The aim of this study was to examine the priming effect of sphingosine 1-phosphate (S1P) on fMLP-activated neutrophils, mainly to detect the neutrophil respiratory burst products, and to investigate the signaling pathway involved in S1P activity. Flow cytometry was used to evaluate the new isolated neutrophil; the superoxide anion output was detected indirectly by cytochrome C reduction in respiratory burst; the dihydro-rhodamine 123 was used to detect the intensity of respiratory burst; the signal transduction pathways of neutrophil respiratory burst were explored by Western blot. The results showed that after pretreated with S1P, the level of superoxide anion released by fMLP-activated neutrophils significantly increased; the Rhodamine 123 mean fluorescence intensity in S1P primed fMLP-activated neutrophils group was significantly higher than that in fMLP treatment group; PI3K and Akt proteins involved in the signal pathway of neutrophil respiratory burst. It is concluded that S1P is a new priming reagent, which primes respiratory burst of fMLP-activated neutrophils; this signal pathway may be that S1P interacts with its receptor, activates PI3K, then activates Akt-transmitting signals through NADPH oxidase, finally results in the respiratory burst.
Cells, Cultured ; Humans ; Lysophospholipids ; metabolism ; NADPH Oxidases ; metabolism ; Neutrophils ; metabolism ; physiology ; Proto-Oncogene Proteins c-akt ; metabolism ; Receptors, Lysosphingolipid ; metabolism ; Respiratory Burst ; Signal Transduction ; Sphingosine ; analogs & derivatives ; metabolism ; Superoxides ; metabolism

OBJECTTo investigate the therapeutic effect of microencapsulated porcine retinal pigmented epithelial cells (RPE-M) transplantation on rat model of Parkinson's disease (PD).

METHODSPrimary porcine RPE cells were harvested by enzyme digestion and expanded in culture medium. Determine the levels of dopamine (DA) and homovanillic acid (HVA) by high performance liquid chromatography electrochemical (HPLC) assay, and the levels of brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF) were detected by ELISA. Alginate-polylysine-alginate (APA) microencapsulated cells were produced by using a high voltage electrostatic system. PD rat model was established by unilateral injection of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB). After that, the RPE-M was transplanted into the corpus striatum of PD rat, and then the rotation test scores were recorded and biochemical changes of the corpus striatum were tested.

RESULTSThe levels of DA, HVA, BDNF and GDNF secreted by RPE were stable in the RPE culture supernatant and were not changed by the microencapsulation. Eighty-three percent rats developed PD by unilateral lesion of 6-OHDA in the MFB. The RPE-M transplantation had therapeutic effect on 33% PD rats.

CONCLUSIONPorcine RPE cells grow actively in vitro and could secrete DA, HVA, BDNF, and GDNF constantly, which does not be affected by the passage culture and the APA miroencapsulation. RPE-M transplantation of may be a curative therapy for PD.

Adrenergic Agents ; toxicity ; Animals ; Brain-Derived Neurotrophic Factor ; metabolism ; Cell Transplantation ; methods ; Cells, Cultured ; Disease Models, Animal ; Dopamine ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells ; metabolism ; transplantation ; Glial Cell Line-Derived Neurotrophic Factor ; metabolism ; Male ; Oxidopamine ; toxicity ; Parkinson Disease ; etiology ; surgery ; Rats ; Rats, Sprague-Dawley ; Retina ; cytology ; Swine ; anatomy & histology ; Time Factors ; Transplantation, Heterologous ; methods ; Tyrosine 3-Monooxygenase ; metabolism

AIM: To explore the value of corneal epithelial thickness parameters in the diagnosis of early keratoconus by Fourier domain optical coherence tomography(FD-OCT).

METHODS: Retrospective control study. Patients with early keratoconus who were admitted to the Department of Ophthalmology of the First Affiliated Hospital of Soochow University from January 2015 to October 2020 and patients who received laser refractive surgery for myopia during the same period were included in this study. Using FD-OCT to measure the cornea in circle centralized by the pupil and with diameter of 9mm in the early-keratoconus group and the control group. This was to obtain the thickness parameter of corneal epithelium with diameter of 7mm, the respective averaged thickness value of corneal epithelium and the respective averaged thickness value of the full corneal layer on each of the 25 divisions. The parameter value differences between the early-keratoconus group and the control group were compared, and the diagnosis value of parameters that hold difference was analyzed.

