OBJECTIVES: The application of photodynamic therapy in solid tumors has attracted increasing attention in recent years, and the efficiency of photosensitizers is a crucial determinant of therapeutic efficacy. This study aims to evaluate the cytotoxic effects of a novel photosensitizer, PEG-MTPABZ-PyC, in photodynamic therapy against gastric cancer cells. METHODS: Gastric cancer MKN45 cells were treated with PEG-MTPABZ-PyC. A high-content live-cell imaging system was used to assess the cellular uptake kinetics and subcellular localization of the photosensitizer. The cytotoxic effects of PEG-MTPABZ-PyC-mediated photodynamic therapy were examined using the cell counting kit-8 (CCK-8) assay and flow cytometry, while the intrinsic cytotoxicity of the photosensitizer alone was verified by the CCK-8 assay. Intracellular reactive oxygen species (ROS) generation after photodynamic therapy was detected using 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA). RESULTS: PEG-MTPABZ-PyC alone exhibited no cytotoxicity toward MKN45 cells, indicating excellent cytocompatibility. The compound efficiently entered cells within 6 hours and localized predominantly in lysosomes. Upon light irradiation, PEG-MTPABZ-PyC-mediated photodynamic therapy induced significant cytotoxicity compared with the control group (P<0.05) and generated abundant intracellular ROS. CONCLUSIONS The novel photosensitizer PEG-MTPABZ-PyC demonstrates potent photodynamic cytotoxicity against gastric cancer cells, showing promising potential for further development in gastric cancer photodynamic therapy.
Humans ; Stomach Neoplasms/drug therapy* ; Photochemotherapy/methods* ; Photosensitizing Agents/pharmacology* ; Cell Line, Tumor ; Polyethylene Glycols/chemistry* ; Reactive Oxygen Species/metabolism* ; Mesoporphyrins/pharmacology*

Objective To observe the effects of ice-water swimming on pathological changes in model gouty rats,and investigate the relevant regulatory mechanism of the purinergic P2X7R receptor.Methods Male Sprague Dawley rats were divided into normal(NORM)and experimental groups including gouty control(GC),ice-water swimming(IWS),and Brilliant Blue G(BBG,a P2X7R inhibitor)groups.Rats in the experimental groups were modeled to simulate hyperuricemia and gouty arthritis by inhibiting uric acid metabolism combined with the Coderre method.Rats in the ice-water swimming group were treated with 5 min of endurance swimming in an ice-water mixture at a depth of about 0.5 m for 0 h and 12 h after modeling by the Coderre method,while rats in the BBG group were injected intraperitoneally with BBG solution once after modeling.Ankle swelling index was calculated using a formula.Serum uric acid levels were detected by uricase assay,and serum levels of the inflammatory factors interleukin(IL)-1β,1L-6,and tumor necrosis factor(TNF)-αwere detected by enzyme-linked immunosorbent assay.The pathological status of the ankle joints was examined by hematoxylin and eosin staining.P2X7R and NLRP3 protein expression levels in synovial tissue were detected by Western blot and immunohistochemistry,respectively.Results Serum uric acid levels and the ankle joint swelling index were significantly higher in the experimental groups compared with the normal group(P＜0.05 or P＜0.01),and the synovial tissues showed different degrees of inflammatory infiltration.The ankle swelling index was significantly higher in the ice-water swimming group compared with the gouty control group at 12 h(P＜0.05).Serum IL-1β,IL-6,and TNF-α levels(P＜0.01)and P2X7R and NLRP3 protein levels in synovial tissues were all significantly elevated(P＜0.05).Histopathology showed that the cartilage surface was broken and the synovial tissue showed severe hyperplasia and erosion,accompanied by numerous inflammatory cell aggregates.There were no significant changes in P2X7R or NLRP3 protein expression or pathology in synovial tissues in the BBG group compared with the gouty control group(P＞0.05),but serum IL-1β,IL-6,and TNF-α levels were all significantly suppressed(P＜0.01).Conclusions Cold stimulation and strenuous exercise simulated by ice-water swimming may exacerbate pathological damage in gouty arthritis via a mechanism related to high P2X7R expression in the joints.

Objective:To establish a TAL gel-clot bacterial endotoxin test for aluminum hydroxide adjuvant.Methods:According to the bacterial endotoxin test in general chapter 1143 of the Chinese Pharmacopoeia 2020,pre-interference test was performed using 3 types of buffer solutions and Ca-Mg additive by orthogonal design.The interference test and sample preparation validation were carried out using different batches of aluminum hydroxide adjuvant from 2 TAL manufacturers.Results:No interference was obtained through pre-interference test.Under the validated conditions,aluminum hydroxide adjuvant combined with phosphate buffered saline and Ca-Mg additive was employed to remove interference in bacteria endotoxin test.Conclusion:The established TAL gel-clot method is applicable to bacterial endotoxin test for aluminum hydroxide adjuvant.

