Objective To screen the targets related to the metabolic enzymes involved in the cholesterol synthesis pathway that inhibits the polarization of macrophages towards M1 phenotype,and verify the intervention effects and underlying mechanisms in colorectal cancer cells.Methods Mouse colorectal cancer MC38 cells were divided into control group(si-NC)and experimental groups(the expression of enzymes in cholesterol synthesis pathway was interfered with siRNA for corresponding targets).RT-qPCR was used to detect the mRNA levels of corresponding targets in MC38 cells after transfection.After peritoneal macrophages were extracted from male C57BL/6 mice(6 weeks old,weighing 13～18 g),the macrophages were then treated with the conditioned media of MC38 cells transfected with different siRNAs for 48 h.RT-qPCR was employed to detect the mRNA levels of IL-1β,IL-6 and TNF-α in the macrophages so as to evaluate the effect of the culture media on the M1 polarization.MC38 cells were divided into control groups(OE-NC and sh-NC),farnesyl-diphosphate farnesyltransferase 1(FDFT1)overexpression group(OE-FDFT1)and FDFT1 knockdown group(sh-FDFT1).RT-qPCR was applied to detect the mRNA expression of FDFT1,and Western blotting was conducted to measure the protein level of FDFT1.C57BL/6 mice were subjected randomly to construct a subcutaneous tumor-bearing model and a model of intraperitoneal metastatic tumor(n=5)respectively.The growth of tumor mass was then measured.Flow cytometry was used to observe the proliferation and apoptosis of MC38 cells,and Trans well assay to detect migration ability of MC38 cells.Five C57BL/6 macrophage-depleted mice(established with injection of clodronate liposome suspension through tail vein)received intraperitoneal implantation to construct a metastasis model,and then the obtained tumor masses were then weighted.Results Compared with MC38 cells after si-NC transfection,the mRNA levels of corresponding targets in MC38 cells in the experimental groups were significantly reduced(P＜0.05).Significant increases were found in the mRNA levels of IL-1β,IL-6 and TNF-α of the macrophages with FDFT1 interference than the control cells(P＜0.05).There were no statistical differences in the proliferation,apoptosis and migration of MC38 cells in the control group(OE-NC and sh-NC)and the cells of the FDFT1 overexpression group and FDFT1 knockdown group(P＞0.05).In both the subcutaneous tumor-bearing model and the model of intraperitoneal metastatic tumor,the mass weight was significantly heavier in the OE-FDFT1 group than the OE-NC group(P＜0.01),and was notably smaller in the sh-FDFT1 group than the sh-NC group(P＜0.01).For the macrophage-depleted mouse tumor model,no remarkable change was observed in the tumor weight between the OE-FDFT1 group and the OE-NC group as well as the sh-FDFT1 group and the sh-NC group.Conclusion FDFT1,the metabolic enzyme in the cholesterol synthesis pathway of colorectal cancer tumor cells,is a potential target for tumor immunotherapy targeting macrophages,which promotes tumor progression by regulating macrophages.

Objective To compare the long-term efficacy of intradiscal injection of autologous platelet-rich plasma(PRP)and intradiscal injection of autologous PRP combined with pulsed radiofrequency(PRF)therapy for discogenic back pain(DBP).Methods Prospective inclusion of 98 DBP patients who received treatment at Hai'an People's Hospital from July 2019 to December 2023 was conducted.The patients were randomly divided into study group(receiving PRP combined with PRF,n=50)and control group(receiving PRP only,n=48).The preoperative and the postoperative 1-,3-,6-,and 12-month baseline data,visual analog scale(VAS)score,Roland-Morris Disability Questionnaire(RMDQ)score,Leeds Neurological Symptoms and Signs Score(LANSS),European Society for Quality of Life Five Dimensional Questionnaire(EQ-5D)score,and the use of non-steroidal anti-inflammatory and analgesic drugs were compared between the two groups.Results There was no statistically significant differences in the baseline data and the incidence of intraoperative and postoperative adverse reactions between the two groups(both P＞0.05).At 12 months after surgery,the VAS,RMDQ,LANSS,EQ-5D scores,and the use of non-steroidal anti-inflammatory and analgesic drugs in all patients were significantly better than those before surgery(all P＜0.05).The postoperative 3-,6-,and 12-month VAS,RMDQ,LANSS,EQ-5D scores,and the analgesic dosage in the study group were significantly better than those in the control group(all P＜0.05),and the percentages of patients with pain relief greater than 50％and greater than 80％were also significantly higher than those in the control group(all P＜0.01).Conclusion for the treatment of DBP,both CT-guided intradiscal injection of autologous PRP and CT-guided intradiscal injection of autologous PRP combined with PRF can significantly improve pain,body function,and quality of life in DBP patients,and significantly reduce the use of non-steroidal anti-inflammatory and analgesic drugs.The efficacy of autologous PRP combined with PRF is remarkably better than that of autologous PRP alone.

