Objective: To investigate the cytotoxic effects of induced pluripotent stem (iPS) cells of anti-mesothelin (MSLN)-chimeric antigen receptor natural killer (CAR-NK) cells (anti-MSLN-iCAR-NK cells) on ovarian epithelial cancer cells. Methods: Twenty cases of ovarian cancer patients who underwent surgical treatment at Henan Provincial People's Hospital from September 2020 to September 2021 were collected, and 20 cases of normal ovarian tissues resected during the same period due to other benign diseases were also collected. (1) Immunohistochemistry and immunofluorescence were used to verify the expression of MSLN protein in ovarian cancer tissues. (2) Fresh ovarian cancer tissues were extracted and cultured to obtain primary ovarian cancer cells. Recombinant lentiviral vectors targeting anti-MSLN-CAR-CD₂₄₄ were constructed and co-cultured with iPS cells to obtain anti-MSLN-iCAR cells. These cells were differentiated into anti-MSLN-iCAR-NK cells using cytokine-induced differentiation method. The cell experiments were divided into three groups: anti-MSLN-iCAR-NK cell group, natural killer (NK) cell group, and control group. (3) Flow cytometry and live cell staining experiment were used to detect the apoptosis of ovarian cancer cells in the three groups. (4) Enzyme-linked immunosorbent assay (ELISA) was used to measure the expression levels of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), granzyme B (GZMB), perforin 1 (PRF1), interleukin (IL)-6, and IL-10 in the three groups of ovarian cancer cells. Results: (1) Immunohistochemistry analysis showed that a positive expression rate of MSLN protein in ovarian cancer tissues of 65% (13/20), while normal ovarian tissues had a positive rate of 30% (6/20). The comparison between the two groups was statistically significant (χ²=4.912, P=0.027). Immunofluorescence analysis revealed that the positive expression rate of MSLN protein in ovarian cancer tissues was 70% (14/20), while normal ovarian tissues had a positive rate of 30% (6/20). The comparison between the two groups was statistically significant (χ²=6.400, P=0.011). (2) Flow cytometry analysis showed that the apoptotic rate of ovarian cancer cells in the anti-MSLN-iCAR-NK cell group was (29.27±0.85)%, while in the NK cell group and control group were (8.44±0.34)% and (6.83±0.26)% respectively. There were statistically significant differences in the comparisons between the three groups (all P<0.01). Live cell staining experiment showed that the ratio of dead cells to live cells in the anti-MSLN-iCAR-NK cell group was (36.3±8.3)%, while in the NK cell group and control group were (5.4±1.4)% and (2.0±1.3)% respectively. There were statistically significant differences in the comparisons between the three groups (all P<0.001). (3) ELISA analysis revealed that the expression levels of IFN-γ, TNF-α, GZMB, PRF1, IL-6, and IL-10 in ovarian cancer cells of the anti-MSLN-iCAR-NK cell group were significantly higher than those in the NK cell group and the control group (all P<0.05). Conclusion: The anti-MSLN-iCAR-NK cells exhibit a strong killing ability against ovarian cancer cells, indicating their potential as a novel immunotherapy approach for ovarian cancer.
Humans ; Female ; Carcinoma, Ovarian Epithelial/metabolism* ; Ovarian Neoplasms/metabolism* ; Interleukin-10/pharmacology* ; Induced Pluripotent Stem Cells/metabolism* ; Iron-Dextran Complex/pharmacology* ; Tumor Necrosis Factor-alpha/metabolism* ; Cell Line, Tumor ; Killer Cells, Natural ; Interleukin-6

Objective To establish a mouse model of exogenous iron overload combined with tuberculosis(TB). Methods C57BL/6N mice were divided into negative control, low-, medium-, and high-dose iron groups and received intraperitoneal injection of iron dextran at 0, 3.75, 7.50, and 15.00 mg/dose(3 times/week for 4 weeks), respectively.After 4 weeks, the organ morphology and body weight of the mice were evaluated.The content of serum iron, ferritin, transferrin, and transferrin receptor was determined by ELISA.Heart, liver, spleen, lung, kidney, and small intestine were analyzed for tissue iron content and iron deposition pathology.
Animals ; Humans ; Iron ; Iron Overload ; Iron-Dextran Complex ; Mice ; Mice, Inbred C57BL ; Tuberculosis

