Objective: To investigate the effect of integrin α5 on the expression of NOD-like receptor thermal protein domain associated protein 3 (NLRP3) in periodontal ligament fibroblasts (PDLFs) within an inflammatory microenvironment. Methods: This study was approved by the Ethics Committee of Laboratory animals. After rat PDLFs were treated with LPS (0.5, 5, and 50 µg/mL) for 24 h, the primary medium was discarded and replaced with serum-free culture medium. After 24 h, the supernatant was collected and mixed with DMEM medium containing 10% exosome-free serum at a volume ratio of 1:1 to obtain conditioned medium (CM). The groups were labeled as the 0.5-CM, 5-CM, and 50-CM groups. In addition, PDLFs cultured in DMEM medium containing 10% exosome-free serum were considered the 0-CM group. PDLFs were cultured with the above CM. In the inhibitor group, PDLFs were cultured in 0-CM containing different concentrations of integrin α5 inhibitor ATN-161 (0, 0.025, 0.25, 2.5, 25, and 250 μg/mL). The effect of CM and integrin α5 inhibitor ATN-161 on cell viability was assessed using the CCK-8 assay. According to the CCK-8 results, in further inhibitor intervention experiments, PDLFs were cultured in 0-CM, 5-CM (without/with 25 μg/mL ATN-161), and 0-CM containing 25 μg/mL ATN-161, which were labeled as the 0-CM, 5-CM, ATN-161+5-CM, and ATN-161 groups, respectively. The expression changes of integrin α5 and NLRP3 were detected using Western blot and qRT-PCR techniques. For in vivo experiments, 24 rats were randomly divided into four groups (n=6). The control group contained healthy rats that received no treatment. The rats in the other three groups were injected with 40 µL of 0-CM containing 25 μg/mL ATN-161 or 5-CM (without or with 25 μg/mL ATN-161) on the palatal side of the left maxillary first molar every three days; these groups were classified as the ATN-161, 5-CM, and ATN-161+5-CM groups, respectively. On the 30th day, the left maxillary tissue of rats was used for Micro-CT, HE staining, and immunohistochemical detection. Results : The CCK-8 assay showed that CM, 25 μg/mL ATN-161, and ATN-161 concentrations below 25 μg/mL had no significant effect on cell viability at 12 h and 24 h (P > 0.05). 50-CM and 25 μg/mL ATN-161 significantly inhibited cell viability at 48 h (P < 0.05). For in vitro experiments, compared to the 0-CM group, both the protein and mRNA levels of integrin α5 and NLRP3 were significantly increased in rat PDLFs in the 5-CM group (P < 0.05). Intervention with 25 μg/mL ATN-161 significantly attenuated the enhancement of 5-CM on the expression of integrin α5 and NLRP3 (P < 0.05). For in vivo experiments, compared to the control group, alveolar bone resorption and periodontal inflammatory cell infiltration were significantly increased in the 5-CM and ATN-161+5-CM groups, and the expression of integrin α5 and NLRP3 was significantly increased (P < 0.01). However, compared to the 5-CM group, the ATN-161+5-CM group had less alveolar bone resorption and fewer periodontal inflammatory cells. Further, the expression of integrin α5 and NLRP3 was significantly reduced (P < 0.01). Conclusion In vitro and in vivo experiments showed that integrin α5 mediated NLRP3 expression in PDLFs under an inflammatory microenvironment. ATN-161 inhibited the expression of integrin α5, thus significantly downregulating the expression of NLRP3, which plays a role in inhibiting inflammation.

