The traditional Chinese medicine （TCM） myocardial infarction （MI） unit is a standardized， regulated， and continuous integrated care unit guided by TCM theory and built upon existing chest pain centers or emergency care units. This unit emphasizes multidisciplinary collaboration and forms a restructured clinical entity without altering current departmental settings， offering comprehensive diagnostic and therapeutic services with full participation of TCM in the treatment of MI. Its core medical model is patient-centered and disease-focused， providing horizontally integrated TCM-based care across multiple specialties and vertically constructing a full-cycle treatment unit for MI， delivering prevention， treatment， and rehabilitation during the acute， stable， and recovery phases. Additionally， the unit establishes a TCM-featured education and prevention mechanism for MI to guide patients in proactive health management， reduce the incidence of myocardial infarction， and improve quality of life.

OBJECTIVE To investigate the inhibitory effect and mechanism of Modified qifang weitong granules on gastric cancer based on in vitro and in vivo experiments. METHODS Human gastric cancer HGC-27 cells were divided into the following groups: control group （treated with fetal bovine serum）， 10% drug-containing serum group， 15% drug-containing serum group， 20% drug-containing serum group， and 5-fluorouracil （5-Fu） group （positive control， 3.90 μg/mL）. After culturing the cells in each group with the corresponding serum/drug solution， their proliferation， migratory and invasive abilities， as well as the cell cycle， were assessed. Additionally， the expression levels of epithelial-mesenchymal transition （EMT）-related proteins [E-cadherin， N-cadherin， and vimentin] in the cells were measured. Logarithmic-phase HGC-27 cells were harvested and subcutaneously injected into the right axillary region of nude mice to establish a subcutaneous xenograft tumor model in nude mice. The successfully modeled tumor-bearing nude mice were randomly divided into model group， low-， medium- and high-dose groups of Modified qifang weitong granules （17.65， 35.29 and 70.58 g/kg， respectively）， and 5-Fu group （25 mg/kg）， with 5 mice in each group. After 14 days of treatment with the corresponding drugs in each group， the histopathological morphology of the tumor tissues in the nude mice was observed. Immunohistochemistry and Western blot assay were employed to detect the expression levels of EMT- related proteins in the tumor tissues of the nude mice. RESULTS In the cell experiment， compared with the control group， the cell proliferation rate， migration rate， number of invasive cells， as well as the expression levels of N-cadherin and vimentin proteins， and the percentage of cells in the G2/M phase were all significantly decreased/reduced in the 15% drug-containing serum group， 20% drug-containing serum group （P＜0.05）. Conversely， the percentage of cells in the G0/G1 phase and the expression level of E- cadherin protein were significantly increased （P＜0.05）. In animal experiment， compared with the model group， the high-dose group of Modified qifang weitong granules exhibited significantly reduced tumor mass and expression levels of N-cadherin and vimentin proteins in the tumor tissues of nude mice （P＜0.05）， while the expression level of E-cadherinprotein in the tumor tissues was significantly increased （P＜0.05）. Additionally， the tumor cells varied in size and showed extensive necrosis. CONCLUSIONS Modified qifang weitong granules effectively inhibit gastric cancer in both in vitro and in vivo models， and the mechanism of action is related to the suppression of EMT.

In general, bioassay-guided fractionation and isolation of bioactive constituents from botanical materials frequently ended up with the reward of a single compound. However, botanical materials typically exert their therapeutic actions through multi-pathway effects due to the intrinsic complex nature of chemical constituents. In addition, the content of bioactive compounds in botanical materials is largely dependent on humidity, temperature, soil, especially geographical origins, from which rapid and accurate identification of plant materials is pressingly needed. These long-standing obstacles collectively impede the deep exploitation and application of these versatile natural sources. To address the challenges, a new paradigm integrating biogravimetric analyses and machine learning-driven origin classification (BAMLOC) was developed. The biogravimetric analyses are based on absolute qHNMR quantification and in vivo zebrafish model-assisted activity index calculation, by which bioactive substance groups jointly responsible for the bioactivities in all fractions are pinpointed before any isolation effort. To differentiate origin-different botanical materials varying in the content of bioactive substance groups, principal component analysis, linear discriminant analysis, and hierarchical cluster analysis in conjunction with supervised support vector machine are employed to classify and predict production areas based on the detection of volatile organic compounds by E-nose and GC-MS. Expanding BAMLOC to Codonopsis Radix enables the identification of polyacetylenes and pyrrolidine alkaloids as the bioactive substance group for immune restoration effect and accurately determines the origins of plants. This study advances the toolbox for the discovery of bioactive compounds from complex mixtures and lays a more definitive foundation for the in-depth utilization of botanical materials.