BACKGROUND: Plasminogen activator inhibitor-1 (PAI-1) plays an important role in the pathophysiology of sepsis, but the exact mechanism remains debatable. In this study, we investigated the associations among the serum levels of PAI-1, the incidence of 4G/5G promoter PAI-1 gene polymorphisms, immunological indicators, and clinical outcomes in septic patients. METHODS: A total of 181 patients aged 18-80 years with sepsis between November 2016 and August 2018 in the intensive care unit in the Xinhua Hospital were recruited in this retrospective study, with 28-day mortality as the primary outcome. The initial serum level of PAI-1 and the presence of rs1799768 single nucleotide polymorphisms (SNPs) were examined. Univariate logistic regression and multivariate analyses were performed to determine the factors associated with different genotypes of PAI-1, serum level of PAI-1, and 28-day mortality. RESULTS: The logistic analysis suggested that a high serum level of PAI-1 was associated with the rs1799768 SNP of PAI-1 (4G/4G and 4G/5G) (Odds ratio [OR]: 2.49; 95% confidence interval [CI]: 1.09, 5.68). Furthermore, a high serum level of PAI-1 strongly influenced 28-day mortality (OR 3.36; 95% CI 1.51, 7.49). The expression and activation of neutrophils (OR 0.96; 95% CI 0.93, 0.99), as well as the changes in the expression patterns of cytokines and chemokine-associated neutrophils (OR: 1.00; 95% CI: 1.00, 1.00), were both regulated by the genotype of PAI-1. CONCLUSIONS Genetic polymorphisms of PAI-1 can influence the serum levels of PAI-1, which might contribute to mortality by affecting neutrophil activity. Thus, patients with severe sepsis might clinically benefit from enhanced neutrophil clearance and the resolution of inflammation via the regulation of PAI-1 expression and activity.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Humans ; Middle Aged ; Young Adult ; Genotype ; Neutrophils ; Plasminogen Activator Inhibitor 1/genetics* ; Polymorphism, Single Nucleotide/genetics* ; Retrospective Studies ; Sepsis/genetics*

Objective:To explore the correlation between the serum 25-(OH)D 3, adiponectin (APN), and chemerin levels of pregnant women with gestational diabetes mellitus (GDM) and insulin resistance. Methods:28 pregnant women with GDM were selected for the study group from May 2020 to December 2021, and 45 pregnant women with normal glucose tolerance were selected for the control group. 25-(OH)D 3, APN, chemerin, islet resistance index (HOMA-IR), fasting blood glucose, fasting insulin, and HbA1c were compared between the two groups. The correlation between 25-(OH)D 3, APN, chemerin, and GDM insulin resistance was analyzed. Results:Fasting blood glucose, fasting insulin, HOMA-IR, and chemerin in the GDM group were higher than those in the control group (all P<0.05), while 25-(OH)D 3 and APN were lower than those in the control group (all P<0.05). There was no statistical difference in HbA1c between the two groups. Logistic regression analysis showed that serum 25-(OH)D 3, APN, and chemerin were all related influencing factors of GDM (all P<0.05). Spearman's correlation analysis showed that serum 25-(OH)D 3 was negatively correlated with fasting blood glucose and HOMA-IR (all P<0.05), chemerin was positively correlated with fasting insulin and HOMA-IR (all P<0.05). ROC curve analysis showed that the AUC of 25-(OH)D 3 was 0.841 (95% CI: 0.746~0.967). AUC of APN was 0.678 (95% CI: 0.545~0.812). AUC of chemerin AUC was 0.360 (95% CI: 0.233~0.487). Conclusions:The levels of 25-(OH)D 3, APN, and chemerin have a certain correlation with the pathogenesis of GDM, which has a certain reference value for the prediction of GDM.

