ObjectiveTo investigate the therapeutic effect of Baihe Wuyaotang （BWT） on non-alcoholic fatty liver disease （NAFLD） and elucidate its underlying mechanism. MethodC57BL/6J mice were randomly assigned to six groups： normal control， model， positive drug （pioglitazone hydrochloride 1.95×10^-3 g·kg^-1）， and low-， medium-， and high-dose BWT （1.3，2.5 and 5.1 g·kg^-1）. Following a 12-week high-fat diet （HFD） inducement， the mice underwent six weeks of therapeutic intervention with twice-daily drug administration. Body weight was monitored weekly throughout the treatment period. At the fifth week， glucose tolerance （GTT） and insulin tolerance （ITT） tests were conducted. Subsequently， the mice were euthanized for the collection of liver tissue and serum， and the subcutaneous adipose tissue （iWAT） and epididymal adipose tissue （eWAT） were weighed. Serum levels of total triglycerides （TG） and liver function indicators，such as alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， were determined. Histological examinations， including oil red O staining， hematoxylin-eosin （HE） staining， Masson staining， and transmission electron microscopy， were performed to evaluate hepatic lipid deposition， pathological morphology， and ultrastructural changes， respectively. Meanwhile， Western blot and real-time quantitative polymerase chain reaction （Real-time PCR） were employed to analyze alterations， at both gene and protein levels， the insulin signaling pathway molecules， including insulin receptor substrate 1/2/protein kinase B/forkhead box gene O1 （IRS1/2/Akt/FoxO1）， glycogen synthesis enzymes phosphoenolpyruvate carboxy kinase （Pepck） and glucose-6-phosphatase （G6Pase）， lipid metabolism-related genes stearoyl-coA desaturase-1 （SCD-1） and carnitine palmitoyltransferase-1 （CPT-1）， fibrosis-associated molecules α-smooth muscle actin （α-SMA）， type Ⅰ collagen （CollagenⅠ）， and the fibrosis canonical signaling pathway transforming growth factor-β₁/drosophila mothers against decapentaplegic protein2/3（TGF-β₁/p-Smad/Smad2/3）， inflammatory factors such as interleukin（IL）-6， IL-8， IL-11， and IL-1β， autophagy markers LC3B Ⅱ/Ⅰ and p62/SQSTM1， and the expression of mammalian target of rapamycin （mTOR）. ResultCompared with the model group， BWT reduced the body weight and liver weight of NAFLD mice（P<0.05， P<0.01）， inhibited liver lipid accumulation， and reduced the weight of white fat： it reduced the weight of eWAT and iWAT（P<0.05， P<0.01） as well as the serum TG content（P<0.05， P<0.01）. BWT improved the liver function as reflected by the reduced ALT and AST content（P<0.05， P<0.01）. It improved liver insulin resistance by upregulating IRS2， p-Akt/Akt， p-FoxO1/FoxO1 expressions（P<0.05）. Besides， it improved glucose and lipid metabolism disorders： it reduced fasting blood glucose and postprandial blood glucose（P<0.05， P<0.01）， improved GTT and ITT（P<0.05， P<0.01）， reduced the expression of Pepck， G6Pase， and SCD-1（P<0.01）， and increased the expression of CPT-1（P<0.01）. The expressions of α-SMA， Collagen1， and TGF-β₁ proteins were down-regulated（P<0.05， P<0.01）， while the expression of p-Smad/Smad2/3 was downregulated（P<0.05）， suggesting BWT reduced liver fibrosis. BWT inhibited inflammation-related factors as it reduced the gene expression of IL-6， IL-8， IL-11 and IL-1β（P<0.01） and it enhanced autophagy by upregulating LC3B Ⅱ/Ⅰ expression（P<0.05）while downregulating the expression of p62/SQSTM1 and mTOR（P<0.05）. ConclusionBWT ameliorates NAFLD by multifaceted improvements， including improving IR and glucose and lipid metabolism， anti-inflammation， anti-fibrosis， and enhancing autophagy. In particular， BWT may enhance liver autophagy by inhibiting the mTOR-mediated signaling pathway.