OBJECTIVE To investigate the mechanism of Yifei xuanfei jiangzhuo formula （YFXF） against vascular dementia （VD）. METHODS The differentially expressed genes of YFXF （YDEGs） were obtained by network pharmacology. High-risk genes were screened from YDEGs by using the nomogram model. The optimal machine learning models in generalized linear， support vector machine， extreme gradient boosting and random forest models were screened based on high-risk genes. VD model rats were established by bilateral common carotid artery occlusion， and were randomly divided into model group and YFXF group （12.18 g/kg， by the total amount of crude drugs）， and sham operation group was established additionally， with 6 rats in each group. The effects of YFXF on behavior （using escape latency and times of crossing platform as indexes）， histopathologic changes of cerebral cortex， and the expression of proteins related to the secreted phosphoprotein 1 （SPP1）/phosphoinositide 3-kinase （PI3K）/protein kinase B （aka Akt） signaling pathway and the mRNA expression of SPP1 in cerebral cortex of VD rats were evaluated. RESULTS A total of 6 YDEGs were obtained， among which SPP1， CCL2， HMOX1 and HSPB1 may be high-risk genes of VD. The generalized linear model based on high-risk genes had the highest prediction accuracy （area under the curve of 0.954）. Compared with the model group， YFXF could significantly shorten the escape latency of VD rats， significantly increase the times of crossing platform （P＜0.05）； improve the pathological damage of cerebral cortex， such as neuronal shrinkage and neuronal necrosis； significantly reduce the expressions of SPP1 protein and mRNA （P＜0.05）， while significantly increase the phosphorylation levels of PI3K and Akt （P＜0.05）. CONCLUSIONS VD high-risk genes SPP1， CCL2， HMOX1 and HSPB1 may be the important targets of YFXF. YFXF may play an anti-VD role by down-regulating the protein and mRNA expressions of SPP1 and activating PI3K/Akt signaling pathway.

The "flipped classroom" teaching model practiced in the teaching ofcurriculum was introduced. Firstly, the roles and responsibilities of teachers were clarified, indicating teachers provided examples and lectures, and a comprehensive assessment system was established. Secondly, the "flipped classroom" teaching model was split into online learning, classroom learning and offline learning. Online learning aimed at forming a study report by a wide search of relevant information, which was submitted to teachers for review and assessment. Classroom learning was designed to communicate study ideas among students and teachers. Offline learning was intended to revise and improve the study report and refined learning methods. Lastly, the teaching practice effects of "flip classroom" were evaluated by comprehensive rating and questionnaire assessment, which assessed the overall performance of students and overall levels of paper; the learning ability was enhanced, and the interest and motivation of learning were also improved. Therefore, "flipped classroom" teaching mode was suitable for the curriculum of, and could be recommended into the teaching practice of related curriculum of acupuncture and tuina.

OBJECTIVETo evaluate the differences in the autophagy activity of alveolar macrophages between patients with different stages of coal workers' pneumoconiosis (CWP).

METHODSA total of 116 coal workers were investigated in the field. Their lung lavage fluid was collected and purified to obtain alveolar macrophages. The morphological characteristics of autophagy were observed by transmission electron microscopy. The expression of autophagy marker (LC3) and autophagy regulators (Beclin1, mTOR, and p-mTOR) was measured by Western blot. The autophagy activity of alveolar macrophages was compared between dust-exposed subjects and patients with stage I, II, and III CWP.

RESULTSThe autophagy activity of alveolar macrophages differed between patients with different stages of CWP, according to transmission electron microscopy. Patients with stage II CWP had significantly higher protein expression of LC3 II/I and Beclin1 in pulmonary macrophages than those with stage ICWP (P < 0.05); patients with stage III CWP had significantly lower protein expression of LC3 II/I and Beclin1 in pulmonary macrophages than those with stage II CWP (P < 0.05), but had significantly higher protein expression of LC3 II/I and Beclin1 than those with stage I CWP (P < 0.05); patients with stage II CWP had a significantly higher protein expression of Beclin1 than the dust-exposed subjects (P < 0.05). Patients with stage II CWP had significantly lower expression of mTOR and p-mTOR in pulmonary macrophages than the dust-exposed subjects and those with stage I CWP (P < 0.05), while patients with stage III CWP had significantly higher expression of mTOR and p-mTOR than those with stage II CWP (P < 0.05).

CONCLUSIONThe autophagy activity of alveolar macrophages varies between patients with different stages of CWP.

