OBJECTIVE To predict and validate the mechanisms of Chaiqi yigan granules （CQYG） improving hepatocellular carcinoma （HCC）. METHODS The signaling pathways of CQYG intervention in HCC were predicted using network pharmacology. A mice model of transplanted hepatocellular carcinoma was established by injecting H22 hepatoma cells into the axilla. Successfully modeled mice were randomly divided into model group （normal saline）， sorafenib group （positive control， 50 mg/kg）， and CQYG low-， medium- and high-dose groups （24.83， 49.66， 99.32 g/kg）， with 10 mice in each group. Mice in each group were administered the corresponding drug solution or normal saline intragastrically， once a day， for 14 consecutive days. After last administration， pathological morphological changes in the tumor tissues of mice were observed in each group. Immunohistochemical staining was performed to detect the expression of the nuclear proliferation antigen Ki-67 in tumor tissues of mice. Western blot assay was used to measure the expression of proteins related to epithelial-mesenchymal transition （EMT） [N-cadherin， E-cadherin， Vimentin， matrix metalloproteinase 7 （MMP7）] and the mitogen-activated protein kinase （MAPK） signaling pathway [p38 MAPK， phosphorylated p38 MAPK， c-Jun N-terminal kinase （JNK）， phosphorylated JNK， extracellular regulated protein kinase 1/2 （ERK1/2）， phosphorylated ERK1/2] in tumor tissue of mice. RESULTS Network pharmacology analysis revealed that metabolic pathways， pathways in cancer， and the MAPK signaling pathway were key signaling pathways through which CQYG exert their anti-hepatocellular carcinoma effects. In animal experiments， the tumor tissues of mice in the model group exhibited dense tumor cells and vigorous growth. Compared with model group， CQYG high-dose group showed a decreased density of tumor cells in the tumor tissues of mice. Moreover， the expression levels of Ki-67， N-cadherin， MMP7 and Vimentin proteins， along with the phosphorylation levels of ERK1/2 and JNK proteins， were all significantly reduced （ P ＜0.05）. The expression level of E-cadherin protein was significantly increased （ P ＜0.05）， the phosphorylation level of p38 MAPK protein was increased， the difference was not statistically significant （ P ＞0.05）. CONCLUSIONS CQYG can inhibit EMT by regulating the MAPK signaling pathway， thereby suppressing tumor cell invasion and metastasis and ultimately exerting a therapeutic effect in improving HCC.

Objective To optimize the culture process of insect cell-baculovirus expression vector system(BEVS) in order to further improve the protein expression of recombinant hepatitis E virus-like particles(HEV-LPs), and to scale up and verify the process.Methods Serum-free suspension culture of Sf9 insect cells was carried out and a 7 L bioreactor was established through shaking flask cascade amplification. The parameters such as viable cell density, culture temperature and recombinant virus inoculation ratio were optimized when the cells were inoculated with recombinant baculovirus. The cell growth status was observed and the expression level of the target protein was analyzed. The target protein was detected by SDSPAGE, Western blot, high-performance liquid chromatography(HPLC) and transmission electron microscope(TEM). The morphology of HEV-LPs and protein yield were integrated, and the suitable expression parameters of recombinant HEV-LPs were determined to prepare three batches of 7 L recombinant hepatitis E vaccine.Results The optimal parameters for the cultivation of insect cells in a 7 L bioreactor were as follows: stirring 90 r/min, pH(6. 2 ± 0. 1), optimal incubation temperature 27 ℃, cell density of recombinant baculovirus inoculation 6. 0 × 10~6 cells/mL, and dissolved oxygen 90%. Under this condition, the relative molecular mass of the expressed HEV-LPs target protein was about 58 000, which had a specific binding with HEV mouse monoclonal antibody. The final yield was 60-70 mg/L, with the purity of more than 95%. Spherical particles with a diameter of about 20 nm were observed under TEM. The HEV-LPs recombinant protein prepared by the optimized parameters had stable yield and uniform particle morphology.Conclusion The culture and expression process of recombinant hepatitis E vaccine was successfully optimized, and the stability of the amplification process was verified, which lays a foundation for the development of recombinant hepatitis E vaccine production process.

