Soluble endoglin和sFlt-1在子痫前期患者血清中的表达

目的探讨可溶性CD105淋巴细胞抗原及可溶性血管内皮生长因子受体1(sFlt-1)在子痫前期患者血清中表达水平的变化及意义。方法采用双抗体夹心酶联免疫吸附(ELISA)法测定子痫前期组、子痫前期各个亚组及正常晚期妊娠组血清sEng及sFlt-1的水平。结果①子痫前期组孕妇血清sEng、sFlt-1水平分别为(5.27±1.05)ng/mL、(391.14±70.25)pg/mL,正常晚期妊娠组分别为(1.26±0.40)ng/mL、(27.57±4.36)pg/mL,两组比较差异有统计学意义(P<0.01);②轻度子痫前期组sEng水平与重度子痫前期组比较差异有统计学意义(P<0.01),两组sFlt-1水平比较差异无统计学意义(P>0.05);③早发型重度子痫前期组sEng水平与晚发型重度子痫前期组比较差异有统计学意义(P<0.01),两组sFlt-1水平比较差异无统计学意义(P>0.05)。结论子痫前期患者血清中sEngs、Flt-1水平显著升高,sEng在早发型重度子痫前期血清中升高更显著,对早期诊断子痫前期有重要意义。

LAIR-1通过阻断JAK2 V617F突变的人HEL细胞JAK/STAT和PI3K/AKT/mTOR信号通路抑制其增殖并促进其凋亡

目的研究人白细胞相关免疫球蛋白样受体1(LAIR-1)对Janus激酶2(JAK2)V617F突变的人急性髓系白血病HEL细胞JAK/信号转导子与转录激活子(STAT)和磷脂酰肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/AKT/mTOR)信号通路的调节作用,以及对细胞增殖和凋亡的影响。方法采用反转录PCR和基因测序鉴定JAK2 V617F突变;应用免疫共沉淀和Western blot法鉴定LAIR-1募集的蛋白酪氨酸磷酸酶(PTP)种类;采用CCK-8法检测HEL细胞的增殖;采用异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双标记结合流式细胞术检测HEL细胞的凋亡率;采用Western blot法检测JAK/STAT和PI3K/AKT/mTOR通路蛋白酪氨酸磷酸化水平及细胞周期蛋白D1(cyclin D1)、Bcl2相关X蛋白(BAX)和B细胞淋巴瘤因子2(Bcl2)的蛋白表达。结果在JAK2 V617F突变的HEL细胞中,LAIR-1与其配体胶原蛋白结合后可募集含Src同源域2磷酸酶2(SHP-2);LAIR-1可以下调HEL细胞JAK2、STAT1、STAT3、STAT5、AKT和mTOR的蛋白酪氨酸磷酸化水平,并能够显著抑制cyclin D1和Bcl2的表达,而对BAX的表达水平未见显著影响;LAIR-1能够明显抑制HEL细胞的增殖,促进HEL细胞凋亡。结论在JAK2 V617F突变的人白血病HEL细胞中,LAIR-1可通过募集SHP-2抑制JAK/STAT和PI3K/AKT/mTOR信号通路的活化,进而抑制HEL细胞的增殖,促进细胞凋亡。

Different expression pattern of serum soluble intercellular adhesion molecules-1 and neutrophilic expression of CD18 in patients with diabetic retinopathy

AIM: To investigate the levels of serum soluble intercellular adhesion molecules-1 (sICAM-1) and neutrophilic expression of CD18 in patients with various stages of diabetic retinopathy and to determine their different expression pattern in the development of diabetic retinopathy(DR). METHODS: Levels of serum sICAM-1 and CD18 on the surface of neutrophile were measured in 41 DR patients, they were classified in three subgroups according to the stage of retinopathy as determined by fund's ophthalmoscopy; 10 control subjects were also studied. sICAM-1 were measured by enzyme-linked immunosorbent assay and CD18 by flow cytometry. RESULTS: The neutrophilic CD18 expression and serum sICAM-1 level were all significantly elevated in all diabetic subgroups compared to control subjects (P <0.01). The differences of CD18 and sICAM-1 among the diabetic subgroups were significant in CD18 but not in sICAM-1. The progression of retinopathy was associated with an increase both in CD18 and in sICAM-1 levels by simple correlation analysis (beta =0.74, P<0.001; beta =0.38, P<0.01, respectively). But stepwise multiple regression analysis revealed that only CD18 Was independent determinant of retinopathy (beta =1.04, P<0.01). CONCLUSION: Our results confirm the contribution of endothelial and neutrophilic activation in the development of DR as indicated by increased levels of CD18 and sICAM-1. However, a direct implication of CD18 and ICAM-1 in the progression of DR can be supported only in the CD18 but not ICAM-1. CD18 and ICAM-1 may play different role in the development of diabetic retinopathy.