Anthracosis ; pathology ; Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; Beclin-1 ; Biomarkers ; Bronchoalveolar Lavage Fluid ; Coal ; Coal Mining ; Dust ; Humans ; Macrophages, Alveolar ; pathology ; Membrane Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism ; Occupational Exposure ; Pneumoconiosis ; pathology

Adenoma ; pathology ; Colonic Neoplasms ; pathology ; Humans ; Intestinal Mucosa ; pathology

Objective To investigate the significance of superoxide dismutase (SOD) and homocysteine (HCY) in patients with acute cerebral infarction and acute cerebral hemorrhage .Methods Serum levels of SOD and HCY were detected in 251 acute cere‐bral infarction patients ,204 acute cerebral hemorrhage patients and 485 healthy subjects .Results Compared with healthy subjects , serum levels of SOD in acute cerebral infarction patients and acute cerebral hemorrhage patients were lower ,but serum levels of HCY were higher (P<0 .05) .There were significant differences of serum SOD and HCY levels among different types of cerebral infarction patients (P<0 .05) .Conclusion There could be important value for determine of SOD and HCY in patients with acute cerebral infarction and acute cerebral hemorrhage .

Objective To investigate the effects of CD147/MMP-9 pathway on early left ventricular remodeling Methods 30 healthy eight-week male SHR were divided into 3 groups (n = 10 for each group). SHR group received tail vein injections of normal saline weekly; CD147 group received CD147 of 600 ng·kg-1 weekly; and CD147+DOX group received CD147 of 600 ng/kg weekly and intragastric administration of DOX ( doxycycline ) of 30 mg/kg daily . 10 healthy eight-week male Wistar-Kyoto rats (WKY group) were treated as SHR group. Echocardiography, myocardial sections microscopy examination (HE and VG stain), and Western blot (for assessing levels of MMP-9, TIMP-1, CD147, and collagen I and Ⅲin myocardial tissues) were performed on day 56. Left ventricular weight index (LVWI)was measured and calculated. Collagen volume fractions (CVF) were obtained by image analysis. Results As compared with WKY group , levels of CD147 , MMP-9 , and MMP-9/TIMP-1 were lower but TIMP-1 and collagenⅠand Ⅲ were significantly higher in SHR group. The abundance of CD147 and MMP-9 protein and the ratio of MMP-9/TIMP-1 were obviously increased in CD147 group than in SHR group (P < 0.05). Levels of CD147, MMP-9, and MMP-9/TIMP-1 did no differ between CD147+DOX group and CD147 group. LVWI and contents of collagenⅠand Ⅲ were obviously declined in CD147 group as compare with SHR group. Cardiomyocyte hypertrophy , partial myocardial fibre rupture , myocyte dissolution and fuzzy myocardial fibre boundaries , more abundant of collagen fibers, and higher CVF were found in SHR group. Cardiac fibrosis was significantly improved after CD147 intervention, but the action was suppressed as DOX was administrated simultaneously. Conclusions Early ventricular remodeling may be involved in the inhibition of CD147/MMP-9 pathway in SHR. Input of CD147 to upregulate the pathway can improve the remodeling.

Objective:To differentiate human embryonic stem cells ( hESCs ) into keratinocytes ( K-hESCs) and analyse the expression characteristics of biomarkers of K-hESCs.Methods: The hESCs of line H9 were seeded on matrigel in mTeSR1 medium.The hESCs were directly differentiated into kerati-nocytes in epithelial differentiation medium with bone morphogenetic protein 4, retinoic acid and N2 sup-plement.The karyotype of K-hESCs was analyzed, comparing the gene expression differences of K-hESCs with human gingival epithelial cells (HGECs), human immortalized oral epithelial cells (HIOECs) and HaCaT by Real-time PCR.Molecular characteristics of the cell differentiation were observed throughout the process by immunocytochemical techniques.Results:H9-hESCs were successfully differentiated into the cells that exhibited characteristics of keratinocytes in epithelial differentiation medium.The karyotype of K-hESCs was 46, XX; and the keratinocyte gene p63 expression in K-hESCs was significantly lower than that in HaCaT ( P<0.05) , but there was no significant difference of p63 expression in K-hESCs, comparing with that in HGECs and HIOECs ( P >0.05 ) .Conclusion: H9-hESCs could be directly differentiated into K-hESCs.The gene expression of K-hESCs was similar to that of epithelial cells in the early stage of monolayer cells differentiation with high proliferative activity.

c-Met receptor tyrosine kinase plays an important role in signaling pathways including cell proliferation, metabolism as well as tumorigenic growth, migration and angiogenesis. c-Met has emerged as an attractive target for cancer therapy. Moreover, the interactive cross-talk between c-Met signaling and several other signaling pathways underlies a key effect for resistance of anti-cancer drugs. Thus, multi-target inhibitors become a new approach to cancer therapy. This paper introduces the c-Met signaling pathway and the resistance of kinase inhibitors caused by the cross-talk between c-Met and other membrane receptors and then will reviews the progress of single-target and multi-target c-Met inhibitors.