Gut microbiota dysbiosis is an avenue for the promotion of atherosclerosis (AS) and this effect is mediated partly via the circulating microbial metabolites. More microbial metabolites related to AS vascular inflammation, and the mechanisms involved need to be clarified urgently. Paeonol (Pae) is an active compound isolated from Paeonia suffruticoas Andr. with anti-AS inflammation effect. However, considering the low oral bioavailability of Pae, it is worth exploring the mechanism by which Pae reduces the harmful metabolites of the gut microbiota to alleviate AS. In this study, ApoE^-/- mice were fed a high-fat diet (HFD) to establish an AS model. AS mice were administrated with Pae (200 or 400 mg·kg^-1) by oral gavage and fecal microbiota transplantation (FMT) was conducted. 16S rDNA sequencing was performed to investigate the composition of the gut microbiota, while metabolomics analysis was used to identify the metabolites in serum and cecal contents. The results indicated that Pae significantly improved AS by regulating gut microbiota composition and microbiota metabolic profile in AS mice. We also identified α-hydroxyisobutyric acid (HIBA) as a harmful microbial metabolite reduced by Pae. HIBA supplementation in drinking water promoted AS inflammation in AS mice. Furthermore, vascular endothelial cells (VECs) were cultured and stimulated by HIBA. We verified that HIBA stimulation increased intracellular ROS levels, thereby inducing VEC inflammation via the TXNIP/NLRP3 pathway. In sum, Pae reduces the production of the microbial metabolite HIBA, thus alleviating the ROS/TXNIP/NLRP3 pathway-mediated endothelial inflammation in AS. Our study innovatively confirms the mechanism by which Pae reduces the harmful metabolites of gut microbiota to alleviate AS and proposes HIBA as a potential biomarker for AS clinical judgment.
Animals ; Mice ; Atherosclerosis/drug therapy* ; Diet, High-Fat ; Endothelial Cells ; Inflammation/drug therapy* ; Mice, Inbred C57BL ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Reactive Oxygen Species

ObjectiveTo explore the anti-tumor effect and mechanism of Shenqi Yiliu prescription in the intervention of pyroptosis. MethodTen male BALB/c mice were randomly selected and assigned to the blank group. The remaining 40 mice underwent the induction of the liver cancer xenograft model. After 5 days of modeling， 40 surviving mice were randomly divided into model group， cisplatin group ［2.5×10^-3 g·kg^-1·（3 d）^-1］， Shenqi Yiliu prescription group （27 g·kg^-1·d^-1）， and a combination group （Shenqi Yiliu prescription group + cisplatin）. The mice in the blank group and the model group were treated with an equal volume of normal saline for 10 days. The general conditions of mice in each group were observed. After the intervention， the tumor weight of the mice was weighed and the tumor inhibition rate was calculated. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in tumor tissues. The levels of mouse liver function indicators， including alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were detected. The TdT-mediated dUTP-biotin nick end labeling （TUNEL） assay was used to detect DNA damage in mouse tumor tissue cells. Immunohistochemistry （IHC）， immunofluorescence （IF）， and Western blot were used to detect the protein expression levels of NOD-like receptor protein 3 （NLRP3）， cysteinyl aspartate-specific protease-1 （Caspase-1）， and gasdermin D （GSDMD） in tumor tissues. The levels of interleukin-1β （IL-1β） and interleukin-18 （IL-18） in tumor tissues were detected by enzyme-linked immunosorbent assay （ELISA）. ResultCompared with the mice in the blank group， those in the model group were in a poor mental state， sleepy， and lazy， and their fur color was dull， with increased levels of serum ALT and AST in liver function tests （P<0.01）. Compared with the model group， the groups with drug intervention showed improved mental state， inhibited tumor growth to varying degrees， and decreased tumor weight， and the tumor inhibition rate in the combination group was the highest （P<0.01）. HE staining showed that the pathological and morphological lesions of the tumor tissues in the model group were significant， while those in all groups with drug intervention were improved to a certain extent. The karyolysis and nuclear rupture in the Shenqi Yiliu prescription group and the combination group were more significant. In the liver function test， the serum ALT and AST levels of mice in the Shenqi Yiliu prescription group and the combination group decreased （P<0.01）， and the inflammatory factors IL-1β and IL-18 in each group with drug intervention decreased （P<0.05， P<0.01）. Among them， the declining trend of IL-1β and IL-18 in the Shenqi Yiliu prescription group was the most significant （P<0.01）. TUNEL staining showed that the positive TUNEL staining in each group with drug intervention decreased after intervention （P<0.05， P<0.01）， especially the cisplatin group and Shenqi Yiliu prescription group （P<0.01）. Western blot， IHC， and IF found that the protein expression levels of NLRP3， Caspase-1， and GSDMD in each group with drug intervention decreased （P<0.05， P<0.01）. Compared with the mice in the cisplatin group， those in the Shenqi Yiliu prescription group and the combination group had better mental state and regular tumor morphology， and the tumor weight of the mice in the combination group decreased （P<0.05）. The levels of ALT and AST in the Shenqi Yiliu prescription group decreased （P<0.05）， and the levels of IL-1β and IL-18 in the Shenqi Yiliu prescription group and the combination group decreased （P<0.05， P<0.01）， especially in the combination group （P<0.01）. The results of IHC showed that the expression of GSDMD protein in the tumor tissues of mice in the combination group was reduced （P<0.01）. IF detection showed that the expression of NLRP3 in the tumor tissues of the Shenqi Yiliu prescription group was reduced （P<0.01）. The results of Western blot showed that the expression level of NLRP3 protein in the Shenqi Yiliu prescription group and the combination group decreased （P<0.01）， and the expression level of Caspase-1 protein in the combination group decreased （P<0.01）. The decrease in GSDMD protein expression was not significant， and the difference was not statistically significant. ConclusionShenqi Yiliu prescription combined with cisplatin has an obvious anti-tumor effect， which may be achieved by down-regulating the NLRP3/Caspase-1/GSDMD inflammatory pyroptosis pathway to inhibit cell pyroptosis， and relieve the inflammatory response in mice with liver cancer.