Objective To compare the changes in biological indicators of human corneal epithe-lial(HCET)cells and rabbit corneas after exposure to different doses of ultraviolet B(UVB)radia-tion,so as to evaluate the impact of UVB radiation on corneal injury effects.Methods In cell exper-iment,HCET cells were divided into groups with radiation doses of 0,6,12,18,and 24 mJ/cm2.The effect of UVB radiation on HCET cell viability was detected using the CCK-8 assay,and the level of intracellular DNA damage was assessed by immunofluorescence.In the animal experiment,15 healthy New Zealand white rabbits(30 eyes)were randomly divided into groups with radiation doses of 0,1.35,2.16,4.32,and 6.48 J/cm2.The UVB exposure time for the radiation groups was 30 minutes per day for 3 consecutive days.Corneal injury was evaluated using methods such as slit-lamp microscopy,sodium fluorescein staining,central corneal thickness measurement,optical coherence tomography(OCT)imaging,and hematoxylin and eosin(HE)staining.Results Compared with the control group,cell viability in the radiation groups gradually decreased,and the level of DNA damage gradually increased with increasing radiation dose.As the radiation dose increased in the radiation groups,the degree of corneal opacity in rabbits gradually worsened,the central corneal area gradu-ally thickened,and OCT revealed high-intensity scattered light signals with the formation of shadow areas.Results from HE staining,immunohistochemistry,Western blot(WB),and sodium fluores-cein staining showed that the 1.35 J/cm2 group caused mild corneal injury,with damage reaching the corneal epithelial layer.In the 2.16 J/cm2 group,the corneal injury presented as dense punctate distribution,with damage extending from the epithelial layer to the superficial stroma.The number of ephrin type-A receptor 2(EphA2)protein-stained cells was relatively small,and the staining was light,showing a weak positive result.In the 4.32 J/cm2 and 6.48 J/cm2 groups,the corneal injury was irreversible,with damage gradually progressing from the corneal epithelial layer and superficial stroma to the endothelial layer.The number of EphA2 protein-stained cells was relatively large,and the staining was dark,showing a strong positive result.Conclusion This study comprehensively e-valuates the dose-dependent injury effects of UVB on HCET cells and New Zealand white rabbit cor-neas through cell and animal experiments.It elucidates that UVB radiation could induce corneal cell DNA damage,promote inflammatory responses,and trigger apoptosis by upregulating γ-phosphoryla-ted histone H2AX(γH2AX)and EphA2.The self-repair ability and process of corneal injury are preliminarily explored,providing a basis for further research on mechanisms of corneal injury caused by ultraviolet radiation and the development of protective drugs.

[Objective]To explore the expression of transcription factor KLF16 in nonalcoholic fatty liver disease(NAFLD)and its effect on lipid metabolism.[Methods]An animal model of NAFLD was constructed in mice induced by a high-fat diet.The mice were divided into normal diet group(ND)and high fat diet group(HFD).NAFLD cell model was constructed by primary mouse liver cells induced by oleic acid.The cells were divided into control group(Control group)and oleic acid induction group(OA group).Real-time fluorescence quantitative PCR(RT-qPCR)and Western blot were used to detect KLF16 expression in NAFLD animal and cell models.In vitro and in vivo models of KLF16 knockdown were constructed by injection of adeno-associated virus(AAV)into mouse tail veins and transient transfection of cell siRNA.Hematoxylin-eosin staining(HE)and other methods were used to detect changes in lipid deposition in NAFLD models be-fore and after KLF16 knockout.RT-qPCR was used to detect the expression of key genes of lipid metabolism in both cellu-lar and animal NAFLD models before and after KLF16 knockdown.Western blot assay was used to detect the expression of endoplasmic reticulum stress protein in NAFLD model before and after KLF16 knockdown.[Results]The expression level of KLF16 was up-regulated in HFD group and OA group,and lipid deposition was increased in OA group after KLF16 was depressed.There was no change in TC level in hepatocytes between groups(P>0.05),and TG level was increased in differ-ent degrees(P<0.05,P<0.001).At the same time,the change of KLF16 expression also caused the change of ER stress protein expression in OA group.[Conclusion]The transcription factor KLF16 may alleviate lipid deposition in nonalcoholic fatty liver disease by endoplasmic reticulum stress.