RESULTS: The Min-e value in early-keratoconus group was significantly lower than that of the control group. On contrary, the value of Std.Dev, Max-Min-e, and Max-e were significantly higher in early-keratoconus group than that in latter one(all P<0.001). The value of Std.Dev and Max-Min-e hold a greater diagnostic efficiency on diagnosis of early keratoconus(AUC=0.937, 0.928), while Max -e value and Min-e value performed a medium diagnostic efficiency(AUC=0.871, 0.797). Analysis on thickness of corneal epithelium on the 25 divisions found that the thickness of corneal epithelium at infratemporal region of inner ring(EIT1)became thinner most obviously in early-keratoconus group, with greatest diagnostic efficiency(AUC=0.827). The average corneal thickness of the 25 zones of the early keratoconus group was significantly thinner than that in the normal control group. Averaged corneal thickness at infratemporal region of inner ring(CIT1)hold a relatively high diagnostic efficiency among the 25 divisions(AUC=0.903).

CONCLUSION: The parameter values of corneal epithelium thickness including Std.Dev, Max-Min-e, Max-e, Min-e, the averaged thickness value of corneal epithelium at infratemporal region of inner ring, and the thickness value of full corneal layer at infratemporal region of inner ring can all be used for diagnosis of early keratoconus.

Abstract?AIM: To investigate mechanism of bradykinin ( BK) on inflammations of retinal pigment epithelium ( RPE) cells.?METHODS: ARPE -19 cells were cultured in vitro, stimulated by 100nM BK for 24h. Cell morphology changes were observed by microscope, and BK receptor localization was detected through cell immunofluorescence. Changes of Ca2+in BK and BR antagonist stimuli were detected by laser scanning confocal microscopy.The expressions of COX-1, COX-2, eNOS and iNOS protein in control group and BK group were detected by Western Blot.?RESULTS: After the stimulation of BK, there was no significant changes of ARPE-19 cells in morphology.Kinin B1 receptors ( B1R ) and B2 receptors ( B2R ) could be detected in ARPE-19 cells.Compared with control group, Ca2+concentrations significantly increased in BK group; in B1R antagonist group and B2R antagonist group Ca2+concentrations increased less than BK group; B1R and B2R antagonist group showed no obvious changes in Ca2+concentrations.Compared with control group, COX-2 and iNOS protein concentrations were significantly increased in BK group (P<0.001).?CONCLUSION:BK induces the increasing expression of COX-2 and iNOS in the cultured ARPE cells through binding with either B1R or B2R.

AIM To investigate the effect of β-cyclodextrin-assisted extraction on Cassiae Semen.METHODS The chemical constituents in aqueous extract and β-cyclodextrin extract determined and analyzed by UPLC and principal component analysis had their antioxidant activities tested by six methods (protective effects on lipid peroxidation injuries induced by spontaneous liver,CCl4,H2 O2,FeSO4-vitamin C,and scavenging effects on DPPH,hydroxyl free radicals).RESULTS Compared with aqueous extract,the component content and antioxidant activity of β-cyclodextrin extract were increased by 10.476％ and 80.88％,respectively.CONCLUSION β-Cyclodextrin can effectively enhance the component extraction rate and antioxidant activity of Cassiae Semen.

Objective To study the clinical efficacy and safety of edaravone in the treatment of traumatic cerebral infarction.Methods A total of 72 patients with traumatic cerebral infarction were randomly divided into the control group (n =36) and the treatment group (n =36).The control group received conventional treatments including intracranial pressure control,nerve nutrition and microcirculation regulation.The patients in treatment group were given intravenous infusion of edaravone 20 mL + 0.9％ NaC1 200 mL on the basis of the conventional treatment,bid,14 d.The changes of hemorheologic parameters were measured and compared between the two groups respectively.The total effective rate were evaluated according to the Glasgow scores,and the safety of clinical medication was evaluated.Results The total effective rate of the treatment group was 91.67％ (33/36 cases) while that of the control group was 72.22％ (26/36 cases),with statistically significant difference between the two groups (P ＜ 0.05).Mter treatment,high shear whole blood viscosity in the treatment group and the control group were (4.73 ± 0.56),(5.36 ± 0.78) mPa · s respectively,while low shear whole blood viscosity were (8.15 ± 1.03),(9.54 ± 1.26) mPa · s,plasma viscosity were (1.41 ± 0.15),(1.76 ± 0.19) mPa · s,red blood cell pressure were (37.15 ± 3.21) ％,(43.17 ± 4.03)％,fibrin deposition were (3.58 ± 0.45),(4.67 ±0.54)g· L-1,and the differences of all the parameters above between the two groups were statistically significant (P ＜ 0.05).The incidence of adverse drug reactions in the treatment group and control group were 5.56％ (1/36 cases) and 2.78％ (2/36 cases) respectively,while no statistical significance was observed (P ＞ 0.05).Conclusion Edaravone is able to improve hemorheology indexes in patients with traumatic brain injury complicated with cerebral infarction.The treatment effect is better than the routine nervous medical treatment,and edaravone has a positive effect on the clinical treatment of traumatic brain injury complicated with cerebral infarction.

