Objective To investigate the role of atherosclerotic monocytes Siglec-1 in stimulating CD4 + and CD8 + T lymphocytes proliferation and activation.Methods Experimental research.Cluster of differentiation antigen 14 (CD14) positive monocytes of 18 acute coronary syndrome (ACS),41 stable angina (SA) and 32 healthy volunteers were separated by magnetic-activated cell sorting.Different concentration of interferon-α (IFN-α,0,2,5,10 ng/ml) were used to up-regulate Siglec-1 and small interfering RNA (siRNA) or blocking antibody were used to down-regulate Siglec-1.Then monocytes were cocultured with CD4 + T/CD8 + T cells from a third healthy volunteer for 5 days.The experiment was designed for 11 groups (n=10 for each group),that was (1) normal CD14,(2) normal CD14 + IFN-α 5 ng/ml,(3) normal CD14 + IFN-α 5 ng/ml + anti-Siglec-1 2 μg/ml,(4) ACS CD14,(5) ACS CD14 + control siRNA group (Mock),(6) ACS CD14 + siRNA 679 40 nmol/L,(7) ACS CD14 + anti-Siglec-1 2 μg/ml,(8) SA CD14,(9) SA CD14 + Mock,(10) SA CD14 + siRNA 679 40 nmol/L and (11) SA CD14 + antiSiglec-1 2 μg/ml.Cell Counting Kit-8 (CCK-8) was used to determine T cells proliferation and ELISA was used to detect Interleukin-2 (IL-2),IL-10,IL-12 and IFN-γ of culture supematant.Data of cytokines detection were expressed as medium (quartiles) and analyzed by nonparametric rank sum test.Results By the blockage of Siglec-1 (group 6),the proliferation of CD4 and CD8 were decreased.Secretion of IL-2,IL-12 and IFN-γ by CD4 cells [67.00(62.50-87.30),0.86(0-1.63),and 47.82(37.60-56.67) pg/ml,respectively] were decreased and IL-10 [56.00(46.25-67.40) pg/ml] was increased compared with those in control group [group 4,213.70 (187.50-275.30),6.87 (4.90-8.93),114.90 (89.50-167.40),and 21.08 (15.70-33.20) pg/ml,respectively,with U =8.50,17.00,8.50,and 87.50,respectively.all P ＜ 0.05].When monocytes Siglec-1 from control group was up-regulated by IFN-α (group 2),secretion of IL-2,IL-12 and IFN-γ [220.44(174.30-312.30),7.90(6.540-10.40) and 143.75(78.20-210.00) pg/ml,respectively] were increased and IL-10 [21.95 (16.30-25.00) pg/ml] was decreased (compared with group 1,with U =89.50,98.00,100.00,and 0,respectively.all P ＜ 0.05).Regulation of Siglec-1 had no role in cytokines production in cocultured CD8 + T cells (all P ＞ 0.05).Conclusions IFN-α can upregulate monocytes Siglec-1.Siglec-1 may participate in the pathogenesis of AS via promoting proliferation of CD4 +/CD8 + T cells and Thl cytokines secretion of CD4 + T cells.

Objective To investigate the enhancing effect of apolipoprotein C3 (APOC3) on THP-1 cell adhesion to aortas of mice.Methods Microsurgery was performed to separate the aorta of C57BL/6 mice in sterile condition.After stimulated by APOC3 (100 △g/ml) in vitro for 16 h,the aorta was allowed to adhere for 1 h with CFSE labeled THP-1 cells (1 ×106/ml).Then the adhesion effect was observed,and the expressions of vascular adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) were detected by immunohistochemical method.Results Adhesion effect of the mice aorta with THP-1 cells in the APOC3 stimulated group was stronger than the control group.Both the expressions of VCAM-1 and ICAM-1 in aortas were increased by APOC3,but the former was significantly up-regulated than the latter.Conclusion Apolipoprotein C3 could enhance THP-1 cell adhesion to aortas of mice.