ObjectiveTo observe the clinical efficacy of acupuncture combined with bloodletting and cupping in the treatment of chronic spontaneous urticaria（CSU） and to explore its potential mechanisms of action. MethodsSeventy CSU patients were randomly divided into loratadine group and acupuncture + bloodletting group， with 35 patients in each group. The loratadine group received oral loratadine tablets， 10 mg once daily in the evening. The acupuncture + bloodletting group received acupuncture at Zhongwan （CV 12）， Guanyuan （CV 4）， Tianshu （ST 25）， Zusanli （ST 36）， Sanyinjiao （SP 6）， Xuehai （SP 10）， Quchi （LI 11）， Hegu （LI 4）， Taichong （LR 3）， Baihui （GV 20）， and Shenting （GV 24）， once daily，along with bloodletting and cupping at Dazhui （GV 14） and Geshu （BL 17）， every other day. Both groups were treated for 4 weeks. The 7-day urticaria activity score（UAS7） was assessed before and after the treatment， and levels of serum immunoglobulin E （IgE）， interleukin-4 （IL-4）， interleukin-5 （IL-5）， eosinophil cationic protein （ECP）， plasma tissue factor （TF）， activated factor Ⅶ （FⅦa）， prothrombin fragment 1+2 （F1+2）， D-dimer （D-D） and complement component 5a （C5a） were detected. ResultsA total of 65 patients were included in the final analysis， 32 in the loratadine group and 33 in the acupuncture + bloodletting group. Before treatment， there was no significant difference in UAS7 score， serum IgE， IL-4， IL-5， ECP levels， or plasma TF， FⅦa， F1+2， D-D， C5a levels between groups （P＞ 0.05）. After treatment， both groups showed significant reductions in UAS7 score， serum IgE， IL-4， IL-5， and plasma TF， FⅦa， F1+2， D-D， and C5a levels compared to those before treatment （P＜0.01）. However， after treatment， there was no significant difference in UAS7 score and serum ECP， IgE， IL-4， IL-5 levels between groups （P＞0.05）. The acupuncture + bloodletting group showed lower plasma TF， FⅦa， F1+2， D-D and C5a levels compared to the loratadine group （P＜0.05 or P＜0.01）. ConclusionAcupuncture combined with bloodletting and cupping can effectively improve the skin symptoms of CSU patients and reduce the levels of inflammatory factors. The potential mechanism of action may involve the regulation of the coagulation-complement-mast cell activation axis， thereby inhibiting mast cell degranulation.

Rapid eye movement sleep behavior disorder （RBD） is a type of parasomnia closely associated with neurodegenerative diseases related to α-synucleinopathies， such as Parkinson disease， dementia with Lewy bodies， and multiple system atrophy， and early diagnosis is of great importance for disease monitoring and intervention.At present， RBD is mainly diagnosed based on video polysomnography （v-PSG） and nocturnal abnormal behaviors， but the application of v-PSG is limited by its high technical demands.Various validated RBD-related scales have become essential tools for auxiliary diagnosis， which provides methods and tools for the diagnosis of RBD and the assessment of disease progression and outcomes.

Humans ; Vascular Stiffness ; Coal Mining ; Occupational Diseases/epidemiology* ; Risk Factors ; Coal ; Miners

OBJECTIVE To investigate the effects of quercetin on mitochondrial energy metabolism function after myocardial ischemia. METHODS H9c2 cells were divided into blank group， model group， quercetin high-dose， medium-dose and low-dose groups （40， 20， 10 μmol/L）， and positive control group （cyclosporine A， 1 μmol/L）. Reactive oxygen species （ROS）， mitochondrial membrane potential （MMP）， openness of mitochondrial permeability transition pore （MPTP）， adenosine triphosphate （ATP）， malondialdehyde （MDA）， lactate dehydrogenase （LDH） and creatine kinase （CK） were observed after cell hypoxia treatment. Rats were randomly assigned into sham operation group， model group， quercetin high-dose， medium-dose and low-dose groups （100， 50， 25 mg/kg）， and positive control group （trimetazidine， 6.3 mg/kg）， with 8 rats in each group. They were given relevant medicine intragastrically， once a day， for 7 consecutive days. After the last medication， myocardial ischemia model was induced by the ligation of the left anterior descending branch of the coronary artery. The contents of LDH， MDA， creatine kinase isoenzyme-MB （CK-MB）， superoxide dismutase （SOD）， complex Ⅰ， complex Ⅳ and ATP in serum were all determined. RESULTS Compared with the model group， ROS fluorescence intensity， openness of MPTP， the contents of CK， LDH and MDA were significantly decreased in quercetin low-dose， medium-dose and high-dose groups， and positive control group， while the contents of MMP and ATP were all increased significantly （P＜0.01）； the contents of CK-MB， LDH and MDA in serum were all decreased significantly in quercetin low-dose， medium-dose and high-dose groups， and positive control group， while the contents of SOD， complex Ⅰ， complex Ⅳ and ATP （except for positive control group） were increased significantly （P＜ 0.05 or P＜0.01）. CONCLUSIONS Quercetin can effectively reduce myocardial hypoxic injury， promote endogenous energy production and improve mitochondrial function after myocardial ischemia.

