Objective:To investigate the effects of Xuebijing injection on gut microbiota and intestinal mechanical barrier in mice with lipopolysaccharide (LPS)-induced sepsis, and analyze the potential mechanism by which Xuebijing injection protects gastrointestinal tract.Methods:Twenty-four healthy and clean grade male C57BL/6N mice were divided into four groups, control group, LPS group, LPS+ 5 μl/g Xuebijing injection group (5 μl/g Xuebijing injection group), and LPS+ 10 μl/g Xuebijing injection group (10 μl/g Xuebijing injection group), with six mice in each group. A mouse model of sepsis was established by intraperitoneal injection of mice with 10 μg/g LPS. At 0 and 12 h after successful modeling, the mice were intraperitoneally injected with 5 or 10 μl/g Xuebijing injection. Blood, ileum, and colon fecal samples were collected 12 h after the second administration. ELISA was used to detect the levels of diamine oxidase (DAO), D-blood lactic acid (D-Lac), TNF-α, and IL-6. HE staining was used to observe the local ileum damage, and Chiu′s score was used to evaluate the degree of intestinal tissue damage. Immunohistochemical staining and Western blot were used to detect the expression of Claudin-1, Occludin, and zona occludins-1(ZO-1) in ileum tissues, followed by semi quantitative analysis. One-way analysis of variance was used for intergroup comparisons, and LSD or Tamhane′s T2 test was used for pairwise comparisons based on the homogeneity of variance. The diversity and species composition of mouse fecal microbiota, and the differences among groups were analyzed using 16S rRNA sequencing.Results:The levels of DAO, D-Lac, TNF-α, and IL-6 in the LPS group were higher than those in the control group (all P<0.000 1). After the intervention with Xuebijing injection, the levels of DAO, D-Lac, TNF-α, and IL-6 decreased (all P<0.05) and showed no significant differences with those in the control group (all P>0.05). Besides, 10 μl/g Xuebijing injection was more effective than 5 μl/g Xuebijing injection in reducing the concentrations (all P<0.05). Chiu′s score was higher in the LPS group than in the control group and the 10 μl/g Xuebijing injection group (both P<0.05). Western blot showed that the expression levels of Occludin, Claudin-1, and ZO-1 in the LPS group were lower than those in the control group (all P<0.01), and Xuebijing injection intervention significantly increased the expression levels of these proteins in a dose-dependent manner as compared with the LPS group (all P<0.000 1). Apart from the expression level of ZO-1, which showed no significant difference between the two Xuebijing injection groups ( P>0.05), the results of immunohistochemical staining were consistent with those of Western blot. The 16S rRNA sequencing results showed that there were differences in the Alpha and Beta diversity indices, and the composition and structure of gut microbiota among the four groups. The structure of gut microbiota in the mice treated with Xuebijing injection was similar to that in the mice of the control group and it was in a dose-dependent manner. Wilcoxon rank sum test showed that there were statistically significant differences in six gut microbiota groups at the phylum level, and 32 gut microbiota groups at the genus level among the mice of four groups (all P<0.05). Conclusions:Xuebijing injection can provide protective effects on the gastrointestinal tract by protecting the structure of gut microbiota and intestinal barrier function, and the protective effect is somewhat correlated with the drug dosage.

Objective:To investigate the effect of risk forewarning hierarchical management on mental state,major adverse cardiovascular events(MACE)and quality of life in patients with acute coronary syndrome after percutane-ous coronary intervention(PCI).Methods:This randomized controlled study enrolled 152 patients with acute coro-nary syndrome who underwent PCI in the First Hospital of Qinhuangdao between July 2021 and June 2023.Patients were randomly divided into control group(n=76,conventional intervention program)and intervention group(n=76,additional risk forewarning hierarchical management).After 3-month intervention,mental state[Hamilton anxiety scale(HAMA)and Hamilton depression scale(HAMD)scores],health literacy[perceived knowledge scale for coronary heart diseases(PKS-CHD)score],quality of life[Seattle angina questionnaire(SAQ)score]and in-cidence of MACE were compared between the two groups.Results:Compared to those in the control group,patients in the intervention group had significantly higher total score of PKS-CHD[(35.08±3.59)points vs.(30.53±3.85)points],SAQ dimensional scores of physical limitation[(15.24±1.73)points vs.(13.26±1.45)points],an-ginal stability[(16.84±1.39)points vs.(15.18±1.48)points],disease perception[(17.00±1.46)points vs.(14.34±1.93)points],anginal frequency[(18.55±1.18)points vs.(16.11±1.39)points]and treatment satis-faction[(17.12±1.45)points vs.(14.04±1.93)points](P＜0.001 all),and significantly lower HAMA score[(11.89±1.64)points vs.(13.55±1.56)points],HAMD score[(12.61±1.76)points vs.(16.42±1.92)points]and the incidence of MACE(8.00％vs.28.38％)(P＜0.001 all).Conclusion:Risk forewarning hierarchical man-agement may effectively relieve patients'negative emotions,improve their health literacy and quality of life,and significantly reduce the risk of MACE in patients with acute coronary syndrome after PCI.