Investigation on the Effects of Soluble Programmed Death-1 (sPD-1) Enhancing Anti-tumor Immune Response

Summary: By using semi-quantitative RT-PCR method, it was found that PD-L1 mRNA but not PD-L2 mRNA was expressed in H22 hepatoma cells and both PD-L1 and PD-L2 mRNAs were expressed in tumor tissues of tumor-bearing mice and upregulated as compared with muscle tissues in normal mice and H22 hepatoma cells. PD-L1 and PD-L2 were also expressed on the surface of the activated T cells. The soluble recombinant sPD-1 expressed from the constructed eukaryotic expression vector could enhance the lysis of tumor cells by lymphocytes stimulated specifically with antigen. The expresssion of sPD-1 by local gene therapy on the inoculation site of H22 hepatoma cells could inhibit the growth of tumor. The results of this study indicate that expression of soluble receptor of negative costimulatory molecules could reduce the inhibitory effect on T cells in tumor microenvironment and enhance the cytotoxicity of T cells on tumor cells. This possibly provides a new method of improving efficacy of tumor gene therapy.

Soluble neuregulin-1(NRG1): a factor promoting peripheral nerve regeneration by affecting Schwann cell activity immediately after injury

Neuregulin-1 (NRG1) is a well known growth factor playing contradictory roles in myelination depending on the existing isoform. Transmembrane NRG1 acts as a promyelinating factor, while the soluble isoform inhibits myelination. In this perspective, we would like to emphasize this conflicting role played by NRG1 isofbrms regarding their roles in myelination, remyelination and the entire process of peripheral nerve regeneration.

Elevated pancreatic enzymes, IgM, soluble interleukin-2 receptor in anti-GADab(+) type 1 diabetes

Type 1 diabetes can be classified into immune-mediated diabetes (type 1A) and idiopathic diabetes, which lacks immunological evidence for beta cell autoimmunity (type 1B). Type 1A diabetes is characterized by the presence of the anti-glutamic acid decarboxylase antibody (anti-GADab). Fulminant type 1 diabetes is classified as type 1B diabetes, and characterized by the absence of anti-GADab, flu-like symptoms, and elevated serum exocrine pancreatic enzymes. We report a type 1 diabetic patient who showed flu-like symptoms, elevated serum exocrine pancreatic enzymes, and an extremely high-titer of anti-GADab, manifesting the characteristics of both type 1A and fulminant type 1 diabetes.

Comparison of p38 MAPK, soluble endoglin and endothelin-1 level in severe preeclampsia and HELLP syndrome patients

Objective: To compare the levels of p38 mitogen-activated protein kinase (MAPK), soluble endoglin and endothelin-1 in the serum of patients with severe preeclampsia, hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome and normal pregnancies. Methods: This study was an observational analytic cross-sectional study performed at Wahidin Sudirohusodo Hospital, Makassar, Indonesia, in the period of 5th February 2016 to 20th January 2017. P38 MAPK, soluble endoglin and endothelin-1 levels of patients with normal pregnancies, severe preeclampsia and HELLP syndrome were measured by enzyme-linked immunoabsorbentassay technique, using kits of human soluble endoglin, endothelin-1 and p38 MAPK, Quantikine immunoassay: R&D System Inc. Results: Level of serum p38 MAPK in HELLP syndrome group was higher than in severe preeclampsia and normal pregnancy groups. Soluble endoglin and endothelin-1 levels in pregnancies with severe preeclampsia and HELLP syndrome were higher than normal pregnancy but there was no significant difference between these two groups (P>0.05). Levels of p38 MAPK, soluble endoglin and endothelin-1 also had a positive linear correlation with systolic and diastolic blood pressures (P<0.05). Conclusions: P38 MAPK in serum may be a marker for evidence of the severe hypoxia and its application may be considered for the diagnosis of HELLP syndrome.