Microglia, originating from primitive macrophages in the yolk sac, serves as both immune system defenders and regulators of homeostasis. These cells exhibit two primary polarization states: conventionally activated(M1)and alternatively activated(M2). The polarization of microglia plays a crucial role in influencing inflammatory disorders, metabolic imbalances, and neural degeneration. This process is implicated in various aspects of ocular diseases, especially age-related macular degeneration(AMD), including inflammation, oxidative stress and pathological angiogenesis. The distinct functional phenotypes of microglia impact disease progression and prognosis. Thus, regulating the polarization or functional phenotype of microglia at different stages of AMD holds promise for personalized therapeutic approaches. This comprehensive review outlines the involvement of microglia polarization in both physiological and pathological conditions, emphasizing its relevance in AMD. The discussion underscores the potential of polarization as a foundation for personalized treatment strategies for AMD.

Objective To observe the effect of extracorporeal shockwave therapy (ESWT) in regulating endothelial cell phosphatase and tensin homolog deleted on chromosome ten (PTEN) expression on lower ex-tremity vascular lesion and its possible mechanism.Methods Twenty-four 2-month-old healthy male SD rats were randomly divided into the three groups:control group (group A),diabetes angiopathy group (group B),diabetes angiopathy+ESWT group (group C).The group B and C were fed with high fat and high sugar and intraperitoneally injected with streptozotocin 60 mg/kg to establish the rat model of diabetes vascular lesion. The group C received ESWT at 1 week (T1),2 weeks (T2),3 weeks (T3) and 4 weeks (T4) after modeling,and the blood stream velocity of rat femoral artery vascular lesion area and vascular internal diameter were measured at T4 by ultrasound.At the end of ESWT,the rats were immediately killed for taking their femoral arteries and gastrocnemius.The structures of the femoral arteries in each group were observed under electron microscopy.Western blot was used to detect the expression levels of PTEN,PI3K and Akt proteins,while qRT-PCR was used to detect the expression levels of PTEN mRNA.Immunofluorescence was used to detect the expression level of CD31 in gastrocnemius muscle .Results The peak systolic flow velocity and end-dias-tolic flow velocity of femoral artery at T4 in group B and C were significantly lower than those in group A (P<0.05),but group C was higher than group B (P<0.05).The internal diameter of femoral artery had no statistical difference among 3 groups (P>0.05).The PTEN expression level in group B and group C was sig-nificantly lower than that in group A (P<0.05),while group C was higher than group B (P<0.05).The ex-pression levels of PI3K and Akt in group B were higher than those in group A (P<0.05),and group C was lower than group B (P<0.05).The PTEN mRNA expression level in group B and group C was significantly lower than that in group A (P<0.05),but group C was higher than group B (P<0.05).Under electron mi-croscopy,it was observed that after ESWT,the endothelial cell damage in group C was obvious when com-pared with group B.The CD31 expression level in group B and group C was significantly lower than that in group A (P<0.05),but group C was higher than group B (P<0.05).Conclusion ESWT could improve the vascular function,increase the peak velocity during systolic period of femoral artery in diabetes rats and im-prove the microvessel density of gastrocnemius muscle by up-regulating PTEN in lower extremity artery and down-regulating PI3K and Akt in diabetes rats.

