This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20％to 0.91％,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100％compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.

Objective To explore the effect of Rhizoma Polygonati(RP)on arterial aging in naturally aging Wistar rats.Methods SPF Wistar rats aged 72 weeks were divided randomly divided into 4 groups:an old group and RP low,medium,and high-dose groups(n=14 rats per group).Another 14 male SPF Wistar rats aged 8～12 weeks were selected as the young group.Rats in the RP high,medium,and low-dose groups were administered with 4,2,and 1 g/kg RP,respectively,by gavage,and rats in the old and young groups were given the same amount of distilled water once a day for 12 weeks.Seven rats from each group were sacrificed under anesthesia at weeks 4 and 12 and aortas were isolated.The relative smooth muscle cell(SMC)and collagen fiber(CF)contents were analyzed,total antioxidant capacity(T-AOC),glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),and malondialdehyde(MDA)levels were measured,and the expression levels of cell cycle-associated proteins in arterial tissue were detected by Western Blot.Results Rats in the old group showed obvious signs of vascular aging but there was no significant changes in arterial vascular tissue indexes in the old group with increased age.Aortas were obviously injured,relative contents of SMC and CF were significantly increased(P＜0.01),T-AOC,SOD,and GSH-Px contents were significantly decreased and MDA was increased(P＜0.01)in the old group compared with the young group at 4 and 8 weeks,and expression levels of cell cycle-associated proteins were significantly up-regulated(P＜0.01).RP intervention significantly decreased the relative SMC and CF contents and MDA levels(P＜0.05 or P＜0.01)and significantly increased T-AOC,SOD,and GSH-Px(P＜0.05 or P＜0.01).Expression levels of cell cycle-associated proteins were also significantly decreased(P＜0.05 or P＜0.01).High-dose RP had the greatest effect.Conclusions Arterial aging is relatively stable in the short term in naturally aging rats.RP could delay arterial aging in naturally aging rats by regulating the level of oxidative stress and the expression of cell cycle-associated proteins.

ObjectiveTo explore the clinical efficacy and safety of Da Chaihutang （DCH） for the treatment of sepsis with Yang syndrome. MethodsA total of 70 patients suffering from sepsis with Yang syndrome were randomly divided into an observation group and a control group， with 35 cases in each group. They both received standard Western medicine treatment. The observation group was additionally given a dose of DCH， which was boiled into 100 mL and taken twice. The control group was additionally given an equal volume and dosage of warm water. The intervention lasted for three days. The 28-day all-cause mortality and the changes in the following indicators before and after intervention were compared between the two groups， including sequential organ failure assessment （SOFA）， acute physiology and chronic health evaluation Ⅱ （APACHE Ⅱ） score，white blood cell （WBC），the percentage of neutrophils （NEU%），C-reactive protein （CRP），procalcitonin （PCT），alanine transaminase （ALT），aspartate transaminase （AST），total bilirubin （TBil），creatinine （Cr），blood urea nitrogen （BUN），acute gastrointestinal injury （AGI） grade，gastrointestinal dysfunction score （GDS），serum intestinal fatty acid-binding protein （iFABP）， citrulline （CR），platelet （PLT），prothrombin time（PT），activated partial thromboplastin time （APTT），fibrinogen （Fib），international normalized ratio （INR），and D-dimer （D-D）. ResultsThere was no significant difference between the two groups regarding 28-day all-cause mortality. After the intervention，SOFA，WBC，PCT，and Cr were significantly decreased， and PLT was significantly increased in the control group （P<0.05）. SOFA，APACHE Ⅱ，NEU%，CRP，PCT，ALT，AST，Cr，BUN，AGI grade，GDS，and serum iFABP and CR were significantly improved in the observation group （P<0.05）. After the intervention，APACHE Ⅱ，PCT，AGI grade，GDS，and serum iFABP in the observation group were significantly lower than those in the control group ，while CR and PLT were higher （P<0.05，P<0.01）. There were significant differences regarding the gap of SOFA，APACHE Ⅱ，AST，TBil，AGI grade，GDS，iFABP，CR， and PLT between the two groups （P<0.05，P<0.01）. There were slight differences regarding PT，APTT，Fib，INR，and D-D between the two groups，which were in the clinical normal range. ConclusionOn the basis of Western medicine， DCH helped to reduce sepsis severity and improved multiple organ dysfunction with high clinical efficacy and safety， but further research on its impact on the prognosis of patients with sepsis is still required.

