This study examined the effect of IL-10 on immunoglobulin-like transcript (ILT4) expression of human monocytic leukemic cell line THP-1, especially the role of the ILT4 promoter activity. ILT4 promoter area was amplified by PCR, and was cloned into the eukaryotic expressing vector pGL3-Basic. The pGL3-ILTP obtained was tested by double endonuclease digestion and sequencing. Then, the recombinant plasmid was transfected into THP-1 cells by using lipofectamine. After culture with IL-10 for 12 h, the mRNA extracted from THP-1 cells was detected by RT-PCR and the protein was detected by FACS. The dual-luciferase reporter assay system was employed to detect the activity of ILT4 promoter with or without IL-10. The results showed that the activity of pGL3-ILTP was significantly increased and was more than ten times that of pGL3-Basic cells. After culture with IL-10 for 12 h, the expression of ILT4 protein and its mean fluorescence intensity (MFI) were increased. Moreover, the mRNA was remarkably higher than that of the control group. Dual-luciferase reporter assay revealed that ILT4 promoter was much more activated after being treated with IL-10. We were led to conclude that pGL3-ILTP containing ILT4 promoter was constructed successfully. The expression of ILT4 could be up-regulated by IL-10 both at the transcriptional and translational level. Furthermore, ILT4 promoter could be much more active after addition of IL-10. This study suggests that IL-10 up-regulates ILT4 expression on monocytes via increasing ILT4 gene promoter activity, which may have implication for inducing transplantation tolerance in clinical practice.

Objective: To investigate the relation between ser um hyaluronic acid (HA) concentration and cryopreservation-reperfusion injury. Methods: The animals were randomly assigned to 3 groups: (1 ) group A: the control; (2) group B: liver allografts were stored in lactated R inger's solution (0℃) for 2 h before implantation; (3) group C:liver allografts were stored in lactated Ringer's solution (0℃) for 4 h before implantation. Th e serum sample and liver specimen were taken up at 2 h and 4 h after transplanta tion to detect the concentration of HA, AST and LDH, and to get pathologic obser vation. Results: Serum HA increased earlier and decreased more s hortly than AST and LDH after transplantation in group A. Serum HA increased sig nificantly in group B and C, much higher than that in group A(P<0.01). The i njury of vascular endothelium and the disorder of hepatic sinuses and hepatic lo b ules were observed in group B and C. In the specimen of 4 h in group C, evident infiltration of inflammatory cell was present. Conclusion: Cryopreservation leads to injury of endothelial cell and reperfusion aggravat es this injury. The serum HA concentration indicates the degree of cold ischemia -reperfusion injury.

A rapid, sensitive, label-free and separation-free analytical method for determination of melamine ( MA) was developed based on surface enhanced Raman scattering ( SERS ) effect of gold nanoparticles. Through tri-sodium citrate reduction method, gold nanoparticles with average diameter of 30 nm were obtained. The melamine detection platform was constructed after self-assembling 4-mercapto phenylboronic acid (4-MPBA) on the surface of gold nanoparticles through Au S covalent bond. When MA existed in solution, 4-MPBA functionalized gold nanoparticles would aggregate because of strong hydrogen bond interaction between MA and 4-MPBA. Moreover, following increase of the concentration of MA, gold nanoparticles would aggregate more intensively and form more hot spots. As a result, Raman signal of 4-MPBA and MA was enhanced greatly. The characteristic Raman peaks of 4-MPBA and MA respectively located at 1076 cm-1 and 715 cm-1 . Hence, the qualitative and quantitative detection for MA were realized based on the ratio value of I715 cm-1 to I1076 cm-1 . The linear range of MA detection was 0 . 1 μmol/L-1. 5 μmol/L. The limit of detection (LOD) reached 0. 02 μmol/L in terms of three times signal to noise.