OBJECTIVE: To analyze the clinical significance of combined newborn hearing and deafness gene screening in Yuncheng area of Shanxi Province. METHODS: Results of audiological examinations, including transient evoked otoacoustic emission and automatic discriminative auditory brainstem evoked potentials, for 6 723 newborns born in Yuncheng area from January 1, 2021 to December 31, 2021, were retrospectively analyzed. Those who failed one of the tests were considered to have failed the examination. A deafness-related gene testing kit was used to detect 15 hot spot variants of common deafness-associated genes in China including GJB2, SLC26A4, GJB3, and mtDNA12S rRNA. Neonates who had passed the audiological examinations and those who had not were compared using a chi-square test. RESULTS: Among the 6 723 neonates, 363 (5.40%) were found to carry variants. These have included 166 cases (2.47%) with GJB2 gene variants, 136 cases (2.03%) with SLC26A4 gene variants, 26 cases (0.39%) with mitochondrial 12S rRNA gene variants, and 33 cases (0.49%) with GJB3 gene variants. Among the 6 723 neonates, 267 had failed initial hearing screening, among which 244 had accepted a re-examination, for which 14 cases (5.73%) had failed again. This has yielded an approximate prevalence of hearing disorder of 0.21% (14/6 723). Among 230 newborns who had passed the re-examination, 10 (4.34%) were found to have carried a variant. By contrast, 4 out of the 14 neonates (28.57%) who had failed the re-examination had carried a variant, and there was a significant difference between the two groups (P < 0.05). CONCLUSION Genetic screening can provide an effective supplement to newborn hearing screening, and the combined screening can provide a best model for the prevention of hearing loss, which can enable early detection of deafness risks, targeted prevention measures, and genetic counseling to provide accurate prognosis for the newborns.
Infant, Newborn ; Humans ; Connexins/genetics* ; Retrospective Studies ; Deafness/genetics* ; Connexin 26/genetics* ; Neonatal Screening/methods* ; Mutation ; Genetic Testing/methods* ; China/epidemiology* ; Hearing ; DNA Mutational Analysis

