Objective To investigate the pharmacological effects of"Yajieshaba"on mice with alcohol-induced liver injury and to investigate the mechanism of the impact of"Yajieshaba"on the regulation of intestinal flora by 16S rDNA technology.Methods Healthy male KM mice were randomly divided into control,model,"Yajieshaba"low,medium,and high dose(0.39,1.17 and 3.51 g·kg-1)groups and Bifendatatum(2.93 mg·kg-1)groups,with eight mice in each group.After one week of pre-administration of"Yajieshaba",a mouse model of acute alcoholic liver injury was established by a single instillation of 56%alcohol,and the levels of AST and ALT in the serum of mice were measured,and the morphological changes of liver histology were observed by HE staining;secondly,faecal DNA was extracted from each group under aseptic operation,and 16S rDNA sequencing and differential analysis by alpha diversity and species composition at the phylum and genus levels were performed.Results The results showed that the biochemical indexes of liver function(ALT and AST)were significantly improved by"Yajieshaba",and the degree of liver damage was significantly reduced by HE staining.At the phylum level,it significantly decreased the abundance of Aspergillus and increased the quantity of Bacteroides;at the genus level,it significantly up-regulated the plenty of Bacteroides and Prevotella and downregulated a lot of Prevotella and Helicobacter.At the genus level,"Yajieshaba"significantly up-regulated the abundance of Bacillus spp.and Prevotella spp.and down-regulated the abundance of Prevotella spp.and Helicobacter spp.Conclusion"Yajieshaba"may play an anti-acute alcoholic liver injury effect by regulating the intestinal flora and metabolites.

Objective:To retrospectively analyze the treatment status of tyrosine kinase inhibitors (TKI) in newly diagnosed patients with chronic myeloid leukemia (CML) in China.Methods:Data of chronic phase (CP) and accelerated phase (AP) CML patients diagnosed from January 2006 to December 2022 from 77 centers, ≥18 years old, and receiving initial imatinib, nilotinib, dasatinib or flumatinib-therapy within 6 months after diagnosis in China with complete data were retrospectively interrogated. The choice of initial TKI, current TKI medications, treatment switch and reasons, treatment responses and outcomes as well as the variables associated with them were analyzed.Results:6 893 patients in CP ( n=6 453, 93.6%) or AP ( n=440, 6.4%) receiving initial imatinib ( n=4 906, 71.2%), nilotinib ( n=1 157, 16.8%), dasatinib ( n=298, 4.3%) or flumatinib ( n=532, 7.2%) -therapy. With the median follow-up of 43 ( IQR 22-75) months, 1 581 (22.9%) patients switched TKI due to resistance ( n=1 055, 15.3%), intolerance ( n=248, 3.6%), pursuit of better efficacy ( n=168, 2.4%), economic or other reasons ( n=110, 1.6%). The frequency of switching TKI in AP patients was significantly-higher than that in CP patients (44.1% vs 21.5%, P<0.001), and more AP patients switched TKI due to resistance than CP patients (75.3% vs 66.1%, P=0.011). Multi-variable analyses showed that male, lower HGB concentration and ELTS intermediate/high-risk cohort were associated with lower cytogenetic and molecular responses rate and poor outcomes in CP patients; higher WBC count and initial the second-generation TKI treatment, the higher response rates; Ph + ACA at diagnosis, poor PFS. However, Sokal intermediate/high-risk cohort was only significantly-associated with lower CCyR and MMR rates and the poor PFS. Lower HGB concentration and larger spleen size were significantly-associated with the lower cytogenetic and molecular response rates in AP patients; initial the second-generation TKI treatment, the higher treatment response rates; lower PLT count, higher blasts and Ph + ACA, poorer TFS; Ph + ACA, poorer OS. Conclusion:At present, the vast majority of newly-diagnosed CML-CP or AP patients could benefit from TKI treatment in the long term with the good treatment responses and survival outcomes.

