OBJECTIVES: To explore the role and potential mechanisms of chitinase-3-like protein 1 (CHI3L1) in coronary artery lesions in a mouse model of Kawasaki disease (KD)-like vasculitis. METHODS: Four-week-old male SPF-grade C57BL/6 mice were randomly divided into a control group and a model group, with 10 mice in each group. The model group mice were intraperitoneally injected with 0.5 mL of lactobacillus casei cell wall extract (LCWE) to establish a mouse model of KD-like vasculitis, while the control group mice were injected with an equal volume of normal saline. The general conditions of the mice were observed on the 3rd, 7th, and 14th day after injection. Changes in coronary artery tissue pathology were observed using hematoxylin-eosin staining. The level of CHI3L1 in mouse serum was measured by enzyme-linked immunosorbent assay. Immunofluorescence staining was used to detect the expression and localization of CHI3L1, von Willebrand factor (vWF), and α-smooth muscle actin (α-SMA) in coronary artery tissue. Western blot analysis was used to detect the expression of CHI3L1, vWF, vascular endothelial cadherin (VE cadherin), Caspase-3, B cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), nuclear factor κB (NF-κB), and phosphorylated NF-κB (p-NF-κB) in coronary artery tissue. RESULTS: The serum level of CHI3L1 in the model group was significantly higher than that in the control group (P<0.05). Compared to the control group, the expression of CHI3L1 in the coronary artery tissue was higher, while the expression of vWF was lower in the model group. The relative expression levels of CHI3L1, Bax, Caspase-3, NF-κB, and p-NF-κB were significantly higher in the model group than in the control group (P<0.05). The relative expression levels of vWF, VE cadherin, and Bcl-2 were lower in the model group than in the control group (P<0.05). CONCLUSIONS In the LCWE-induced mouse model of KD-like vasculitis, the expression levels of CHI3L1 in serum and coronary arteries increase, and it may play a role in coronary artery lesions through endothelial cell apoptosis mediated by inflammatory reactions.
Male ; Animals ; Mice ; Mucocutaneous Lymph Node Syndrome/pathology* ; Coronary Vessels/pathology* ; NF-kappa B ; Caspase 3/metabolism* ; bcl-2-Associated X Protein/metabolism* ; Chitinase-3-Like Protein 1 ; von Willebrand Factor/metabolism* ; Mice, Inbred C57BL ; Cadherins

As a special kind of in vitro diagnostic devices（IVDs）, laboratory developed tests（LDTs） are of great significance to the development of clinical laboratories. This study aims to explore the regulatory requirements ideas of LDTs. By introducing the development of LDTs and the changing of regulatory requirements in the United States, combing the current regulatory framework and discussing relevant ideas in the regulatory requirements of LDTs.
Clinical Laboratory Services ; Laboratories ; Reagent Kits, Diagnostic ; United States ; United States Food and Drug Administration

AIM To establish the HPLC fingerprints of Kangfuxin Liquid (extract of Periplaneta americana L.) and to determine the contents of six constituents.METHODS The analysis of this drug was performed on a TOSOH TSK-GEL ODS column (250 mm × 4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.07％ acetic acid) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 280 nm.RESULTS There were twenty-four common peaks in the fingerprints of ten batches of samples (Ⅰ-Ⅹ) with the similarities of 0.932-0.993 (except for sample Ⅰ).Uracil,hypoxanthine,xanthine,inosine,protocatechuic acid and Cyclo (Gly-Tyr) showed good linear relationships within the ranges of 3.460-173.0,3.960-198.0,3.596-179.8,1.338-66.9,3.672-183.6 and 3.552-177.6 μg/mL,whose average recoveries (RSDS) were99.8％ (2.65％),98.0％ (2.55％),99.7％ (1.59％),100.7％ (2.80％),102.0％ (2.09％) and 99.6％ (1.88％),respectively.CONCLUSION This accurate,stable and simple method can be used for the quality control of Kangfuxin Liquid.

Objective:To investigate the technique and efficacy of PTES for treatment of L5/S1 disc herniation.Methods:PTES was performed on 52 cases of L5/S1 herniations without spinal instability and central spinal canal stenosis,including 24 cases of high iliac crest,from November 2012 to April 2013.The operation duration,frequency of intraoperative fluoroscopy,blood loss and hospitalization days were recorded.Leg pain was evaluated by using the visual analog scale(VAS)Preoperatively and immediately,1 week,1 month,2 months,3 months,6 months,1 year and 2 years after surgery.The results were determined to be excellent,good,fair,or poor according to the Macnab classification,and complications were observed at 2-year follow-up.Objective:The mean operation duration was(56.3 ±11.5)min per segment.The median frequency of intraoperatively fluoroscopy was 5(3-14)times.The median blood loss was 5(2-20)mL.The median hospital stay was 3(2-4)days.The average postoperative follow-up was(26.2±2.0)months.The median preoperative VAS score of leg pain was 9(6-10),1(0-3)immediately after the operation and 0(0-3)2 years after operation,and the differences were statistically significant(P<0.001).There were 3 cases of lower limb rebound pain 1 week after operation,which were relieved within 2 months after operation.The rate of excellent and good curative effect was 98.1％(51/52)2 years after operation.No complications such as nerve injury,infection,abdominal organ damage and rupture of large vessels occurred.No recurrence occurred.Conclusions:PTES for L5/S1 disc herniation including the cases with high iliac crest is an easy,effective and safe technique.The method has the advantages of simple positioning,easy puncture,simple steps and less fluoroscopy,and the learning curve is not steep for surgeons.