PURPOSE: In the present study, the dependence of intraocular pressure (IOP) on body position was compared between pseudophakic and phakic eyes after cataract surgery performed at least 3 months prior. METHODS: IOP was measured in 214 patients over 40 years of age who received cataract surgery at least 3 months prior. The present study included 104 patients who did not have any other ocular disease which could influence visual acuity or IOP. The IOP was measured in 4 different positions, sitting, supine, right decubitus, and left decubitus by a single skilled researcher using Icare tonometer. In addition, IOP was compared between the phakic and pseudophakic eyes, specifically, the dependent eye in the decubitus position. RESULTS: The comparison between the phakic and pseudophakic eyes in patients after cataract surgery in a single eye showed IOP was not different between the sitting and supine positions; however, in the decubitus position, IOP in the dependent eye was significantly higher than the non-dependent eye (p = 0.001). Additionally, regardless of lens status, IOP was significantly higher in the dependent eye than non-dependent eye. CONCLUSIONS: Both body position and lens status can affect IOP. After cataract surgery, regardless of lens status, IOP was higher in the dependent eye than non-dependent eye of patients in the decubitus position.
Cataract* ; Glaucoma ; Humans ; Intraocular Pressure* ; Iron-Dextran Complex ; Supine Position ; Visual Acuity

PURPOSE: Rebound tonometer has been used to measure the intraocular pressure (IOP) in the supine as well as normal upright positions. We investigated the reliability of IOP measurements using the rebound tonometer in the upright and supine positions. METHODS: IOP was measured in 30 patients (60 eyes) with open-angle glaucoma who had no history of ocular surgery and no anterior segment pathology, in both the upright and supine positions using rebound tonometer (IcarePRO; Icare Finland Oy, Finland). The average IOP value after 6 measurements was recorded. We measured IOP repeatedly until 3 reliable values within normal limits of the measurement's variation were obtained. We calculated the intraclass correlation coefficient (ICC), coefficient of variation, and number of repeated measurements necessary to obtain 3 reliable IOP values in each position as measured by one examiner. RESULTS: ICC values for IOP measurements were 0.852 (95% confidence interval [CI], 0.784-0.903; p < 0.001) in the upright position and 0.684 (95% CI, 0.563-0.784; p < 0.027) in the supine position. Coefficient of variation was 8.7 +/- 0.1% in the upright position and 24.0 +/- 0.1% in the supine position. An average of 3.3 times of repeated measurements in the upright position and 6.2 times in the supine position were necessary to obtain 3 reliable IOP values within the normal range of standard deviation. CONCLUSIONS: When measuring IOP using the IcarePRO rebound tonometer, the measurement reliability was different between the upright and supine positions. Reproducibility of IOP measurements was lower in the upright than the supine position.
Finland ; Glaucoma, Open-Angle ; Humans ; Intraocular Pressure ; Iron-Dextran Complex ; Pathology ; Reference Values ; Supine Position*

PURPOSE: To compare the accuracy and reliability of intraocular pressure (IOP) measurements in enucleated porcine eyes using the Icare PRO in the upright and horizontal positions. METHODS: We designed an enucleated porcine eye model whose anterior chamber was cannulated with a 30-gauge needle, connected in parallel to a pneumatic pressure device. The reference pressure was manipulated by changing the air pressure from 70 to 10 mm Hg at 10 mm Hg intervals, and the IOP of porcine eyes was measured with the Icare PRO at each pressure. Correlation analysis, comparison using the Bland-Altman plot and Wilcoxon signed rank test, was performed to assess the accuracy of IOP measurements. Intraclass correlation coefficients were calculated to assess the intra-observer variability in the upright and horizontal positions, respectively. RESULTS: The IOP value in both upright and horizontal positions was well correlated with the reference pressure (r = 0.992 and 0.985, respectively). The Bland-Altman plot showed good agreement between the two positions. However, all IOP values in both positions were lower than the reference pressures. The IOP values in the horizontal position were significantly lower than those in the upright position at the a reference pressure of 50 mm Hg or greater. Values of intraclass correlation coefficient ranged from 0.911 to 0.984 when measured in the upright position and from 0.707 to 0.914 in the horizontal position. CONCLUSIONS: IOP measurements of Icare PRO in porcine eyes were remarkably lower than reference pressures controlled by the pneumatic method even though they showed a good correlation with reference values. The higher was the reference pressure, the greater was the degree of underestimation of IOP measurement in both positions. This trend was more pronounced in the horizontal position, and the reliability of IOP measurements was also lower than that in the upright position.
Air Pressure ; Anterior Chamber ; Intraocular Pressure ; Iron-Dextran Complex* ; Needles ; Observer Variation ; Reference Values