Objective To investigate the association between body mass index(BMI),sex hormone and single nucleotide polymorphisms(SNPs)of follicle-stimulating hormone receptor(FSHR)gene rs2268361 and rs2349415 and its correlation with the risk of polycystic ovary syndrome(PCOS).Methods Peripheral blood was collected from 213 PCOS patients and 207 healthy controls,attending the Department of Reproductive Medicine at the First Hospital of Shanxi Medical University,and 32 follicular fluids were randomly collected from each of the PCOS and control groups from March to August 2021.Calculation of BMI of the PCOS and control groups;The levels of follicle-stimulating hormone(FSH),luteinizing hormone(LH),estradiol(E2),testosterone(T),progesterone(P)and prolactin(PRL)in peripheral blood of the two groups were detected by immunochemiluminescence method.Polymerase chain reaction(PCR)and high-resolution melting curve(HRM)were used to analyze the polymorphisms of rs2268361 and rs2349415 in FSHR of the two groups.Quantitative real-time PCR was used to detect the expression of FSHR gene mRNA in peripheral blood and ovarian granulosa cells.Results There was a strong positive correlation between LH and LH/FSH(r=0.88,P<0.05);The levels of BMI,E2,LH,LH/FSH and T in PCOS group were significantly higher than those in control group(P<0.05);FSH level was significantly lower than that of control group(P<0.001).HRM analysis showed the frequencies of CC,CT and TT genotypes at rs2349415 were 55.9%,34.3%and 9.8%in PCOS group and 68.6%,23.2%and 8.2%in control group,respectively.The frequencies of C and T alleles were 73.0%and 27.0%in PCOS group and 80.2%and 19.8%in control group,respectively.There were significant differences in genotype frequencies and allele frequencies between the two groups(P<0.05);The expression level of FSHR mRNA was higher in ovarian granulosa cells in PCOS group than in control group(P=0.004),the expression level of FSHR mRNA in rs2349415 TT genotype was higher than that in CC(P=0.002)and CT(P=0.035)genotype.Conclusion High levels of BMI, LH, E2 and T allele of rs2349415 increased the risk of PCOS.

BACKGROUND:Some patients with cervical spondylosis have not been fully corrected sagittal position balance after cervical surgery,and this continuous sagittal position imbalance may be an important reason for the poor long-term clinical outcome of patients. OBJECTIVE:To analyze the correlation between the cervical sagittal position balance parameters and their changes and the clinical efficacy of patients in the unbalanced state after anterior cervical decompression and fusion and to explore the necessity of surgical correction of sagittal balance in order to improve the clinical effect in the later stage. METHODS:A retrospective analysis was performed on 125 patients with cervical spondylosis who underwent anterior cervical decompression and fusion in the Department of Spinal Surgery of Affiliated Hospital of Southwest Medical University from July 2019 to July 2022.Follow-up patients had good postoperative recovery(neck disability index score less than 10%one week after surgery)and had complete follow-up data.According to the axial vertical distance(C2-7 SVA)in sagittal position one week after surgery,patients were divided into type I imbalance group(C2-7 SVA loss≤5 mm,n=27),type Ⅱ imbalance group(C2-7 SVA loss>5 mm,and≤10 mm,n=19),and type Ⅲ imbalance group(C2-7 SVA loss>10 mm,n=12),and non-unbalanced group(C2-7 SVA in the normal range,n=67).The changes of visual analog scale score and neck disability index were compared among groups postoperatively and the last follow-up,as well as the changes of imaging sagittal balance parameters C2-7 cobb angle,C2-7 SVA value,neck inclination angle,T1 inclination angle,and thoracic entrance angle.The correlation between the late clinical effect and postoperative cervical sagittal disequilibrium was explored. RESULTS AND CONCLUSION:(1)There was no statistical difference in general data among the four groups(P>0.05).All patients underwent successful surgery without serious complications and postoperative wound infection.The follow-up time was more than 1 year.(2)There was no significant difference in preoperative symptom score and clinical efficacy one week after surgery(P>0.05).At the last follow-up,pain visual analog scale score,neck disability index and C2-7 SVA were lower than those before surgery but higher than those one week after surgery(P<0.05).C2-7 cobb angle was increased compared with those before operation(P<0.05).T1 inclination angle was decreased compared with those before operation(P<0.05).(3)Pearson correlation test showed that the change of neck disability index was positively correlated with the change of C2-7 SVA(P<0.05).(4)It is indicated that anterior cervical decompression and fusion is effective in the treatment of cervical spondylosis,and can effectively relieve the symptoms of patients.Patients with more severe cervical sagittal disequilibrium after surgery had worse curative effect in the later period.Continuous sagittal disequilibrium in patients with cervical spondylosis after surgery is an important cause of poor curative effect in the later stage.Clinicians should pay more attention to the correction of cervical sagittal balance before and during surgery,formulate surgical strategies and plans according to sagittal balance parameters before surgery,and correct C2-7 SVA intraoperatively to the normal range.