Objective:To screening differentially expressed genes (DEGs) in proliferative diabetic retinopathy (DR) patients to provide new biological therapeutic targets for proliferative DR (PDR) therapy.Methods:A basic research. A total of 3 PDR patients (group PDR) and 3 non-diabetic patients (control group) were enrolled in the study in Tianjin Medical University Eye Hospital in October 2020. In addition, 40 cases of PDR and non-diabetic patients were selected and divided into PDR validation group and control validation group. Peripheral blood validation test was performed in PDR validation group and control validation group; RNA sequencing was performed in PDR group and control group. Transcriptomics (RNAseq) sequencing technology was used to screen DEG in PDR group and control group. The selected DEGs were analyzed by gene ontology (GO) function enrichment analysis, signal pathway enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) and protein-protein interaction network (PPI). The gene expression database was used to find the high-throughput data related to PDR, and multi queue comparison analysis was carried out. The target genes of differentially expressed miRNAs were predicted through targetscan platform, so as to clearly screen the correlation between DEG and PDR. Reverse transcription polymerase chain reaction and Western blot were used to verify the expression of DEG mRNA and protein related to PDR. The relative expression of PDR related DEG mRNA and protein between PDR validation group and control validation group were compared by paired t-test. Results:A total of 1 337 DEGs were screened by RNAseq sequencing in the peripheral blood of patients with PDR, of which 419 genes were up-regulated and 918 down-regulated. Among them, direct inhibitor of apoptosis protein-binding protein with low isoelectric point ( DIABLO), zinc finger and BTB domain containing 10 ( ZBTB10), polo-like kinases 3 ( PLK3), regulatory subunit 1 ( PIK3R1) and B cell translocation gene 3 (BTG3) were differentially expressed in PDR patients. The function of GO was enriched from the analysis of molecular function, biological process and cellular composition. The results showed that DIABLO, ZBTB10, PLK3, PIK3R1, BTG3 were involved in the pathological process related to PDR. KEGG enrichment analysis showed that glucose metabolic pathways such as extracellular matrix receptors, cytokine regulatory pathway, p53 signal pathway and galactose metabolism may be involved in the process of differential genes. The analysis of PPI protein interaction network showed that the larger the DEG-associated protein node, the greater the number of associated nodes. Among them, DIABLO, ZBTB10, PLK3, PIK3R1 and BTG3 played significant roles in the formation of the action network. By comparing and analyzing the existing high-throughput data related to diabetic retinopathy in Gene Expression Omnibus database and predicting by Targetscan platform, it was found that some significant differences in miRNA reported in aqueous humor, vitreous fluid and plasma of DR patients can be regulated by the differential genes found in this study. Compared with the control verification group, the relative expressions of DIABLO, ZBTB10, PLK3, PIK3R1 mRNA and protein in peripheral blood of the PDR verification group were up-regulated, and the relative expression of BTG3 mRNA and protein was down-regulated. Conclusion:DIABLO, ZBTB10, PLK3, PIK3R1 and BTG3 are DEGs in patients with PDR, and they can participate in the disease process by regulating the biological processes of cell proliferation, fibrosis and oxidative stress.

Blast injury of the chest injury is the most common wound in modern war trauma and terrorist attacks, and is also the most fatal type of whole body explosion injury. Most patients with severe blast injury of the chest die in the early stage before hospitalization or during transportation, so first aid is critically important. At present, there exist widespread problems such as non-standard treatment and large difference in curative effect, while there lacks clinical treatment standards for blast injury of the chest. According to the principles of scientificity, practicality and advancement, the Trauma Society of Chinese Medical Association has formulated the guidance of classification, pre-hospital first aid, in-hospital treatment and major injury management strategies for blast injury of the chest, aiming to provide reference for clinical diagnosis and treatment.