Objective To investigate the involvement of heme oxygenase (HO-1) in PM2.5 induced toxic responses in human umbilical vein endothelial cells (HUVECs).Methods The experiment groups are as follows:(1) control group; (2) PM2.5 groups:the cells were cultured with various concentrations of PM2.5 (200,400,800 μg/ml) for 24 h and 400 μg/ml was chosen for the main study; (3) PM2.5 + Trion group:the cells were pre-treated by 10 μmol/L Trion [a scavenger of reactive oxygen species(ROS)] for 1 h before PM2.5 (400 μg/ml) treatment for 24 h; (4) PM2.5 +ZnPP group:the cells were pretreated by HO-1 inhibitor ZnPP (10 μmol/L) for 1 h before treatment with PM2.5 (400 μg/ml) for 24 h.MTT assay was used to detect cell viability.Reverse transcription polymerase chain reaction (RT-PCR) and indirect immunofluorescence assay were used to determine the mRNA and protein expressions of HO-1.Fluorescence labeling probe method was used to measure intracellular ROS level and flow cytometry was used for cell apoptosis.Colorimetric assay was used to detect intracellular caspase-3 activity.Results Compared with control,PM2.5 significantly decreased cell viability,increased intracellular ROS,cell apoptosis and caspase-3 activity (all P ＜ 0.05),these effects were significantly attenuated in PM2.5 + Tiron group while enhanced in PM2.5 + ZnPP group (all P ＜ 0.05 vs.PM2.5 group).PM2.5 upregulated HO-1 mRNA and protein expressions in HUVECs which was downregulated in both PM2.5 + Tiron group and PM2.5 + ZnPP group.Conclusion PM2.5 could induce oxidative injury through increasing ROS production via modulating HO-1 mRNA and protein expressions,the injury could be aggravated with inhibition of the activity of HO-1 suggesting a potential protective role of HO-1 against PM2.5 induced oxidative stress in HUVECs.

Background Roux-en-Y gastric bypass (GBP) is the main surgical procedure used in type 2 diabetes.The objective of this study was to evaluate the different types of GBP in treatment of type 2 diabetes.Methods Patients with type 2 diabetes were randomly divided into two groups:those who underwent gastrojejunal loop anastomosis bypass and those who underwent gastrojejunal Roux-en-Y bypass.Blood glucose alterations,operation time,and operation complicatiors were observed.Results Gastrojejunal loop anastomosis bypass and gastrojejunal Roux-en-Y bypass were both effective in the treatment of selected patients with type 2 diabetes.Compared with gastrojejunal Roux-en-Y bypass,gastrojejunal loop anastomosis bypass had the advantages of easier implementation,shorter operation time,and fewer operation complications.Conclusions Gastrojejunal loop anastomosis is effective in treatment of type 2 diabetes.It is safe,easy to implement,and worthy of clinical popularization.

Background Acute lung injury (ALI) and end-stage acute respiratory distress syndrome (ARDS) were among the most common causes of death in intensive care units.The activation of an inflammatory response and the damage of pulmonary epithelium and endotheliumwerethe hallmark of ALI/ARDS.Recent studies had demonstrated the importance of mesenchymal stem cells (MSCs) in maintaining the normal pulmonary endothelial and epithelial function as well as participating in modulating the inflammatory response and they are involved in epithelial and endothelial repair after injury.Here,our study demonstrates MSCs therapeutic potential in a rat model of ALI/ARDS.Methods Bone marrow derived MSCs were obtained from Sprague-Dawley (SD) rats and their differential potential was verified.ALl was induced in rats byoleic acid (OA),and MSCs were transplanted intravenously.The lung injury and the concentration of cytokines in plasma and lung tissue extracts were assessed at 8 hours,24 hours and 48 hours after OA-injection.Results The histological appearance and water content in rat lung tissue were significantly improved at different time points in rats treated with MSCs.The concentration of tumor necrosis factor-α and intercellular adhesion molecular-1 in rats plasma and lung tissue extracts were significantly inhibited after intravenous transplantation of MSCs,whereas interleukin-10 was significantly higher after MSCs transplantation at 8 hours,24 hours and 48 hours after OA-challenge.Conclusions Intravenous transplantation of MSCs could maintain the integrity of the pulmonary alveolar-capillary barrier and modulate the inflammatory response to attenuate the experimental ALI/ARDS.Transplantation of MSCs could be a novel cell-based therapeutic strategy for prevention and treatment of ALI/ARDS.