Mucin-type O-glycosylation is one of the most common post-translational modifications in proteins,capable of altering protein conformation and biological functions.It plays a crucial role in biological processes such as cell signaling,cell adhesion,and immune responses.Polypeptide N-acetylgalactosaminyltransferase 3(GALNT3),as the initiating enzyme of mucin-type O-glycosylation,is of paramount importance in maintaining the homeostasis of human cells and tissues.Dysfunction of GALNT3 has been found to play a role in various diseases,such as calcium-phosphorus metabolism disorders and atherosclerosis.Additionally,GALNT3 is abnormally expressed in several types of tumors,including colorectal cancer,lung cancer,and ovarian cancer.Its expression is associated with the clinical pathological features of patients and poor prognosis,making it a potential biomarker for early tumor diagnosis and prognosis evaluation.Further research shows that GALNT3 can both regulate glycosylation levels to reduce adhesion between tumor cells and activate multiple metabolism-related pathways,promoting tumor cell invasion and metastasis.This review summarizes the role of GALNT3 in the development of malignant tumors and discusses the prospects and challenges of developing anti-tumor drugs targeting GALNT3.

Objective:To investigate the influencing factors of prolonged postoperative ileus (PPOI) in patients undergoing pancreaticoduodenectomy (PD) during hospitalization.Methods:The data of 339 patients underwent PD admitted to the Department of Hepatobiliary Surgery of the Second Hospital of Hebei Medical University from January 2018 to September 2023 were retrospectively analyzed, including 204 males and 135 females, aged (60.6±11.2) years. Among the 339 patients, 112 (33.0%) had pancreatic tumors, 94 (27.7%) had Vater ampullary tumors, 82 (24.2%) had common bile duct tumors, and 51 (15.0%) had duodenal tumors. A total of 339 patients with PPOI were included in the PPOI group ( n=43) and those without PPOI were included in the control group ( n=296). The two groups were compared in terms of age, PD operation (open or laparoscopic), gastrojejunostomy (retrocolic or antecolic gastrojejunostomy), grade B or C pancreatic fistula, hypokalemia, and postoperative use of patient-controlled intravenous analgesia (PCIA). The index comparing P<0.05 between the two groups was further included in the multivariate logistic regression analysis to analyze the influencing factors of PPOI in PD patients. Results:There were statistically significant differences in age >70 years, PD operation, gastrojejunostomy, grade B or C pancreatic fistula, hypokalemia, and postoperative use of PCIA between the two groups (all P<0.05). Multivariate logistic regression analysis showed grade B or C pancreatic fistula ( OR=3.17, 95% CI: 1.48-6.82), open surgery ( OR=2.90, 95% CI: 1.35-6.24), retrocolic gastrojejunostomy ( OR=2.47, 95% CI: 1.23-4.95), postoperative usage of PCIA ( OR=2.61, 95% CI: 1.21-5.62), age >70 years ( OR=2.47, 95% CI: 1.71-5.19) had a high risk of PPOI during postoperative hospitalization (all P<0.05). Conclusion:Postoperative grade B or C pancreatic fistula, open surgery, retrocolic gastrojejunostomy (compares with antecolic gastrojejunostomy), postoperative using PCIA, and age >70 years are independent risk factors for PPOI in patients undergoing PD during postoperative hospitalization.

Humans ; Parkinson Disease/diagnosis* ; Machine Learning

Humans ; Atrial Fibrillation/surgery* ; Left Atrial Appendage Closure ; Heart Atria/surgery* ; Catheter Ablation/methods* ; Atrial Appendage/surgery* ; Treatment Outcome