Objective:To investigate the effect of risk forewarning hierarchical management on mental state,major adverse cardiovascular events(MACE)and quality of life in patients with acute coronary syndrome after percutane-ous coronary intervention(PCI).Methods:This randomized controlled study enrolled 152 patients with acute coro-nary syndrome who underwent PCI in the First Hospital of Qinhuangdao between July 2021 and June 2023.Patients were randomly divided into control group(n=76,conventional intervention program)and intervention group(n=76,additional risk forewarning hierarchical management).After 3-month intervention,mental state[Hamilton anxiety scale(HAMA)and Hamilton depression scale(HAMD)scores],health literacy[perceived knowledge scale for coronary heart diseases(PKS-CHD)score],quality of life[Seattle angina questionnaire(SAQ)score]and in-cidence of MACE were compared between the two groups.Results:Compared to those in the control group,patients in the intervention group had significantly higher total score of PKS-CHD[(35.08±3.59)points vs.(30.53±3.85)points],SAQ dimensional scores of physical limitation[(15.24±1.73)points vs.(13.26±1.45)points],an-ginal stability[(16.84±1.39)points vs.(15.18±1.48)points],disease perception[(17.00±1.46)points vs.(14.34±1.93)points],anginal frequency[(18.55±1.18)points vs.(16.11±1.39)points]and treatment satis-faction[(17.12±1.45)points vs.(14.04±1.93)points](P＜0.001 all),and significantly lower HAMA score[(11.89±1.64)points vs.(13.55±1.56)points],HAMD score[(12.61±1.76)points vs.(16.42±1.92)points]and the incidence of MACE(8.00％vs.28.38％)(P＜0.001 all).Conclusion:Risk forewarning hierarchical man-agement may effectively relieve patients'negative emotions,improve their health literacy and quality of life,and significantly reduce the risk of MACE in patients with acute coronary syndrome after PCI.

Objective:To investigate the effects of Xuebijing injection on gut microbiota and intestinal mechanical barrier in mice with lipopolysaccharide (LPS)-induced sepsis, and analyze the potential mechanism by which Xuebijing injection protects gastrointestinal tract.Methods:Twenty-four healthy and clean grade male C57BL/6N mice were divided into four groups, control group, LPS group, LPS+ 5 μl/g Xuebijing injection group (5 μl/g Xuebijing injection group), and LPS+ 10 μl/g Xuebijing injection group (10 μl/g Xuebijing injection group), with six mice in each group. A mouse model of sepsis was established by intraperitoneal injection of mice with 10 μg/g LPS. At 0 and 12 h after successful modeling, the mice were intraperitoneally injected with 5 or 10 μl/g Xuebijing injection. Blood, ileum, and colon fecal samples were collected 12 h after the second administration. ELISA was used to detect the levels of diamine oxidase (DAO), D-blood lactic acid (D-Lac), TNF-α, and IL-6. HE staining was used to observe the local ileum damage, and Chiu′s score was used to evaluate the degree of intestinal tissue damage. Immunohistochemical staining and Western blot were used to detect the expression of Claudin-1, Occludin, and zona occludins-1(ZO-1) in ileum tissues, followed by semi quantitative analysis. One-way analysis of variance was used for intergroup comparisons, and LSD or Tamhane′s T2 test was used for pairwise comparisons based on the homogeneity of variance. The diversity and species composition of mouse fecal microbiota, and the differences among groups were analyzed using 16S rRNA sequencing.Results:The levels of DAO, D-Lac, TNF-α, and IL-6 in the LPS group were higher than those in the control group (all P<0.000 1). After the intervention with Xuebijing injection, the levels of DAO, D-Lac, TNF-α, and IL-6 decreased (all P<0.05) and showed no significant differences with those in the control group (all P>0.05). Besides, 10 μl/g Xuebijing injection was more effective than 5 μl/g Xuebijing injection in reducing the concentrations (all P<0.05). Chiu′s score was higher in the LPS group than in the control group and the 10 μl/g Xuebijing injection group (both P<0.05). Western blot showed that the expression levels of Occludin, Claudin-1, and ZO-1 in the LPS group were lower than those in the control group (all P<0.01), and Xuebijing injection intervention significantly increased the expression levels of these proteins in a dose-dependent manner as compared with the LPS group (all P<0.000 1). Apart from the expression level of ZO-1, which showed no significant difference between the two Xuebijing injection groups ( P>0.05), the results of immunohistochemical staining were consistent with those of Western blot. The 16S rRNA sequencing results showed that there were differences in the Alpha and Beta diversity indices, and the composition and structure of gut microbiota among the four groups. The structure of gut microbiota in the mice treated with Xuebijing injection was similar to that in the mice of the control group and it was in a dose-dependent manner. Wilcoxon rank sum test showed that there were statistically significant differences in six gut microbiota groups at the phylum level, and 32 gut microbiota groups at the genus level among the mice of four groups (all P<0.05). Conclusions:Xuebijing injection can provide protective effects on the gastrointestinal tract by protecting the structure of gut microbiota and intestinal barrier function, and the protective effect is somewhat correlated with the drug dosage.