Objective To observe the association between adiponectin and small dense low-density lipoprotein (sLDL-c) in coronary artery disease (CAD) patients.Furthermore,we sought to determine the association between single nucleotide polymorphisms (SNP) rs1501299 ( + 276G/T ), rs266729 (-11365C/G) and the incidence of CAD.Methods Consecutive subjects with chest discomfort were examined by coronary angiography and divided into non-CAD [ n =250,147 male,mean age (60.26 ±7.52) years] and CAD [n =267,153 male,mean age (60.79 ±9.63) years] groups.Blood samples were collected from all participants following an overnight fasting for at least 12 hours.Plasma adiponectin levels were measured by competitive enzyme-linked immunosorbent assay (ELISA).The serum levels of sLDL-C and oxidized low-density lipoprotein (ox-LDL) were determined by ELISA.Genotypes in rs1501299 and rs266729 of the adiponeetin were determined by polymerase chain reaetion ( PCR ).Results 1.The adiponectin levels were significantly lower [ ( 306.17 ± 74.52 ) mg/L vs.( 321.78 ± 86.28 ) mg/L ],whereas sLDL-C and ox-LDL levels were significantly higher [ ( 276.30 ± 45.55 ) ng/L vs.( 249.00 ±32.02) ng/L and (545.06 ± 115.46 ) μg/L vs.(497.74 ± 106.09 ) μg/L,P ＜ 0.05 ] in CAD group than non-CAD group.2.Adiponectin level was negatively associated with sLDL-C,whereas sLDL-C positively correlated with ox-LDL in all subjects.3.Genotype distribution and allele frequencies of rs1501299 and rs266729 were similar between CAD and non-CAD subjects and not related to the serum levels of adiponectin, sLDL-C and ox-LDL.Conclusions Reduced adiponectin and increased sLDL-C were independent risk factors for coronary artery disease.Genetic polymorphisms in rs1501299 and rs266729 were not linked with coronary artery disease.

Objective To investigate the relationship between dietary patterns and hypertension in Hanzhong city of Shaanxi province.Methods A cross-sectional survey on dietarypattern and hypertension together with a semi-quantitative food frequency questionnaire was conducted among rural residents aged from 18 to 80 in Hanzhong of Shaanxi province in 2010.Factor analysis by SPSS was used to identify food patterns based on the frequency of food.Logistic regression was used to analyze the relationship between dietary patterns and hypertension.Results 2929 rural residents were investigated with an average age as 50.0 years old and average schooling-years as 6.8.The prevalence rate of hypertension was 34.3％,with 35.4％ in males and 33.7％ in females.Among 29 kinds of food under investigation,only 11 kinds with the frequencies of consumption more than 1 time per week.Four main factors were derived as four dietary patterns by factor analysis and they were diversity dietary pattern,alcohol dietary pattern,beverage dietary pattern and simple dietary pattern.Among them,the balanced dietary pattern was significantly associated with hypertension.All the participants were categorized into 4 groups (Q1-Q4) according to their factor scores,quartile with Q1 as a reference.The residents who were more closer to having balance dietary pattern,were under less risk of suffering from hypertension after controlling for age,sex,BMI,education and income.Compared to Q1,the Q4 had lower risk of hypertension (OR=0.59,95％CI:0.44-0.77) for the balanced pattern.Conclusion Dietary pattern could be related to the prevalence of hypertension.A rational diet with diversity of foods should be suggested as one of the major measures for the prevention of hypertension.

OBJECTIVETo explore the biological characteristic of third-party-derived tolerogenic DC(tDC) and the influence of third-party-derived tDC on acute graft-versus-host-disease (aGVHD) following allogeneic bone marrow transplantation (allo-BMT) in mice.