Objective:To observe the effect of bone morphogenetic protein 4 (BMP4) on the proliferation and migration of human retinal microvascular endothelial cells (hRMEC) under oxidative stress.Methods:The hRMEC cultured in vitro were divided into control group, 4-hydroxynonenal (HNE) treatment group (4-HNE group), 4-HNE+BMP4 group (BMP4 group). Cell culture medium of 4-HNE treatment group was added with 10 μmmol/L 4-HNE; cell culture of BMP4 group was cultured with 10 μmmol/L 4-HNE, and after stimulation for 6 h, 100 ng/ml recombinant human BMP4 was added. The effects of 4-HNE and BMP4 on hRMEC viability was detected by thiazole blue colorimetric method. The effects of 4-HNE and BMP4 on cell migration was determined by cell scratch test. The relative expression of BMP4 mRNA in the cells of the control group and 4-HNE treatment group and the mRNA expression of the control group, the fibronectin (FN) of BMP4 group, laminin (Laminin), α-smooth muscle contractile protein (α-SMA), and collagen type Ⅰ (Collagen Ⅰ), vascular endothelial growth factor (VEGF), and connective tissue growth factor (CTGF) were detected by real-time quantitative polymerase chain reaction (qRT-PCR). Western blot was used to detect the relative expression of BMP4 protein in the control group and 4-HNE group. The control group and 4-HNE group were compared by t test. Results:Compared with the control group, cell viability ( t=12.73, 16.26, P=0.000 2, <0.000 1), cell migration rate ( t=28.17, 37.48, P<0.000 1, <0.000 1) in 4-HNE group and BMP4 group were significantly increased, and the difference was statistically significant; the relative expression of BMP4 mRNA and protein in the 4-HNE group was significantly increased, and the difference was statistically significant ( t=16.36, 69.35, P=0.000 1, <0.000 1). The qRT-PCR test results showed that compared with the control group, the relative expression of VEGF, FN, Laminin, α-SMA, Collagen Ⅰ, and CTGF mRNA in the cells of the BMP4 group was significantly increased, and the difference was statistically significant ( t=10.61, 17.00, 14.85, 7.78, 12.02, 10.61, P=0.0004, <0.000 1, 0.000 1, 0.001 5, 0.000 1, 0.000 4). Conclusion:BMP4 can induce the proliferation and migration of hRMEC; it can also regulate the expression of angiogenesis factors and fibrosis-related factors in hRMEC.

Objective:To investigate the difference of metabolomics between acute heart failure (AHF) patients and control. To find and validate new metabolic biomarkers.Methods:This was a single-center case-control study which included 89 acute heart failure patients admitted to the emergency department of Henan Provincial People's Hospital from January 2018 to June 2019. Eighty people without heart failure and diastolic dysfunction were enrolled as control group whose age and sex were matched to the study group. The fasting blood samples were collected from femoral arterial. Qualitative and quantitative analyses of plasma metabolites were performed in 2 groups by high performance liquid chromatography tandem mass spectrometry (UHPLC-MS), Orthogonal partial least squares-discriminant analysis (OPLS-DA) model and ROC curve method were applied.Results:Compared with the control group, we found that AHF group had higher likelihood to groups with coronary heart disease (37% vs. 7%, P<0.001), hypertension (58% vs. 28%, P<0.001), diabetes (33% vs. 18%, P=0.033), atrial fibrillation (24% vs. 4%, P<0.001), smoking history (42% vs. 18%, P=0.001), and that AHF group had higher creatinine level [(121.6 ± 78.4) vs. (69.0 ± 21.0), P<0.001], higher urea level [(11.5 ± 7.6) vs. (6.2 ± 2.0), P<0.001], higher heart rate [(92 ± 23) vs. (78 ± 14), P<0.001], hypoproteinemia [(32.4 ± 5 .2) vs. (40.4 ± 2.2), P<0.001], and significantly increased BNP level [(4 200 ± 5 229) vs. (100 ± 68), P<0.001], lower left ventricular ejection fraction[(45 ± 8) vs. (57 ± 6), P<0.001], low serum sodium level ( P<0.001). The metabolites of AHF group were significantly different from those of the control group. The metabolites involved amino acids, fatty acids, lipids, nucleosides and their derivatives. Adenine, N-acetyl-L-glutamic, pseudouridine and Gamma-Glutamylcysteine had certain diagnostic value for AHF comparing to control. The AUC were 0.995, 0.932, 0.920 and 0.900. And the AUC value for BNP diagnosis of AHF is 0.978. Conclusions:There were significant differences in metabolism between AHF group and control group including multiple substances. Adenine, N-acetyl-L-glutamic, pseudouridine and Gamma-Glutamylcysteine has similar diagnostic value compared with BNP for diagnosing AHF.