This study was aimed at comparing performance differences among three metagenomic sequencing platforms,MGISEQ-2000,Illumina NextSeq 2000,and Ion GeneStudio S5 Plus,to optimize the sequencing process for trace samples.The three sequencing platforms were used to perform high-throughput sequencing on DNA standards and simulated samples.Through analysis of the quality of raw data and microbial detection capabilities,systematic differences among platforms were compared.The sequencing results were optimized for trace samples by incorporation of exogenous nucleic acids during the li-brary preparation process.In terms of data output per batch and base quality,MGISEQ-2000 surpassed the other two plat-forms.Illumina NextSeq 2000 had the lowest proportion of duplicate reads,whereas Ion GeneStudio S5 Plus had the highest proportion,and significant differences were observed across platforms(P＜0.001).In sequencing uniformity,MGISEQ-2000 and Illumina NextSeq 2000 were superior to Ion GeneStudio S5 Plus.MGISEQ-2000 provided a substantial advantage in microbial detection capability(P＜0.001),but the advantage diminished with decreasing bacterial fluid concentration.Ion GeneStudio S5 Plus had the shortest duration for single-batch sequencing.Moreo-ver,for trace samples with DNA content ≤0.05 ng,the experi-mental group(with added exogenous nucleic acids)achieved a higher number of reads than the control group(without exogenous nucleic acids),with a 11.09±8.03 fold increase.In conclu-sion,the different sequencing platforms each had advantages and disadvantages,thus allowing researchers to choose the appro-priate platform according to specific needs.Furthermore,the addition of exogenous nucleic acids improved the microorganism detection efficiency,and provided better support for subsequent diagnosis and evaluation of results.

Objective:To discover the relationship between the RNF213 gene and acute myeloid leukemia(AML),and explore the effect of RNF213 on the proliferation and apoptosis of THP-1 cells.Methods:Analyze the expression of RNF213 gene in AML and its relationship with prognosis through the GEPIA database.Collecting 30 AML patients and non-tumor hematological patients who went to the Affiliated Hospital of Zunyi Medical University from January 2017 to January 2022.RT-qPCR and Western blot were used to detect the expression levels of RNF213 mRNA and protein.Perform survival of patients was analysed by Kaplan-Meier.Meanwhile,the expression levels of RNF213 mRNA and protein were detected in AML cell lines(THP-1,OCI-AML2).CRISPR-Cas9 was used to knockdown the RNF213 gene in THP-1 cells;flow cytometry was used to detect apoptosis rate of cell.CCK-8 and colony formation assay were used to detect cell proliferation.Western blot was used to detect the expression level of Cleaved-Caspase 3 protein.Results:Compared with the control group,the expression level of RNF213 in AML patients was significantly increased,and patients with high expression of RNF213 have a worse prgnosis.Higher expression level of RNF213 protein in THP-1 cells.After knocking down the RNF213 gene of THP-1 cells,cell proliferation was significantly reduced,and the apoptosis rate and expression of apoptosis related protein Cleared-Caspase3 were significantly increased.Conclusion:AML patients have high expression of RNF213,and the prognosis of high expression patients is poor.The RNF213 gene affects AML cell proliferation and apoptosis,and may be a prognostic marker and potential therapeutic target for AML.