Objective:To analyze the epidemiological and clinical features of patients with visceral leishmaniasis (VL), and to improve the understanding of the disease and standardize the diagnosis and treatment.Methods:The clinical data and diagnosis and treatment process of 62 patients with VL admitted to The First Hospital of Lanzhou University from January 2010 to December 2020 were retrospectively analyzed. The clinical symptoms and laboratory data, including aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and procalcitonin (PCT) were compared between children and adults. Chi-square test was used for statistical analysis.Results:Of the 62 patients, 31(50.0%) patients were males, 38(61.3%) patients were children and 24(38.7%) were adults. There were 32 cases (51.6%) living in Longnan City, and 11 cases (17.7%) living in Gannan Tibetan Autonomous Prefecture. Forty-seven cases (75.8%) had a delay over 30 days from onset to diagnosis. All patients had fever and chills, 34 cases (54.8%) had fatigue and poor appetite, 30(48.4%) had cough and expectoration, 10 cases (16.1%) had headache and dizziness, 45 cases (72.6%) had splenomegaly, 40 cases (64.5%) had anemia, and 29 cases (46.8%) had hepatomegaly. There were 32 cases (51.6%) accompanied with respiratory infection, 11 cases (17.7%) accompanied with hemophagocytic lymphohistiocytosis and 10 cases (16.1%) accompanied with abnormal liver function. Of the 60 patients who received treatment, 14 were recurrent cases who had received at least one course of pentavalent antimony monotherapy before admission. Forty-six patients received standard antimony monotherapy, 14 patients received standard antimony combined with amphotericin B therapy. Thirteen patients discontinued amphotericin B due to impaired renal function, and 50 patients were followed up for half a year without recurrence. The proportions of splenomegaly and hepatomegaly in children were 86.8%(33/38) and 65.8%(25/38), respectively, and the corresponding numbers in adults were 50.0%(12/24) and 16.7%(4/24), respectively. The differences were statistically significant ( χ2=10.03 and 14.26, respectively, both P<0.050). Thirty-three cases (86.8%) in children and 14 cases (58.3%) in adults had a delay over 30 days from onset to diagnosis. The difference was statistically significant ( χ2=6.52, P=0.011). The proportions of patients who had elevated AST, LDH and PCT were 76.3%(29/38), 94.3%(33/35) and 73.9%(17/23) in children, respectively, while 45.8%(11/24), 71.4%(15/21) and 5/17 in adults, respectively, and the differences were all statistically significant ( χ2=5.97, 3.89 and 7.82, respectively, all P<0.050). Conclusions:Patients with VL are tend to have complications and be severe. Early diagnosis and standardized treatment are important measures to improve the prognosis. Patients who have failed antimony treatment can be treated with amphotericin B or combination therapy, but kidney injury and other adverse reactions should be closely monitored.

Objective: To establish a diagnostic prediction model for esophageal squamous cell carcinoma (ESCC) and search the potential biomarkers of ESCC. Methods: Serum samples from 59 patients with ESCC and 57 healthy controls were collected, and randomly divided into the training group (44 patients and 42 healthy controls) and validation group (15 patients and 15 healthy controls). Serum proteins/peptides were extracted and purified with weak cation-exchange chromatography Magnetic Beads (WCX-MB), and detected by the matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). Then the differentially expressed proteins/peptides were screened out, and a diagnostic prediction model for ESCC was established and preliminarily validated. Results: The ClinProTools software identified 31 differential peptide peaks (P＜0.05), among which 18 peaks had significant difference (P＜0.01). Compared with healthy controls, 8 peaks were up-regulated in ESCC patients, while 10 peaks were down-regulated. Among them, the areas under the receiver operating characteristics (ROC) curve (AUC ROC ) of m/z 2 660.84 and m/z 5 336.49 peaks were 0.95 and 0.91, respectively, and their expressions were up-regulated in ESCC patients. The validation results showed that the accuracy, sensitivity and specificity of the diagnostic prediction model established by the genetic algorithm (GA) were 93.10%, 92.90% and 93.30%, respectively. Conclusion The established diagnostic prediction model may be used for the auxiliary diagnosis of ESCC. Two peptide peaks of m/z 2 660.84 and m/z 5 336.49 may be the potential biomarkers of ESCC.

Objective To evaluate the efficacy and safety of TurboHawk plaque resection system in the treatment of lower extremity arteriosclerosis obliterans.Methods Clinical data of 36 patients with atherosclerotic occlusion of lower extremity treated with TurboHawk from January 2016 to August 2017 were retrospectively collected.The characteristics of lesion,improvement of symptoms,ankle brachial index (ABI) and postoperative complications were analyzed.The measurement data were expressed as ((x) ± s),All patients were reviewed every 3 months after operation,followed up for 3-18 months,with an average of (9 ± 0.5) months.And the ABI comparison before and after treatment were performed by paired t test;the counting data were expressed as rate (％),and the comparison of different stages was performed by chi-square test.Results In 36 patients,the technical success rate was 100％,and the symptoms of lower limbs were significantly improved.The ABI (0.85 ±0.07) and ABI (0.75 ±0.10) were significantly better in 3 days and in 3 months after operation than before operation (0.29 ±0.10)(t =37.76,P ＜0.001).Postoperative complications occurred in 2 cases,in one case,the artery dissection was covered with a bare scaffold,and in the other case,the blood vessel was ruptured after resection,and was closured by using balloon angioplasty.Conclusion TurboHawk plaque resection system is an effective,less traumatic and safer option for the treatment of lower extremity arteriosclerosis obliterans.