Objective:To determine the erythromycin resistance of Bordetella pertussis isolates and their ptxP1 and ptxP3 phenotypic composition and compare clinical manifestations of children with pertussis caused by the two types of strains. Methods:This was a cross-sectional study, the pertussis cases diagnosed using bacterial culture from January 2019 to December 2022 in Beijing Children′s Hospital and the First People′s Hospital of Wuhu were collected.Any suspected Bordetella pertussis colonies were identified by the slide agglutination test.The susceptibility of isolates to erythromycin was detected by the E-test and K-B test.The ptxP gene was amplified by polymerase chain reaction and sequenced to determine its genotype. t-test, Mann-Whitney U-test, Chi-square test and Fisher′s exact test were use to statistical analysis. Results:A total of 192 strains of Bordetella pertussis were identified, including 188 (97.9%) erythromycin-resistant strains.Among the 188 strains, 30.3%(57/188) belonged to the ptxP1 genotype and 69.7%(131/188) belonged to the ptxP3 genotype.In children aged below 1 year old, the incidence of paroxysmal cough caused by infection with the ptxP3 strain was higher than that with the ptxP1 strain (57.1% vs.29.4%, P<0.05), and children infected with the ptxP3 strain were more likely to develop apnea or asphyxia (23.8% vs.17.6%), post-tussive vomiting (44.4% vs.32.4%), whooping cough (72.0% vs.50.0%) and pneumonia or bronchitis (85.7% vs.73.5%) compared to those infected with the ptxP1 strain, but the differences were not statistically significant(all P>0.05). In children aged 1 year old and above, the white blood cell count of children infected with the ptxP1 strain was higher than that of infections with the ptxP3 strain [13.5(9.9, 24.5)×10 9/L, 10.3 (7.0, 16.4)×10 9/L, P<0.05], and children infected with the ptxP1 strain were more likely to contract other pathogen infections than those infected with the ptxP3 strain (17.4% vs.4.4%, P>0.05). Conclusions:ptxP3 erythromycin-resistant Bordetella pertussis has become the main pathogen of pertussis.Infants with pertussis caused by the ptxP3 erythromycin-resistant strain show more significant manifestations and a higher possibility of severe symptoms than those infected with the ptxP1 erythromycin-resistant strain.

BACKGROUND:As a dominant breed pig in southwest China,the southern Yunnan small-ear pig has been widely used as an experimental animal in the basic research of other disciplines,but there are still no reports on its application in anterior cruciate ligament reconstruction. OBJECTIVE:To establish a large animal model of the southern Yunnan small-ear pig with anterior cruciate ligament with autologous Achilles tendon was established. METHODS:Twenty adult female Yunnan small-ear pigs were equally randomized into two groups.In the autologous Achilles tendon group,the right knee anterior cruciate ligament was reconstructed with autologous Achilles tendon as a graft,while in the sham-operated group,a similar operation was performed on the right knee without any treatment of the anterior cruciate ligament.General conditions of each pig were observed and recorded before and 12 months after surgery.Ligaments and grafts were taken for gross observation and MAS scoring.Hematoxylin-eosin staining was performed to observe morphological characteristics of ligaments.The staining and arrangement of type I and type Ⅲ collagen were evaluated by immunohistochemistry.Transmission electron microscopy was used to observe the type,size,diameter,ratio,and distribution of collagen fibers in ligaments. RESULTS AND CONCLUSION:All animals had normal diet and activity,good wound healing,no obvious inflammatory reaction,no local purulent infection,and no significant changes in mental and urinary conditions compared with those before surgery.The reconstructed cruciate ligament of the knee was intact,with no stiffness and normal range of motion.Both the anterior drawer and Lachman tests were negative.Gross observation of the graft:12 months after surgery,the grafts was in good position,with good integrity,obvious tension,ligament color close to the original anterior cruciate ligament,and complete surface synovial coverage.Most of the intraarticular ligaments in the autologous Achilles tendon group were defined as MAS I type and a few were defined as MAS Ⅱ type.In the sham-operated group,the intraarticular ligament was defined as MAS I type.Hematoxylin-eosin staining indicated that,12 months after surgery,collagen fibers in the autologous Achilles tendon group began to appear bundled,isotropic,and uniformly arranged,with more obvious isotropic corrugations,and the nuclei were mainly linear or spindle-shaped,which were similar to those in normal anterior cruciate ligament tissue of the sham-operated group.Immunohistochemistry results indicated that,12 months after surgery,there was a higher expression of type I collagen and significantly less expression of type Ⅲ collagen in the reconstructed anterior cruciate ligament in the autologous Achilles tendon group.The degree of type I and type Ⅲ staining was similar in the two groups.Under the transmission electron microscope,the diameter,arrangement and density of collagen fibers in the reconstructed anterior cruciate ligament of the autologous Achilles tendon group were similar to those of the original anterior cruciate ligament at 12 months after surgery,indicating that the ligament remodeling process had been basically completed in the autologous Achilles tendon group at 12 months after surgery.Through a comprehensive evaluation of animal general conditions,ligament general view,MAS score,hematoxylin-eosin staining,immunohistochemistry,and transmission electron microscopy observation,we successfully established a large animal model of anterior cruciate ligament reconstruction using autogenous Achilles tendon in southern Yunnan small-ear pigs,with good morphological,histological and ultrastructural results.