ObjectiveTo investigate the impact of early intervention with Yishen Huazhuo prescription （YHP） on the learning and memory of accelerated aging model mice， as well as its underlying mechanism. MethodForty-eight 3-month-old male SAMP8 mice were randomly assigned into four groups， including the model group， low-dose YHP group， high-dose YHP group， and donepezil group. Additionally， 24 SAMR1 mice of the same age were divided into a control group and a YHP treatment control group， each consisting of 12 mice. The YHP groups received YHP at doses of 6.24 g·kg^-1 and 12.48 g·kg^-1， while the donepezil group was treated with donepezil at a dose of 0.65 mg·kg^-1. The model group and control groups were given physiological saline. The mice were gavaged once daily for a duration of four weeks. Spatial learning and memory abilities of mice were assessed using the Morris water maze test. Immunofluorescence staining was employed to evaluate neuronal density as well as expression levels of M1 microglial （MG） polarization marker inducible nitric oxide synthase （iNOS） and M2 MG polarization marker arginase-1 （Arg-1） in the hippocampus region. Enzyme-linked immunosorbent assay （ELISA） was used to measure serum levels of pro-inflammatory factor interleukin 1β （IL-1β） and anti-inflammatory factor transforming growth factor-β₁ （TGF-β₁）. Furthermore， Western blot analysis was conducted to determine expressions of amyloid β peptide1-42 （Aβ_1-42） along with triggering receptor expressed on myeloid cells 2 （TREM2）/nuclear factor kappa B （NF-κB） signaling pathway-related proteins TREM2， phospho （p）-NF-κB p65， and phospho-inhibitory kappa B kinase β （IKKβ） in the hippocampus. ResultCompared with the control group， the model group exhibited a significantly prolonged escape latency （P<0.01）， a significant reduction in neuron-specific nuclear protein （NeuN） expression in the hippocampus， a significant increase in iNOS expression in MG， and a significant decrease in Arg-1 expression. The serum IL-1β content was significantly increased， while the TGF-β₁ content was significantly decreased. Additionally， there was a significant decrease in TREM2 expression in the hippocampus and significant increases in p-NF-κB p65， p-IKKβ， and Aβ_1-42 expressions （P<0.05， P<0.01）. However， no significant changes were observed in escape latency， times of crossing the platform， and hippocampal NeuN expression in the YHP treatment control group. Conversely， iNOS expression in MG as well as the hippocampal p-NF-κB p65， p-IKKβ， and Aβ_1-42 expressions were significantly decreased. Furthermore， TREM2 expression was significantly increased （P<0.05， P<0.01）. In comparison to the model group， the low-dose YHP group showed a significantly shortened escape latency and an increased number of crossing the platform （P<0.05， P<0.01）. In the high-dose YHP group， the escape latency was significantly shortened （P<0.05）. In the low-dose YHP group， high-dose YHP group， the expression of NeuN in the hippocampus was significantly increased， the expression of iNOS in MG was significantly decreased， and the expression of Arg-l was significantly increased. The serum IL-1β content was significantly decreased， while the TGF-β₁ content was significantly increased. Furthermore， the expression of TREM2 in the hippocampus was significantly increased， and the expressions of p-NF-κB p65， p-IKKβ， and Aβ_1-42 were significantly decreased （P<0.01）. ConclusionEarly YHP intervention may promote the transformation of hippocampal MG from M1 to M2 by regulating the TREM2/NF-κB signaling pathway， reduce the release of neuroinflammatory factors， protect hippocampal neurons， and reduce the deposition of Aβ_1-42， and finally delay the occurrence of learning and memory decline in SAMP8 mice.