PURPOSE: To evaluate changes in intraocular pressure (IOP) according to position using a portable rebound tonometer. METHODS: We measured the IOP values of 20 healthy volunteers (40 eyes) in the sitting, supine, right lateral decubitus and left decubitus positions with a portable rebound tonometer, and then analyzed using the Wilcoxon signed rank test. IOP in sitting position was also measured with a non-contact tonometer and a Goldmann applanation tonometer, and analyzed with Kruskal-Wallis test and Spearman correlation analysis. Agreement among the 3 tonometers was calculated using the Bland-Altman method. RESULTS: The IOP measured with rebound tonometer in the supine position was significantly higher than in the sitting position (p = 0.002). However, there was no significant difference in IOP between the supine and decubitus positions. In the decubitus position, there was no significant difference in IOP between the dependent and non-dependent eyes. IOP measurement using the rebound tonometer showed positive correlation with that of the noncontact and Goldmann applanation tonometers. CONCLUSIONS: In normal subjects, IOP measurement obtained with a rebound tonometer in the supine position was significantly higher than in the sitting position, but there was no significant difference in IOP between the supine and decubitus positions. A rebound tonometer may be useful for patients whose intraocular pressure measurement with Goldmann applanation tonometer or non-contact tonometer is impossible. When using a portable rebound tonometer in bed-ridden or pediatric patients, we should pay attention to the interpretation of IOP in the supine position.
Healthy Volunteers ; Humans ; Intraocular Pressure* ; Iron-Dextran Complex* ; Supine Position

PURPOSE: This study was carried out to evaluate the postural intraocular pressure (IOP) change in Trendelenburg, reverse Trendelenburg, and supine positions in healthy young males. METHODS: We measured the IOP values of 5 healthy young male volunteers (10 eyes) using an Icare PRO rebound tonometer in sitting, Trendelenburg, reverse Trendelenburg, and supine positions. RESULTS: The mean IOP in the supine position (18.63 mm Hg) was significantly higher (p < 0.01) than in the sitting position (15.31 mm Hg). When maintaining the Trendelenburg position, IOP gradually increased. CONCLUSIONS: In our study, the Trendelenburg position significantly increased the IOP compared to that in the supine position. The effects of increased IOP should be considered in situations that require Trendelenburg positioning, such as exercise or surgery.
Head-Down Tilt ; Humans ; Intraocular Pressure* ; Iron-Dextran Complex ; Male ; Supine Position ; Volunteers

PURPOSE: To compare the intraocular pressure (IOP) measured by portable rebound tonometer and TonoPen applanation tonometer with pressure measured by Goldmann applanation tonometer (GAT) and analyze the factors affecting IOP disagreement between tonometers. METHODS: In a prospective study of 463 eyes, IOP was measured with Icare Pro(R) rebound tonometer, TonoPen AVIA(R) applanation tonometer, and GAT. Bland-Altman plot, intraclass correlation coefficient, Pearson's correlation analysis, and multiple regression analysis were performed to evaluate the agreement of IOP measured by each tonometer and the factors affecting the measurements. RESULTS: The IOP values measured by Icare Pro(R) and TonoPen AVIA(R) were consistently higher than those measured by GAT, but showed no significant differences with those measured by GAT (p = 0.307 and 0.114, respectively). In Bland-Altman plot, the IOP values measured by Icare Pro(R) and TonoPen AVIA(R) exhibited excellent agreement with those measured by GAT. Both Icare Pro(R)/GAT and TonoPen AVIA(R)/GAT differences increased with younger age (p = 0.041 and 0.049, respectively) and higher central corneal thickness (p = 0.019 and 0.035, respectively). CONCLUSIONS: IOPs measured by portable Icare Pro(R) rebound tonometer and TonoPen AVIA(R) applanation tonometer were significantly correlated with IOP measured by GAT. Therefore, such instruments can be useful when measuring IOP with GAT is difficult. However, central corneal thickness and age should be considered when measuring IOP with portable tonometers.
Intraocular Pressure* ; Iron-Dextran Complex ; Prospective Studies

