ObjectiveTo investigate the mechanism of modified Si Junzitang and Shashen Maidong Tang ［Yiqi Yangyin Jiedu prescription （YQYYJD）］ in enhancing the sensitivity of cisplatin in epidermal growth factor receptor tyrosine kinase inhibitor （EGFR-TKI）-resistant lung adenocarcinoma cells based on aerobic glycolysis. MethodsThe effects of different concentrations of YQYYJD （0， 2， 3， 4， 5， 6， 7， 8 g·L^-1） and cisplatin （0， 3， 6， 9， 12， 15， 18， 21， 24， 27 mg·L^-1） on the proliferation and activity of PC9/GR cells were detected by the cell counting kit-8 （CCK-8） assay after 24 hours of intervention. The half-maximal inhibitory concentration （IC₅₀） for PC9/GR cells was calculated to determine the concentrations used in subsequent experiments. PC9/GR cells were divided into blank group （complete medium）， YQYYJD group （5 g·L^-1）， cisplatin group （12 mg·L^-1）， and combined group （YQYYJD 5 g·L^-1 + cisplatin 12 mg·L^-1）. After 24 hours of intervention， cell viability was measured using CCK-8 assay. Cell proliferation was assessed by colony formation assay， and cell migration was evaluated by scratch and Transwell assays. Glucose consumption， lactate production， and adenosine triphosphate （ATP） levels were measured by colorimetric assays. The expression levels of glycolysis-related proteins， including hexokinase 2 （HK2）， phosphofructokinase P （PFKP）， pyruvate kinase M2 （PKM2）， lactate dehydrogenase A （LDHA）， glucose transporter 1 （GLUT1）， and monocarboxylate transporter 4 （MCT4）， were determined by Western blot. ResultsBoth YQYYJD and cisplatin inhibited the viability of PC9/GR cells in a concentration-dependent manner. The IC₅₀ of PC9/GR cells for YQYYJD and cisplatin were 5.15 g·L^-1 and 12.91 mg·L^-1， respectively. In terms of cell proliferation， compared with the blank group， the cell survival rate and the number of colonies formed in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group showed a further significant reduction in cell survival rate and colony formation （P<0.01）. In terms of cell migration， compared with the blank group， the cell migration rate and the number of cells passing through the Transwell membrane in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group exhibited a further significant reduction in cell migration rate and the number of cells passing through the Transwell membrane （P<0.01）. In terms of glycolysis， compared with the blank group， glucose consumption， lactate production， and ATP levels in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group showed a further significant reduction in glucose consumption， lactate production， and ATP levels （P<0.05）. Compared with the blank group， the protein expression levels of HK2， PFKP， PKM2， and LDHA in the YQYYJD， cisplatin， and combined groups were significantly decreased （P<0.01）. The combined group showed a further significant reduction in the expression levels of these proteins compared with the YQYYJD and cisplatin groups （P<0.01）. No significant differences were observed in the protein expression levels of GLUT1 and MCT4 among the groups. ConclusionYQYYJD can synergistically inhibit the proliferation and migration of PC9/GR cells and enhance their sensitivity to cisplatin. The mechanism may be related to the downregulation of the expression of glycolysis-related rate-limiting enzymes， including HK2， PFKP， PKM2， and LDHA， thereby inhibiting glycolysis.

