Objective To investigate the role of lipopolysaccharide(LPS)in regulating the inflammatory response of neutrophil through the leucine-rich α-2 glycoprotein 1(LRG1)/Rho-associated protein kinase(ROCK1)signaling.Methods HL-60 cells were treated with 1 μmol/L all-trans retinoic acid(ATRA)and 12.5 μL/mL dimethyl sulfoxide(DMSO)for 72 h and 96 h,and the morphological changes were observed by Wright-Giemsa staining.The expression of CD11b was detected by flow cytometry.LPS induced the activation of dHL-60 and human peripheral blood neutrophils.The transcription and secretion levels of LRG1,ROCK1 and inflammatory cytokines were detected by qPCR and ELISA,respectively.The expression levels of LRG1 and ROCK1 after the activation of dHL-60 were detected by Western blotting.Furthermore,dHL-60 was treated with the recombinant protein LRG1 and ROCK1 inhibitor Y-27632;the transcription levels of inflammatory cytokines were detected by qPCR.Results Neutrophils were activated by LPS.The expression levels of LRG1 and ROCK1 were significantly increased,and the transcription levels of inflammatory cytokines were significantly increased.The recombinant protein LRG1 activated dHL-60 in vitro,and the transcription levels of ROCK1 and inflammatory cytokines were significantly increased.Using the ROCK1 inhibitor Y-27632,the production levels of inflammatory cytokines were significantly reduced.Conclusion LPS can regulate the production levels of neutrophil inflammatory cytokines through activating the LRG1/ROCK1 signaling,thus exacerbating the inflammatory response.

Objective To explore the value of controlling nutritional status(CONUT)score and related inflammatory indicators in predicting the prognosis of clear cell renal cell carcinoma(ccRCC)patients,to provide a reference for better clinical assessment and individualized treatment plan.Methods A retrospective study was conducted on 132 patients with ccRCC admitted to four comprehensive hospitals in Yibin during 2010 and 2018.Patients'medical and follow-up records were collected,and receiver operating characteristic(ROC)curve was drawn to analyze the area under the curve(AUC)and optimal cut-off value of CONUT score and related indicators.Survival curve was plotted with Kaplan-Meier method,and the influencing factors of prognosis were analyzed with Log-rank test and Cox regression.Results During the follow-up of 62(53,71)months,37(28.03％)deaths occurred.The disease-specific survival(DSS)and progression-free survival(PFS)were 51(41,58)and 46(35,56)months,respectively.The 3-year and 5-year DSS and PFS were 84.09％and 71.97％,and 75.00％and 71.97％,respectively.The AUC of CONUT score,neutrophil to lymphocyte ratio(NLR),platelet to lymphocyte ratio(PLR),and lymphocyte to monocyte ratio(LMR)were 0.980,0.905,0.899 and 0.884,respectively,with the optimal cut-off values of 3.50,3.19,89.07 and 3.56,respectively.Cox regression showed that CONUT score(HR=0.042,95％CI:0.013-0.140)and PLR(HR=0.182,95％CI:0.045-0.744)were associated with DSS;CONUT score(HR=0.029,95％CI:0.010-0.086)and LMR(HR=2.984,95％CI:1.227-7.258)were associated with PFS.Conclusion The prognosis of ccRCC patients is related to their nutritional,immune,and inflammatory status.CONUT score and inflammatory factors(PLR,LMR)may be important predictors of DSS and PFS.