Objective:To explore the clinical correlation and prognostic value of the Albumin-Bilirubin(ALBI)score in children with secondary hemophagocytic syndrome(sHLH).Methods:A retrospective analysis was conducted on the data of children's sHLH cases clearly diagnosed in the Affiliated Hospital of Zunyi Medical University from January 2012 to March 2023.Survival analysis was conducted according to the ALBI classification.Spearman correlation analysis was conducted between the ALBI score and clinical indicators.The Receiver Operating Characteristic(ROC)curve was used to evaluate the ALBI score,select the best cutoff value,and evaluate the accuracy of prognostic prediction value.Kaplan-Meier method was used to draw the survival curve.Log-rank method was used to compare the differences of survival curve between groups.Cox regression was used for prognostic analysis and restricted cubic spline curves used to calculate the relationship between ALBI scores and the risk of death in children with sHLH.Results:A total of 128 children with sHLH were included in this study,with a median age of 38(13.25,84)months.There were 70 males(54.69％)and 58 females(45.31％).The survival analysis results of ALBI grading showed that the survival rate of HLH patients with ALBI grade 3 was significantly lower than those with ALBI grades 1 and 2.Spearman correlation analysis results showed that ALBI score was positively correlated with splenomegaly,respiratory failure,disseminated intravascular coagulation(DIC),pulmonary hemorrhage,gastrointestinal hemorrhage,central nervous system involvement,ALT,AST,TG,LDH,PT,APTT,and SF(the correlation coefficients are:r=0.181,0.362,0.332,0.221,0.351,0.347,0.391,0.563,0.180,0.448,0.483,0.37,0.356),and was negatively correlated with HB,PLT,and FIB(the correlation coefficients are:r=-0.321,-0.316,-0.423),but was not significantly correlated with EBV infection,fungal infection,hepatomegaly,and ANC(P＞0.05).Using the ROC curve,the cutoff value of ALBI was-1.76.Single factor Cox regression analysis results showed that HB＜90 g/L,ALT ≥ 80 U/L,AST≥200 U/L,LDH ≥1 000 U/L,PT ≥20 s,APTT≥40 s,FIB＜1.5 g/L,ALBI ≥-1.76,combined pulmonary hemorrhage,DIC,central nervous system involvement,gastrointestinal bleeding,and not using blood purification may be the prognostic risk factors for children with sHLH(P＜0.05).Multivariate Cox regression results showed that FIB＜1.5 g/L(HR=2.119,95％CI:1.028-4.368),ALBI≥-1.76(HR=2.452,95％CI:1.233-4.875),and central nervous system involvement(HR=4.674,95％CI:2.486-8.789)were independent risk factors affecting prognosis,while blood purification(HR=0.306,95％CI:0.153-0.612)was an independent protective factor for prognosis.The application of restricted cubic splines shows that the risk of death increases with the increase of ALBI score.The area under the ROC curve(AUC)of the ALBI score for predicting the risk of 1-week,2-week,4-week,and overall mortality were 0.825,0.807,0.700,and 0.693,respectively,indicating good predictive performance for early mortality risk.According to subgroup analysis results of clinical manifestations,compared with the ALBI＜-1.76 group,ALBI≥-1.76 was associated with age ≤2 years,EBV infection,HLH-1994/2004 treatment,concomitant respiratory failure,and ANC≤1.0 × 109/L,HB＜90 g/L,PLT＜100 × 109/L,TG≥3.0 mmol/L,LDH ≥ 1 000 U/L,APTT≥40 s,and FIB＜1.5 g/L(P＜0.05).Conclusion:The ALBI score is related to the clinical characteristics and laboratory indicators of sHLH,and can be used as a beneficial indicator for assessing the prognostic risk of sHLH in children.It has good accuracy and clinical application value in predicting the prognosis of sHLH in children.