Objective To explore the safety of Yttrium-90 resin microsphere selective internal radiation therapy (⁹⁰Y-SIRT) on malignant liver tumors. Methods A retrospective analysis was conducted on 64 patients with malignant liver tumors who underwent ⁹⁰Y-SIRT from February 2023 to November 2024 at Weifang People’s Hospital. The clinical characteristics of the patients and the occurrence of adverse reactions after treatment were analyzed to assess the safety of ⁹⁰Y-SIRT. Results Among the 64 patients, there were 52 males (81.25%) and 12 females (18.75%); the average age was (56.29±11.08) years. Seven patients (10.94%) had tumors with maximum diameter of less than 5 cm, 38 patients (59.38%) had tumors with maximum diameter of 5-10 cm, and 19 patients (29.68%) had tumors with maximum diameter of greater than 10 cm. There were 47 cases (73.44%) of solitary lesions and 17 cases (26.56%) of multiple lesions; 53 cases (82.81%) were primary liver cancers and 11 cases (17.19%) were metastatic liver cancers. Of the 64 patients, 63 successfully completed the Technetium-99m macroaggregated albumin (^99mTc-MAA) perfusion test and received the ⁹⁰Y-SIRT; one patient received ⁹⁰Y-SIRT after the second ^99mTc-MAA perfusion test due to a work error. The most common adverse reactions included grade 1 alanine aminotransferase (ALT) elevation in 26 cases (40.62%) and grade 2 in 2 cases (9.37%), grade 1 aspartate aminotransferase (AST) elevation in 27 cases (42.18%) and grade 2 in 7 cases (10.93%); grade 1 nausea in 17 cases (26.56%) and grade 2 in 6 cases (9.37%); grade 1 abdominal pain in 12 cases (18.75%), grade 2 in 5 cases (7.81%), and grade 3 in 1 case (1.56%); grade 1 vomiting in 11 cases (17.18%), grade 2 in 5 cases (7.81%), and grade 3 in 1 case (1.56%). Conclusion The adverse reactions of ⁹⁰Y-SIRT for treating malignant liver tumors are mild, indicating good safety.

Dry eye is a chronic ocular surface disease caused by multiple factors. It is caused by the instability of tear film and the imbalance of the microenvironment of ocular surface, and may be accompanied by ocular surface inflammation, damage, and abnormal nerve sensation. The instability of tear film is its core characteristic. Mucin is an important component of the tear film and plays a role in stabilizing the tear film. The reduction of its secretion and the change of its structure lead to the occurrence and development of dry eye. The intracellular Ca2+ signal is the key to controlling the secretion of water and enzymes by exocrine glands. A decrease in the Ca2+ signal can cause dry eye. Conjunctival goblet cells are the main cells that secrete mucin. By activating the intracellular PLC-IP3-Ca2+ pathway, RyRs pathway, cAMP signaling pathway, P2X receptor, BLT1 and ChemR23 receptors, cholinergic receptor, and ALX signaling pathway, the content of Ca2+ can be increased, and the replenishment of mucin granules can be accelerated, thereby relieving the symptoms of dry eye. The Ca2+ signaling pathway may be an important target for the treatment of dry eye. This article reviews the role of mucin in dry eye and the influence of the Ca2+ signal on the secretion of mucin by conjunctival goblet cells.

The Janus kinase/signal transducers and activators of transcription (JAK-STAT) control natural killer (NK) cells development and cytotoxic functions, however, whether long non-coding RNAs (lncRNAs) are involved in this pathway remains unknown. We found that miR155HG was elevated in activated NK cells and promoted their proliferation and effector functions in both NK92 and induced-pluripotent stem cells (iPSCs)-derived NK (iPSC-NK) cells, without reliance on its derived miR-155 and micropeptide P155. Mechanistically, miR155HG bound to miR-6756 and relieved its repression of JAK3 expression, thereby promoting the JAK-STAT pathway and enhancing NK cell proliferation and function. Further investigations disclosed that upon cytokine stimulation, STAT3 directly interacts with miR155HG promoter and induces miR155HG transcription. Collectively, we identify a miR155HG-mediated positive feedback loop of the JAK-STAT signaling. Our study will also provide a power target regarding miR155HG for improving NK cell generation and effector function in the field of NK cell adoptive transfer therapy against cancer, especially iPSC-derived NK cells.

