Objective:To investigate the anti-inflammatory activity of alcohol extract of Polyrhachis dives(PDAEs)against rheumatoid arthritis(RA)in vitro and in vivo.Methods:In vivo and in vitro anti-RA inflammatory response of PDAEs was investigated using a rat model of collagenous arthritis induced by bovine type Ⅱ collagen and an LPS-induced RAW264.7 cell inflammatory model.Results:PDAEs inhibited the polarization of M1 type macrophages in vivo and in vitro,reduced expressions of TNF-α,IL-1β,IL-6,iNOS,and promoted polarization of M2 type macrophages,enhanced expressions of anti-inflammatory cytokines,such as IL-10 and TGF-β,so as to achieve the anti-inflammatory effect.The experiments in vivo also showed that PDAEs had the immunomodulatory effect,the potential mechanism may be the regulation of Th17/Treg balance by regulating the expression of PD-1 and TGF-β,thus correcting the over-strong autoimmune response.Conclusion:PDAEs may reduce the inflammatory reaction of RA through anti-inflam-matory and immunomodulatory effects.

AIM:This study aims to investigate the sensitizing effects of lycorine(Lyc)in combination with cisplatin(Cis)on human osteosarcoma cells and to explore the underlying mechanisms of action.METHODS:Human osteosarcoma cell lines MG63,HOS,and cisplatin-resistant HOS/DDP cells were utilized to evaluate the effects of Lyc and cisplatin,both alone and in combination,on cell viability using the CCK8 assay.The clonogenic assay was performed to assess cell proliferation capacity,while the scratch assay evaluated the drugs' effects on cell migration.Reactive oxygen species(ROS)levels were measured using a ROS assay kit,and changes in intracellular glutathione(GSH)levels were assessed with a GSH/oxidized glutathione(GSSG)assay kit.The mitochondrial membrane potential was analyzed via JC-1 staining to determine the drugs' effects on mitochondrial function.Intracellular iron(Ⅱ)content changes were detected us-ing FerrOrange,a fluorescence probe,and cellular malondialdehyde(MDA)levels were measured using an MDA assay kit.RT-qPCR was employed to evaluate the expression levels of key genes related to ferroptosis,and Western blot analysis was conducted to detect changes in the protein expression levels of Yes-associated protein 1(YAP1),acyl-CoA synthetase long-chain family member 4(ACSL4),glutathione peroxidase 4(GPX4),heme oxygenase-1(HO-1),and transferrin re-ceptor(TFRC).RESULTS:Both Lyc and cisplatin effectively inhibited the proliferation of human osteosarcoma cells.Notably,the combination of Lyc and cisplatin led to a more substantial reduction in cell viability,proliferation,and migra-tion abilities in MG63,HOS,and HOS/DDP cells compared to cisplatin alone.Additionally,this combination significant-ly increased ROS levels while decreasing GSH content,indicating mitochondrial damage and elevated iron(Ⅱ)and MDA levels.RT-qPCR results revealed that the combination treatment more significantly downregulated ferroptosis-promoting genes and upregulated ferroptosis-inhibiting genes compared to cisplatin treatment alone(P<0.05).Western blot results showed a slight decrease in GPX4 protein expression following Lyc and cisplatin treatment,while expression levels of YAP1,TFRC,ACSL4,and HO-1 were significantly increased(P<0.05).CONCLUSION:Lyc enhances the sensitivi-ty of MG63,HOS,and HOS/DDP cells to cisplatin by promoting ferroptosis through the YAP1/TFRC signaling pathway.