PURPOSE: To compare Icare rebound tonometer (IRT) and Goldmann applanation tonometer (GAT) and investigate the clinical usefulness of IRT. METHODS: In a retrospective study of 131 eyes with glaucoma and glaucoma suspect, intraocular pressure (IOP) was measured with IRT and GAT. The correlation between IRT and GAT and the influence of refractive error (spherical equivalent; SE) and central corneal thickness (CCT) were analyzed. RESULTS: A distinct correlation between IRT and GAT was found. IOP measured with GAT was 2.23 +/- 3.16 mm Hg higher than with IRT (p < 0.001). As CCT increased, IOP was measured higher with both GAT and IRT. IOP in myopic eyes (SE > or = -5 diopter) was measured relatively lower than in hyperopic eyes (SE < -5 diopter) with GAT (+1.50 +/- 0.68 mm Hg) and IRT (+1.88 +/- 0.75 mm Hg). CONCLUSIONS: IRT showed strong correlation with GAT, but IOP measured approximately 2 mm Hg lower. The IOP tends to measure lower at low CCT and in myopic eyes.
Eye ; Glaucoma ; Intraocular Pressure ; Iron-Dextran Complex ; Manometry ; Peptides ; Refractive Errors ; Retrospective Studies

OBJECTIVETo establish a mouse model of iron overload by intraperitoneal injection of iron dextran and investigate the impact of iron overload on bone marrow hematopoiesis.

METHODSA total of 40 C57BL/6 mice were divided into control group, low-dose iron group (12.5 mg/ml), middle-dose iron group (25 mg/ml), and high-dose iron group (50 mg/ml). The control group received normal saline (0.2 ml), and the rest were injected with intraperitoneal iron dextran every three days for six weeks. Iron overload was confirmed by observing the bone marrow, hepatic, and splenic iron deposits and the bone marrow labile iron pool. In addition, peripheral blood and bone marrow mononuclear cells were counted and the hematopoietic function was assessed.

RESULTSIron deposits in bone marrow, liver, and spleen were markedly increased in the mouse models. Bone marrow iron was deposited mostly within the matrix with no significant difference in expression of labile iron pool.Compared with control group, the ability of hematopoietic colony-forming in three interventional groups were decreased significantly (P<0.05). Bone marrow mononuclear cells counts showed no significant difference. The amounts of peripheral blood cells (white blood cells, red blood cells, platelets, and hemoglobin) in different iron groups showed no significant difference among these groups;although the platelets were decreased slightly in low-dose iron group [(780.7±39.60)×10(9)/L], middle dose iron group [(676.2±21.43)×10(9)/L], and high-dose iron group [(587.3±19.67)×10(9)/L] when compared with the control group [(926.0±28.23)×10(9)/L], there was no significant difference(P>0.05).

CONCLUSIONSThe iron-overloaded mouse model was successfully established by intraperitoneal administration of iron dextran. Iron overload can damage the hepatic, splenic, and bone marrow hematopoietic function, although no significant difference was observed in peripheral blood count.

Animals ; Bone Marrow ; drug effects ; physiopathology ; Disease Models, Animal ; Hematopoiesis ; drug effects ; Iron Overload ; chemically induced ; physiopathology ; Iron-Dextran Complex ; administration & dosage ; toxicity ; Male ; Mice ; Mice, Inbred C57BL ; Spleen ; drug effects