Objective To prepare glucose transporter 1(Glut1)-targeted doxorubicin(DOX)-loaded liposomes(doxorubicin/polyphyllin H-liposomes,DOX/ppH-LPs)using polyphyllin H(ppH)instead of cholesterol as the liposomal membrane material,and to investigate their in vitro synergistic anti-tumor activity against non-small cell lung cancer(NSCLC).Methods DOX/ppH-LPs were prepared using thin-film hydration,and the formulation was optimized by single-factor investigation.The optimized DOX/ppH-LPs were characterized for morphology,particle size,polydispersity index(PDI),and zeta potential with transmission electron microscopy(TEM)and dynamic light scattering(DLS).Drug loading DL%was determined by high-performance liquid chromatography(HPLC).The storage stability was evaluated by observing in PBS at 4℃for 7 d,and the serum stability was observed in DMEM containing 10%fetal bovine serum(FBS)at 37℃for 48 h.In vitro drug release was studied in PBS at pH 7.4 and pH 5.0 values,respectively.Human NSCLC A549 cells were subjected as the model,MTT assay was performed to detect the proliferation inhibition by DOX/ppH-LPs at different concentrations(0.5,5.0,15.0 μg/mL)and the control group(ppH+DOX/LPs,a physical mixture of free ppH and DOX-loaded liposomes).Fluorescence microscopy was used to observe cellular uptake of DOX/ppH-LPs and DOX/LPs(containing 5 μg/mL DOX)at 15 min and 2 h.Live/dead cell staining was applied to assess apoptosis/necrosis induced by formulations(15 μg/mL DOX)after 48 h incubation.Transwell assay was conducted to evaluate inhibitory effect on cell migration and invasion,and the targeting property and in vitro synergistic anti-NSCLC activity of DOX/ppH-LPs were then comprehensively evaluated.Results The optimal formulation of DOX/ppH-LPs was determined as hydration temperature at 50℃,6 mg DOX,2 mg ppH,and 24 mg lecithin.The prepared DOX/ppH-LPs were in spherical shape,uniform distribution,and at an average particle size of 145.13±22.14 nm,a PDI of 0.15±0.05,a zeta potential of-23.92±1.73 mV,and a DL of 10.13±0.71%for DOX and(1.22±0.21)%for ppH.DOX/ppH-LPs maintained stable particle size,PDI,and exhibited significantly unchanged zeta potential after storage in PBS at 4℃for 7 d or incubation in DMEM containing 10%FBS at 37℃for 48 h,demonstrating excellent physical and serum stability.Both liposomes showed slow release at pH 7.4 value,while drug release was significantly accelerated at pH 5.0 value(P<0.05),indicating pH-sensitive release characteristics.MTT assay revealed that DOX/ppH-LPs exerted significantly stronger cytotoxicity against A549 cells than the ppH+DOX/LPs control group(P<0.05).Compared with ppH+DOX/LPs,DOX/ppH-LPs showed remarkably enhanced cellular uptake in A549 cells(P<0.05),with more DOX localized in the nucleus.Live/dead cell staining showed that at the same DOX concentration(15 μg/mL),the proportion of apoptotic/necrotic cells induced by DOX/ppH-LPs was significantly higher than that of the DOX/LPs control group.Transwell assay demonstrated that there were significantly less cells migrating and invading through the membrane in the DOX/ppH-LPs group than the ppH+DOX/LPs group.Conclusion Glut1-targeted doxorubicin-loaded liposomes(DOX/ppH-LPs)constructed by substituting cholesterol with ppH can target NSCLC cells,significantly enhance the in vitro synergistic anti-NSCLC activity of DOX and ppH.

Objective: To investigate the prevalence and influencing factors of latent tuberculosis infection (LTBI) among students close contacts diagnosed with pulmonary tuberculosis in Baoshan District, Shanghai, so as to provide the basis for the prevention and control of pulmonary tuberculosis among students. Methods: Pulmonary tuberculosis cases identified among students or teaching staff were selected as index cases through the Surveillance System of Chinese Disease Prevention and Control Information System, school reports, notification of Centers for Disease Control and Prevention from April 2021 to November 2023, and student close contacts in their schools were selected as research subjects. Demographic information, lifestyle and Mycobacterium tuberculosis test results were collected through questionnaires surveys and pulmonary tuberculosis screening. LTBI was defined as a positive Mycobacterium tuberculosis test result with the exclusion of active pulmonary tuberculosis. The influencing factors for LTBI among student close contacts were identified using a multivariable logistic regression model. Results: A total of 1 212 student close contacts were included, with 651 males and 561 females, resulting in a gender ratio of 1.16︰1. The mean age was (18.48±4.33) years. Among them, 32 cases were detected with LTBI, yielding a detection rate of 2.64%. Higher LTBI detection rates were observed among students who shared the same dormitory with pulmonary tuberculosis cases (9.26%), attended private schools (5.54%), lived on campus (3.54%), and obtained meals through take-out services (6.52%). Multivariable logistic regression analysis showed that sharing the same dormitory with pulmonary tuberculosis cases (OR=3.604, 95%CI: 1.256-10.338), attending a private school (OR=2.327, 95%CI: 1.083-5.003), and three meals a day through canteens (OR=9.561, 95%CI: 2.029-45.057) or through take-out services (OR=19.222, 95%CI: 3.528-104.732) were associated with a higher risk of LTBI. Conclusion The close contacts of students with pulmonary tuberculosis in Baoshan District are at risk of LTBI, which is mainly affected by the degree of contact with tuberculosis cases, the strength of school epidemic prevention and control, and students' eating habits.