Lactic acid bacteria(LAB)were isolated and cultivated from the intestinal contents of rabbits by MRS-CaCO3 solid medium.Identification was achieved through morphological observa-tion,Gram staining,physiological and biochemical characterisation,16S rDNA sequence analysis,and ERIC-PCR analysis.Strains displaying typical Lactobacilli characteristics were exanimated for their biological characteristics,resistance properties,adherence capacity in vitro,colonization abili-ty in vivo,and safety profile.In this study,a total of four strains of Lactobacillus reuteri were iso-lated from rabbits,all of which exhibited typical biological characteristics of LAB.These strains demonstrated inhibitory effects on common pathogenic bacteria in the gastrointestinal tract,with the primary inhibitory substance being bacteriocin.Furthermore,they showed sensitivity to chlor-amphenicol,rifampicin,and erythromycin,and displayed a degree of tolerance to gastrointestinal conditions and high temperature.These stains were capable of successful colonization in rabbits with a higher degree of safety.This study lays a foundation for the development of LAB prepara-tions for the prevention and treatment of rabbit intestinal diseases.

Posttranslational modifications of antibody products affect their stability,charge distribution,and drug activity and are thus a critical quality attribute.The comprehensive mapping of antibody modifications and different charge isomers(CIs)is of utmost importance,but is challenging.We intended to quanti-tatively characterize the posttranslational modification status of CIs of antibody drugs and explore the impact of posttranslational modifications on charge heterogeneity.The CIs of antibodies were fraction-ated by strong cation exchange chromatography and verified by capillary isoelectric focusing-whole column imaging detection,followed by stepwise structural characterization at three levels.First,the differences between CIs were explored at the intact protein level using a top-down mass spectrometry approach;this showed differences in glycoforms and deamidation status.Second,at the peptide level,common modifications of oxidation,deamidation,and glycosylation were identified.Peptide mapping showed nonuniform deamidation and glycoform distribution among CIs.In total,10 N-glycoforms were detected by peptide mapping.Finally,an in-depth analysis of glycan variants of CIs was performed through the detection of enriched glycopeptides.Qualitative and quantitative analyses demonstrated the dynamics of 24 N-glycoforms.The results revealed that sialic acid modification is a critical factor ac-counting for charge heterogeneity,which is otherwise missed in peptide mapping and intact molecular weight analyses.This study demonstrated the importance of the comprehensive analyses of antibody CIs and provides a reference method for the quality control of biopharmaceutical analysis.

Head and neck squamous cell carcinoma (HNSCC) still lacks effective targeted treatment. Therefore, exploring novel and robust molecular targets is critical for improving the clinical outcome of HNSCC. Here, we reported that the expression levels of family with sequence similarity 64, member A (FAM64A) were significantly higher in HNSCC tissues and cell lines. In addition, FAM64A overexpression was found to be strongly associated with an unfavorable prognosis of HNSCC. Both in vitro and in vivo evidence showed that FAM64A depletion suppressed the malignant activities of HNSCC cells, and vice versa. Moreover, we found that the FAM64A level was progressively increased from normal to dysplastic to cancerous tissues in a carcinogenic 4-nitroquinoline-1-oxide mouse model. Mechanistically, a physical interaction was found between FAM64A and forkhead box protein M1 (FOXM1) in HNSCC cells. FAM64A promoted HNSCC tumorigenesis not only by enhancing the transcriptional activity of FOXM1, but also, more importantly, by modulating FOXM1 expression via the autoregulation loop. Furthermore, a positive correlation between FAM64A and FOXM1 was found in multiple independent cohorts. Taken together, our findings reveal a previously unknown mechanism behind the activation of FOXM1 in HNSCC, and FAM64A might be a promising molecular therapeutic target for treating HNSCC.
Animals ; Carcinogenesis ; Cell Line, Tumor ; Cell Proliferation ; Cell Transformation, Neoplastic ; Head and Neck Neoplasms/genetics* ; Homeostasis ; Mice ; Squamous Cell Carcinoma of Head and Neck