Objective:To evaluate the role of interleukin-6 (IL-6) and CD4 + T-lymphocytopenia in assessing the severity and prognosis of coronavirus disease 2019 (COVID-19). Methods:A prospective observational study was conducted. Forty-five patients with COVID-19 admitted to Henan Provincial People's Hospital from January 13 to March 13, 2020 were enrolled and divided into normal group (13 cases), severe group (20 cases), critically severe group (12 cases) according to the severity of the disease. A total of 15 healthy subjects receiving physical examinations during the same period were collected as the healthy control group. Clinical data were collected to compare the clinical characteristics, general test results, IL-6 and CD4 + T-lymphocytopenia levels of patients in different disease severity groups and healthy control group. The receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of each indicator for the severity of COVID-19. Multivariate Cox regression analysis was used to analyze the risk factors affecting the prognosis of COVID-19 patients, and Kaplan-Meier survival curve analysis was performed. Results:The age of the critically severe group was significantly higher than that of the severe and normal groups (years old: 66.91±17.01 vs. 59.35±18.07, 40.23±12.61, both P < 0.05), and the negative conversion time of the 2019 novel coronavirus (2019-nCoV) was significantly longer than that of the severe and normal groups (days: 19.00±10.66 vs. 18.00±7.18, 9.31±3.49, both P < 0.05). With the increase of the severity of disease, white blood cell count (WBC), C-reactive protein (CRP), calcitonin (PCT), total bilirubin (TBil), troponin I (TnI), IL-6, D-dimer and other indicators were significantly increased, while lymphocyte count (LYM), platelet count (PLT), CD4 +, CD8 +, oxygenation index (PaO 2/FiO 2) were significantly decreased (all P < 0.01). ROC curve showed that PaO 2/FiO 2, IL-6 and CD4 + had certain predictive value for disease severity of COVID-19, the area under the ROC curve (AUC) of them were 0.903, 0.871, 0.689, and the 95% confidence interval (95% CI) were 0.806-0.949, 0.769-0.974, 0.542-0.853; the best cut-off values were 196.00 mmHg (1 mmHg = 0.133 kPa), 6.02 ng/L, 355 cells/μL, respectively; the sensitivity were 73.3%, 99.3%, 73.3%, and the specificity were 96.6%, 62.1%, 65.5%, respectively. Multivariate Cox regression analysis showed that age, PaO 2/FiO 2, high IL-6 and low CD4 + (IL-6≥6.02 ng/L and CD4 + < 355 cells/μL) were independent risk factors affecting the prognosis of COVID-19 [hazard ratio ( HR) was 1.077, 0.053 and 3.490, respectively, all P < 0.05]. Kaplan-Meier survival analysis showed that when both high IL-6 and low CD4 + (IL-6≥6.02 ng/L and CD4 + < 355 cells/μL) were present, the mean time of adverse prognosis was (20.53±5.71) days; when increased IL-6 and decreased CD4 + were inconsistent, the mean time of adverse prognosis was (53.21±3.16) days. Conclusions:The levels of IL-6 and CD4 + T-lymphocytopenia are closely related to the severity of COVID-19 disease. When IL-6 ≥ 6.02 ng/L and CD4 + < 355 cells/μL occur simultaneously, the prognosis is poor.

Objective To observe the effects of exosomes derived from rat mesenchymal stem cells (MSC-exosomes) on the rat experimental autoimmune uveitis (EAU) model.Methods Twelve Lewis rats were randomly divided into experimental group and control group by random number table,with 6 rats in each group.Rats in the experimental group were established with EAU model,100 μl of MSC-exosomes (50 μg) were periocular injected on the 9th day after modeling while the control rats were injected with the same volume of phosphate buffer.At different time points after modeling,the retinal structure was observed by hematoxylin and eosin (HE) staining,and the clinical and pathological manifestations were evaluated.T cells from the two groups were analyzed by flow cytometry,Immunohistochemical staining was used to observe the expression of macrophage surface marker CD68.The effect of MSC-exosomes on T cells was measured by lymphocyte proliferation assays.And flow cytometry was used to detect Th 1,Th 17 and regulatory T cells Variety.Electroretinogram (ERG) was used to evaluate the retinal function.Data were compared between the two groups using the t test.Results HE staining showed that the retina structure of the experimental group was more complete than that of the control group on the 15th day after modeling.Immunohistochemical staining showed that the positive expression of CD68 in the experimental group was significantly less than that in the control group.On the 15th day after modeling,the retinal pathological score of the experimental group was lower than that of the control group.On the 9th to 13th day after modeling,compared to the control group,the average clinical scores of the retina in the experimental group were lower,and the difference was statistically significant (t=3.665,3.21,3.181,4.121,3.227;P＜0.01).The results of T cell proliferation assay showed that exosomes (1.0,10.0 μg/ml) inhibited the proliferation of T cells under different concentrations of R16 (1,10,30 μg/ml),and the difference was statistically significant (F=1 1.630,4.188,6.011;P＜0.05).The results of flow cytometry showed that the number ofThl,Th17 and Treg cell subsets in the experimental group was decreased compared with the control group,and the difference was statistically significant (t=7.374,4.525,6.910;P＜0.01).There was no difference in the proportion of cells in the T cells and lymph nodes (t=1.126,0.493,0.178;P=0.286,0.632,0.862).The results of ERG showed that,compared with the control group,the amplitudes of 0.01,3.0 cd/m2 a wave and b wave of the experiment group were all increased on the 15th day after modeling,and the differences were statistically significant (t=3.604,4.178,4.551,2.566,P＜0.05).Conclusions MSC-exosomes can reduce the clinical and pathological manifestations of EAU,protect retinal function,reduce ocular macrophage infiltration,down-regulate the proportion of inflammatory cells in the eye,and inhibit T cell proliferation.