ObjectiveTo explore the effects of different emulsion mixtures and emulsification methods on the inflammation severity in an experimental autoimmune uveitis （EAU） model in rats， and to analyze the characteristics of the current EAU model. MethodEAU was induced in Lewis rats by subcutaneous injection of interphotoreceptor retinoid-binding protein （IRBP） 1177-1191 emulsified with Freund's complete adjuvant （CFA）， with or without intraperitoneal injection of pertussis toxin （PTX）. Slit lamp examination， HE staining， and optical coherence tomography were used to evaluate factors affecting EAU modeling， including different doses of the emulsion mixture （IRBP1177-1191， PTX， and inactivated Mycobacterium tuberculosis） and four different emulsification methods. The classification， characteristics， modeling methods， advantages， and disadvantages of EAU animal models were summarized and analyzed based on the clinical diagnostic criteria and syndrome characteristics of chronic uveitis in both traditional Chinese medicine （TCM） and western medicine， to evaluate the consistency between TCM and western medical syndromes. ResultIncreasing the dose of inactivated M. tuberculosis and antigen peptide in the emulsion mixture exacerbated the anterior segment inflammation in EAU rats. Increasing the injection of PTX also exacerbated anterior segment inflammation and increased retinal thickness in EAU rats. The severity of the EAU model was closely related to the emulsification method used. All four emulsification methods successfully induced EAU in rats. Comparatively， the ultrasonic cell disruptor and T10 basic disperser achieved successful emulsification in a short time. The degree of emulsification of the mixture also influenced the severity of the EAU model in rats. The existing EAU animal model shows a high degree of consistency with western medical diagnoses and the main ocular syndromes in TCM. ConclusionIRBP1177-1191， PTX， inactivated M. tuberculosis， and emulsification methods can affect the severity of the EAU model through different pathways. The existing EAU animal models can simulate the clinical characteristics of western medicine well but lack the etiology， pathogenesis， and syndrome characteristics of TCM. Therefore， it is necessary to construct an EAU animal model that combines disease and syndrome characteristics.

Objective To investigate the expression of long non-coding RNA(lncRNA)LINC02695 in human retinal microvascular endothelial cells(HRMECs)in high glucose(HG)environment and its effect on the proliferation,migration and neovascularization of HRMECs.Methods HRMECs was divided into four groups:the normal glucose(NG)group(5.5 mmol/L),the HG group(30.0 mmol/L),the HG+LINC02695 silenced group(HG+si-LINC02695),and the HG+silenced control group(HG+si-NC).Real-time quantita-tive fluorescent PCR(qPCR)was used to detect the expression of LINC02695 and vascular endothelial growth factor(VEGF)mRNA in HRMECs of each group.The cell proliferation of each group was measured by Cell Counting Kit-8(CCK-8)method.The migration ability of cells in each group was detected by Transwell as-say.The tube forming ability of cells in each group was detected by tube forming experiment.Results The qPCR results showed that compared with the NG group,LINC02695 and VEGF were highly expressed in the HG group(P＜0.05).Compared with the HG group,VEGF mRNA expression level in the HG+si-LINC02695 group was significantly decreased(P＜0.05).The results of CCK-8 experiment showed that the proliferation ability of the HG group was significantly enhanced compared with the NG group(P＜0.05).Compared with the HG group,the cell proliferation ability of the HG+si-LINC02695 group was significantly decreased(P＜0.05).The results of Transwell experiment showed that the cell migration ability of the HG group was significantly increased compared with the NG group(P＜0.05).Compared with the HG group,the cell migration ability of the HG+si-LINC02695 group was significantly decreased(P＜0.05).The results of tube formation experiment showed that compared with the NG group,the tube formation ability of the HG group was significantly increased(P＜0.05).Compared with the HG group,canalization ability of cells in the HG+si-LINC02695 group was significantly decreased(P＜0.05).Conclusion LINC02695 may be involved in promoting the proliferation,migration and angiogenesis of HRMECs induced by HG.

