Objective To evaluate the repeatability and agreement of two optical biometers (Lenstar LS900 (R) and SW-9000) for ocular biometry in Chinese adolescents.Methods A prospective study was conducted which included 65 ametropic patients,with an average age of (11.45 ± 2.67) years (age ranging from 8 to 18 years).The ocular biometry for right eyeball was performed with Lenstar LS900 (R) and SW-9000 respectively,followed by evaluation of the repeatability of the two biometers using one-way analysis of variance,and the agreement of the two instruments using the Bland-Altman plot.Results The repeatability of parameters measured by Lenstar LS900 (R),including axial length (AL),K value in the flattest meridian (K1),K value in the steepest meridian (K2),central corneal thickness (CCT),anterior depth (AD),lens thickness (LT),pupil diameter (PD),was well,and all intraclass correlation coefficient (ICC) ＞ 0.9;the repeatability of white to white (WTW) was inferior to other parameters,but it was still ＞0.88.The repeatability of AL,K1,K2,CCT measured by SW-9000 was good,with their ICC ＞ 0.9,but the repeatability of other parameters was poor.The parameters with good repeatability including AL,K1,K2,CCT measured by SW-9000 and Lenstar LS900 (R) were compared respectively,and the results showed that AL and CCT examined by SW-9000 were slightly longer than those measured by Lenstar LS900 (R),and the difference was statistically significant (all P ＜ 0.05).However,there was no significant difference about K1,K2 (all P＞0.05).Moreover,the AL,K1,K2 and CCT measured by the two instruments had close linear correlation (all r ＞0.97,all P ＜0.01).The BlandAltman plot showed that 95％ LoA (limits of agreement) of AL was (-0.057 to 0.133) mm,K1 was (-0.456 to 0.369) D,K2 was (-0.388 to 0.549) D and CCT was (-3.483 to 8.016) μm.Conclusion Biometric parameters including AL,K1,K2,CCT measured by Lenstar LS900 (R) and SW-9000 have good repeatability in the adolescents aged 8-18 years and they are highly correlated;meanwhile,the agreement of AL,K1,K2,CCT measured by SW-9000 with Lenstar LS900 (R) is acceptable in clinical practices.

Objective: To investigate the effect of human umbilical cord mesenchymal stem cells derived exosomes (hUC-MSC-exo) on the phenotype of peripheral blood macrophages from rabbit autoimmune dry eye and the expression of related cytokines. Methods: The hUC-MSCs were isolated and characterized.Exosomes derived from hUC-MSCs were extracted by ultracentrifugation and observed directly using electronic microscopy.Specific markers of exosomes were analyzed by Western blot.Six rabbits were randomly divided into the normal control group and the dry eye group by using the random number table method, 3 rabbits for each group.Rabbit model of autoimmune dry eye was established in the dry eye group, and the lacrimal glands were collected for quantitative real-time PCR (qRT-PCR) at the 8th week.In vitro, activated peripheral blood mononuclear cells (PBMCs) from rabbit autoimmune dry eye model were incubated with hUC-MSC-exo or phosphate buffered saline (PBS). After 48 hours, cells from the hUC-MSC-exo group and the PBS control group were collected.The mRNA expression levels of related cytokine genes and subpopulation-related marker genes in macrophages were quantified by qRT-PCR.The use and care of the animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.This study protocol was approved by Ethic Committee of Tianjin Medical University Eye Hospital (No.TJYY20181217001). Written informed consent was obtained from each family before obtaining umbilical cord. Results: Exosomes derived from hUC-MSCs had typical morphology and specific markers.qRT-PCR results showed that, the relative expression quantity of M1 macrophages phenotypic molecular nitric oxide synthase 2 (NOS2) mRNA and inflammatory factor tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) mRNA in the lachrymal organization in the dry eye model group was significantly higher than those in the normal control group (3.06±1.00 vs. 1.00±0.03, 2.77±0.72 vs. 1.01±0.02 and 1.30±0.08 vs. 1.01±0.01, respectively), the relative expression quantity of M2 macrophages phenotypic molecular arginase 1 (Arg1), CD206 and IL-10 mRNA in the lachrymal organization in the dry eye model group was significantly lower than those in the normal control group (0.55±0.07 vs. 1.00±0.00, 0.60±0.13 vs.1.00±0.00, 0.65±0.14 vs. 1.01±0.01, respectively), with significant differences between them (all at P<0.05). In vitro, the relative expression quantity of M1 macrophages phenotypic molecular NOS2 mRNA and inflammatory factor TNF-α, IL-1β mRNA in PBMCs in the hUC-MSC-exo group was significantly lower than those in the PBS control group (0.59±0.08 vs.0.98±0.03, 0.56±0.07 vs. 1.03±0.11, 0.47±0.04 vs.1.00±0.08)(all at P<0.05); the relative expression quantity of M2 macrophages phenotypic molecular Arg1, CD206 mRNA and anti-inflammatory cytokine TGF-β, IL-10 and IL-4 mRNA in PBMCs in the hUC-MSC-exo group was significantly higher than those in the PBS control group (2.13±0.28 vs. 1.10±0.17, 1.32±0.03 vs. 1.01±0.06, 1.53±0.20 vs. 1.05±0.10, 1.47±0.08 vs.0.98±0.03, 1.51±0.16 vs. 1.01±0.03), with significant differences between them (all at P<0.05). Conclusions hUC-MSC-exo can polarize peripheral blood macrophages toward immune-suppressive M2-like phenotype, inhibit the production of pro-inflammatory cytokines TNF-α and IL-1β, and meanwhile increase the expression of anti-inflammatory factors IL-10 and TGF-β.