Objective To explore the effects of melatonin (MT) on the expression of interleukin (IL)-10,interleukin-6 and interleukin-8 as well as the inflammatory reaction and nerve repair after acute spinal cord injury (SCI).Methods One hundred and eight Sprague-Dawley rats were randomly divided into a spinal cord injury group (group A),an MT treatment group (group B) and a sham operation group (group C),each with 36 rats.SCI models were established in the rats of groups A and B using a version of Allen's weight drop method (50gcf at the T12 level).Group C had removal of the lamina only.Ten minutes later,group A was injected with 5％ ethanol in saline (the MT solvent) and group B with 100 mg/kg of melatonin preparation.At 6,12,18 and 24 hours,IL-6,IL-8 and IL-10 levels in serum were detected in 6 rats of each group.At 18 hours post-surgery,spinal cord specimens were taken from 6 rats of each group for hematoxylin eosin staining,morphological examination and immunohistochemical SP detection of IL-10 expression.Results The specimens of group A showed inflammatory reaction and ulceration at 48 h; groups B and C had no ulcers.Group B showed the highest levels of IL-10 in serum and IL-10 mRNA in the spinal cord,while group C showed the lowest level.The differences were statistically significant.Group A had the highest levels of IL-6 and IL-8 and group C had the lowest.The difference between group B and groups A and C was significant.The morpho-logical observation showed that after melatonin treatment the IL-10 levels in the spinal cord's central canal and around the gray matter improved.Conclusions Melatonin can improve nerve lipid peroxidation and inflammatory reaction in the treatment of spinal cord injury by increasing IL-10 expression and inhibiting IL-6 and IL-8 expression.

Aim: To develop a practical synthetic route of raltegravir, a drug for HIV treatment. Methods: Raltegravir was synthesized through an eight-step process including aminonitrile formation, protection with benzyloxy-carbonyl group, conversion of the nitrile to the amidoxime, cyclization to form hydroxypyrimidinone, N-methyla-tion, amidation with microwave-assistance, deprotection, amidation with acyl chloride. Results: The overall yield of the eight-step synthesis is about 12. 0% and the structure of the target compound was confirmed by ~1H NMR, ~(13)C NMR, LR-MS and HR-MS. Conclusion: The reported synthetic process of raltegravir highlights the advantages in terms of readily available starting materials, convenient operation and low cost.

Objective Inflammation plays a critical role in the presence , development and maintenance of atrial fibrillation ( AF) , but it remains unclear what factors induce inflammation in AF patients , especially in those with valvular heart disease ( VHD) . The aim of this paper was to investigate the role of the shedding of Syndecan -4 in left atrial inflammation in patients with valvular atrial fibrillation. Methods Sixty VHD patients scheduled for valvuloplasty or valve replacement surgery were divided into three groups of equal number:sinus rhythm (SR), paroxysmal atrial fibrillation (PaAF), and persistent atrial fibrillation (PeAF).Another 10 pa-tients with congenital heart disease but no valve damage and atrial fibrillation were included in a control group .Baseline clinical data were recorded and tissues were obtained from the left atrial appendage during operation .The expressions of iNOS , HMGB1, and Syn-decan-4 in the left atrium were detected by Western blot , and the pathological changes of the left atrial tissue observed by HE staining . Results Western blot analysis was performed to detect expression levels of proteins .The iNOS level was significantly higher in pa-tients from the paroxysmal AF group (1.61 ±0.10) and persistent AF group (1.67 ±0.08) than those from sinus group (1.06 ± 0.11) and control group (1.02 ±0.12), as was the protein level of HMGB1 (0.63 ±0.05, 0.95 ±0.10, 0.45 ±0.07 and 0.46 ± 0.06 in paroxysmal AF group, persistent AF group, sinus group and controlgroup respectively ).Inflammatory cell infiltration in-creased, while syndecan 4 was down-regulated in AF groups.All these comparisons were significant (P<0.05). Conclusion The decreased expression of Syndecan-4 and enhanced inflammatory response in the left atrial tissue indicate that the shedding of Synde-can-4 may play a role in the presence and development of inflamma-tion in the left atrium .