Objective To establish orthotopic and ectopic pancreatic cancer models in C57BL/6N mice with normal immune function using in vivo imaging technology for visual characterization.Methods Orthotopic and ectopic pancreatic cancer models were established in Kunming mice by injecting a small volume of cell suspension containing firefly luciferase-expressing Panc02-luciferase pancreatic cancer cells into the head of the pancreas or the right axillary region.In vivo imaging technology was used to optimize the modeling method and timing in Kunming mice.Subsequently,the same method was applied to C57BL/6N mice using wild-type Panc02 pancreatic cancer cells to establish orthotopic and ectopic pancreatic cancer models with intact immune function.Key parameters,including body weight,inoculation positive rate,tumor growth time,tumor volume,and pathological characteristics across different organs,were compared be-tween the orthotopic and ectopic models in C57BL/6N mice to evaluate the applicability of these models.Results Both the small animal in vivo imaging experiments in Kunming mice and the tumor growth observation in C57BL/6N mice demonstrated that the construction periods for orthotopic and ectopic pancreatic cancer models were 20 days,with survival rates exceeding 90%.The inoculation positive rates in C57BL/6N mice were 92.3%for the orthotopic model and 78.6%for the ectopic model.On day 20 post-inoculation,the tumor volumes were(117.04±109.56)mm3 for the orthotopic model and(155.68±168.73)mm3 for the ectopic model,indicating high model success rates and consistent tumor growth.HE staining revealed pathological mitotic figures and poorly differentiated tumor tissues in both models of C57BL/6N mice,with no evidence of metastasis to other organs.Conclusions Orthotopic and ectopic pancreatic cancer models in immu-nocompetent mice were successfully developed in this study,mimicking early-stage pancreatic cancer characteristics.These models pro-vide a reliable platform for screening early diagnostic biomarkers and evaluating therapeutic interventions for pancreatic cancer.

Objective To investigate the diagnostic value of endoscopic sign of light blue crest(LBC)capsuling papillary lesion(LCPL)for high-risk intestinal metaplasia(IM).Methods A total of 314 patients(352 biopsy specimens)who underwent endoscopic examination and biopsy in Department of Gastroenterology of Army Medical Center of PLA from January 2021 to June 2023 were recruited,and HE and HID-AB staining(the golden standard of high-risk IM)were apllied to detect the histological types and IM types.The samples were subsequently divided into chronic inflammation group,low-risk IM group,high-risk IM group,well-differentiated intestinal-type gastric cancer group,and poorly-differentiated intestinal-type gastric cancer group.The positive rate of LCPL in each group and its diagnostic efficacy were analyzed based on endoscopic images of the biopsy sites.Logistic regression analysis was used to investigate the relationship between LCPL sign and high-risk IM,as well as the clinical and pathological features associated with LCPL sign.Receiver operating characteristic(ROC)curve was plotted to evaluate the diagnostic efficacy of LCPL for high-risk IM,using indicators such as sensitivity,specificity,Youden index and area under the curve(AUC).Results The positive rate of the LCPL sign in high-risk IM group was 75.70%,significantly higher than that of the other groups(all P<0.001).Logistic regression analysis showed that LCPL sign was significantly correlated with high-risk IM(OR=30.286,95%CI:13.528～67.804,P<0.001).When the sign was employed in diagnosing high-risk IM,the sensitivity was 69.84%,the specificity was 93.75%,the Youden's index was 0.636,and the AUC value was 0.818(95%CI:0.773～0.857).Besides sensitivity,all above parameters of LCPL sign showed significantly better diagnostic efficacy than those of traditional LBC sign,which is used as a sign for diagnosing IM(P<0.001).Moreover,recognition of LCPL sign was not easily affected by age(OR=1.130,95%CI:0.709～1.800,P=0.607),lesion site(Angular incisure:OR=2.360,95%CI:0.732～7.613,P=0.151;Autrum:OR=2.257,95%CI:0.756～6.744,P=0.145),and presence of peptic ulcers(OR=1.085,95%CI:0.208～5.652,P=0.923).Significantly,94.12%of positive and 66.94%of negative LCPL signs could be rapidly recognized within 3 s(OR=4.536,95%CI:1.372～14.997,P=0.013).Conclusion LCPL sign shows high efficacy and potential clinical application value for high-risk IM in gastric mucosa of endoscopic diagnosis.

Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering, which limit sequence and functional diversity, thereby constraining their therapeutic and application potential. Protein design tools have significantly advanced the creation of novel protein sequences, structures, and functions. However, further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability. Here, we extended an evolution-based design protocol to create a novel minibinder, BindHer, against the human epidermal growth factor receptor 2 (HER2). It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity. Radiolabeling experiments with ^99mTc, ⁶⁸Ga, and ¹⁸F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models, with minimal nonspecific liver absorption, outperforming scaffolds designed through traditional engineering. These findings highlight a new rational approach to automated protein design, offering significant potential for large-scale applications in therapeutic mini-protein development.

