白细胞介素-12家族在缺血性脑卒中发生发展中的作用机制研究

目前研究表明炎症反应和免疫功能异常对缺血性脑卒中的发生发展起重要作用,发现白细胞介素(IL)-12家族参与多种慢性炎症性疾病,对动脉硬化形成起关键作用,其可增强免疫反应和(或)抑制免疫功能。本文对IL-12家族的细胞因子在缺血性脑卒中中的作用现状进行简述,可能为缺血性脑卒中患者的预后判断和治疗方面提供新的参考。

白细胞介素-12促进氧化应激参与干燥综合征肝脏病变

目的:探讨白细胞介素-12(IL-12)对干燥综合征小鼠肝脏病变及氧化应激通路的影响,阐明干燥综合征肝脏病变的可能机制。方法:分别检测NOD、IL-12基因敲除(IL-12KO)NOD和C57BL/6(B6)小鼠的唾液流率、肝功能、肝脏病理、IL-12水平等。取不同浓度IL-12及JAK2、TYK2抑制剂作用于小鼠肝癌细胞。测定各组小鼠血清及细胞培养上清的氧化应激指标。结果:与其他两组小鼠相比,NOD小鼠GOT和GPT显著升高(P<0.05),血清GSH-PX、SOD和CAT降低(P<0.05)。不同浓度IL-12处理后,小鼠肝癌细胞CAT、GSH、GSH-PX和SOD有下降趋势,MDA增加。JAK2/TYK2抑制剂能逆转IL-12对肝癌细胞SOD、GSH和MDA的调节(P<0.05)。结论:IL-12可能通过促进干燥综合征肝脏细胞的氧化应激,进而参与干燥综合征肝脏病变过程。

术前血清VEGF和IL-12水平对原发性肝癌腹腔镜肝切除术预后的影响

目的分析原发性肝癌患者血清血管内皮生长因子(VEGF)、白细胞介素12(IL-12)水平对腹腔镜肝切除术预后的影响。方法回顾性收集2020年1月至2022年6月在首都医科大学附属北京佑安医院接受腹腔镜肝切除术治疗的原发性肝癌患者的临床资料,根据其预后结局分为预后不良组(80例)与预后良好组(80例)。所有患者一般资料、实验室检查资料均完整。分析术前血清VEGF、IL-12水平对原发性肝癌腹腔镜肝切除术治疗预后的影响。结果预后不良组与预后良好组间血清VEGF、IL-12、甲胎蛋白(AFP)水平比较差异均有统计学意义(P<0.05);Logistic回归分析发现,腹腔镜肝切除术前患者高水平血清VEGF、AFP可能是术后预后不良的风险因子(OR>1,P<0.05),而术前高水平血清IL-12可能是其保护因子(OR<1,P<0.05);受试者工作特征曲线(ROC)显示,原发性肝癌患者接受腹腔镜肝切除术治疗前血清VEGF、IL-12水平预测手术预后的曲线下面积(AUC)均>0.80,血清VEGF和IL-12联合预测的AUC为0.842。结论原发性肝癌患者术前血清VEGF、IL-12水平单独或联合均可以较好地预测腹腔镜肝切除术预后,高水平的VEGF以及低水平的IL-12均提示较差的预后。

子痫前期患者血清白细胞介素-12、基质细胞衍生因子-1的表达及其与不良妊娠结局的相关性

目的观察子痫前期患者血清白细胞介素-12(IL-12)、基质细胞衍生因子-1(SDF-1)的表达,并分析二者与不良妊娠结局的相关性。方法前瞻性纳入2019年6月至2021年6月南阳市中心医院收治的96例子痫前期患者作为研究对象,所有患者均随访至分娩结束后30 d,根据本次妊娠最终是否发生不良妊娠结局分组,全部患者于入院时检测血清IL-12、SDF-1水平,以点二列分析子痫前期患者血清IL-12、SDF-1水平与不良妊娠结局的相关性。结果96例子痫前期患者中无失访病例,随访至产后30 d,其中发生不良妊娠结局患者25例,占比为26.04%;发生组子痫前期患者血清IL-12、SDF-1水平高于未发生组,差异有统计学意义(P<0.05);经点二列相关性检验,结果显示,子痫前期患者血清IL-12、SDF-1水平与不良妊娠结局呈正相关(r=0.640、0.656,P<0.001);经logistic回归分析,结果显示,高血清IL-12水平、高SDF-1水平是不良妊娠结局的风险因子(OR>1,P<0.05)。结论子痫前期患者血清IL-12、SDF-1水平可能与不良妊娠结局存在一定关系,且高血清IL-12、高SDF-1水平子痫前期患者不良妊娠结局发生风险较高。