Objective:To observe the effect of Xuebijing injection on sepsis combined with acute gastrointestinal injury (AGI), and analyze the risk factors of sepsis combined with AGI.Methods:A retrospective cohort study was conducted. Patients with non-gastrointestinal origin admitted to the department of intensive care medicine of the First Hospital of Qinhuangdao from May 1, 2021 to October 30, 2023 were enrolled. The baseline data, source of sepsis infection, vital signs, acute physiology and chronic health evaluationⅡ(APACHEⅡ), sequential organ failure assessment (SOFA), laboratory tests, comorbidities, interventions during treatment, and the 28-day prognosis were collected. The patients were divided into Xuebijing group and non-Xuebijing group according to whether Xuebijing injection was used or not. According to whether AGI was merged or not, patients were divided into merged AGI group and non-merged AGI group. The main observational indexes were the difference in the incidence of AGI between the Xuebijing group and non-Xuebijing group and the difference in the magnitude of the decline in procalcitonin (PCT), C-reactive protein (CRP), and white blood cell count (WBC) at 7 days after admission, and the difference in the 28-day morbidity and mortality. Risk factors for AGI in septic patients were explored by univariate analysis, and statistically significant indicators were screened and included in binary Logistic regression analysis to determine independent risk factors.Results:A total of 129 patients with sepsis of non-gastrointestinal origin were enrolled, including 57 patients in the Xuebijing group and 72 patients in the non-Xuebijing group. Among 129 patients, 80 patients in the merged AGI group and 49 patients in the non-merged AGI group. There were no statistically significant differences between Xuebijing group and non-Xuebijing group in gender, age, body mass index (BMI), underlying disease, source of infection, vital sign, APACHEⅡscore, SOFA score, and clinical intervention, and there were no statistically significant differences in laboratory tests except for aspartate aminotransferase (AST) and blood urea nitrogen (BUN). The incidence of AGI was significantly lower in the Xuebijing group than that in the non-Xuebijing group [50.87% (29/57) vs. 70.83% (51/72), P < 0.05], and the 28-day mortality was slightly lower than that in the non-Xuebijing group [24.56% (14/57) vs. 30.56% (22/72), P > 0.05]. In the Xuebijing group, the decreases in CRP, PCT and WBC at 7 days after admission were greater than those in the non-Xuebijing group, with statistically significant differences in the decreases of CRP and PCT [CRP (mg/L): 47.12±67.34 vs. 7.76±111.03, PCT (μg/L): 14.08 (-1.22, 50.40) vs. 2.94 (-1.27, 14.80), all P < 0.05]. Univariate analysis showed that the use of acid suppressants, the use of analgesic sedation, the non-use of Xuebijing injections, pulmonary infections, and urinary tract infections were the risk factors for the development of AGI in patients with sepsis. Binary Logistic regression analysis further showed that the use of acid suppressants [odds ratio ( OR) = 2.450, 95% confidence interval (95% CI) was 1.021-5.883, P = 0.045], use of analgesic sedatives ( OR = 2.521, 95% CI was 1.074-5.918, P = 0.034), and urinary tract infection ( OR = 4.011, 95% CI was 1.085-14.831, P = 0.037) were independent risk factors for sepsis combined with AGI, in which the use of Xuebijing injection was a protective factor ( OR = 0.315, 95% CI was 0.137-0.726, P = 0.007). Conclusions:Xuebijing injection reduced the incidence of AGI in patients with non-gastrointestinal sepsis. PCT and CRP decreased more markedly than in patients who did not use Xuebijing injection. The use of acid-suppressing agents, analgesic and sedative agents, and urinary tract infections were independent risk factors for sepsis in combination with AGI, while the use of Xuebijing injection is a protective factor.

Objective To establish the UPLC fingerprint chromatogram combined with chemometric analysis for the quality evaluation of classical formula Linggui Zhugan Decoction.Methods SHIMADZU Shim-Pack GIST C18 column(100 mm×2.1 mm,2.0 μm)was used with acetonitrile-0.1%phosphoric acid aqueous solution as mobile phase,gradient elution;flow rate was 0.2 mL/min;the detection wavelength was 266 nm for the first 30 minutes and 235 nm for the last 36 minutes;the column temperature was 30℃.The UPLC fingerprint of Linggui Zhugan Decoction was established by Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012.130723 version),and the common peak was determined and the similarity evaluation was carried out.Based on the peak area determination results of the common peak of the fingerprint,the quality of different batches of Linggui Zhugan Decoction was evaluated by chemometrics such as clustering analysis and principal component analysis.Results A total of 24 common peaks were confirmed and 14 components were identified by using reference substances.The similarity of 10 batches of Linggui Zhugan Decoction samples was greater than 0.950,which could be divided into two categories by chemometrics,and the principal component 1-4 were the main factors affecting its quality evaluation.OPLS-DA identified 6 differential markers.Conclusion The fingerprint research method established in the study is simple,reliable and reproducible.Through the method of fingerprint combined with chemometrics analysis,the differences between Linggui Zhugan Decoction from different origins of medicinal materials are identified,which provides a reference for the internal quality evaluation of Linggui Zhugan Decoction.