Objective To discuss the technical advantages of MRI image fusion technology in guiding percutaneous balloon compression(PBC)for trigeminal neuralgia(TN).Methods The clinical data of 13 patients with TN,who received MRI image fusion technology-guided PBC from November 2022 to July 2023,were retrospectively analyzed.The MRI images of the trigeminal nerve obtained one week before surgery were fused with the intraoperative DynaCT images of the skull base so as to simultaneously display the Meckel s cave and foramen ovale,and under the 3-D view the optimal puncture path was determined.The needle was positioned at the entry point of the skin,than the skin was cut open with a sharp surgical blade and the needle was inserted to the foramen ovale area to a predetermined depth.Lateral skull base fluoroscopy and DynaCT scan were used to check that the puncture needle tip was placed into the foramen ovale.Than the puncture needle was replaced by a fine guiding-needle and it was pushed into the Meckel's cave.Under the dual guidance of lateral fluoroscopy and fusion image,the balloon was push forward and was filled with iodine contrast media to compress the trigeminal ganglion within the Meckel's cave.After completion of the treatment,the balloon and puncture needle were removed and manual oppression was applied on the face to achieve hemostasis.Results Immediately after PBC,the pain was relieved in all patients.No permanent or serious complications occurred.One patient had a relapse 3 months after PBC and a second PBC procedure had to be carried out.No obvious pain recurrence was observed in the remaining patients during follow-up period.During the surgery,the mean number of foramen ovale puncturing was(1.31±0.46)times,the mean X-ray exposure time was(8.64±5.66)min,and the mean cumulative dose of X-ray was(570.29±257.15)mGy.After PBC,12 patients(92.31％)developed facial numbness and one patient(7.69％)developed facial pain,all of which were healed after treatment.Conclusion MRI image fusion technology can improve the visualization and accuracy of PBC procedure.It can also reduce the number of puncturing,decrease the radiation exposure dose,and improve the surgical ability of young doctors.Therefore,MRI image fusion technology should be further developed and applied in clinical practice.

Objective To establish orthotopic and ectopic pancreatic cancer models in C57BL/6N mice with normal immune function using in vivo imaging technology for visual characterization.Methods Orthotopic and ectopic pancreatic cancer models were established in Kunming mice by injecting a small volume of cell suspension containing firefly luciferase-expressing Panc02-luciferase pancreatic cancer cells into the head of the pancreas or the right axillary region.In vivo imaging technology was used to optimize the modeling method and timing in Kunming mice.Subsequently,the same method was applied to C57BL/6N mice using wild-type Panc02 pancreatic cancer cells to establish orthotopic and ectopic pancreatic cancer models with intact immune function.Key parameters,including body weight,inoculation positive rate,tumor growth time,tumor volume,and pathological characteristics across different organs,were compared be-tween the orthotopic and ectopic models in C57BL/6N mice to evaluate the applicability of these models.Results Both the small animal in vivo imaging experiments in Kunming mice and the tumor growth observation in C57BL/6N mice demonstrated that the construction periods for orthotopic and ectopic pancreatic cancer models were 20 days,with survival rates exceeding 90%.The inoculation positive rates in C57BL/6N mice were 92.3%for the orthotopic model and 78.6%for the ectopic model.On day 20 post-inoculation,the tumor volumes were(117.04±109.56)mm3 for the orthotopic model and(155.68±168.73)mm3 for the ectopic model,indicating high model success rates and consistent tumor growth.HE staining revealed pathological mitotic figures and poorly differentiated tumor tissues in both models of C57BL/6N mice,with no evidence of metastasis to other organs.Conclusions Orthotopic and ectopic pancreatic cancer models in immu-nocompetent mice were successfully developed in this study,mimicking early-stage pancreatic cancer characteristics.These models pro-vide a reliable platform for screening early diagnostic biomarkers and evaluating therapeutic interventions for pancreatic cancer.

