quartiles, and P₃₃, P₅₀, P₆₇ and P₇₅ were used as data cut points for sensitivity analysis. Results The proportion of adverse pregnancy history in the case group (32.46%) was higher than that in the control group (12.23%) (P < 0.001). Among 160 subjects with≥2 pregnancies, the proportion of adverse pregnancy history in the case group (57.81%) was higher than that in the control group (17.71%) (P < 0.001). The results of χ² test stratified by pregnancy for different PAH-DNA adducts levels between the two groups showed that the PAH-DNA adducts level was associated with missed abortion in subjects with≥2 pregnancies (χ²=10.14, P=0.017). Being further stratified by adverse pregnancy history, the PAH-DNA adducts level in subjects with no adverse pregnancy history was associated with missed abortion (χ²=9.70, P=0.021). The results of logistic regression analysis showed that adverse pregnancy history (OR=5.88, 95%CI: 2.79−12.39) and PAH-DNA adducts (OR=3.01, 95%CI: 1.22−7.40) increased the risk of missed abortion, but no interaction between them was found. The relative excess risk of interaction (RERI), the attributable percentage of interaction (AP), and the synergy index (SI) and its 95%CI were 0.60 (95%CI: −0.58−1.77), 0.74 (95%CI: −0.83−2.30), and 0.20 (95%CI: 0.01−5.43), respectively. Conclusions Adverse pregnancy history and PAH-DNA adducts in pregnant women may increase the risk of missed abortion. The effect of the interaction between them on the occurrence of missed abortion is not supported by the current study.

Objective To evaluate the transdermal safety of lanthanum (La) in sunscreen and jellyfish sting protective lotion, establish a microwave digestion-inductively coupled plasma opticalemission spectroscopy (ICP-OES) method for determination of lanthanum (La) in rat’s whole blood. Methods The whole blood samples were digested by microwave and analyzed by inductively coupled plasma emission spectrometer (ICP-OES). Using 333.749 nm as the analysis line, the content of La in rat whole blood was determined. Results The correlation linearity of the standard curve of this method was good (r>0.9994), the detection limit of the method was 0.0025 μg/ml, the limit of quantification was 0.0077 μg/ml, the precision was less than 3%, and the recovery rate was between 94.9% and 102.0%. Conclusion The ICP-OES method based on microwave digestion is stable and reliable, and can provide an important basis for the study of the transdermal safety of lanthanum.

One major strategy to generate genetically modified mouse models is gene targeting in mouse embryonic stem (ES) cells, which is used to produce gene-targeted mice for wide applications in biomedicine. However, a major bottleneck in this approach is that the robustness of germline transmission of gene-targeted ES cells can be significantly reduced by their genetic and epigenetic instability after long-term culturing, which impairs the efficiency and robustness of mouse model generation. Recently, we have established a new type of pluripotent cells termed extended pluripotent stem (EPS) cells, which have superior developmental potency and robust germline competence compared to conventional mouse ES cells. In this study, we demonstrate that mouse EPS cells well maintain developmental potency and genetic stability after long-term passage. Based on gene targeting in mouse EPS cells, we established a new approach to directly and rapidly generate gene-targeted mouse models through tetraploid complementation, which could be accomplished in approximately 2 months. Importantly, using this approach, we successfully constructed mouse models in which the human interleukin 3 (IL3) or interleukin 6 (IL6) gene was knocked into its corresponding locus in the mouse genome. Our study demonstrates the feasibility of using mouse EPS cells to rapidly generate mouse models by gene targeting, which have great application potential in biomedical research.