Objective To establish rat models of polycystic ovary syndrome(PCOS) induced by different methods,to assess the serum levels of several related hormones,to examine the morphological changes in the ovaries,and to discuss their significance.Methods Letrozol,sodium prasterone sulfate,or sodium prasterone sulfate combined with human chorionic gonadotropin were used to establish rat models of PCOS.Radioimmunoassay was used to measure the serum levels of hteinizing hormone(LH),follicle-stimulating hormone(FSH),estrogen(E_2),progesterone(P),testosterone(T),prolactin(PRL),and insulin(INS).HE staining was used to examine the morphological changes of the ovaries.Results Comparing with the normal group A,the serum FSH was increased and the serum progesterone was reduced in the group B,with a statistically significant difference(P<0.05).The serum testosterone was significantly higher in the group B than in the group A(P<0.01).The levels of serum sex hormones and insulin were not significantly different in the group D and C(P>0.05).In comparison with the group C,the levels of serum testosterone and LH/FSH ratio was significantly increased in the group E.(P<0.05).Comparing with the group D,the serum levels of progesterone and testosterone were significantly increased in the group E(P<0.05).The ovaries in the rats of groups A and C showed almost a normal histyology,with mature follicles and dominant follicles.Polycystic changes were observed only in the ovaries of groups B,D and E.Conclusion At the aspect of affecting the level of sex hormones in serum and changing the ovarian morphology.adopting letrozol tablets or sodium prasterone sulfate combined with HCG to induce rat PCO model is more close to clinic manifestations and meets the criteria of PCO animals.In the rat PCOS models induced with letrozol or with sodium prasterone sulfate combined with HCG,either the serum levels of sex hormones and ovarian histology are quite similar to those of human clinical appearance,and may well meet the modeling requirements for future experimental studies of polycystic ovary syndrome.

BACKGROUND: Dietary intervention may be the most available, safe, economic and efficient approach to treat spinal cord injury. OBJECTIVE: To observe the effects of dietary restriction in the form of every-other-day fasting (EODF) on the pathological changes and functional recovery of rats with clip-compression injury of the spinal cord. METHODS: The models of spinal cord injury were established by a medical aneurysm clip in 36 Sprague-Dawley rats, and model rats were then randomized into four groups: group A with spinal cord injury and EODF, group B with spinal cord injury, group C with sham operation and EODF, and group D with sham operation. Food intake and body mass were observed. Motor functional recovery in rats was assessed by Basso, Beattie, Bresnahan scores at preoperative 1 day and postoperative 1 day, 1, 2, 4, 6, 8, 10, and 12 weeks. At 12 weeks after operation, the morphological changes of the spinal cord were observed through hematoxylin-eosin staining. RESULTS AND CONCLUSION: The Basso, Beattie, Bresnahan score in the group B was decreased to the lowest at postoperative 1 day, and gradually increased with time. At 8-12 weeks after operation, the scores in the group A were significantly superior to those in the group B. At 12 weeks after operation, hematoxylin-eosin staining showed less lesion of the spinal cord in the group A than the group B. These results indicate that EODF can improve the motor functional of rats with spinal cord injury, and exerts effective protection on the injured spinal cord.

Objective To detect the number of peripheral blood Treg cells and secreted IL-35 expression level in the patients with rheumatoid arthritis(RA) and to explore their correlation with RA occurrence.Methods Peripheral blood was collected from 45 cases of RA,22 cases of osteoarthritis(OA) and 26 persons undergoing healthy physical examination(control group).The number of CD4+ CD25+ foxp3+ regulatory T cells was determined by flow cytometry,while plasma IL-35 level was determined by ELISA.Then the relationship between Treg and expressed IL-35 with clinical indicators was analyzed.Results The percentage of peripheral blood Treg ceils to total CD4+T cells in the RA group was (5.65 ± 2.33)％,which was significantly increased compared with (4.12 ± 1.75) ％ in the control group(P＜0.05);the difference between the OA group and RA group was not statistically significant(P=0.086).The average fluorescence intensity of foxp3 had no statistical difference among 3 groups(P＞0.05).The higher the DAS28 score,the lower the peripheral blood Treg cells number and foxp3 fluorescence intensity.The plasma IL-35 level in the RA group [(34.22± 14.35)ng/L] was significantly lower than that in the OA group[(78.63± 24.58)ng/L] and control group [(67.56±25.43)ng/L],the difference was statistically significant(P＜0.05).The Treg number in RA patients was negatively correlated with ESR and DAS28 score(r=-0.223,-0.343,P=0.023,0.011),but had no correlation with rheumatoid factor,C-reactive protein and anti-CCP antibody.Conclusion The peripheral blood Treg cells number in RA patients is elevated,while the IL-35 level is decreased,the negative regulation ability in the patients with Treg cell function deficit is attenuated.