子痫前期患者血清白细胞介素-12、胎盘生长因子表达及其与胎盘早剥的关系

目的 观察子痫前期患者血清白细胞介素-12(IL-12)、胎盘生长因子(PLGF)的表达,分析其与胎盘早剥(PA)的关系。方法 选择2018年2月至2020年2月郑州人民医院收治的395例子痫前期患者作为研究对象,入院时测定血清IL-12、PLGF水平,随访至分娩,统计PA发生情况,比较两组一般资料与实验室指标水平,分析血清IL-12、PLGF水平与子痫前期患者PA的关系。结果 本研究395例子痫前期患者中,19例发生PA,PA发生率为4.81%(19/395);发生PA的患者子痫前期程度重于无PA的患者,入院时舒张压水平、血清IL-12水平高于无PA的患者,入院时血清PLGF水平低于无PA的患者(P<0.05);经logistic回归分析检验,结果显示,入院时舒张压、IL-12、PLGF与子痫前期患者发生PA有关(P<0.05);森林图显示,在子痫前期患者PA相关因素中,入院时IL-12水平与PA的关联度最强;绘制受试者工作特征(ROC)曲线发现,入院时血清IL-12、PLGF水平预测子痫前期患者PA风险的曲线下面积(AUC)分别为0.896、0.875,AUC均>0.80,有一定预测价值;绘制决策曲线分析(DCA)曲线,结果显示,最大净受益率为0.048,在阈值0~0.72范围内,入院时血清IL-12、PLGF联合预测子痫前期患者PA的净受益率优于单纯某一指标。结论 子痫前期患者血清IL-12表达越高或血清PLGF表达越低时,PA风险越高,入院时血清IL-12、PLGF水平预测子痫前期患者PA具有一定的价值。

男性尖锐湿疣患者血清IL-12、IL-10水平及其对复发的预测价值

目的探讨血清白细胞介素(IL)-12、IL-10在男性尖锐湿疣(CA)患者CO_(2)激光联合5-氨基酮戊酸光动力(ALA-PDT)治疗前后的变化及对复发的预测价值。方法选取2020年1月至2021年6月于安阳地区医院接受治疗并随访至2022年1月的112例CA患者作为研究对象,均接受CO_(2)激光联合ALA-PDT治疗,对比治疗前后的血清IL-12、IL-10水平,并根据患者治疗后是否复发分为复发组(n=23)与未复发组(n=89),分析一般资料与血清IL-12、IL-10的相关性;采用受试者工作特征(ROC)曲线分析IL-12、IL-10预测复发的效能。结果112例CA患者治愈率为90.18%,不良反应发生率为1.79%,复发率为20.54%,且治疗后血清IL-12高于治疗前,IL-10低于治疗前(P<0.05)。复发组与未复发组在年龄、婚姻情况、吸烟史、饮酒史、受教育程度、正常使用安全套等方面差异无统计学意义(P>0.05),而在皮损数量、皮损部位、人乳头状瘤病毒(HPV)感染情况、治疗前IL-12、IL-10方面差异有统计学意义(P<0.05)。logistic回归分析显示,多个皮损部位、多重HPV感染、皮损数量增加及治疗前血清IL-12下降、IL-10升高是影响CA患者复发的独立危险因素(P<0.05)。Pearson相关性分析显示,皮损数量与IL-10呈正相关(P<0.05);Spearman相关性分析显示,HPV感染情况、皮损部位与IL-12呈负相关,与IL-10呈正相关(P<0.05)。ROC曲线分析显示,IL-12、IL-10预测CA复发的曲线下面积(AUC)分别为0.920、0.709,敏感度分别为95.70%、91.30%,特异度分别为89.90%、50.60%。结论血清IL-12、IL-10在CA患者中呈异常表达,CO_(2)激光联合ALA-PDT治疗有利于降低IL-10和提高IL-12水平,并且动态监测IL-12、IL-10有利于为预测CA复发提供重要参考。

An effective vaccine against colon cancer in mice:Use of recombinant adenovirus interleukin-12 transduced dendritic cells