Objective To investigate the effect of Shenfu injection on the expressions of endoplasmic reticulum stress-related factors oxygen-regulated protein 150 (ORP150),X-box binding protein 1 (XBP1),and C/EBP homologous protein (CHOP) mRNAs in cerebral cortical neurons after hypoxic-ischemic brain damage (HIBD) in neonatal rats.Methods The 7 day-newborn Sprague-Dawley rats were randomly divided into sham operation,normal saline and Shenfu treatment (Shenfu injection,10 ml/kg per day,peritoneal injection,for 3 days) groups.They were redivided into 3 h,6 h,12 h,24 h,3 d and 7 d subgroups at different time points after modeling (n =8 in each group).A HIBD model of the neonatal rats was induced.Reverse transcription-polymerase chain reaction was used to detect the expressions of ORP150,XBP1,and CHOP mRNAs in rat cerebral cortex.Results The expressions of ORP150,XBP1 and CHOP mRNAs of the sham operation group was very week.The expressions of ORP150,XBP1 and CHOP mRNAs in the normal saline group and the Shenfu treatment group were significantly upregulated compared to those in the sham operation group at 3 h after modeling (all P ＜0.05); the expressions of XBP1 and CHOP mRNAs reached the peak at 6 and 12 h,respectively.Then they decreased mildly and closed to the level in the sham operation group at day 7.The expressions of XBP1 and CHOP mRNAs in the Shenfu treatment group at 12,24 h and day 3 after modeling were significantly lower than those in the normal saline group (all P ＜0.05).The XBP1 mRNA expression was significantly positively correlated with the CHOP mRNA expression in the normal saline group (r =0.649,P ＜0.05).Conclusions HIBD in neonatal rats induces endoplasmic reticulum stress response.ORP150,XBP1 and CHOP may be involved in the delayed neuronal damage process after HIBD in neonatal rats.Shenfu injection downregulates the expression of XBP1 and CHOP mRNAs and inhibits endoplasmic reticulum stress response.

Objective This study was conducted to establish a stable and highly efficient method for isolation and purification of pancreatic islets from NOD mice and to evaluate their characteristics in vitro and in vivo. Methods The is-lets were isolated from mouse pancreas using modified collagenase digestion and Ficoll density gradient centrifugation. The endocrine secretory function was assessed by insulin secretion in either low or high dose glucose stimulation. To evaluate the function of the graft,body weight and blood glucose were monitored,and IVGTT was performed. In addition,to assess sur-vival of the implanted islets,Pathology using HE staining and insulin immunostaining of the graft were performed. Results The average islet yield was 116 ± 12 islets/pancreas and purity was higher than 90%. Compared with islets from Kunming mice,the islets isolated from NOD mice were poorly responsive to glucose challenge. Blood glucose levels and body weight changes of the islet-transplanted diabetic mice were significantly improved compared with the sham-operated mice. In addi-tion,blood glucose levels in vivo after an IVGTT also significantly improved. However,these improvements were only main-tained for 2 weeks. Furthermore,HE staining and immunostaining assays demonstrated that there were insulin-positive cell clusters and lymphocyte infiltration in the graft-bearing kidney. Conclusions A large number of quality islets can be isola-ted and purified from NOD mice by using the modified mouse islet isolation method, which can be used to develop thera-peutic strategies to protect transplanted islets from rejection and autoimmune attack.