AIM: To report the long- term clinical outcomes of accelerated trans-epithelial corneal cross-linking ( CXL ) protocols using KXL System ( Avedro, USA ) in the treatment of progressive keratoconus. · METHODS: Totally 52 patients ( 102 eyes ) with progressive keratoconus between December 2014 and February 2017 [ maximum keratometry values ( Kmax) ≤60.0D, minimum corneal thickness(Thk) ≥400m] were treated with an accelerate trans-epithelial CXL protocol (UV-A irradiation intensity 45mW/cm2 with a total fluence of 7. 2J/cm2 ) using KXL system ( Avedro, USA ) in Southwest Hospital. The average follow-up time was 11. 65mo (range: 9-26mo). Uncorrected distance visual acuity ( UDVA) , corrected distance visual acuity ( CDVA) , intra- ocular pressure ( IOP ) , slit-lamp microscope examination, Kmax and average keratometry values ( AveK ) , corneal stromal demarcation line depth and endothelial cell density ( ECD) were evaluated. ·RESULTS:The 52 patients (102 eyes) were included in this research, male 36 (70 eyes) and female 16 (32 eyes), average age was 19. 5±4. 6 years. Preoperative CDVA was 0. 84±0. 89 (LogMAR), postoperative CDVA was 0. 69±0. 72 ( P = 0. 398 ). Preoperative UDVA was 1. 02 ± 0. 62 (LogMAR), postoperative UDVA was 0. 85 ± 0. 59 ( P =0. 154 ). Preoperative IOP was 12. 95 ± 4. 40mmHg, postoperative IOP was 11.92±3. 66mmHg (P=0. 272). No statistical difference (P=0. 552) has been found between preoperative and postoperative ECD. Nevertheless, on the Sirius anterior system ( Sirius, CSO, Itlay) , significant statistical difference (P=0. 017) was confirmed between preoperative Kmax ( 50. 83 ± 3. 48D ) and postoperative Kmax (52. 05±3. 63D). Meanwhile, the postoperative Avek (47.74±2. 51D) was significantly lower (P=0. 041) than the preoperative Avek ( 48. 73 ± 4. 33D ). The average corneal stromal demarcation line depth ( 192 ± 23. 6μm ) was detected by the anterior segment OCT. No statistical difference ( P = 0. 816 ) has been found between preoperative and postoperative Thk. No severe complication was observed in all cases. ·CONCLUSION: Accelerated trans-epithelial CXL was effective in decreasing keratometry values for progressive keratoconus in this research, and the outcomes remained stable during the follow-up time. No endothelium damage or other severe complications were observed in this clinical research. The accelerated trans-epithelial CXL is as effective as the standard CXL.

Objective To understand the effect of hyaluronic acid (HA) on the proliferation and apoptosis of chondrocytes cultured in vitro with Kashin-Beck disease(KBD) to provide the experimental evidences for treating KBD diseases with HA. Methods The articular cartilage samples collected from KBD patients were selected according to Diagnosis for Kaschin-Beck Disease(GB 16003-1995). And the normal cartilage samples were collected from victims of incidence (control). Chandrocytes were separated and cultured in vitro. Then varying dosages of HA were administered to chondrocytes and individed into 0,100,500 mg/L group, according to HA doages. The effect of HA on the proliferation and apoptosis of chondrocytes cultured/n vitro both KBD and the controls were investigated by methyl thiazolyl tetrazolium(MTT), Annexin V/PI staining on 2~(nd), 4~(th), 6~(th) day. Results In the control group, 500 mg/L group(0.140 ± 0.049) promoted chondrocyte proliferation significantly than 0 mg/L group (0.116 ± 0.021 ) at the 4~(th) day(P < 0.05), similar phenomenon was observed in KBD group in the 6~(th) day between 500 and 0 mg/L group(0.179 ± 0.081,0.128 ± 0.017, P< 0.05). In the KBD group, compared with 0 mg/L (12.860 ± 2.159), both 100 and 500 mg/L( 10.458 ± 1.143,7.877 ± 1.346) inhibited chondrocyte apoptosis rate (P < 0.05). In control, apoptosis rate of 500 mg/L group(4.045 ± 1.204) descreased compared with 0 mg/L group (7.128 ± 1.244, P < 0.05). Conclusion HA can promote the proliferation and inhibit the apoptosis of KBD chondrocytes cultured in vitro, and 500 mg/L HA play more effective role than that of 100 mg/L in promoting proliferation and inhibiting poptosis.

OBJECTIVETo study the effect of astragaloside IV on the expression of cytokines in bone mesenchymal stem cells (MSCs) in rats.

METHODSMSCs were isolated from Wistar rats by the method of adhesive cultiration and clone, and then their biological activities were assessed using indirect immunofluorescence. Proliferation of MSCs stimulated with astragaloside IV was ascertained by the MTT method. Expression of cytokines was ascertained using RT-PCR in MSCs with astragaloside IV stimulation or not.