Objective: To explore the effect of hyperbaric oxygen preconditioning with different frequency on the survival rate of flap and ischemia-reperfusion injury in rats after transplantation, and to explore the best preconditioning conditions to improve the survival rate of rat flaps after transplantation. Methods: Thirty-six Sprague Dawley rats were randomly divided into four groups according to the random number table method, 9 groups in each group.Four groups of rats were pretreated with hyperbaric oxygen pretreatment for 0, 2, 4, and 6 days before the operation, control group, pretreatment 2 d group, pretreatment 4 d group, and pretreatment 6 d group. Taking the midline of the back of the rat as the axis, an ultra-long random flap with a pedicle at the tail end and about 1 cm from the superior iliac spine was designed and cut to a size of 10.0 cm×2.5 cm. The survival of the flaps in each group was observed and the final survival area and survival rate of the flaps were measured on the 7th day after surgery. On the 7th day after operation, the tissue was taken at a distance of 5 cm from the pedicle, and the histopathology was observed; The content of superoxide dismutase (SOD) and malondialdehyde (MDA) in flap tissue was detected by immunohistochemistry, and the expression rate of positive cells in each group was calculated. Immunofluorescence was used to detect the expression of interleukin-6 (IL-6) in the flap tissue. Results: On the 7th day after the operation, the survival area and survival rate of the transplanted flaps in the hyperbaric oxygen pretreatment group were significantly higher than those in the control group (P<0.05), and the pretreatment 4 d and 6 d groups were significantly higher than the pretreated 2 d group (P<0.05), but there was no significant difference between the pretreated 4 d group and the 6 d group (P=0.095). Pathological observation on the 7th day after operation showed that there was some necrosis in the control group, the vascular cells in the pretreated 2 d group had more vascular structures, and more neovascularization was observed in the pretreated 4 d group. The inflammatory cells were the least in the 6 d pretreatment group, and the neovascularization was the same as the pretreatment 4 d group. The absorbance A value of SOD in the control group was 0.009 7±0.000 3, and the positive expression rate was 20%, which was significantly lower than that in the hyperbaric oxygen pretreatment group. The difference was statistically significant (P<0.05). However, the absorbance A value of MDA in the control group was 0.055 1±0.003 0, and the positive expression rate was 55%, which was significantly higher than that in the hyperbaric oxygen pretreatment group. The difference was statistically significant (P<0.05). Among them, the SOD absorbance A value of the pretreated 2 d group was 0.023 8±0.003 0, and the positive expression rate was 30%, which was lower than the pretreatment 4 d group (absorbance A value 0.046 9±0.003 0, positive expression rate 35%) and 6 d group (absorbance A value 0.047 2±0.003 6, positive expression rate 40%), The MDA absorbance A value of the pretreated 2 d group was 0.037 2±0.003 2, and the positive expression rate was 30%, which was higher than the pretreatment 4 d group (absorbance A value 0.014 7±0.002 4, positive expression rate 5%) and 6 d group (absorbance A value 0.017 0±0.001 8, positive expression rate 10%), the difference was statistically significant (P<0.05). However, there was no significant difference in the expression of SOD and MDA between the pretreated 4 d group and the pretreated 6 d group (P>0.05). The expression of IL-6 in the hyperbaric oxygen pretreatment group was significantly lower than that in the control group (absorbance A value 44.937 0±0.594 0), the difference was statistically significant (P<0.05). The absorbance A value in the pretreated 2 d group was 41.698 0±0.724 0, which was significantly higher than the pretreatment 4 d group (absorbance A value 34.049 0±0.323 0) and 6 d group (absorbance A value 33.524 0±0.639 0). The difference was statistically significant (P<0.05). There was no significant difference in the expression of the pretreated 4 d group compared with the 6 d group (P=0.068). Conclusions Hyperbaric oxygen preconditioning can significantly promote the survival of rat flaps after transplantation. Preoperative hyperbaric oxygen preconditioning once daily for 4 consecutive days, can enhance the tolerance of flap tissue to ischemia and anoxia and reduce tissue ischemia-reperfusion injury.

To develop a clinical diagnosis technique for bluetongue virus infection, we established serotype-specific methods to detect serotype 4 of bluetongue virus (BTV-4). Two monoclonal antibodies (mAbs) against VP2 protein of BTV-4, named 4A-1G7 and 4B-1B6, were used as competitive antibodies in the competitive enzyme-linked immunosorbent assays (C-ELISA). We detected 50 negative serum samples from sheep, goats and cattle by C-ELISA. The cut-off values of 4A-1G7 and 4B-1B6 mAbs were 49% and 40%, respectively. The results of the sensitivity, specificity and repeatability by detecting standard positive serum, were consistent with the general standard of Office International Des Epizooties. Furthermore, serum samples of BTV-4, BTV-18 and BTV-20 infection could be screened out through the combined C-ELISAs by 4A-1G7 and 4B-1B6 mAbs. Thus, this technique may diagnose BTV-4, BTV-18 and BTV-20 infections.