Objective To investigate the changes in serum level of microparticles (EMPs) in mice with ventilator-induced lung injury (VILI), and explore its significance in VILI. Methods Forty-eight grade SPF male C57BL/6J mice were randomly divided into two groups, with 24 mice in each group: the mice in mechanical ventilation (MV) group were given high tidal volume (VT 30 mL/kg) MV for 4 hours after tracheal intubation, and those in spontaneous breathing group were spontaneously breathed for 4 hours. The apical blood of 12 mice in each group were collected, and serum levels of interleukins (IL-1β, IL-6) and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA), and serum EMPs levels were determined by flow cytometer. The correlations between EMPs and IL-1β, IL-6, and TNF-α were analyzed by linear regression analysis. The lung tissues of other 12 mice in each group were harvested, and wet/dry weight (W/D) ratio was assessed. After hematoxylin-eosin (HE) staining, the morphological changes in lung tissue were observed under light microscope. After double staining of uranium acetate and lead citrate, the ultrastructural changes in lung tissue were observed with electron microscope. Results Compared with spontaneous breathing group, the levels of lung W/D ratio in MV group was significantly increased (5.47±0.14 vs. 4.34±0.11), the levels of IL-1β, IL-6, TNF-α and EMPs were also significantly increased [IL-1β (ng/L): 42.4±4.4 vs. 7.7±3.6, IL-6 (ng/L): 1 239.5±66.3 vs. 21.7±4.6, TNF-α (ng/L):237.6±25.8 vs. 37.1±19.1, EMPs (cells/μL): 28.6±1.8 vs. 5.9±1.8, all P < 0.01]. It was shown by correlation analysis that EMPs were positively related with IL-1β, IL-6, and TNF-α (r value was 0.968, 0.932, 0.945, respectively, all P = 0.000). It was shown by fitting linear regression analysis that when EMPs increased by 1 cell/μL, IL-1β increased by 2.4 ng/L [95% confidence interval (95%CI) = 1.9-2.8, P < 0.001], IL-6 increased by 34.5 ng/L (95%CI = 25.1-44.0, P < 0.001), and TNF-α increased by 13.6 ng/L (95%CI = 10.3-16.9,P < 0.001). It was shown by light microscope that the structure of lung tissue and alveolar of mice in spontaneous breathing group appeared normal, while the shrinks of alveolar and disappearance of alveolar architecture were found in MV group. It was shown by electron microscopy that alveolar wall edema and thickening and broken alveolar septa were found in MV group, by contrast, the structure of alveolar was normal in spontaneous breathing group. Conclusion 30 mL/kg VT ventilation for 4 hours could induce VILI with increase in EMPs, suggesting EMPs closely related to VILI, and EMPs level may be putative biomarker of VILI.

OBJECTIVETo explore the effect and safety of topical propranolol hydrochloride gel for treatment of infantile hemangioma. METHODS Thirty nude mice (BALA/c, nu/nu) were divided into three groups, experimental group, control group and normal group. Human hemangioma endothelial cells cultured in vitro were injected subcutaneously in experimental group and control group to establish infantile hemangioma model. Topical propranolol hydrochloride gel was applied on the surface of the hemangioman in experimental group and normal group. Tumor volumn change and the skin situations (edema, erythema, ulceration) were observed at different periods. 45 days after cell injection, the mice were killed and plasma concentration was detected in the experimental group and the control group by high performance liquid chromatography with evaporative light scattering detector, and tumors were subjected to histopathologic examination and immunohistochemistry for CD31 and CD34. The correlation between volumes and plasma concentration was statistically analyzed with SPSS 13.0 paired samples t test with α = 0.05 as statistical standard.

RESULTSAt 45 days, the volume of the tumor in control group was (366.57 ± 17.08) mm³, which has a significant difference as compared to the experimental group (13.36 ± 2.09) mm³ (P < 0.05); and the plasma concentration was (16.83 ± 1.53) ng/ml in experimental group, and (18.42 ± 2.21) ng/ ml in normal group (P > 0.05 ). Topical propranolol hydrochloride gel (3%) has no irritation to nude mice's skin.

CONCLUSIONSTopical application of 3% propranolol hydrochloride gel is effective and safe for the treatment of infantile hemangioma.

Animals ; Gels ; administration & dosage ; Hemangioma ; drug therapy ; pathology ; Humans ; Immunohistochemistry ; Mice ; Mice, Nude ; Propranolol ; administration & dosage ; Skin Neoplasms ; drug therapy ; pathology ; Tumor Burden ; drug effects