Objective To explore the correlation between blood biochemical indexes and blood pressure levels among college students in Kunming City.Methods In November 2021,a cluster sampling method was used to survey 4,781 college students at a university in Yunnan Province.Data collected included height,weight and blood pressure,and blood samples were collected for blood biochemical testing.The Mann-Whitney U test and Kruskal-Wallis H test were used to compare the distribution differences of hypertension among college students with different demographic characteristics,while a generalized linear model was used to analyze the correlation between blood biochemical indicators and blood pressure levels.Results The prevalence rate of hypertension among college students was 8.45%(404/4 781).After adjusting confounding variables,blood glucose(GLU,β=1.48,95%CI:0.75～2.20)and serum Total Protein(TP,β=0.25,95%CI:0.19～0.32)were found to be correlated with systolic blood pressure(SBP)levels(all P<0.05);blood glucose(GLU,β=1.25,95%CI:0.64～1.86)and serum total protein(TP,β=0.28,95%CI:0.23～0.34)were correlated with diastolic blood pressure(DBP)level(P<0.05).Stratified group analysis by sex showed that,after controlling for confounding factors,TP(β=0.32,95%CI:0.18～0.45)was correlated with SBP levels in male students;TP(β=0.32,95%CI:0.21～0.43)was correlated with DBP levels in male students(P<0.05).GLU(β=2.18,95%CI:1.29～3.07)and TP(β=0.23,95%CI:0.15～0.31)were correlated with SBP levels in female students;GLU(β=1.48,95%CI:0.73～2.24)and TP(β=0.26,95%CI:0.20～0.33)were correlated with DBP levels in female students(all P<0.05).Conclusion Our findings suggest that blood glucose and serum total protein level are correlated with blood pressure levels in female college students,while serum total protein level are correlated with blood pressure levels in male college students.

This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20％to 0.91％,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100％compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.

Objective To explore the effect of Rhizoma Polygonati(RP)on arterial aging in naturally aging Wistar rats.Methods SPF Wistar rats aged 72 weeks were divided randomly divided into 4 groups:an old group and RP low,medium,and high-dose groups(n=14 rats per group).Another 14 male SPF Wistar rats aged 8～12 weeks were selected as the young group.Rats in the RP high,medium,and low-dose groups were administered with 4,2,and 1 g/kg RP,respectively,by gavage,and rats in the old and young groups were given the same amount of distilled water once a day for 12 weeks.Seven rats from each group were sacrificed under anesthesia at weeks 4 and 12 and aortas were isolated.The relative smooth muscle cell(SMC)and collagen fiber(CF)contents were analyzed,total antioxidant capacity(T-AOC),glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),and malondialdehyde(MDA)levels were measured,and the expression levels of cell cycle-associated proteins in arterial tissue were detected by Western Blot.Results Rats in the old group showed obvious signs of vascular aging but there was no significant changes in arterial vascular tissue indexes in the old group with increased age.Aortas were obviously injured,relative contents of SMC and CF were significantly increased(P＜0.01),T-AOC,SOD,and GSH-Px contents were significantly decreased and MDA was increased(P＜0.01)in the old group compared with the young group at 4 and 8 weeks,and expression levels of cell cycle-associated proteins were significantly up-regulated(P＜0.01).RP intervention significantly decreased the relative SMC and CF contents and MDA levels(P＜0.05 or P＜0.01)and significantly increased T-AOC,SOD,and GSH-Px(P＜0.05 or P＜0.01).Expression levels of cell cycle-associated proteins were also significantly decreased(P＜0.05 or P＜0.01).High-dose RP had the greatest effect.Conclusions Arterial aging is relatively stable in the short term in naturally aging rats.RP could delay arterial aging in naturally aging rats by regulating the level of oxidative stress and the expression of cell cycle-associated proteins.