【Objective】 To investigate the relationship between the initial serum ammonia level and the risk of ICU and hospital mortalities in critically ill patients without hepatic disease. 【Methods】 A retrospective cohort study was conducted among patients admitted to the eICU Collaborative Research Database (eICU-CRD) for a single admission who had serum ammonia test records within 48 hours of the first ICU admission and had no hepatic disease. The age, sex, ethnicity, Acute Physiologic and Chronic Health Evaluation Ⅳ score (APACHE Ⅳ score), treatment methods, complications, and outcomes were extracted. Univariable and multivariable Logistic regression were used to analyze the relationship between serum ammonia level and the risk of mortality. Interactions were used to analyze whether the relationship between serum ammonia level and the risk of mortality differed in subgroups of APACHE Ⅳ scores, age, sex, and ethnicity; subgroup analyses were made. 【Results】 A total of 1 674 patients were included. The multivariable Logistic regression showed that for every 10 μg/dL increase in ammonia, the risk of ICU death increased by 6.9% (OR=1.069, 95% CI: 1.036-1.104), and the risk of hospital death increased by 4.6% (OR=1.046, 95% CI: 1.017-1.076). The risk of ICU death was 1.7 times greater in patients with initial ammonia level of 49-82 μg/dL than in those with <49 μg/dL (OR=1.700, 95% CI: 1.165-2.482), the risk of ICU death was 2.862 times greater in patients with a level of ≥82 μg/dL compared to those with <49 μg/dL (OR=2.862, 95% CI: 1.792-4.570), and the risk of hospital death was 1.844 times higher in the ≥82 μg/dL group than in the <49 μg/dL group (OR=1.844, 95% CI: 1.213-2.804). There were no significant differences between initial ammonia level and the risk of mortalities in different subgroups of APACHEⅣ scores, age, sex, or ethnicity. 【Conclusion】 In critically ill patients without hepatic disease, elevated initial serum ammonia level after ICU admission is associated with a high risk of ICU and hospital mortality.

Objective:To explore the application value of Oxford nanopore technologies (ONT) in the diagnosis and treatment of periprosthetic joint infection (PJI).Methods:A prospective analysis was conducted on 32 patients with PJI admitted to the joint department of Xi'an Honghui Hospital from October 2021 to March 2023, who met the 2018 PJI diagnostic criteria of the American Skeletal Infection Society (MSIS), including 15 males and 17 females with an average age of 63.93±8.93 years. 32 revision patients who did not meet the 2018 MSIS PJI criteria during the same period were collected as controls (non PJI group), including 13 males and 19 females with an average age of 65.53±8.54 years. All patients underwent joint fluid puncture before or during surgery, and the specimens were tested by ONT, metagenomic next generation sequencing (mNGS), and general microbial culture. The receiver operating characteristic (ROC) curves were drawn for both groups, and the sensitivity, specificity, positive predictive value, negative predictive value, and Youden index of the three detection techniques were calculated and compared to evaluate the detection efficiency of different detection methods in PJI.Results:Among the 32 patients with PJI, 30 were positive for ONT, with a total of 30 pathogenic bacteria detected, and the detection time was 22.37±8.36 h. 31 were positive for mNGS, with a total of 33 bacterial species detected, and the detection time was 46.25±9.36 h. 17 were positive for microbial culture, with a total of 8 bacterial species detected, and the detection time was 96.23±15.62 h. Among the 32 patients with non PJI group, 1 was positive for ONT and 5 were positive for mNGS, with a total of 1 and 3 bacterial species detected, respectively. The results of microbial culture were all negative. The detection time and area under the curve (AUC) of ONT and mNGS were 22.37±8.36 h and 0.953[95% CI (0.901, 1.006)], 46.25±9.36 h and 0.906[95% CI (0.835, 0.977)], respectively, which were better than those of microbial culture 96.23±15.62 h and 0.766[95% CI (0.678, 0.853)], and the difference was statistically significant ( P<0.05). The sensitivity of ONT, mNGS, and microbial culture were 0.938, 0.969, and 0.531, respectively, and the specificity was 0.969, 0.844, and 1.000, respectively. The Jordan index was 0.906, 0.813, and 0.531, respectively. Conclusion:ONT testing has higher diagnostic efficacy than mNGS and microbial culture in the diagnosis of PJI, and also has advantages in detection time. It also suggests that some PJI are not caused by a single microbial infection.