ObjectiveTo compare the differences in sleep structure characteristics between adolescents with depressive disorder and adolescents with bipolar disorder， and to explore the impact of sleep indicators and other factors on the suicide risk of adolescents with affective disorder. MethodsThe medical records of adolescents with depressive disorder （n=97） and bipolar disorder （n=52） who met the International Classification of Diseases， tenth edition （ICD-10） and hospitalized in the Affiliated Brain Hospital of Guangzhou Medical University from January 1， 2019 to June 30， 2021 were retrospectively reviewed. Data including age， gender， body mass index （BMI）， psychiatric diagnosis， the Nurses' Global Assessment of Suicide Risk （NGASR） score and polysomnography （PSG） results of the patients were collected. Then patients were divided into two groups according to NGASR score， scored 0~5 were in the low risk group （n=32） and scored above 5 were in the high risk group （n=117）. Meantime， the PSG data of normal adolescents （n=80） in the previous literature were collected as the control group. Thereafter， a multiple linear regression model was established to explore the related factors affecting suicide risk in adolescents with affective disorder. ResultsThe sleep efficiency and the proportion of stage N2 sleep in high risk group were lower than those in low risk group （Z=-2.138， -2.520， P<0.05）. The total sleep time， N2 sleep duration and rapid eye movement （REM） sleep time in depression group were less than those in bipolar group （t=-2.822， -3.087， -2.277， P<0.05 or 0.01）. The proportion of REM sleep in depression group and bipolar group were lower than those in control group （t=-2.369， -2.069， P<0.05）. Linear regression analysis denoted that the factors affecting the suicide risk in adolescents with affective disorder included stage N1 sleep duration （β=0.019， P<0.05）， gender （male vs. female， β=-4.051， P<0.01） and psychiatric diagnosis （bipolar disorder vs. depressive disorder， β=-1.429， P<0.05）. ConclusionIn contrast to adolescents with bipolar disorder， the sleep structure of adolescents with depressive disorder is characterized by poor sleep continuity and less light sleep. Furthermore， the N1 sleep duration， female gender and diagnosis of depressive disorder are risk factors affecting the suicide in adolescents with affective disorder.

ObjectiveTo explore the factors associated with the risk of violent behaviour in inpatients with acute mental disorders， and to provide references for early detection and intervention of violent behaviour in patients with acute mental disorders. MethodsBased on the medical record system of the Affiliated Brain Hospital of Guangzhou Medical University， 1 107 inpatients with acute mental disorders from January to December 2016 were selected， all of whom met the diagnostic criteria of International Classification of Diseases， tenth edition （ICD-10）. At admission， the risk assessment tools were used to assess the risk level of violent behaviour of inpatients， and 8 variables containing general demographic data and clinical data were selected to explore the factors associated with the risk of violent behaviour in inpatients with acute mental disorders. Thereafter， Logistic regression analysis was performed to screen the factors affecting the high risk of violent behaviour among inpatients. ResultsAmong the 1 107 inpatients with acute mental disorders， 357 （32.25%） patients were at high risk of violence. Regression analysis showed that gender of male （OR=1.747， 95% CI： 1.303~2.342）， manic episodes （OR=2.018， 95% CI： 1.310~3.108） and emergency admission （OR=4.244， 95% CI： 3.083~5.840） were risk factors affecting the high risk of violent behaviour of inpatients. Among different types of mental disorders， patients with depressive disorder had a relatively low risk of violent behaviour （OR=0.397， 95% CI： 0.233~0.677）. ConclusionAmong inpatients with acute mental disorders， patients of emergency admission， being male and manic episodes are more likely to be at high risk of violent behaviour.