Objective: To analyze the impact of parental sports concept and behavior on physical activity in preschool children, so as to provide a foundation for future guidance on fostering childrens physical activity within the family context. Methods: From November to December 2020, a clustered convenience sampling method was employed to conduct surveys, and a total of 283 children were selectal from one kindergarten each in Beijing, Shenyang, and Xian. Participating children wore ActiGraph GT9X accelerometers continuously for one week to collect data on different intensity levels of physical activity. Physical Activity afterschool Questionnaire for Preschooler (P-PAQ) was utilized to assess parental sports concept and behavior. The gender differences in physical activity level and physical activity compliance rate were analyzed by using ttest, Mann-Whitney U test, and Chisquare test; and the relationship between parental exercise concepts and behaviors and physical activity of preschool children was analyzed using Spearman rank correlation analysis and multiple linear regression analysis. Results: Parental sports concept was significantly positively correlated with average daily moderatetovigorous physical activity (MVPA) and total physical activity (TPA) in children (r=0.12-0.16, P<0.05). Parental sports behavior was significantly positively correlated with childrens average daily TPA (r=0.25, P<0.05). Multiple linear regression revealed that parental sports concept was positively correlated with average daily MVPA and TPA in both boys and girls (B=0.65-0.83), while parental sports behavior only was positively correlated with boys average daily MVPA and TPA (B=0.24-0.25)(P<0.05). Conclusions Parental sports concept and behavior can impact physical activity levels in preschool children, exhibiting gender differences. Future guidance on physical activity in family upbringing should consider both parental sports concept and behavior, and pay attention to the influence of childrens gender.

Objectives:To discuss the inhibitory effect of Lactobacillus reuteri on the replication of rotavirus(RV)strain SA11 in vivo and in vitro,and to clarify its effect on the expression of related immune factors.Methods:For in vitro experiments,Lactobacillus reuteri was cultured and identified,and the standard curve and growth curve were plotted to screen the optimal time and concentration for Lactobacillus reuteri cultivation.The cells were infected with Lactobacillus reuteri at the concentrations of 5×108,10×108,50×108,100×108,200×108,and 500×108 CFU·mL-1,and the surival rates of Caco-2 cells were detected by trypan blue staining method.Various concentrations of Lactobacillus reuteri were co-incubated with RV in vitro and applied to the Caco-2 cells.The cells were divided into negative control group(NC group),positive control group(PC group),and 107,108,109,and 1010 CFU·mL-1 Lactobacillus reuteri groups.Immunofluorescence focus method was used to detect the viral titers in the Caco-2 cells after treated with Lactobacillus reuteri and real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the copy numbers of RV VP6 gene in the Caco-2 cells after treated with various concentrations of Lactobacillus reuteri.In in vivo experiments,25 litters of SPF suckling mice were divided into control group,RV group(infected with SA11 strain),Ab-NC group(treated with antibiotic to deplete gut microbiota),Ab-RV group(depleting gut microbiota and then infected with SA11 strain),and Ab-Lac-RV group(depleting gut microbiota,treated with Lactobacillus reuteri,and then infected with SA11 strain).The fecal samples were collected on days 2,4,6,8,and 10 gavage,colon tissue sample were collected on day 4 of and RT-qPCR method was used to detect the copy numbers of RV VP6 gene in feces and the mRNA expression levels of interleukin(IL)-1β,IL-8,IL-10,interferon-γ(IFN-γ),and tumor necrosis factor-α(TNF-α)in colon tissue of the suckling mice in vartious groups.Results:The Lactobacillus reuteri grew well,with round,smooth,and milky white convex colonies and neat edges.After Gram staining,the bacteria appeared purple,irregular,and square-shaped rods.16SrDNA sequencing showed 99%sequence homology,indicating successful activation of Lactobacillus reuteri.The number of live Lactobacillus reuteri was linearly related to the absorbance(A)value,and the standard curve for regression analysis was Y=0.437 5X+0.000 6,R2=0.999 4.During the 0-2 h cultivation period,the bacteria were at the logarithmic growth phase with slow growth;from 2-14 h,the bacteria grew rapidly and stabilized at 14-16 h,reaching the growth rate peak at 16 h,after which they entered the decline phase.Infection with Lactobacillus reuteri at concentrations of 5×108,10×108,50×108,100×108,and 200×108 CFU·mL-1 resulted in the survival rates of Caco-2 cells were all>90%,so these concentrations were selected for the further experiments.Compared with PC group,the copy numbers of RV VP6 gene in the Caco-2 cells in 5×108,10×108,50×108,100×108,and 200×108 CFU·mL-1 Lactobacillus reuteri groups were significantly decreased(P<0.01).Compared with PC group,the viral titers in the Caco-2 cells in 107,108,109,and 1010 CFU·mL-1 Lactobacillus reuteri groups were significantly decreased(P<0.01).Compared with control group,the numbers of gut microbiota colonies in Ab-NC,Ab-RV,and Ab-Lac-RV groups were significantly decreased,indicating successful depletion of gut microbiota in the suckling mice.On days 2 and 4 after gavage,the RV VP6 gene copy number in the feces in Ab-RV group was significantly lower than that in RV group(P<0.05).On days 4,6,8,and 10 after gavage,the RV VP6 gene copy number in the feces in Ab-Lac-RV group was significantly lower than that in Ab-RV group(P<0.05).Compared with control group,the expression levels of IL-1β,IL-10,IFN-γ,and TNF-α mRNA in colon tissue in Ab-RV and Ab-Lac-RV groups were significantly increased(P<0.05 or P<0.01),while the expression level of IL-8 mRNA was significantly decreased(P<0.05),and the expression level of IL-10 mRNA in colon tissue in Ab-LAC-RV group was significantly increased(P<0.01).Conclusion:Lactobacillus reuteri may inhibit the RV replication by upregulating the expressions of IL-1β,IL-10,IFN-γ,and TNF-α mRNA and downregulating the expression of IL-8 mRNA.