Objective:To explore any effect of pulsed electromagnetic field (PEMF) stimulation on intervertebral disc degeneration (IDD).Methods:Forty Sprague-Dawley rats were randomly divided into a control group, an IDD model group, a PEMF group and an observation group. An IDD model was induced in all except those in the control group. Both the PEMF and observation groups were given PEMF stimulation, while the latter was additionally injected with the A2AR agonist CGS-21680. Eight weeks after the modelling any pathological changes in the morphology of the rats′ intervertebral disc tissues were evaluated using saffron solid green staining. The expression of A2AR, cyclic adenosine phosphate (cAMP), protein kinase A (PKA), cysteine aspartate proteolytic enzyme-3 (Caspase-3), type II collagen (COL-II) and matrix metalloproteinase-3 (MMP3) in the intervertebral discs were evaluated.Results:The nucleus pulposus had shrunk, while fibrous tissues and chondrocytes had increased in the IDD model group. In the observation group the nucleus pulposus was intact and of basically normal shape. A2AR mRNA and protein levels were higher in the intervertebral disc tissue of the model group than among the control group, on average, while the levels in the observation group were significantly higher than in the other groups. In the PEMF and observation groups cAMP and PKA mRNA were significantly higher than in the IDD model group. The p38 MAPK and P-P38 MAPK levels of the IDD model group and its average P-P38 MAPK/p38 MAPK ratio were significantly higher than in the control group. In the PEMF and observation groups those indices had decreased to varying degrees, with those of the observation group significantly lower than among the model and PEMF groups on average, except for the p38 MAPK values. Caspase-3 and its mRNA were significantly higher in the model group than in the control group, on average, and those values were significantly lower in the PEMF and observation groups than in the IDD model group. The average MMP3 contents of the IDD model group had increased significantly compared with the control group, while the Col-Ⅱ level had decreased significantly. Compared with the IDD model group, the MMP3 level had decreased but Col-Ⅱ expression had increased in both the PEMF and observation groups, with significant differences between the IDD model and observation groups.Conclusions:The activation of the p38 MAPK signaling pathway by inflammatory factors to induce apoptosis is one of the important reasons for the aggravation of IDD lesions. PEMF combined with A2AR agonists can activate the A2AR/cAMP/PKA signaling pathway, inhibit p38 MAPK phosphorylation, reduce apoptosis of nucleus pulposus cells, and relieve IDD damage.

Objective:To explore the potential clinical value of 99Tc m-methoxyisobutylisonitrile(MIBI) SPECT/CT muscle imaging in the diagnosis of cervical dystonia (CD). Methods:From January 2021 to April 2022, 50 patients with CD (14 males, 36 females; age (45.8±12.5) years) who were treated in Second Affiliated Hospital of Soochow University were prospectively included. The 99Tc m-MIBI SPECT/CT muscle imaging results of 400 pieces of muscle (bilateral sternocleidomastoid, musculus scapulae, splenius capitis and musculus trapezius; each of 100 pieces) in 50 patients were analyzed and divided into the dystonic muscle group and normal muscle group according to the electromyography (EMG). Toronto western spasmodic torticollis rating scale (TWSTRS) score, SUV max and target-to-background ratio (TBR) of single superficial cervical muscle and total cervical muscle, and EMG diagnosis results were obtained before botulinum toxin injection. ROC curves of SUV max and TBR of dystonic muscles were constructed to determine AUCs and the difference was compared by Delong test. Differences of SUV max and TBR between 2 groups were analyzed by Mann-Whitney U test. Spearman rank correlation analysis was used to analyze the correlation of SUV max, TBR and TWSTRS scores of total cervical muscle. Results:There were 205 pieces of muscle in dystonic muscle group and 195 pieces of muscle in normal muscle group. The uptake of 99Tc m-MIBI in dystonic muscle was significantly increased in CD patients, and the non-whole uptake of 99Tc m-MIBI was increased in some dystonic muscles, which was manifested as uneven uptake of the whole muscle. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of visual analysis were 95.12%(195/205), 75.90%(148/195), 85.75%(343/400), 85.58%(195/242) and 93.67%(148/158), respectively. There were significant differences of SUV max (1.74(1.42, 2.12) vs 0.92(0.81, 0.99)) and TBR (2.55(1.92, 3.27) vs 1.44(1.22, 1.73)) between the dystonic muscle group and the normal muscle group ( z value: -15.29, -12.69, both P<0.001). The diagnostic efficiency of SUV max in dystonic muscle was better than TBR (AUC: 0.942 vs 0.867; z=5.03, P<0.001). SUV max, TBR and TWSTRS score in the neck muscles of patients with CD showed positive correlation ( rs values: 0.44, 0.45, both P<0.001). Conclusion:99Tc m -MIBI SPECT/CT muscle imaging is a good diagnostic method for dystonic muscle in patients with CD.