OBJECTIVE: To analyze the clinical features and genetic variants of two patients from a pedigree affected with Smith-Lemli-Opitz syndrome and explore their genotype-phenotype correlation. METHODS: Clinical data and family history of the pedigree were collected. Whole exome sequencing was carried out to identify the potential variants. Suspected variants were verified by Sanger sequencing of the family members. RESULTS: The proband and her sister both presented with feeding difficulty, facial dysmorphism, seizures, and mental and speech retardation. The third child of this family presented with feeding difficulty, poor weight gain and severe malnutrition after birth. He had died of unknown cause at 6 months without genetic testing. The fourth child was a healthy boy. Genetic testing showed that both the proband and her sister have carried c.127G>T (p.Val43Phe) and c.820_825del (p.Asn274_Val275del) compound heterozygous variants of the DHCR7 gene (NM_001360.2), but the fourth child carried neither of the variants. The two variants were unreported in the literature and disease-related databases, and were not included in the 1000G and gnomAD databases. The c.820_825del variant may affect the sterol-sensitive region of the DHCR7 protein, which can lead to deletion of two amino acids at positions 247 and 275, causing truncation of the DHCR7 protein. It is speculated that this may affect the stability of protein's spatial conformation, thereby decrease the activity of the enzyme. The c.127G>T variant may affect the first transmembrane region of the protein, which is involved in the transmembrane transport of proteins. Multiple software predicted it to be harmful. Conservation analysis suggested that the three amino acids all locate in a highly conserved region of the protein. In consideration of the clinical phenotype, family history and result of genetic testing, we speculated that both patients had Smith-Lemli-Opitz syndrome due to variants of the DHCR7 gene. CONCLUSION This pedigree has enriched the phenotypic and genotypic data of Smith-Lemli-Opitz syndrome, which clarified the genetic etiology of the patients and provided a basis for genetic counseling of this pedigree.
China ; Female ; Genetic Testing ; Humans ; Male ; Mutation ; Oxidoreductases Acting on CH-CH Group Donors/genetics* ; Pedigree ; Smith-Lemli-Opitz Syndrome/genetics*

Objective To get well-informed of the current status of emergency training in primary hospitals in Sichuan Province and to find weaknesses in the system in order to provide scientific basis for emergency knowledge and skills training.Methods A stratified random cluster sampling method as adopted and an anonymous questionnaire survey method was conducted to investigate the basic circumstances of the current training and the training needs of 1 000 emergency medical staff in 9 regions throughout Sichuan Province.Results The scope of the survey covered emergency doctors,nurses and technicians in hospitals of Grade Ⅱ,Grade Ⅲ and below.A total of 1 000 surveys were sent out and 983 were recovered.The questionnaire completion rate was 98.3％.The respondents mainly worked for more than 10 years,most of which with Bachelor degree,junior and intermediate titles.Results of the survey showed,63.1％ Most respondents have only one or two training opportunities per year.The training methods are mainly continuing education (59.0％,580 trips),training organized within the unit (58.5％,575 trips),and training organized by health administrative units (39.3％,386 trips).The main contents of the training are:basic professional skills learning (66.7％,655 trips),basic professional theoretical study (59.2％,582 trips),and new emergency technology (42.5％,418 trips),only 32.0％ of the respondents believe that these methods meet the practical needs.The main factors that influence participation in the training were:not be arranged by employer (36.2％,356 trips);the training was different from actual work (31.6％,311 trips);no replacement for work so they couldn't join training (29.8％,293 trips).The survey showed that the most desirable way of training is:practical application (60.1％,591 trips) and the most desirable training content is:emergency first aid knowledge (73.7％,724 trips).There were significant differences in case discussion,technical observation,academic lectures,special study trainingand learning methods in hospitals of different levels (P＜0.05),but there was no significant difference in the choice of professional practice methods (P＞0.05),and the selection of training contents was in emergency first aid knowledge,general medical knowledge and prevention and treatment of chronic diseases.There was no statistical difference in the selection of training contents of knowledge,infectious disease knowledge and department management knowledge (P＞0.05).There were significant differences in the choice of nursing knowledge training (P＜0.05).Conclusion The current situation showed a lack of first-aid knowledge and skills training for emergency personnel in Sichuan Province.The present situation of skill training can't satisfy the need of their desire to participate in training.It is urgent to develop a standardized,systematic and scientific training mode to improve the emergency first aid ability of primary medical and emergency personnel.

OBJECTIVE: To identify differentially expressed genes in peripheral blood mononuclear cells between patients with continuous mild-to-moderate asthma and healthy controls using mRNA microarray in order to explore the underlying signaling pathways and clarify the roles of CD4+ T cells in the pathogenesis of asthma. METHODS: Global transcriptomic profiles of the CD4+ T cells were defined by using Agilent Sure Print G3 Human GE 8×60K microarray. Enrichment pathways were analyzed with Ingenuity Pathway Analysis (IPA) software. RESULTS: Compared with controls, 805 genes were up-regulated, 192 were down-regulated in asthma patients. Among these, the expression of 38 annotated genes have varied by 4 times or more. Expression of CD300A was inversely proportional to the absolute value of eosinophils (r=-0.89, P=0.02) as well as the proportion of eosinophils (r=-0.94, P=0.004), while CSF1R was inversely proportional to PD20 (r=-0.83, P=0.04) and AQLQ (r=-0.88, P=0.02) by correlation analysis. CONCLUSION Numerous pathophysiological pathways may be involved in the pathogenesis of asthma. Above findings have provided a basis for the delineation the pathogenesis of asthma.
Antigens, CD ; genetics ; Asthma ; immunology ; CD4-Positive T-Lymphocytes ; cytology ; Case-Control Studies ; Eosinophils ; Gene Expression Profiling ; Humans ; Leukocytes, Mononuclear ; Oligonucleotide Array Sequence Analysis ; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; Receptors, Immunologic ; genetics ; Transcriptome