AIM:This study aims to investigate the sensitizing effects of lycorine(Lyc)in combination with cisplatin(Cis)on human osteosarcoma cells and to explore the underlying mechanisms of action.METHODS:Human osteosarcoma cell lines MG63,HOS,and cisplatin-resistant HOS/DDP cells were utilized to evaluate the effects of Lyc and cisplatin,both alone and in combination,on cell viability using the CCK8 assay.The clonogenic assay was performed to assess cell proliferation capacity,while the scratch assay evaluated the drugs' effects on cell migration.Reactive oxygen species(ROS)levels were measured using a ROS assay kit,and changes in intracellular glutathione(GSH)levels were assessed with a GSH/oxidized glutathione(GSSG)assay kit.The mitochondrial membrane potential was analyzed via JC-1 staining to determine the drugs' effects on mitochondrial function.Intracellular iron(Ⅱ)content changes were detected us-ing FerrOrange,a fluorescence probe,and cellular malondialdehyde(MDA)levels were measured using an MDA assay kit.RT-qPCR was employed to evaluate the expression levels of key genes related to ferroptosis,and Western blot analysis was conducted to detect changes in the protein expression levels of Yes-associated protein 1(YAP1),acyl-CoA synthetase long-chain family member 4(ACSL4),glutathione peroxidase 4(GPX4),heme oxygenase-1(HO-1),and transferrin re-ceptor(TFRC).RESULTS:Both Lyc and cisplatin effectively inhibited the proliferation of human osteosarcoma cells.Notably,the combination of Lyc and cisplatin led to a more substantial reduction in cell viability,proliferation,and migra-tion abilities in MG63,HOS,and HOS/DDP cells compared to cisplatin alone.Additionally,this combination significant-ly increased ROS levels while decreasing GSH content,indicating mitochondrial damage and elevated iron(Ⅱ)and MDA levels.RT-qPCR results revealed that the combination treatment more significantly downregulated ferroptosis-promoting genes and upregulated ferroptosis-inhibiting genes compared to cisplatin treatment alone(P<0.05).Western blot results showed a slight decrease in GPX4 protein expression following Lyc and cisplatin treatment,while expression levels of YAP1,TFRC,ACSL4,and HO-1 were significantly increased(P<0.05).CONCLUSION:Lyc enhances the sensitivi-ty of MG63,HOS,and HOS/DDP cells to cisplatin by promoting ferroptosis through the YAP1/TFRC signaling pathway.

Objective:To investigate the anti-inflammatory activity of alcohol extract of Polyrhachis dives(PDAEs)against rheumatoid arthritis(RA)in vitro and in vivo.Methods:In vivo and in vitro anti-RA inflammatory response of PDAEs was investigated using a rat model of collagenous arthritis induced by bovine type Ⅱ collagen and an LPS-induced RAW264.7 cell inflammatory model.Results:PDAEs inhibited the polarization of M1 type macrophages in vivo and in vitro,reduced expressions of TNF-α,IL-1β,IL-6,iNOS,and promoted polarization of M2 type macrophages,enhanced expressions of anti-inflammatory cytokines,such as IL-10 and TGF-β,so as to achieve the anti-inflammatory effect.The experiments in vivo also showed that PDAEs had the immunomodulatory effect,the potential mechanism may be the regulation of Th17/Treg balance by regulating the expression of PD-1 and TGF-β,thus correcting the over-strong autoimmune response.Conclusion:PDAEs may reduce the inflammatory reaction of RA through anti-inflam-matory and immunomodulatory effects.

【Objective】 To evaluate the screening efficacy and practical value of the portable microfluidic biochemical analyzer in the detection of blood donors before blood donation. 【Methods】 Blood donor samples, clinical blood samples and constant quality control products were collected. Referring to the documents of ISO15189 and National Health Industry Standard, the precision and accuracy of hemoglobin (Hb) and alanine aminotransferase (ALT) were verified and compared with other detection systems. 【Results】 The MS200 biochemistry instrument has an intra-batch precision of 1.40% to 1.46%, inter-batch precision of 1.91% to 1.94%, and correctness bias of -0.9% to -1.3% for Hb test, and an intra-batch precision of 3.77% to 4.86%, inter-batch precision of 4.92% to 6.02%, and correctness bias of -3.0% to -4.8% for ALT test, which were within the range of quality requirements of industry standard. Comparison of Hb test results between MS200 biochemistry and Hb201 analyser on 1 189 peripheral blood samples from donors showed no statistically significant difference (P>0.05). 65 samples showed positive correlation between MS200 biochemistry and XS-900i automated hematology analyzer on Hb test results (R²=0.986, P=0.000). Correlation analysis of all the results of ALT detection by MS200 biochemical analyzer and AU480 biochemical analyzer in 1 065 samples showed a positive correlation (R²=0.965, P=0.000). The elevated ALT samples did not affect the Hb test results, and the samples with abnormal Hb value did not affect the ALT test results, with no interference between the two items in the detection. 【Conclusion】 The MS200 biochemical analyzer based on microfluidic technology has reliable methodological performance and can meet the need of pre-donation testing.