OBJECTIVE To conduct a rapid qualitative study on the chemical constituents of the methanol extract in Thamnolia subuliformis (Ehrh.) W.Culb. by UHPLC-Q-Exactive Focus-MS/MS. METHODS The chromatographic separation was performed on a Waters Acquity UPLC BEH C18(2.1 mm×50 mm, 1.7 μm) column with acetonitrile(A)-0.1% formic acid aqueous solution(B) as mobile phase for gradient elution, the flow rate was 0.3 mL·min–1 and the column temperature was 30 ℃. Mass spectrometry was performed using an electrospray ionization and data was collected in negative ion modes, and the detection range was m/z 80−1 200. The chemical constituents in Thamnolia subuliformis (Ehrh.) W.Culb. were identified rapidly and comprehensively based on the accurate relative molecular and combined with literature data and reference substances. RESULTS A total of 39 chemical constituents were speculatively identified, including 9 mono-substituted phenyl rings, 11 depsides, 5 depsidones, 2 dibenzofuran, 10 lipids, and 2 others. CONCLUSION The chemical constituents in the Thamnolia subuliformis (Ehrh.) W.Culb. can be identified accurately and rapidly by this method. Among them, 3 compounds(β-resorcylic acid, usnic acid, atranorin) are unambiguously identified by comparing with reference standards, 19 compounds are found from Thamnolia subuliformis (Ehrh.) W.Culb. for the first time. This paper can provide the important basis for study on pharmacodynamic material and substitute development of Thamnolia subuliformis (Ehrh.) W.Culb..

Objective To explore the relationship between serum microRNA-124(miR-124),CD146,angio-poietin-like protein 2(Angptl2)and the stability of carotid atherosclerosis(CAS)plaque in patients with a-cute cerebral infarction(ACI),and to provide reference for early prevention and treatment of patients with ACI.Methods A total of 191 patients with ACI admitted in Handan Central Hospital from January 2020 to February 2023 were selected as ACI group,and another 61 healthy volunteers who were underwent physical examination during the same period were selected as control group.The patients with ACI were divided into unstable plaque group(56 cases),stable plaque group(71 cases),and non plaque group(64 cases)based on carotid color doppler ultrasound results.The serum miR-124 expression levels of all subjects were detected by real time fluorescence quantitative polymerase chain reaction(RT-qPCR),and the serum CD146 and Angptl2 levels were detected by enzyme-linked immunosorbent assay(ELISA).The influencing factors of the instabili-ty of CAS plaque in patients with ACI was analyzed by univariate and multivariate Logistic regression.The predictive value of serum miR-124,CD146 combined with Angptl2 for the instability of CAS plaque in patients with ACI was analyzed by the receiver operating characteristic(ROC)curve.Results The serum CD146 and Angptl2 levels in ACI group were higher than those in control group(P＜0.05),and the miR-124 expression level was lower than that in control group(P＜0.05).Univariate analysis showed that the stability of CAS plaques in ACI patients was correlated with age,hypertension,hyperlipidemia,fibrinogen(FIB),serum C-reac-tive protein(CRP),serum cystatin C(CyC),CD146,Angptl2 and miR-124(P＜0.05).Multivariate Logistic regression analysis showed that the decrease of serum miR-124,the increase of CD146,the increase of Angptl2 and the combination of hyperlipidemia were risk factors for CAS plaque stability in ACI patients(P＜0.05).The area under ROC curve(AUC)of serum miR-124,CD146,Angptl2 and the combination of the three indi-cators to predict CAS plaque instability in ACI patients were 0.741,0.719,0.781 and 0.834,respectively.Conclusion The serum miR-124 expression level,CD146 and Angptl2 levels are the influencing factors of CAS plaque instability in ACI patients,which may be involved in the formation and development of CAS plaque in ACI patients.The combined detection of the three factors has a good predictive effect on CAS plaque instability in ACI patients.