AIM: To investigate the effect of a vaccine with recombinant adenovirus interleukin-12 (AdVIL-12) transduced dendritic cells (DCs) against colon cancer in mice. METHODS: DCs and AdVIL-12 were incubated together at different time intervals and at different doses. Supernatant was collected and tested for IL-12 by enzyme-linked immunosorbent assay (ELISA). In order to determine whether tumor cell lysate-pulsed (TP) AdVIL-12/DCs enhance therapeutic potential in the established tumor model,CT26 colon tumor cells were implanted subcutaneously (s.c.) in the midflank of nave BALB/c mice. Tumor-bearing mice were injected with a vaccination of CT26 TP AdVIL-12/DCs on d 3 and 10. As a protective colon tumor model,nave BALB/c mice were immunized s.c. in their abdomens with CT26 TP AdVIL-12/DCs twice at seven day intervals. After the immunization on d 7,the mice were challenged with a lethal dose of CT26 tumor cells and survival times were evaluated. Subsequently,cytotoxic T lymphocyte (CTL) activity and interferon gamma (IFNγ) secretion was evaluated in the immunized mice,and assayed CTL ex vivo. RESULTS: Murine DCs were retrovirally transduced with AdVIL-12 effi ciency,and the AdVIL-12 transduced DCs secreted a high level of IL-12 (AdVIL-12/DCs,615.27 ± 42.3 pg/mL vs DCs,46.32 ± 7.29 pg/mL,P < 0.05). Vaccination with CT26 TP AdVIL-12/DCs could enhance anti-tumor immunity against CT26 colon tumor in murine therapeutic models (tumor volume on d 19: CT26 TP AdVIL-12/DCs 107 ± 42 mm3 vs CT26 TP DCs 383 ± 65 mm3,P < 0.05) and protective models. Moreover,the CT26 TP AdVIL-12/DC vaccination enhances tumor-specific CTL activity,producing high levels of IFNγ in immunized mice. Ex vivo primed T cells with AdVIL-12/DCs were able to induce more effective CTL activity than in primed T cells with CT26 TP/DCs (E:T = 100:1,69.49% ± 6.11% specif ic lysis vs 37.44% ± 4.32% specifi c lysis,P < 0.05). CONCLUSION: Vaccination with recombinant AdVIL-12 transduced DC pulsed tumor cell lysate enhance anti-tumor immunity specifi c to colon cancer in mice.

Expression and significance of intratumoral interleukin-12 and interleukin-18 in human gastric carcinoma

AIM: To explore the effect of intratumoral expressions of interleukin-12 (IL-12) and interleukin-18 (IL-18) on clinical features, angiogenesis and prognosis of gastric carcinoma. METHODS: The expressions of IL-12 and IL-18 from 50 samples of gastric cancer tissue were analyzed by immunohistochemistry, and microvessel density (MVD) was determined with microscopic imaging analysis system. RESULTS: The positive expression rates of IL-12 and IL-18 were 44% (22/50) and 26% (13/50), respectively. IL-12 was significantly associated with pathologic differentiation, depth of invasion, lymph node metastasis, distant metastasis, and TNM stage, and IL-18 was closely related to distant metastasis. Intratumoral IL-12 and IL-18 expressions were not statistically related to MVD scoring. IL-12-positive patients survived significantly longer than those with IL-12-negative tumors, but there was no significant difference between IL-18-positive patients and IL-18-negative ones. The multivariate analysis with Cox proportional hazard model revealed IL-12, MVD and T stage were independent prognostic factors. CONCLUSION: The positive expressions of IL-12 and IL-18 can play an important role in progression and metastasis of gastric cancer, and IL-12 might be an independent factor of poor prognosis in gastric carcinoma.

Interleukin-12 and Th1 immune response in Crohn’s disease: Pathogenetic relevance and therapeutic inplication

Crohn’s disease (CD) and ulcerative colitis (UC) are chronic inflammatory disorders of the gastrointestinal tract that share clinical and pathological characteristics. The most accredited hypothesis is that both CD and UC result from a deregulated mucosal immune response to normal constituents of the gut microflora. Evidence, however, indicates that the main pathological processes in these two diseases are distinct. In CD, the tissue- damaging inflammatory reaction is driven by activated type 1 helper T-cell (Th1), whereas a humoral response predominates in UC. Consistently, a marked accumulation of macrophages making interleukin (IL)-12, the major Th1-inducing factor, is seen in CD but not in UC mucosa. Preliminary studies also indicate that administration of a monoclonal antibody blocking the IL-12/p40 subunit can be useful to induce and maintain clinical remission in CD patients. Notably, the recently described IL-23 shares the p40 subunit with IL-12, raising the possibility that the clinical benefit of the anti-IL-12/p40 antibody in CD may also be due to the neutralization of IL-23 activity. This review summarizes the current information on the expression and functional role of IL-12 and IL- 12-associated signaling pathways both in patients with CD and experimental models of colitis, thus emphasizing major differences between IL-12 and IL-23 activity on the development of intestinal inflammation.