Monocyte chemotactic protein-1 and soluble adhesion molecules as possible prognostic markers of the efficacy of antiviral treatment in chronic hepatitis C

AIM:To explain the role of Monocyte chemotactic protein-1 (MCP-1) and soluble adhesion molecules in chronic hepatitis C during the treatment of interferon alpha (IFNα) 2 b and ribavirin (RBV). METHODS:Concentrations of MCP-1,soluble adhesion molecules intercellular adhesion molecule-1 (sICAM-1),sP- selectin,interleukin (IL) 6,and IL10 in serum were estimated in the group of 40 patients with chronic hepatitis C treated with IFNalpha2 b and RBV in 0,16,32,48 wk of the therapy, RESULTS:In chronic hepatitis C,before and during the treatment,the serum levels of MCP-1 and sP-selectin in responders were similar to those of healthy subjects.In non- responders (NR),MCP-1 increased in the course of IFNc^+RBV treatment,differences were statistically significant as compared to responders.MCP-1 correlated statistically with the activity of periportal inflammation (r=0.35,P<0.05) but not with staging of liver fibrosis,sICAM-1 positively correlated with inflammatory activity and fibrosis in NR.sP-selectin did not correlate with histological findings in the liver.The MCP-1 correlated with the soluble form of sP-selectin concentrations (r= 6,P<0.001) and with IL-10 level in NR (r=0.4,P<0.05).There was no correlation observed between the concentration of MCP-1 and sICAM-1,IL-6 during the treatment. CONCLUSION:MCP-1 concentration may be a prognostic marker of the efficacy of IFN+RBV therapy in patients with chronic hepatitis C.

Changes of soluble intercellular adhesion molecule-1 in the serum from patients with acute cerebral infarction

objective: To observe the changes of soluble intercellular adhesion molecule-1(sICAM-1) in the serum of patients with acute cerebral infarctlon (ACI) and their clinical significance. Methods: The concen-tration of sICAM-1 in the serum of 91 patients with ACI was determined with ELISA and then the results were compared wlth those of 43 patients with cerebral hemorrhage and 30 healthy individuals. Results: In the 24th hour after infarction. the concentration of sICAMu-1 in the serum was significantly higher in patients with ACI than in patients with cerebral hemorrhage and normal controls (P< 0. 01). In the patients with ACI, the concentration exhibited an decreasing tendency in the period from the 24th hour to the 14th day andwas correlated with the focal size of cerebral infarction. During the first 14 days after infarction, the concen-tration was significantly higher in the patients with the complication of infection than in those without. Con-clusion: sICAM-1 is closely correlated with clinical manifestation of ACI.

The interleukin-1 receptor antagonist (IL-1-Ra) and soluble tumor necrosis factor receptor I (sTNF RI) in periodontal disease

The course and severity of periodontitis can be significantly affected by bacterial virulence as well as host immunity dysfunction. Periodontal tissue destruction has been proved to result from cascade of cytokines synthesized by reactive cells upon stimulation by pathogenic bacteria and lipopolysaccharides within their cell membranes. The clinical use of genetically programmed cells, producing substances blocking IL-1, based on recombinant IL-1 antagonist, as well as cytokines activating fibroblasts and osteoblasts to regenerate the destroyed periodontal tissue could prove alternative to the conventional treatment. Another cytokine of interest in respect to periodontitis ethiopathogenesis is soluble tumor necrosis factor receptor I (sTNF RI). Observation of soluble TNF receptors as physiologic inhibitors of TNF led to its administration in therapeutic process as well as in therapy selected cases of aggressive periodontitis.