Objective:To investigate the clinical features and prognosis of hypertrophic cardiomyopathy（HCM）in children.Methods:The clinical data of 184 children diagnosed as HCM,who visited Beijing Children's Hospital of Capital Medical University from January 1，2006 to August 31，2022 were retrospectively analyzed.The children were divided into primary HCM group and secondary HCM group according to the etiology of HCM，and their clinical characteristics and prognosis were compared.Results:A total of 184 children[115（62.50%）males and 69(37.50%) females] with HCM were included.The median age at first diagnosis was 4.54（0.50，10.25）years.Among the participants，141（76.63%）cases had primary HCM，and 43（23.37%）cases had secondary HCM.Compared with the patients in primary HCM group，the patients in secondary HCM group had lower age of onset，with a higher proportion of the children under 1 year of age.Most cases had atypical symptoms，including a higher proportion of first-onset heart failure，higher proportion of enlarged cardiac shadow on chest radiograph，and lower left ventricular ejection fraction in the secondary HCM group（ P<0.05）.The proportion of older children，ratio of ventricular septal thickness to left ventricular posterior wall thickness，and the detection rate of left ventricular outflow tract obstruction in the primary HCM group was higher than those in patients with secondary HCM group（ P<0.05）.The survival curve for two groups showed that the secondary HCM group had significantly lower 1-year，2-year，3-year，5-year，and 10-year survival rates than those in the primary HCM group（ P<0.05）. Conclusion:The clinical manifestations of HCM in children are heterogenous due to different etiology，and it is necessary to actively clarify the etiology，improve risk assessment，and provide personalized management and treatment method.

The study is designed to observe the mechanism of Tongfu Xiefei Guanchang Solution(TFXF) in the treatment of acute lung injury(ALI) in rats by improving intestinal barrier and intestinal flora structure via p38 mitogen-activated protein kinase(p38 MAPK)/myosin light chain kinase(MLCK) signaling pathway. Sixty SPF-grade Wistar rats were randomly divided into the control(CON) group, lipopolysaccharide(LPS) group(7.5 mg·kg~(-1)), LPS + dexamethasone(DEX) group(3.5 mg·kg~(-1)), LPS + high-dose(HD)-TFXF group(14.74 g·kg~(-1)), LPS + middle-dose(MD)-TFXF group(7.37 g·kg~(-1)), and LPS + low-dose(LD)-TFXF group(3.69 g·kg~(-1)). ALI model of the rat was established by intraperitoneal injection of LPS. The lactate dehydrogenase(LDH) activity and total protein concentration in the bronchoalveolar lavage fluid(BALF) were measured; tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) levels in lung and colon tissue of rats were detected by enzyme linked immunosorbent assay(ELISA). Hematoxylin-eosin(HE) staining was used to observe the pathological expression in the lung and colon tissue of rats. The mRNA expression of p38 MAPK, TNF-α, and IL-1β in rat lung tissue was determined by real-time fluorescence quantitative polymerase chain reaction(real-time PCR). Western blot was used to detect the protein expression related to the p38 MAPK/MLCK signaling pathway in the colon tissue of rats. 16S rRNA sequencing was used to detect changes in the composition and content of intestinal flora in rats, and correlation analyses were performed to explore the regulatory role of intestinal flora in improving ALI in rats. The results showed that compared with those in the LPS group, the histopathological scores of lung and colon tissue, LDH activity, and total protein concentration in BALF were significantly reduced in rats in all groups after drug administration. Except for the LPS + LD-TFXF group, the remaining groups significantly reduced the levels of TNF-α and IL-1β in the lung and colon tissue of rats. The protein expressions of phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK)/p38, phosphorylated myosin light chain(p-MLC)/myosin light chain 2(MLC2), and MLCK in colon tissue of rats in each drug administration group were significantly decreased. The mRNA expression levels of p38 MAPK, TNF-α, and IL-1β were significantly reduced in the LPS + HD-TFXF group. 16S rRNA sequencing results showed that the abundance of intestinal flora was significantly higher in the LPS + HD-TFXF group, and intestinal floras including Sobs, Shannon, and Npshannon were significantly higher. The β-diversity distribution of intestinal flora tends toward the CON group, and the abundance of Firmicutes was significantly higher. The abundance of Proteobacteria was significantly reduced; the abundance of Bacteroides was significantly reduced, and the abundance of Ruminococcus was significantly higher. The main species differences were Blautia, Roseburia＿sp＿499, and Butyricicoccus. TNF-α and IL-1β of lung tissue were negatively correlated with Muribaculaceae, unclassified norank＿f＿Eubacterium＿coprostanoligenes, and Ruminococcus and positively correlated with Bacteroides. Meanwhile, TNF-α and IL-1β of colon tissue were negatively correlated with unclassified norank＿f＿Eubacterium＿coprostanoligenes and Ruminococcus and positively correlated with Bacteroides. The predicted biological function of the flora was related to the biosynthesis of secondary metabolites, amino acid biosynthesis, sugar metabolism, and oxidative phosphorylation. The above studies show that TFXF can repair lung and colon tissue structure and regulate inflammatory factor levels by modulating the abundance and diversity of intestinal flora species in ALI rats. Its mechanism of action in ameliorating ALI in rats may be related to the inhibition of inflammation, improvement of intestinal mucosal permeability, and maintenance of intestinal flora homeostasis and barrier through the p38 MAPK/MLCK signaling pathway.
Animals ; Acute Lung Injury/genetics* ; Rats ; p38 Mitogen-Activated Protein Kinases/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Myosin-Light-Chain Kinase/genetics* ; Male ; Gastrointestinal Microbiome/drug effects* ; Rats, Wistar ; Signal Transduction/drug effects* ; Interleukin-1beta/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Lung/metabolism* ; Intestinal Mucosa/metabolism* ; Humans