Objective To investigate the relationships of nutritional status and length of stay (LOS) with phase angle (PA) in patients with severe craniocerebral injury. Methods A total of 100 patients [Glasgow Coma Scale (GCS) score ≤8 points] admitted outside the department of neurology were selected as study objects. Body composition analysis and blood samples were used to determine the intracellular and extracellular water content, skeletal muscle, PA, prognostic nutritional index (PNI) and LOS. Patients were divided into low PA group (n=42) and normal PA group (n=58) based on PA values. Body composition related indexes, PNI and LOS were compared between the two groups. The correlation between PA and each index was analyzed. The influencing factors of malnutrition were analyzed. The influencing factors of malnutrition were analyzed. Results Intracellular water, total body water, body cell content and PNI in the low PA group were significantly lower, and LOS was significantly longer than that in the normal PA group (P < 0.05). PA was positively correlated with intracellular water, extracellular water, total body water, skeletal muscle, body cell content, bone mineral content, basal metabolic rate and PNI (P < 0.05), while PA was negatively correlated with LOS (P < 0.05). PA (OR=5.441, P=0.001, 95%CI, 2.011 to 14.719) and LOS (OR=8.373, P < 0.001, 95%CI, 3.079 to 22.765) were the influential factors in the occurrence of malnutrition. Conclusion PA is significantly correlated with nutritional status and LOS in patients with severe craniocerebral injury.

The many-banded krait, Bungarus multicinctus, has been recorded as the animal resource of JinQianBaiHuaShe in the Chinese Pharmacopoeia. Characterization of its venoms classified chief phyla of modern animal neurotoxins. However, the evolutionary origin and diversification of its neurotoxins as well as biosynthesis of its active compounds remain largely unknown due to the lack of its high-quality genome. Here, we present the 1.58 Gbp genome of B. multicinctus assembled into 18 chromosomes with contig/scaffold N50 of 7.53 Mbp/149.8 Mbp. Major bungarotoxin-coding genes were clustered within genome by family and found to be associated with ancient local duplications. The truncation of glycosylphosphatidylinositol anchor in the 3'-terminal of a LY6E paralog released modern three-finger toxins (3FTxs) from membrane tethering before the Colubroidea divergence. Subsequent expansion and mutations diversified and recruited these 3FTxs. After the cobra/krait divergence, the modern unit-B of β-bungarotoxin emerged with an extra cysteine residue. A subsequent point substitution in unit-A enabled the β-bungarotoxin covalent linkage. The B. multicinctus gene expression, chromatin topological organization, and histone modification characteristics were featured by transcriptome, proteome, chromatin conformation capture sequencing, and ChIP-seq. The results highlighted that venom production was under a sophisticated regulation. Our findings provide new insights into snake neurotoxin research, meanwhile will facilitate antivenom development, toxin-driven drug discovery and the quality control of JinQianBaiHuaShe.