Placenta Growth Factor and Soluble Fms-Like Tyrosine Kinase 1 in Preeclampsia and Normotensive Pregnant Nigerian Women

Background: Preeclampsia (PE) is still one of the leading causes of maternal/perinatal morbidity/mortality in Nigeria. Imbalance between placenta growth factor (PLGF) and soluble fms-like tyrosine kinase 1 (sFlt1) has been reportedly present both before and after the manifestation of PE;however, Nigerian data regarding these angiogenesis-related substances are lacking. We here attempted to determine the maternal serum level of PLGF and sFlt1 and sFlt1/PLGF ratio in PE vs. non-PE women in Lagos State University Teaching Hospital, Nigeria. Methods: An observational cross-sectional study was made on 75 women with PE and 75 age-gestational-age matched women without PE, as case and control, respectively. Levels of sFlt-1, PIGF and the sFlt-1: PIGF ratio was compared between the two. Results: Serum levels of Flt-1 and sFlt1/PIGF ratio were significantly higher in PE patients (6581.86 ± 865.75, and 146.42 ± 92.43) than in the normotensive control (4584.52 ± 1479.6 and 11.60 ± 6.42). PIGF was significantly lower in PE patients (70.14 ± 51.03) than the normotensives (494.06 ± 475.8). There were positive and negative correlations between the sFlt-1 and PLGF respectively and mean arterial blood pressure. Conclusion: Serum sFlt-1, sFlt1/PIGF ratio was significantly higher and PIGF levels were significantly lower in PE than normotensive control in Nigerian population.

Elevated soluble 4-1BB is associated with serum markers of hepatitis B virus in patients with chronic hepatitis B

BACKGROUND Previous studies have suggested that the costimulatory molecule 4-1BB plays pivotal roles in regulating immunity during chronic viral infection.However,up to now,there are few studies about 4-1BB in chronic hepatitis B(CHB).AIM To clarify this issue,we report our comprehensive study results on the expression levels of 4-1BB in patients with CHB.METHODS From September 2018 to June 2019,a total of 64 patients with CHB were recruited from the Department of Hepatology,The First Hospital of Jilin University.Peripheral blood samples were collected from 52 treatment-naïve and 12 entecavir-treated patients with CHB as well as 37 healthy donors(including 24 healthy adults and 13 healthy children).The levels of soluble 4-1BB(s4-1BB)in plasma were measured by ELISA.4-1BB mRNA expression in peripheral blood mononuclear cells was detected by real-time quantitative PCR.RESULTS The s4-1BB levels in the plasma of patients with CHB were significantly higher than those in healthy adults(94.390±7.393 ng/mL vs 8.875±0.914 ng/mL,P<0.001).In addition,the s4-1BB level in plasma was significantly increased in patients with a higher viral load and a disease flare up.However,there were no significant differences between treatment-naïve and entecavir-treated patients.Interestingly,among treatment-naïve patients with CHB,the levels of s4-1BB in plasma had a significant positive correlation with hepatitis B surface antigen,hepatitis B virus DNA,hepatitis B e antigen,and triglyceride levels(r=0.748,P<0.001;r=0.406,P=0.004;r=0.356,P=0.019 and r=-0.469,P=0.007,respectively).The 4-1BB mRNA expression was higher in the peripheral blood mononuclear cells of patients with CHB than in the peripheral blood mononuclear cells of healthy adults,but the difference was not statistically significant.CONCLUSION These results suggest that the levels of s4-1BB may be associated with pathogenesis of hepatitis B virus and therefore may be a promising biomarker for disease progression.

SIMULTANEOUS DETERMINATION OF FAT SOLUBLE VITAMINS A, D_3, E AND K_1 IN FORTIFIED MILK POWDERS BY HPLC

A high performance liquid chromatographic method for the simultaneous determination of four fat soluble vitamins: retinol (vitamin A), cholecalciferol(vitamin D), tocopherol(vitamin E) and phylloquinone(vitamin K_1) in fortified milk powders and egg yolk has been developed. The method requires enzymatic hydrolysis of lipid component of the sample as a pretreatment. Several factors which influence the enzymatic hydrolysis were studied-Separation was achieved using μ-Bondapak C-18 column(3.9×300mm), 98% methanol as mobile phase, a double channel detection was selected; vitamins D_3 E_1 K_1 were detected by UV spectrophotometry (265 nm) first, then vitamin A by fluorometry (EX 325nm, EM 480mm). The retention times of vitamin A_1 D_3, E and K_1 4.87, 9.00, 10.58 and 15.45 min respectively. Detection limit were 0.64, 0.25, 0.50 and 0.07 ng; and the recoveries were 90.5%～103.6% 90.0%～95.6%, 91.7%～98.8%, 91.5%～98.6%, respectively. The vitamins A, D_0, E, K_1 contents in foods were determined satisfactorily.