Objective:To investigate the role of microRNA-338-3p (miR-338-3p) in regulating the generation and function of interphotoreceptor retinoid-binding protein (IRBP) 1-20-specific T helper 17 (Th17) cells in experimental autoimmune uveitis (EAU). Methods:Bone marrow cells were flushed from the femurs and tibiae of wild-type C57BL/6 mice and cultured in the presence of granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin-4 (IL-4)to differentiate into bone marrow-derived dendritic cells (BMDCs). On day 5 after induction, immature BMDCs were collected and divided into miR-338-3p mimics transfection group and mimics negative control transfection group, then transfected with miR-338-3p mimics or negative mimics according to grouping.Twenty-four hours after transfection, the BMDCs were stimulated with 100 ng/ml of lipopolysaccharide to mature.Relative expression levels of miR-338-3p, IL-6, IL-23 and IL-1β mRNA in BMDCs of the two groups were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The EAU model was established with IRBP 1-20, incomplete Freund adjuvant and mycobacterium tuberculosis (H37Ra) in mice.On day 13 after modeling, T cells were isolated from the mice spleen or draining lymph nodes and co-cultured with miR-338-3p mimics or negative control mimics-transfected BMDCs under Th17-polarizing conditions.Concentration of IL-17 in the supernatant was detected by ELISA.Relative expression levels of retinoic acid receptor-related orphan nuclear receptor γt (RORγt) and IL-17 mRNA were analyzed by qRT-PCR.The proportion of IL-17 + cells among T cells co-cultured with BMDCs was assessed by flow cytometry.To further verify the role of miR-338-3p in dendritic cells on Th17 cells, BMDCs transfected with miR-338-3p inhibitor or control inhibitor were co-cultured with T cells isolated from spleen or draining lymph nodes of EAU mice.Concentration of IL-17 in the supernatant was detected by ELISA.The use and care of the animals complied with Regulations for the Administration of Affairs Concerning Experiment Animals by State Science and Technology Commission.The study protocol was approved by the Institutional Animal Care and Use Committee of Tianjin Medical University (No.TJYY2019110117). Results:Relative expression level of miR-338-3p in BMDCs was significantly increased in the miR-338-3p mimics transfection group than the mimics negative control group ( t=6.861, P=0.002). In T cells co-cultured with miR-338-3p mimics-transfected BMDCs, the relative expression levels of RORγt and IL-17 mRNA were 1.34±0.16 and 1.33±0.16, which were significantly higher than 1.00±0.01 and 1.00±0.01 in the mimics negative control group ( t=3.632, P=0.022; t=3.681, P=0.021). ELISA showed that the concentration of IL-17 in the supernatant was (5 941.00±452.40)pg/ml in the miR-338-3p mimics transfection group, which was significantly higher than (4 299.00±348.30)pg/ml in the mimics negative control group ( t=4.979, P=0.008), and IL-17 concentration in the supernatant was (3 092.00±200.90)pg/ml in the miR-338-3p inhibitor transfection group, which was lower than (4 063.00±131.50)pg/ml in the inhibitor negative control group ( t=7.005, P=0.002). The proportion of IL-17 + cells among T cells was (8.03±1.35)% in the miR-338-3p mimics transfection group, which was significantly higher than (4.52±0.73)% in the mimics negative control group ( t=3.968, P=0.017). The relative expression levels of IL-6, IL-23, and IL-1β mRNA were 2.23±0.21, 2.21±0.56, 2.32±0.43, respectively in the miR-338-3p mimics transfection group, which were significantly higher than 1.00±0.06, 1.00±0.07, 1.01±0.15 in the mimics negative control group ( t=10.290, P=0.001; t=3.747, P=0.020; t=5.280, P=0.006). Conclusions:Overexpression of miR-338-3p in BMDCs can promote the IRBP 1-20-specific Th17 cell response by increasing the expression of Th17-polarizing cytokines including IL-6, IL-1β and IL-23.