Animals ; SARS-CoV-2 ; Antibodies, Neutralizing ; Macaca mulatta ; COVID-19/prevention & control* ; Antibodies, Viral ; Vaccines

【Objective】 To retrospectively analyze the efficacy of low dose apheresis platelet prophylactic infusion and explore its feasibility. 【Methods】 A total of 392 inpatients with platelet transfusion in our hospital from November 2020 to September 2021 were selected. The conventional dose (1 therapeutic dose) of apheresis platelet transfusion was set as the control group, and the low dose (0.5 therapeutic dose) as the experimental group. Platelet count before and after infusion, platelet elevation value (△PLT) and 24 h platelet count correction increase index (CCI) were observed, and the efficacy of low-dose platelet infusion was analyzed by disease type and gender. 【Results】 The △PLT value and 24h CCI effective infusion rate in control group were higher than those in experimental group: (16±16) ×10⁹ vs (7±10) ×10^9, 71.94% vs 60.46%, P<0.05. The △PLT value of the control group was about 1.2-3.5 times that of the experimental group, and the effective rate was about 1-1.4 times. In control group, the △PLT (×10⁹) was AML (20±14) >AA (14±14) >ALL (13±12) >NHL (9±8) >MDS (7±6). In the experimental group, the △PLT (×10⁹) was AA (11±18) >AML (8±8) >ALL (5±7) >NHL (5±7) >MDS (6±16). The 24h CCI was AML(163/188, 86.70%)>AA(23/32, 71.88%)>ALL(65/98, 66.33%)>MDS(9/17, 52.94%)>NHL(12/22, 51.55%) in the control group, and AML(133/188, 70.74%)>AA(19/32, 59.38%)>NHL(12/22, 51.55%)>ALL(47/98, 47.96%)>MDS(8/17, 47.06%) in the experimental group. The effective infusion rates of AML and ALL2 in the experimental groups were 70.74% (133/188) and 47.96% (47/98), respectively, significantly lower than 86.7% (163/188) and 66.33% (65/98) in the control group(P<0.05). No significant difference was noticed in the effective infusion rate between the experimental group and the control group for other diseases (P>0.05). 【Conclusion】 Low-dose apheresis platelet prophylactic infusion can alleviate the between supply shortage, with an effective infusion rate of 60.46% (236/392), which has certain clinical application value. Patients with AML, AA or ALL were recommended with low dose platelets, while patients with MDS and NHL were not recommended.

【Objective】 To retrospectively analyze the safety and product quality of NGL XCF 3000 blood cell separator for collecting platelet-rich plasma (PRP) in 256 cases, so as to provide reference for safe collection and product quality control of PRP. 【Methods】 The data of 256 patients receiving PRP treatment in our hospital from June 2021 to June 2022 were statistically analyzed, and the differences in the collection time, circulating blood volume and the occurrence of adverse reactions to blood donation were analyzed when NGL XCF 3000 was used to collect autologous PRP among patients of different genders, ages and platelet counts. The differences in platelet content, red blood cell(RBC) contamination and white blood cell(WBC) residues in PRP products were analized. 【Results】 1) There were no significant differences in collection time, circulating blood volume and collection volume among patients of different genders, ages and platelet counts (P<0.05). 2) The contents of WBC, RBC and platelet were not significantly different between male and female patients after collection (P<0.05); 3) The WBC contents increased with the increase of age, and the WBC residue in the elder group[ 56 to78 years old, (0.64±0.41) ×10⁹/L] was significantly higher than that in the younger group[group 1,18 to 40 years old, (0.50±0.35)×10⁹/L], with significant difference was Statistically significant (P<0.05). 4) The residues of WBCs and RBCs in in low platelet group [group 1, (100-150)×10⁹/L] were higher than those in other platelet count groups, and the difference was Statistically significant (P<0.05), and the platelet count in this product was significantly lower than that in other platelet count groups (P<0.05). Conclusion The NGL XCF 3000 blood cell separator is safe and stable for PRP collection in patients with different genders, ages and platelet counts of (100-450)×10⁹/L, and the PRP products collected can meet clinical therapeutic needs.