Objective To investigate the effects of Epigallocatechin gallate (EGCG) on cardiac protection in diabetic rats and the expression of TGF-1/Smad3 signaling pathway. Methods The influence of clean level 40 SD rats, weight 200-220 g, divided into four random groups:control (Sham) group, diabetic cardiomyopathy model (DC) group, EGCG group, and metformin positive control group (Met).Post 8 weeks of high-fat-diet administration, the rats were injected intraperitoneally with STZ to establish the diabetes cardiomyopathy model. Upon successful model establishment, the EGCG group was intraperitoneally injected with EGCG and the cardiac function of the rats was measured after 28 days of drug administration. Then, the pathological results of the myocardial tissue were analyzed. Triglyceride (TG), total cholesterol (TC), glycosylated hemoglobin (HbA1 c), and blood glucose (FBG) concentrations were also measured. Further, the concentrations of superoxide dismutases (SOD), malondialdehyde (MDA), catalase (CAT), and glutathione peroxidase (GPX) in serum were measured by ELISA. The expression of TGF-β1 and Smad3 in kidney tissues of the rats was measured by Western blotting analysis. Results EGCG could reduce the glucose, lipid, and MDA levels in the blood of the diabetic rats, enhance cardiac systolic and diastolic functions, inhibit TGF-β1 and Smad3 protein expression, enhance the activity of SOD, CAT and GPX, and reduce myocardial tissue fibrosis. Conclusion EGCG can protect diabetic rat hearts by improving metabolic disorder, and its mechanism may be related to the oxidative-stress mediated by the TGF-β1/Smad3 signaling pathway.

Objective To investigate the effect and mechanism of suberoylanilide hydroxamic acid (SAHA) on the fear extinction in mice with chronic social defeat stress (SD). Methods Fifty-six male C57BL/6J mice aged 7-8 weeks were randomly divided into control group,social defeat group,control-SAHA group and social defeat-SAHA group to investigate the effect of SAHA and social defeat group,social defeat-AAV BDNF group and social defeat-AAV blank group to investigate the effect of BDNF. Fear extinction in mice was evaluated by fear conditioning test (FC). The levels of BDNF and HDAC2 in mice hippocampus were detected by Western blot (WB). The expression of BDNF-overexpressing virus in hippocampus of mice was detected by immunofluorescence assay. Results (1) Compared with control group,fear extinction in the social defeat group was significantly decreased (P<0. 05). Compared with control group, the level of HDAC2(0. 50±0. 02) in the social defeat group was significantly increased (P<0. 001),while the level of BDNF(0. 16 ± 0. 03) was significantly decreased (P<0. 001) in the social defeat group. ( 2) After using SAHA,fear extinction of mice significantly improved (P<0. 05). Compared with control group,the level of HDAC2 (0. 26±0. 02) in the control-SAHA group was significantly decreased(P<0. 001),and the level of BDNF (0. 40±0. 03) was significantly increased (P<0. 001). Compared with social defeat group,the level of HDAC2 (0. 39±0. 03) in the social defeat-SAHA group was significantly decreased (P<0. 001),and the lev-el of BDNF (0. 28±0. 01) was significantly increased (P<0. 001). (3)After injection BDNF-overexpressing virus,fear extinction was significantly improved(P<0. 05). Conclusion SAHA can enhance fear extinction in mice with chronic social defeat stress and its mechanism may be related to the up-regulation of BDNF ex-pression in hippocampus by inhibiting HDAC2 in hippocampal.

De novo variants (DNVs) are one of the most significant contributors to severe earlyonset genetic disorders such as autism spectrum disorder, intellectual disability, and other developmental and neuropsychiatric (DNP) disorders. Presently, a plethora of DNVs have been identified using next-generation sequencing, and many efforts have been made to understand their impact at the gene level. However, there has been little exploration of the effects at the isoform level. The brain contains a high level of alternative splicing and regulation, and exhibits a more divergent splicing program than other tissues. Therefore, it is crucial to explore variants at the transcriptional regulation level to better interpret the mechanisms underlying DNP disorders. To facilitate a better usage and improve the isoform-level interpretation of variants, we developed NeuroPsychiatric Mutation Knowledge Base (PsyMuKB). It contains a comprehensive, carefully curated list of DNVs with transcriptional and translational annotations to enable identification of isoformspecific mutations. PsyMuKB allows a flexible search of genes or variants and provides both table-based descriptions and associated visualizations, such as expression, transcript genomic structures, protein interactions, and the mutation sites mapped on the protein structures. It also provides an easy-to-use web interface, allowing users to rapidly visualize the locations and characteristics of mutations and the expression patterns of the impacted genes and isoforms. PsyMuKB thus constitutes a valuable resource for identifying tissue-specific DNVs for further functional studies of related disorders. PsyMuKB is freely accessible at http://psymukb.net.