Objective To explore the role of clinical application of enteral nutrition sequential therapy in early enteral nutrition support by comparison with enteral nutrition non-sequential therapy in critically ill patients with cerebrovascular diseases.Methods A total of 62 patients were randomly (random number) divided into sequential group and conventional (non-sequential) grouThe comparisons of tolerance for enteral nutrition support,levels of prealbumin,the mechanically ventilated time and mortality rate in 28-day between two groups were carried out.Results The tolerance of sequential group was superior to that of conventional group (P＜0.05).The higher level of prealbumin and the shorter mechanical ventilation time were observed in sequential group compared with conventional group (P＜0.01).Compared with conventional group,the patients in sequential group had lower mortality rate in 28 days (P＜0.05).Conclusions Sequential therapy is beneficial to the implementation of early enteral nutrition support in patients with severe cerebrovascular disease,reducing the nutritional adverse events,and improves the prognosis.

Objective To evaluate the association between the decrease of the perioperative serum carcinoembryonic antigen (CEA) level during perioperative period and the prognosis in patients with colon cancer after a curative resection.Methods Retrospective analysis was conducted to evaluate the relationship between preoperative serum CEA level and different clinicopathologic features in 605 cases who underwent a curative resection for colon cancer from January 2006 to April 2011.According to the preoperative serum CEA level,the patients were divided into two groups:≤5 ng/ml and ＞5 ng/ml group.The critical value of the CEA decreasing rate in preoperative serum CEA ＞ 5 ng/ml group was calculated,and the relationship between this critical value and survival rate was then analyzed.Univariate and multivariate models were used to detect the risk factors of overall survival rate (OS) and disease free survival (DFS) in preoperative serum CEA ＞ 5 ng/ml patients.Results The preoperative serum CEA levels were significantly associated with lymphatic invasion (x2 =14.122,P＜0.001),T stages (x2 =40.153,P ＜0.001),N stages (x2 =22.721,P ＜0.001) and pathological stages (x2 =38.576,P ＜ 0.001),except for sex (x2 =0.453,P =0.501),age (x2 =0.195,P =0.659) and histological stages (x2 =6.135,P =0.112).The critical values of CEA decreasing rate for OS and DFS were 48.95％ and 50.81％ in preoperative serum CEA ＞5 ng/ml group respectively.There were significant differences of 5-year OS (31.37％ vs.76.63％,x2 =43.235,P ＜ 0.001) and 5-year DFS (27.69％ vs.72.10％,x2 =55.561,P ＜0.001) between patients after operation whose CEA decreasing rate were lower than critical value and those whose were higher.Univariate analysis showed that the decreasing rate of CEA was an influence factor for OS (x2 =43.235,P ＜ 0.001) and DFS (x2 =55.561,P ＜ 0.001) of preoperative serum CEA ＞ 5 ng/ml patients.The N stages and pathological stages were both related to OS (x2 =14.683,P＜0.001;x2 =12.295,P＜0.001) and DFS (x2 =16.212,P＜0.001;x2 =13.704,P＜0.001)respectively.Multivariate model showed that the decreasing rate of CEA level and N stages were both associated withOS (x2=18.885,P＜0.001;x2 =7.523,P＜0.001) and DFS (x2 =19.275,P＜0.001;x2 =6.997,P ＜ 0.001) of preoperative serum CEA ＞ 5 ng/ml patients.Conclusion A high decreasing rate of serum CEA level after operation in colon cancer patients who have high CEA levels before the curative resection can be a protective factor for prognosis,especially when the decreasing rate is higher than the critical value.