Selective deletion of the soluble Colony-Stimulating Factor 1 isoform in vivo prevents estrogen-deficiency bone loss in mice

Neutralizing CSF1 in vivo completely prevents ovariectomy （OVX）-induced bone loss in mice. There are two isoforms of CSF1, soluble （sCSF1）, and membrane-bound （mCSF1）, but their individual biological functions are unclear. It had been previously reported that mCSF1 knockout （K/O） and wild type （Wt） female mice experience the same degree of bone loss following OVX. In Wt mice the expression of sCSF1 was elevated fourfold in skeletal tissue following OVX while expression of mCSF1 was unchanged. To examine the role of sCSF1 in OVX-induced bone loss, mice were engineered in which sCSF1 was not expressed but expression of mCSF1 was unaffected （sCSF1 K/O）. Isoform-specific reverse transcription PCR confirmed the absence of transcripts for sCSF1 in bone tissue isolated from these animals and no circulating CSF1 was detected by ELISA. Surprisingly, there were no significant differences in bone mineral density （BMD） between sCSF1 K/O mice and Wt controls as assessed by dual-energy X-ray absorptiometry and micro-CT. However, one month after OVX, femoral, spinal and total BMD had declined by 11.2%, 8.9%, and 8.7% respectively in OVX-Wt animals as compared to Sham-OVX. In contrast OVX sCSF1 K/O mice showed changes of ＋0.1%, - 2.4%, and ＋2.3% at the same 3 sites compared to Sham-OVX sCSF1 K/O mice. These data indicate important non-redundant functions for the two isoforms of CSF1 and suggest that sCSF1, but not mCSF1, plays a key role in estrogen-deficiency bone loss.

Hydroaminomethylation of 1-Dodecene Catalyzed by Water-soluble Rhodium Complex

The hydroaminomethylation of 1-dodecene catalyzed by water soluble rhodium complex RhCl(CO)(TPPTS)2 in the presence of surfactant CTAB was investigated. High reactivity and selectivity for tertiary amine were achieved under relatively mild conditions.

Relationship of plasma homocysteine, soluble intercellular adhesion molecule-1, high mobility group box 1 protein with carotid intima-media thickness in elderly patients with type 2 diabetes mellitus

Objective:To explore the relationship of plasma homocysteine(Hcy),soluble intercellular adhesion molecule-1(sICAM-1)and high mobility group box 1 protein(HMGB1)with carotid intima-media thickness(c-IMT)in elderly patients with type 2 diabetes mellitus.Methods:A total of 100 elderly patients who were diagnosed as type 2 diabetes mellitus in Baogang Hospital of Inner Mongolia from June 2017 to May 2020 were chosen as research objects.According to c-IMT,they were divided into the normal group(n=35),the mild to moderate group(n=41)and the severe group(n=24).The expression levels of plasma Hcy,sICAM-1 and HMGB1 were compared between groups respectively.Pearson’s correlation coefficient was used to analyze the relationship of plasma Hcy,sICAM-1,HMGB1 with c-IMT.Results:The comparison in plasma Hcy,sICAM-1,HMGB1 and c-IMT among the three groups of patients was of statistical significance(p<.05).The results of correlation analysis showed that the expression levels of plasma Hcy,sICAM-1 and HMGB1 were positively correlated with c-IMT in elderly patients with type 2 diabetes mellitus(r=.627,.598,.614;p<.05).Conclusions:The expression levels of plasma Hcy,sICAM-1 and HMGB1 are abnormally increased in elderly patients with type 2 diabetes mellitus,and related to c-IMT,which can provide a strong evidence for clinical diagnosis and treatment by detecting their levels in clinical practice.