Objective:To search,evaluate and summarize the best evidence about nasal mucosa pressure injury caused by nasal feeding tube at home and abroad,and provide evidence-based basis for clinical practice.Methods:According to the"6S"model of evidence resources,literatures related to nasal mucosal stress injury were systematically searched from UpToDate,Guidelines International Network,Joanna Briggs Institute Evidence-based Healthcare Center,Web of Science,Cochrane Library,CNKI,Wanfang database and other related nutrition professional websites.The search period was from the establishment of the database to February 2024.Four researchers independently evaluated the literature quality,and after consulting the opinions of evidence-based experts,evidence was extracted and summarized for literatures that met the quality standards.Results:A total of 12 studies were included,including 5 guidelines,4 expert consensus,1 clinical decision,1 systematic review and 1 evidence summary,covering 24 pieces of evidence on 5 topics,including team building,nasal mucosa evaluation,nasal mucosa care,placement and maintenance of nasal feeding tube,monitoring and education.Conclusions:The evidence of prevention and nursing of nasal mucosal pressure injury caused by nasal feeding tube can be applied to clinical practice,promote the standardized management of nasal feeding tube and improve the implementation effect of enteral nutrition.

Objective:To investigate the risk factors affecting the occurrence of sudden cardiac death（SCD）of hypertrophic cardiomyopathy（HCM）in children.Methods:This was a retrospective study.A total of 184 children with HCM who admitted to Beijing Children's Hospital，Capital Medical University from 1 January，2006 to 31 August，2022 were collected for clinical data and follow-up information.Risk factors affecting the occurrence of SCD in children with HCM were analyzed by Cox regression.Results:A total of 184 children with HCM were included in this study，including 69（37.50%）female cases and 115（62.50%）male cases.Cox regression showed that age less than one year（ HR=6.232，95% CI 2.858-13.591），female gender（ HR=2.547，95% CI 1.460-4.444），family history（ HR=2.622，95% CI 1.468-4.683），pathological Q waves in the electrocardiogram（ HR=2.290，95% CI 1.285-4.082），QRS fragmentation waves（ HR=3.526，95% CI 1.786-6.963），combined multiple arrhythmias（ HR=2.218，95% CI 1.136-4.333），high interventricular septal thickness on echocardiography（ HR=1.055，95% CI 1.008-1.105）,and high left ventricular posterior wall thickness（ HR=1.060，95% CI 1.026 -1.096）were risk factors for endpoint events in children. Conclusion:Age，sex，family history，electrocardiogram changes（pathological Q wave，QRS fragmentation wave，combined multiple arrhythmia），interventricular septum thickness and left ventricular posterior wall thickness are independent risk factors affecting the occurrence of SCD of HCM in children.