METHODStDC from bone marrow cells of D1 mice was cultured with low doses of GM-CSF, IL-10 and TGF-β1D1. The phenotype, expression of cytokines and function associated molecules were identified with FACS and RT-PCR. Mixed lymphocyte reaction was applied to analyze the influence of third-party-derived tDC on allo-CD4(+)T cells proliferation in vitro. Different doses of D1-tDC were adoptive transferred in the aGVHD model in allogeneic BMT which B6 mice as donors and D2 mice as recipients. Survival time, clinical GVHD score and the levels of Th1/2 cytokines in serum were monitored after allo-BMT using the aGVHD model as control.

RESULTStDC expressed lower levels of MHC II and co-stimulatory molecules, such as CD80, CD86 and CD40, even when stimulated by LPS. The results by RT-PCR indicated that tDC expressed low levels of IL-12p40 and high levels of immunosuppressive molecules, such as IL-10, TGF-β, Fas Ligand, indoleamine 2, 3-dioxygenase (IDO) and arginase. In the allogeneic MLR, third-party tDC suppressed allo-CD4(+)T cells proliferation, which was relative to the dose of tDC. In the B6→D2 mouse model, all aGVHD mice died within 18 days. Remarkably, if 10(4) third-party tDC were transferred, 60% mice survived at least 60 days. When the doses of tDC were reduced to 10(3) cells, only 20% of mice survived day 60, and when increased tDC to 10(5), all of the mice died within day 37 after allo-BMT. The cytokine levels in serum indicated that 10(4) tDC-treated mice secreted in vivo high level of IL-10 21d after BMT (P < 0.05), the levels of IL-10 in 10(3), 10(4) and 10(5) tDC-treated mice were (114.23 ± 7.78), (646.18 ± 212.02), (121.97 ± 10.47) ng/L, respectively.

CONCLUSIONThird-party tDC could suppress allo-CD4(+)T cells proliferation in vitro and prevent aGVHD in allogeneic BMT mode, which may be mediated by modulating tolerogenic cytokines secretion, such as IL-10. And this effect was associated with the dose of tDC. Adoptive therapy by transfusing third-party tDC cultured with low doses of GM-CSF, IL-10 and TGF-β1 could significantly prolong the survival of recipients and prevent aGVHD in allogeneic BMT.

Animals ; Bone Marrow Transplantation ; adverse effects ; CD4-Positive T-Lymphocytes ; cytology ; Cell Proliferation ; Dendritic Cells ; cytology ; immunology ; metabolism ; Graft vs Host Disease ; prevention & control ; Interleukin-10 ; immunology ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Transforming Growth Factor beta1 ; immunology ; Transplantation, Homologous

This study was purposed to investigate the expression of c-fes gene in leukemia patients and its clinical significance. The expression of c-fes mRNA in bone marrow cells from 121 cases of acute and chronic leukemia patients, and the expression of c-fes mRNA in peripheral blood mononuclear cells of 20 normal persons were detected by real time-quantitative reverse transcription polymerase chain reaction (RQ-PCR). The results showed that the level of c-fes mRNA in AML patients was higher than that in normal controls [(48.017 +/- 57.170) x 10(-3) vs (0.152 +/- 0.398) x 10(-3)] (p < 0.0001); but there was no significant differences of level of c-fes mRNA between samples of ALL and normal controls(0.047 +/- 0.068) x 10(-3) vs(0.152 +/- 0.398) x 10(-3) (p>0.05); the level of c-fes mRNA in CML patients was higher than that in normal persons (21.605 +/- 24.818) x 10(-3) vs (0.152 +/- 0.398) x 10(-3) (p < 0.0001). The positive expression rate of c-fes gene in CML-CP patients (80%) was higher than that in CML-AP patients (66.7%) and CML-BP (28.6%) patients. In AML patients, c-fes gene was expressed higher in M(2) (80.77%) and M(3) (92.86%) patients. The remission rate of AML (except M(3))patients who had expression of c-fes gene was 81.08%, which was higher than that of patients with no expression of c-fes gene (40.00%). It is concluded that c-fes gene expression was found in myeloid leukemias, whereas low or no expression in lymphocytic leukemias. The differentiation of myelocytic cells may be related to c-fes gene. All AML (except M(3))patients with high level of c-fes mRNA may get good prognosis.
Adult ; Case-Control Studies ; Female ; Gene Expression ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Prognosis ; Proto-Oncogene Proteins c-fes ; genetics ; RNA, Messenger ; genetics