肿瘤患者外周血中抑制性受体LAIR-1表达升高

目的:研究LAIR-1在肿瘤患者中的表达,探讨其在抗肿瘤免疫应答中的作用。方法:应用夹心ELISA检测肿瘤患者血清中可溶型sLAIR-1的水平;应用免疫荧光染色和流式细胞术分析,观察LAIR-1在患者PBMC中NK细胞、CD4+T细胞、CD8+T细胞和B细胞上的表达。结果:肿瘤患者血清sLAIR-1水平为(4.6±3.2)μg/L,明显高于正常人血清sLAIR-1水平(3.9±3.0)μg/L,P<0.05。NK细胞,CD4+T细胞及CD8+T细胞表达LAIR-1水平上调。肺癌患者CD4/CD8比值明显低于正常人,B细胞百分率明显高于正常人。NK细胞、CD4+T细胞CD8+T细胞中LAIR-1的表达阳性率高于对照组。结论:在肿瘤患者LAIR-1分子表达水平高于正常人。肿瘤患者抑制性受体LAIR-1表达水平的上调可能与肿瘤的免疫逃逸有关。

人LAIR-1/CD305基因启动子的生物信息学分析

目的：研究人LAIR-1／CD305启动子及其5′上游调控序列。方法：通过检索NCBI中的人类基因组数据库，获得LAIR-1的转录本序列及翻译起始位点上游2500bp的序列。利用Promoter 2．0等软件预测LAIR-1的启动子序列，然后利用MatInspector等软件对启动子序列的转录因子结合位点和启动子功能模块进行预测。结果：LAIR-1基因的核心启动子区位于翻译起始密码子上游的600～200bp区域内。在转录调控区发现了一些重要的转录因子结合位点，并预测到13种启动子功能模块。结论：LAIR-1基因的表达可能受到多种转录因子和5′端上游调控序列的调控。

LAIR-1/LAIR-2(CD305/CD306)推定配体分布的研究

目的:研究LAIR-1(CD305)及LAIR-2(CD306)推定配体在多种细胞系上的分布,为鉴定LAIR-1/LAIR-2推定配体提供了实验依据。方法:建立稳定表达LAIR-1-Fc和LAIR-2-Fc融合蛋白的细胞系。以纯化的融合蛋白进行活细胞免疫荧光染色,用流式细胞仪检测LAIR-1及LAIR-2推定配体在多种细胞系膜表面的表达,并比较LAIR-1和LAIR-2推定配体的分布及可能结合的位点。结果:LAIR-1推定配体及LAIR-2推定配体均在人羊膜来源的上皮细胞系WISH上高表达,在人黑色素瘤细胞系C32、人胚肾上皮细胞系293T及人脐静脉内皮细胞系ECV304上有一定程度的表达。LAIR-1-Fc和LAIR-2-Fc融合蛋白与推定配体的结合,可分别被可同时识别LAIR-2和LAIR-1的单克隆抗体(mAb)FMU-LAIR-2.2和FMU-LAIR-2.1阻断。结论:LAIR-1推定配体及LAIR-2推定配体的分布基本一致,主要表达于WISH、C32、293T和ECV304细胞系。LAIR-1和LAIR-2可能拥有共同的配体。

9.1C3/LAIR-1的真核表达、纯化及单克隆抗体的制备和鉴定

目的 表达并纯化 9.1C3/LAIR 1胞膜外区与人的IgG1Fc段的融合蛋白 ,制备针对 9.1C3/LAIR 1胞膜外区的单克隆抗体 (mAb)。方法 构建真核表达载体 pIg/ 3C 9.1C3/LAIR 1,表达并纯化 9.1C3/LAIR 1 Fc融合蛋白。以 9.1C3/LAIR 1 Fc融合蛋白免疫BALB/c小鼠 ,制备。以间接免疫荧光染色和流式细胞术鉴定mAb的特异性 ,以竞争结合实验鉴定mAb识别LAIR 1的表位。结果表达并纯化了 9.1C3/LAIR 1 hIgFc融合蛋白 ,以其免疫BALB/c小鼠 ,获得 1株针对 9.1C3/LAIR 1胞膜外区的mAb 4H7。4H7对HL 6 0细胞和经 9.1C3/LAIR 1cDNA转染的COS7细胞上的表位识别 ,与已报道的抗 9.1C3mAb体不同。结论 成功地构建了 9.1C3/LAIR 1胞膜外区基因的真核表达载体 ,并表达纯化了 9.1C3/LAIR 1 hIgFc融合蛋白。获得一株针对 9.1C3/LAIR 1胞膜外区的mAb ,为进一步研究9.1C3/LAIR