Objective:To investigate the epidemiological and clinical features of patients with hemorrhagic fever with renal syndrome (HFRS) from 2017 to 2019.Methods:Seventy-five patients with HFRS from the Department of Infectious Diseases, The First Affiliated Hospital of Anhui Medical University during January 1, 2017 to December 31, 2019 were included. The data of epidemiology, clinical symptoms, blood routine, urine routine, serum creatinine, liver function and other laboratory examination indexes were retrospectively analyzed. The measurement data with skewness distribution were expressed by M( QR) and compared by nonparametric test. Multivariate logistic regression analysis was used to analyze disease-related risk factors. Results:The 75 patients were mainly located in the western and northern regions of Anhui Province. A total of 37 cases (49.3%) were infected during November, December and January next year. Fifty-four (72.0%) patients were farmers and 10(13.3%) patients had a clear history of rodent contact. Only 19(25.3%) patients had typical clinical manifestations of "three red and three pain" . Fifty-eight (77.3%) patients had elevated white blood cell count, 67(89.3%) patients had decreased platelet count, 55(73.3%) patients had urinary protein + + + , 65(86.7%) patients had abnormal urinary occult blood, and 67(89.3%) patients had elevated serum creatinine. The serum creatinine and potassium levels in 31 severe and critical patients were 495(301) μmol/L and 4.14(0.77) mmol/L, respectively, which were both higher than those in 44 mild and moderate patients (235(289) μmol/L and 3.65(1.02) mmol/L, respectively). The differences were both statistically significant ( Z=-3.187 and -2.796, respectively, both P<0.01). Multivariate logistic regression analysis showed that serum creatinine (odds ratio ( OR)=1.005, 95% confidence interval ( CI)1.002-1.008) and serum potassium ( OR=2.632, 95% CI 1.098-6.313) were independent risk factors for disease severity. All patients received comprehensive medical treatment, and 27 patients received renal replacement therapy. Sixty-eight patients had good prognosis and four patients died. Conclusions:HFRS is still common in the rural area in winter and spring. Patients with atypical clinical manifestations and severe and critical patients should be intensively monitored.

Objective To investigate nutritional knowledge,attitude,and practice (K-A-P) of medical staff in the nerve-related departments in Hebei province,with the aim of determining a basis for the effective education of nutrition.Methods A multicenter,observational study was utilized to conduct a questionnaire survey.642 medical staff in nerve-related departments from 15 hospitals in 11 prefecture level cities was surveyed from September 2016 to March 2017 in Hebei Province.Results were statistically analyzed.Results The overall scores of nutrition knowledge,attitude,and practice were 34.88±22.90,69.14± 16.75 and 42.86± 18.22,respectively.There were positive correlations between nutrition knowledge and attitude (r=0.150,P＜ 0.001),knowledge and practice (r=0.152,P＜0.001),and attitude and practice (r =0.432,P＜0.001).The nutrition knowledge score of male medical staff was higher than that of female (t =3.546,P＜0.01) however,there was no significant difference between males and females in nutrition attitude and practice scores (t =0.685,P=0.494;t=1.649,P=0.091).In addition,the nutrition knowledge scores of physicians and medical technicians were significantly higher than that of nurses (F=46.89,P＜0.01) while the nutrition practice scores of physicians were significantly higher than both medical technicians and nurses (F=7.85,P＜0.01).It was also found that nutrition attitude scores of senior and secondary medical staff were higher than junior medical staff (F=5.116,P=0.002) and nutrition practice scores of senior and secondary senior medical staff were higher than that of junior medical staff (F=4.074,P=0.007).Furthermore,it was found that nutrition knoledge scores of undergraduate,master and doctoral degree staff were higher than that of junior college staff (F=15.617,P＜0.001) and nutrition attitude score of undergraduate and master degree staff were significantly higher than that of junior college staff (F =4.11,P =0.003).It was also found that the nutrition knowledge scores of medical staff with less than 5 years of work experience were significantly higher than those with more than 5 years of work experience (F=3.312,P =0.02).However,the scores of nutrition attitude of medical staff with more than 15 years of work experience were significantly higher than those with less than 15 years of work experience (F=6.356,P＜0.001) and the scores of nutrition practice of medical staff with 10-15 years of work experience were higher than that of less than 5 years and 5-10 years of work experience (F=2.959,P=0.032).It was also identified that the top three ways to obtain nutrition knowledge were learning at work (68.85％),learning at university (68.69％) and reading books and journals (57.79％).Conclusion The medical staff of nerve-related departments in Hebei province has positive attitudes towards nutrition treatments.However,their mastery of nutritional knowledge and practice could and should be improved further.Therefore,it is necessary to strengthen the nutrition education for medical staff of the nerve profession.