【Objective】 To retrospectively analyze the effect of AB platelet secondary compatible transfusion on the efficacy of matched platelet transfusion. 【Methods】 A total of 2 276 cases of platelet transfusion in our hospital were selected from November 2020 to September 2021, including 2 068 ABO matched platelet transfusions and 206 AB platelet secondary compatible transfusions. 117 cases were selected for the first occasion of AB platelet secondary compatible transfusion. The controls were matched transfusion receipts before given AB platelet secondary compatible transfusion, and the experimental group was given matched transfusion after AB platelet secondary compatible transfusion (take the first, second and third transfusion as group 1, 2 and 3, respectively). The platelet count(Plt), platelet elevation (△Plt) and 24 h Plt correction increase index (CCI) of patients before and after platelet transfusion were used as observation indexes to analyze the effect of AB type mis-matched platelet transfusion on the efficacy of matched platelet preventive transfusion by gender, blood type and disease type. 【Results】 Among the 2 276 platelet transfusions, and the △Plt of matched platelet transfusions was significantly higher than that of AB type secondary compatible transfusions, with the mean values at (14±15)×10⁹/L and (11±14)×10⁹/L(P<0.05). The △Plt of female patients was higher than that of male patients (15±16)×10⁹/L vs (13±14)×10⁹/L(P < 0.05). The △Plt values of MDS, NHL and MM were (9±14) ×10⁹/L, (10±12) ×10⁹/L and (8±11) ×10⁹/L, respectively, which were significantly lower than the average value (P < 0.05). For 117 cases of AB platelet secondary compatible transfusion: the Δ Plt was significantly lower than that of the control group and the experimental group, respectively at (9±12) ×10⁹/L, (13±13) ×10⁹/L and (13±12) ×10⁹/L(P<0.05). The effective rate of 24 h CCI was 52.14%, significantly lower than the control group and experimental group (71.59% vs 71.83%), P<0.05; no significant difference was noticed in △ Plt value and 24 h CCI between the experimental group and the control group(P>0.05). The △Plt of the experimental group 3 was significantly lower compared with the experimental group 1, (10±13) ×10⁹/L vs (14±12) ×10⁹/L, P<0.05, and the effective rate decreased from 73.12%(68/93)to 58.70%(27/46), P>0.05. No significant difference in △Plt and the number of effective 24 h CCI cases was found between the experimental group and the control group in terms of gender, blood type and disease type (P>0.05). 【Conclusion】 The △Plt and the effective rate of AB platelet secondary compatible transfusion were lower than those of matched platelet transfusions, and has no significant effect on short term(less than 10 days) matched platelet transfusion.

Acidosis, regardless of hypoxia involvement, is recognized as a chronic and harsh tumor microenvironment (TME) that educates malignant cells to thrive and metastasize. Although overwhelming evidence supports an acidic environment as a driver or ubiquitous hallmark of cancer progression, the unrevealed core mechanisms underlying the direct effect of acidification on tumorigenesis have hindered the discovery of novel therapeutic targets and clinical therapy. Here, chemical-induced and transgenic mouse models for colon, liver and lung cancer were established, respectively. miR-7 and TGF-β2 expressions were examined in clinical tissues (n = 184). RNA-seq, miRNA-seq, proteomics, biosynthesis analyses and functional studies were performed to validate the mechanisms involved in the acidic TME-induced lung cancer metastasis. Our data show that lung cancer is sensitive to the increased acidification of TME, and acidic TME-induced lung cancer metastasis via inhibition of miR-7-5p. TGF-β2 is a direct target of miR-7-5p. The reduced expression of miR-7-5p subsequently increases the expression of TGF-β2 which enhances the metastatic potential of the lung cancer. Indeed, overexpression of miR-7-5p reduces the acidic pH-enhanced lung cancer metastasis. Furthermore, the human lung tumor samples also show a reduced miR-7-5p expression but an elevated level of activated TGF-β2; the expressions of both miR-7-5p and TGF-β2 are correlated with patients' survival. We are the first to identify the role of the miR-7/TGF-β2 axis in acidic pH-enhanced lung cancer metastasis. Our study not only delineates how acidification directly affects tumorigenesis, but also suggests miR-7 is a novel reliable biomarker for acidic TME and a novel therapeutic target for non-small cell lung cancer (NSCLC) treatment. Our study opens an avenue to explore the pH-sensitive subcellular components as novel therapeutic targets for cancer treatment.