RESULTSMSCs were effectively isolated and purified in vitro, and had expression of many cytokines except IL-3, such as stem cell factor (SCF), thrombopoietin (TPO), granulocyte macrophage colony stimulating factor (GM-CSF) and transforming growth factor (TGF-beta1). Astragaloside IV stimulation promoted MSCs proliferation, and 200 mg/mL astragaloside IV treatment produced a peak effect 72 hrs after culture. The SCF expression in MSCs stimulated with astragaloside IV increased significantly compared with that in MSCs without astragaloside IV stimulation.

CONCLUSIONSAstragaloside IV may promote MSCs proliferation and increase SCF secretion in vitro.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Proliferation ; drug effects ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; secretion ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Saponins ; pharmacology ; Stem Cell Factor ; biosynthesis ; genetics ; secretion ; Triterpenes ; pharmacology

OBJECTIVETo establish a diagnostic model of protein fingerprint pattern in the cerebrospinal fluid (CSF) for non-small-cell lung cancer (NSCLC) patients with brain metastases.

METHODSThe CSF samples were obtained from 29 NSCLC patients with brain metastasis, 23 non-tumor patients and 10 early-stage NSCLC patients without brain metastases for analysis of the protein expression profiles using surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS). The data were then analyzed by Biomarker Wizard software, and the tree analysis patterns were generated using the decision-tree model in Biomarker Patterns software. The diagnostic model was tested for its clinical application.

RESULTSFive protein peaks were identified showing differential expression between patients with brain metastases and those without brain metastases. Combination of the 3 protein peaks (m/z: 8698.00, 1215.32 and 1245.70) could discriminate these two samples with a sensitivity of 100.00% (29/29) and a specificity of 100.00% (23/23). Five proteins were differentially expressed between the NSCLC patients with brain metastases and the non-tumor patients. With one protein peak (m/z: 6050.00), these two samples could be discriminated with a sensitivity of 90.00% (9/10) and a specificity of 78.26% (18/23).

CONCLUSIONThe established diagnostic model of CSF protein fingerprint pattern provides high sensitivity and specificity in the diagnosis of NSCLC with brain metastasis.

Adult ; Aged ; Brain Neoplasms ; cerebrospinal fluid ; diagnosis ; secondary ; Carcinoma, Non-Small-Cell Lung ; cerebrospinal fluid ; diagnosis ; pathology ; secondary ; Cerebrospinal Fluid Proteins ; genetics ; Decision Trees ; Early Detection of Cancer ; Female ; Gene Expression Profiling ; Humans ; Lung Neoplasms ; cerebrospinal fluid ; diagnosis ; pathology ; Male ; Middle Aged ; Peptide Mapping ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Aim To investigate the role of P2X7 recep-tor and its mediated NLRP3 inflammatory signaling pathway in alcohol-induced liver injury. Methods The acute alcoholic liver injury model was established by NIAAA method, and thirty C57BL/6 male mice were randomly divided into three groups (n =10):control group, model group, A438079 group, The three groups were processed as follows in the last week:control group and model group: given an equal dose of saline intraperitoneal injection(about 0.2 mL/only) once a day. According to the weight of the mice, A438079 group was given intraperitoneally injection by 200 μmol·kg-1of A-438079 (prepared at 7 g·L-1 of A438079,about 0.2 mL/only) once a day. And it was given a single 31.5% alcohol solution by intragas-tric administration on the last day of the morning,with the dose of 10 mL·kg-1. Nine hours later alanine aminotransferase (ALT), aspartate aminotransferase (AST),cholesterol(TCHO),triglyceride(TG) were measured by orbital blood in mice. HE staining was used to observe the pathological changes of the liver. Immunohistochemical method was applied to detect the expression of P2X7R in liver tissues. Western blot was employed to detect the levels of P2X7R, NLRP3, ASC, IL-1β and IL-18 in liver tissues. Results Compared with control group,the levels of ALT,AST, TG and TCHO in model group were significantly en-hanced, and the liver injury was obvious. Compared with model group, the levels of ALT, AST, TG and TCHO in A438079 group significantly decreased. Compared with control group, the expressions of P2X7, NLRP3, ASC, IL-1β, IL-18 in model group were significantly higher than those in control group. Compared with model group, the expression levels of P2X7, NLRP3, ASC, IL-1β and IL-18 in A438079 group significantly decreased. Conclusion Alcohol-induced liver injury may be associated with P2X7R-NLRP3 signaling pathway.