Objective: The aim of the current study is to compare the short-term clinical outcomes between Billroth-I reconstruction using an overlap method and delta-shaped anastomosis in totally laparoscopic distal gastrectomy (TLDG). Method: A retrospective cohort study was performed. The following inclusion criteria were applied: (1) Preoperative gastroscopy and CT confirmed that the tumor is located in the antrum of the stomach, and the biopsy suggested adenocarcinoma; (2) Chest, abdomen and pelvis enhanced CT showed no evidence of distant metastasis; (3) Preoperative gastric reconstruction CT or endoscopic ultrasonography suggested that the clinical stage of the tumor is stage I-III. (4) During the operation, the tumor position was confirmed to be located in the antrum of the stomach by nanocarbon injection or gastroscope; (5) Complete laparoscopic radical gastrectomy for distal gastrectomy, and the gastrointestinal reconstruction was performed by delta-shaped anastomosis or overlap anastomosis. And the following exclusion criteria were applied: (1) History of gastric surgery; (2) Patients who cannot tolerate laparoscopic surgery because of comorbidities. Finally, data on 43 consecutive patients who underwent TLDG with Billroth-I reconstruction between January 2016 and November 2018 in Peking Union Medical College Hospital were retrospectively reviewed. Patients were divided into those who underwent Billroth-I reconstruction using an overlap method (n=20) or using delta-shaped anastomosis (n=23). The demographic and clinical characteristics and perioperative data of the two groups were analyzed. Measurement data that conformed to the normal distribution were expressed as the mean ± s, and differences between groups were compared using Student′s t-test; comparisons between the counting data groups were performed using the χ² test or the continuously corrected χ² test. Results: The demographic and clinical characteristics were similar between the delta-shaped group and the overlap group (P>0.05). There was no significant difference between groups regarding operation time [(185.9±22.8) minutes vs. (184.0±25.8) minutes, t=0.260, P=0.796], blood loss [(50.9±36.0) ml vs. (47.0±30.8) ml, t=0.375, P=0.709], number of stapler reloads used for anastomosis (5.1±0.3 vs. 5.2±0.6, t=-0.465, P=0.651), time to flatus [(3.3±0.9) days vs. (3.6±0.9) days, t=-1.067, P=0.292) and postoperative hospitalization [(8.8±3.1) days vs. (10.4±3.8) days,t=-1.494, P=0.143]. As for the delta-shaped group and the overlap group, the anastomotic leakage rate was 4.3% (1/23) and 0 (χ²=0.000, P=1.000), respectively. The incidence of anastomotic bleeding was 4.3% (1/23) and 5.0% (1/20) (χ²=0.000, P=1.000), while the incidence of intra-abdominal hemorrhage was 4.3% (1/23) and 0 (χ²=0.000, P=1.000). The incidence of gastric emptying disorders was 4.3% (1/23) and 30.0% (6/20), respectively (χ²=3.454, P=0.063). All complications were cured after conservative treatment or symptomatic treatment. Conclusion The overlap method for Billroth-I reconstruction is safe and feasible.

Objective:To investigate the role of miR-483-5p in adrenocortical cancer (ACC) and its possible mechanism.Methods:Quantitative real-time polymerase chain reaction was conducted to estimate the expression of miR-483-5p and CDK15 in ACC tissues and cell lines. The effects of miR-483-5p on proliferation were determined in vitro using CCK-8 proliferation assays, the changes of invasion of ACC cells were examined by Transwell. The molecular mechanism underlying the relevance between miR-483-5p and CDK15 was confirmed by luciferase assay and rescue assays.Results:We found a relatively higher miR-483-5p (2.36±1.02 vs 1.09±0.43) and lower CDK15 (0.57±0.26 vs 1.06±0.32) expression in ACC specimens and cell lines. CDK15 was verified as a direct target of miR-483-5p by luciferase assay. over-expression of miR-483-5p promoted proliferation (24 h: 0.26±0.03 vs 0.23±0.04, 48 h: 0.56±0.05 vs 0.41±0.03, 72 h: 0.73±0.04 vs 0.59±0.03) and invasion (95.78±4.66 vs 23.89±2.52) by down-regulating CDK15 expression.Conclusion:miR-483-5p plays a tumorigenesis role in ACC progression by down-regulating CDK15 expression, which may lead to a novel insight to the potential biomarker and novel therapeutic strategies for ACC.