Objective To investigate the role of Wnt signaling pathway hypofunction mediated by dephosphorylation ofβ-catenin in the impaired wound healing of type 1 diabetic rats. Methods The back skin defect wounds were produced in rats with type 1 diabetes. These rats were divided into control, diabetes, lithium chloride treatment, and epidermal growth factor ( EGF) treatment groups. The situation of back wound healing, the ratio ofβ-catenin positive cells,β-catenin, phosphorylatedβ-catenin, and vascular endothelial growth factor ( VEGF) levels were detected. Results Compared to diabetes group, the wound granulation tissue was more mature, wound healing time was shorter, and healing rate, as well as the ratio ofβ-catenin positive cells, dephosphorylatedβ-catenin, and VEGF levels, were higher in normal group, lithium chloride treatment group, and EGF treatment group ( P<0. 05 or P<0. 01). Conclusion The hypofunction of Wnt signaling pathway is involved in the process of wound healing in type 1 diabetic rats, of which the dephosphorylation ofβ-catenin is the key point. EGF may play a beneficial role in the wound healing of type 1 diabetic rat models via Wnt pathway.

OBJECTIVETo study the role of outer membrane protein T (OmpT) in the pathogenesis of uropathogenic Escherichia.coli.

METHODSIn cultured human bladder epithelial cell line 5637, we examined the adhesion ability of wild-type (CFT073), ompT gene knockout (COTD), and revertant (pST) strains of E.coli to the cells and the extracellular matrix (ECM). The expressions of the adhesion gene iha and virulence gene iroN were detected by real-time PCR. Murine models of urinary tract infection with the 3 strains were established to evaluate the bacterial burden of the bladder and kidney tissue and bacterial counts in blood. We also detected the expressions of interleukin-6 (IL-6) and IL-8 in the bladder and kidney tissues of the mice.

RESULTThe COTD strain showed a significantly lower cell adhesion rate than CFT073 strain [(4.62∓0.39)% vs (8.81∓1.13)%, P<0.05] with also a lower ECM-adhesion rate [(4.95∓0.59)% vs (8.85∓0.79)%, P<0.05]. The mRNA expressions of iha and iroN in CFT073 strain were 2.1 and 3.8 times that of COTD strain. In the mouse model, the mean bacterial load of CFT073 strain in the bladder tissue was 6.36∓0.06, significantly greater than that of COTD (6.01∓0.07) and revertant (6.29∓0.06) strains (P<0.05); the bacterial load of CFT073 strain in the kidney tissue was also significantly higher than that of COTD strain (6.25∓0.05 vs 5.87∓0.06, P<0.05). In mice infected with the wild-type, knockout, and revertant strains, the detection rates of IL-6, which were identical to those of IL-8, in the inflammatory bladder and kidney tissues were 60%, 12.5%, and 50%, respectively.

CONCLUSIONSOmpT may regulate the expression of the adhesion gene iha and the transferrin gene iroN to affect the adhesion of uropathogenic E.coli to host cells.

Animals ; Bacterial Adhesion ; Bacterial Load ; Bacterial Outer Membrane Proteins ; metabolism ; Cell Line, Tumor ; Escherichia coli Infections ; pathology ; Escherichia coli Proteins ; metabolism ; Gene Knockout Techniques ; Humans ; Inflammation ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Kidney ; microbiology ; Mice ; Peptide Hydrolases ; metabolism ; Receptors, Cell Surface ; metabolism ; Urinary Bladder ; microbiology ; Urinary Tract Infections ; microbiology ; pathology ; Uropathogenic Escherichia coli ; pathogenicity