OBJECTIVE:To optimize extraction technology of protein from Cryptotympana pustulata and investigate its in vitro antioxidant activity,so as to provide reference for further research of protein from C. pustulata. METHODS:Using extraction amount of protein as response value,based on single factor test,Box-Behnken response surface methodology was used to optimize the ratio of liquid to material,ultraonic time and extraction times.Validation test was conducted. Using Vitamin C(VC)as positive control, in vitro antioxidant activity of protein from C. pustulata was evaluated by using scavenging rate of 1, 1-diphenyl-2-picrylhydrazyl(DPPH)and 2,2′-azino-bis(3-ethyl benzothiazoline-6-sulfonic acid)(ABTS)free radical as index. RESULTS:The optimal extraction condition of protein from C.pustulata was as follows as the ratio of liquid to material 28:1(mL/g), ultrasonic time of 65 min,extracting for twice. In validation test,average extraction amount of protein from C. pustulata was 65.45 mg/g(RSD=1.68%,n=3),relative error of which to predicted value was 5.48%. The protein from C. pustulata showed strong scavenging effect on ABTS and DPPH free radicals. When the concentration of protein from C. slough was 0.2 mg/mL,and the scavenging rate of it to ABTS free radicals was 97%,the effect of which was similar to VC.The protein from C. pustulata showed weak scavenging ability to DPPH free radical,IC50was 0.96 mg/mL,the effect of which was not as good as VC. CONCLUSIONS:The extraction technology of protein from C. pustulata optimized by Box-Behnken response surface methodology shows high accuracy and good reliability.The protein from C.pustulata shows certain antioxidant activity in vitro.

OBJECTIVE：To study the difference of dissolution performance of chemical component and anticonvulsant effect of Bombyx mori decoction and powder taken with water，and to provide reference for the selection of application forms of B. mori. METHODS：The yield of dry extract was determined for decoction and powder biomimetic gastric juice of B. mori. HPLC method was used to detect the content of ammonium oxalate in decoction and powder biomimetic gastric juice of B. mori. The content of protein in decoction and powder biomimetic gastric juice of B. mori was determined with bicinchoninic acid（BCA）method. Mice was divided into normal group（1％Sodium carboxymethylcellulose solution），model group（1％Sodium carboxymethylcellulose solution），positive group （phenytoin，2 mg/kg） and B. mori decoction and powder suspensions high-dose，medium-dose and low-dose groups（0.75，1.5，3 g/kg by crude drug）according to random number table，with 20 mice in each group. After 60 min of intragastric administration，electric stimulation was conducted，and the rate of convulsion in mice was recorded. RESULTS：The yields of dry extract were 22.08％ and 26.40％ in decoction and powder gastric juice of B. mori （P＜0.05）；the contents of ammonium oxalate were 11.22％ and 16.83％ （P＜0.05），and the contents of protein was 3.39％ and 4.92％ （P＜0.01）. Compared with normal group，convulsion rate of mice was increased significantly in model group （P＜0.01）；compared with model group，convulsion rates of mice were decreased significantly in administration groups （P＜0.05 or P＜0.01），and the convulsion rate of mice in B. mori powder suspensions group was lower than decoction group. CONCLUSIONS：The dissolution performance of the chemical component from gastric juice of B. mori powder is better than that of decoction， and the anticonvulsant effect of B. mori powder is better than decoction.