BACKGROUND:After the internal fixation of cannulated screws in femoral neck fractures,because the affected limb is often unable to bear weight in the short term and the implants with high stiffness have a stress shielding effect on the fracture end,it is easy to cause osteoporosis of the affected limb and changes in the biomechanical distribution of the proximal femur,the incidence of osteonecrosis of the femoral head is high after surgery.At present,few studies have been conducted on the biomechanical effects of osteoporosis at the proximal end of the femur occurring after femoral neck fracture surgery on femoral neck fracture treated with cannulated screws. OBJECTIVE:Using finite element analysis,to investigate the biomechanical effects of osteoporosis occurring after femoral neck fracture surgery on femoral neck fracture treated with cannulated screws and explore the role of biomechanical factors in osteonecrosis of the femoral head. METHODS:Based on the obtained CT scan data of the femur in a patient with a femoral neck fracture,a proximal femoral model for internal fixation for femoral neck fracture was established by Mimics 19.0,3-Matic,UG 11.0,Hypermesh 14.0,and Abaqus software.One finite element model of the proximal femur without osteoporosis and three finite element models of the proximal femur with osteoporosis were analyzed using Abaqus software.The stress,contact pressure,displacement peak and cloud map under different components of the four models were measured and analyzed,and the internal stress changes and distribution of the femoral head were compared and analyzed. RESULTS AND CONCLUSION:The stresses and contact pressures of the femoral head and lower anterior cannulated screws varied more with the degree of osteoporosis.The peak displacement of the four models increased slowly with the degree of osteoporosis.By one-way analysis of variance,there was no significant effect of the degree of osteoporosis on the peak stress,contact pressure,and displacement of the different components.The internal stress distribution of the femoral head changed with the degree of osteoporosis.Changes in the biomechanical environment of the proximal femur have an important impact on osteonecrosis of the femoral head.

Objective:To obtain hepatitis B virus capsid-like particles (CLPs) displaying B-cell linear epitopes of SARS-CoV-2 spike protein and evaluate their immunogenicity.Methods:Four recombinant plasmids expressing fusion proteins (M1-HBc, S1-57-HBc, S14P5-HBc and S21P2-HBc) were constructed by separately replacing codon of alanine at position 80 of hepatitis B virus core protein (HBc) with four genes coding for four B-cell linear epitopes (M1, S1-57, S14P5 and S21P2). These four recombinant proteins were expressed in E. coli BL21 Star (DE3) strains. The expression products were identified using SDS-PAGE, Western blot and native agarose gel electrophoresis (NAGE). CLPs were purified by sucrose density gradient ultracentrifugation, verified for antigenicity by Western blot and used to immunize BALB/c mice. Serum antibody titers were detected by ELISA. Results:The recombinant fusion proteins M1-HBc and S1-57-HBc self-assembled into M1-CLP and S1-57-CLP. The titer of antibody against S1-57 polypeptide in S1-57-CLP-immunized mouse serum approached 1∶1 000 000.Conclusions:Hepatitis B virus CLPs displaying SARS-CoV-2 M1 or S1-57 linear epitopes are successfully expressed in a prokaryotic system and purified. S1-57-CLP has good immunogenicity. This study provides a new idea for the development of novel diagnostic reagents and vaccines for SARS-CoV-2.