Objective To investigate the role of antigen-processing machinery (APM) component defects in HLA class Ⅰ antigen down-regulation in laryngeal squamous cell carcinoma (SCC) and to assess the clinical significance of these defects. Methods Fifty-one formalin-fixed,paraffin-embedded SCC specimens were examined for the expressions of APM component transporter associated with antigen processing (TAP1) and low molecular weight polypeptide (LMP-7) and HLA class Ⅰ antigen by immunohistochemistry.Results HLA class Ⅰ antigens,TAP-1 and LMP-7 expressions were downregulated in 56.9％ (29/51),39.2 ％ (20/51) and 45.1％ (23/51) of the tested specimens respectively,whereas HLA class Ⅰ antigens,TAP-1 and LMP-7 expressions lost in 21.6 ％ ( 11/51 ),33.3％ ( 17/51 )and 27.5 ％ ( 14/51 ) of the tested specimens respectively.TAP-1 and LMP-7 expressions were significantly correlated with HLA class Ⅰ antigen expression (r =0.460,P < 0.05 and r =0.685,P < 0.05,respectively).HLA class Ⅰ antigens down-regulation was significantly correlated with T stage( x2 =8.61,P ＜ 0.05).Both TAP-1 and LMP-7 down-regulations were significantly correlated with T stage ( x2 valueswere 9.72 and 8.97 respectively,P ＜ 0.05 ) and TNM stage( x2 values were 9.18 and 7.70 respectively,P ＜0.05 ).TAP-1,LMP-7 and HLA class Ⅰ antigen down-regulations were significantly associated with reduced patients' overall survival ( P ＜ 0.05 ) and disease-free survival ( P ＜ 0.05 ).Multivariate analysis showed lymph node metastasis,recurrence and HLA class I antigen down-regulation were unfavorable prognostic factors(P ＜ 0.05).Conclusions Down-regulated expressions of HLA class Ⅰ antigen and APM component TAP-1 and LMP-7 occur frequently in laryngeal squamouss cell carcinoma,by which cancer cells could avoid immune surveillance,while HLA class Ⅰ antigen down-regulation is a major contributing factor to tumour progression and mortality.

Objective To explore the mechanism of fine particulate matter (PM2.5) induced endothelial injury and the efficacy and mechanism of ginsenoside Rg1 on the inhibition of endothelium injuries in human endothelial cells exposured to PM2.5.Methods Human umbilical vein endothelial cells (HUVECs) were stimulated with various concentrations PM2.5 (0.1,0.2,0.4,0.8 mg/ml) and PM2.5 at concentration 0.8 mg/ml induced significant endothelial injury and was chosen for the main study in the presence or absence of Rg1 (0.04 mg/ml).After 24 h treatment,cell growth A value was detected through MTT,intracellular reactive oxygen species (ROS) level through fluorescence labeling probe method and HO-1,Nrf2 mRNA expression was detected by RT-PCR.Results The cell A value was significantly lower while the ROS fluorescence gray value and the average optical density ratio of HO-1 were significantly higher in PM2.5 group than in the control group (all P ＜ 0.05).The average optical density ratio of Nrf2 was similar between PM2.5 group and control group (P ＞ 0.05).The A value and the average optical density ratio of HO-1 were significantly higher while the ROS fluorescence gray value was significantly lower in co-treated PM2.5 (0.8mg/ml) + Rgl (0.04 mg/ml) group than in the PM2.5 (0.8 mg/ml) group (all P ＜ 0.05).Conclusion PM2.5 could induce human endothelial cells injury by increasing oxidative stress which could be attenuated by ginsenoside Rg1.

Objective To analyse the feasibility of detecting F1 antibody to Yersinia pestis in flushing fluid of heart blood of Rhombomys opimus with enzyme linked immunosorbent assay(ELISA) method and its application value in surveillance of the disease. Methods Serum, flushing fluid of heart blood and infusion fluid of liver and spleen of Rhombomys opimus, which were caught by capture in the plague focus of Zunger basin in 2007, were taken to carry out detection for F1 antibodies to Yersinia pestis with ELISA method. The data were processed with SPSS 17.0. Results Positive rate and average titer of serum were 12.35%(11/162) and 25.35, of flushing fluid of heart blood were 10.49%(17/162) and 23.75 and of the infusion fluid of liver and spleen 6.79%(17/162) and 2240,respectively. No statistical difference was found in positive detection rate when it was compared between serum and flushing fluid of heart blood(χ2 = 1.333, P ＞ 0.05), but it was obviously different between serum and infusion fluid of liver and spleen(χ2 = 7.111, P ＜ 0.01 ) and between flushing fluid of heart blood and infusion fluid of liver and spleen(x2 = 6.250, P ＜ 0.05). There was a significant difference in average titer between serum, flushing fluid of heart blood and infusion fluid of liver and spleen(t = 2.290, 3.612, P ＜ 0.05 or ＜ 0.01 ). The plague F1 antibody positive coincidence rate of serum and flushing fluid of heart blood was 85.0%(17/20), of serum and infusion fluid of liver and spleen was 55.0% (11/20), and of flushing fluid of heart blood and infusion fluid of liver and spleen was 64.7%(11/17). Conclusions The ELISA method can detect Fl antibody in flushing fluid of heart blood,and the method is feasible in plague surveillance.