BACKGROUNDLaparoscopic cholecystectomy has been widely used in clinical practice during the recent decades; however, the effects of pneumoperitoneum and the surgery on the coagulation system are largely unknown. This clinical study aimed to observe any possible effects of pneumoperitoneum and the surgery on the coagulation system of patients.

METHODSThis was a prospective observational study. The inclusion criteria included (1) patients with chronic cholecystitis and/or cholecystic polyps and (2) patients in the relief stage of acute cholecystitis. The exclusion criteria included (1) patients in the episodic stage of acute cholecystitis and those complicated with cholangiolithiasis; (2) patients with concomitant hematologic diseases, damages to the liver function, malignant tumors or immune system diseases, or patients complicated with thrombotic or hemorrhagic disorders; and (3) patients who had taken anticoagulant medication within a week before surgery. Fifty patients who were hospitalized into our department for elective laparoscopic cholecystectomy between November 2011 and February 2013 were eligible and enrolled into this study. Of the 50 patients, 22 were male and 28 female. The age of the patients ranged from 29 to 78 (mean 56.7±11.5) years. The surgery for each of the 50 patients was performed with the same equipment and conditions. The surgeries for all the patients were performed under general anesthesia with the patients in a 30-degree head-up tilted posture, and the pressure of pneumoperitoneum was maintained at 13 mmHg. Venous blood specimens were taken from each patient before and at the end of pneumoperitoneum (i.e., 0 hour after surgery) and at 8 hours after surgery for determination of prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fib), thrombin time (TT), and D-dimer (DD). The results of the determinations of these parameters were compared.

RESULTS(1) All the patients recovered well without any complications. (2) The pre-pneumoperitoneum values of the parameters of coagulation had normalized. (3) The PT values slightly increased (P > 0.05) at the end of pneumoperitoneum (i.e., 0 hour after surgery) and decreased by 0.5 seconds at 8 hours after surgery as compared to the pre-pneumoperitoneum values (P < 0.05). (4) APTT at 0 and 8 hours decreased by 1.4 seconds (P > 0.05) and 3.7 seconds (P < 0.05) respectively as compared to pre-pneumoperitoneum values, while the difference between the APTT values at 0 and 8 hours after surgery was not statistically significant (P > 0.05). (5) FIB determined at 0 hour post-operation increased by 0.1 g/L as compared to pre-pneumoperitoneum values (P > 0.05); however, the FIB values at 8 hours after operation increased by 1.2 g/L as compared to the pre-pneumoperitoneum values (P < 0.05), and increased by 1.1 g/L as compared to 0 hour post-operation (P < 0.05). (6) The TT values obtained at 0 and 8 hours post-operation were not significantly different as compared to the pre-pneumoperitoneum values (P > 0.05). (7) The DD values gradually increased after operation; as compared to pre-pneumoperitoneum values, DD at 0 and 8 hours after operation increased by 210.8 ng/ml and 525.9 ng/ml respectively (P < 0.05) and DD at 8 hours after operation increased by 315.1 ng/ml as compared to 0 hour post-operation (P < 0.05).

CONCLUSIONSThe pneumoperitoneum for laparoscopic cholecycstectomy may lead to postoperative hypercoagulation in the patients, and thereby may increase the risks for development of postoperative thrombosis; Patients may have risks for occurrence of thrombosis within 8 hours after the operation, to which attention should be paid in favor of preventing thrombosis.

Adult ; Aged ; Aged, 80 and over ; Blood Coagulation ; physiology ; Cholecystectomy, Laparoscopic ; Female ; Humans ; Male ; Middle Aged ; Partial Thromboplastin Time ; Pneumoperitoneum ; surgery ; Prospective Studies