白细胞介素-12 通过抑制STAT3 途径调控非小细胞肺癌细胞生物学行为及免疫逃逸因子分泌的研究

目的探索白细胞介素-12(IL-12)对非小细胞肺癌细胞生物学行为及免疫逃逸因子分泌的影响。方法收集126例非小细胞肺癌患者癌组织及癌旁正常组织,通过实时定量PCR检测IL-12、STAT3 mRNA的表达水平。构建人肺腺癌A549细胞模型,分为空白对照组、腺病毒对照组、IL-12腺病毒组、质粒对照组、STAT3质粒组、IL-12与STAT3共转染组。比较各组细胞增殖及凋亡情况,采用ELISA法检测细胞培养液中IL-2、INF-γ、TNF-α的分泌量,采用Western blot法检测IL-12、STAT3、pSTAT3蛋白表达水平。结果肺癌组织较癌旁组织IL-12 mRNA表达量降低、STAT3 mRNA表达量增高。IL-12腺病毒组较各对照组细胞增殖及活力降低、凋亡增加。STAT3质粒组细胞增殖及凋亡结果则与之相反。IL-12、STAT3共转染组细胞增殖、活力以及凋亡情况介于IL-12腺病毒组与STAT3质粒组之间。此外,IL-12腺病毒组细胞培养液中IL-12、TNF-α、IFN-γ含量明显高于STAT3质粒组,而STAT3质粒组STAT3、p-STAT3蛋白表达水平显著高于在IL-12腺病毒组。结论IL-12对非小细胞肺癌细胞具有抑制增殖、降低活力及诱导凋亡的能力,其作用机制可能通过抑制STAT3途径以及免疫逃逸而实现。

Interleukin-12 as a Genetic Adjuvant Enhances Hepatitis C Virus NS3 DNA Vaccine Immunogenicity

Hepatitis C virus (HCV) chronic infection is a worldwide health problem, and numerous efforts have been invested to develop novel vaccines. An efficient vaccine requires broad immune response induction against viral proteins. To achieve this goal, we constructed a DNA vaccine expressing nonstructural 3 (NS3) gene (pcDNA3.1-HCV-NS3) and assessed the immune response in C57BL/6 mice. In this study, the NS3 gene was amplified with a nested-reverse transcriptase-polymerase chain reaction (RT-PCR) method using sera of HCV-infected patients with genotype 1a. The resulting NS3 gene was subcloned into a pcDNA3.1 eukaryotic expression vector, and gene expression was detected by western blot. The resultant DNA vaccine was co-administered with interleukin-12 (IL-12) as an adjuvant to female C57BL/6 mice. After the final immunizations, lymphocyte proliferation, cytotoxicity, and cytokine levels were assessed to measure immune responses. Our data suggest that co-administration of HCV NS3 DNA vaccine with IL-12 induces production of significant levels of both IL-4 and interferon (IFN)-γ (p<0.05). Cytotoxicity and lymphocyte proliferation responses of vaccinated mice were significantly increased compared to control (p<0.05). Collectively, our results demonstrated that co-administration of HCV NS3 and IL-12 displayed strong immunogenicity in a murine model.

Study of recombinant human interleukin-12 for treatment of complications after radiotherapy for tumor patients

AIM To evaluate the treatment effects of recombinant human interleukin-12(rh IL-12) on radiotherapy complications, such as severe myelosuppression or pancytopenia, the decline or imbalance of immune function, etc.METHODS The patients received high-dose and short-course precise radiotherapy, such as Cyber knife and image-guided radiotherapy(IGRT), which can cause myelosuppression or pancytopenia and immune function decline within a short time. One-hundred subjects were enrolled in the study, and 50 were randomized to a treatment group which used rh IL-12 and 50 were randomized to a control group which used symptomatic and supportive therapy after radiotherapy. The 50 subjects in the treatment group were further divided into five subgroups and intervenedwith rh IL-12 at a dose of 50, 100, 150, 200 or 250 ng/kg respectively. The dose-effect relationship was observed. RESULTS Rh IL-12 significantly attenuated the decrease of peripheral blood cells in the treatment group, and immune function was improved after treatment. Due to the different radiation doses, there was a fluctuation within 12 h after treatment but mostly showing an increasing trend. As to the clinical manifestations, 2 patients in the 250 ng/kg subgroup showed low fever after administration, 1 patient in the 200 ng/kg subgroup and 2 patients in the 250 ng/kg subgroup showed mild impairment of liver function during the observation period.CONCLUSION Rh IL-12 has effective therapeutic and protective effects on complications following radiotherapy, such as the decline of blood cells, myelosuppression and the decline or imbalance of immune function, which indicated good prospects for development and application.

Interleukin 12/interleukin 23 pathway: Biological basis and therapeutic effect in patients with Crohn's disease

Considering that both innate and adaptive immune responses are involved in the pathogenesis of Crohn's disease(CD), novel therapeutic options have significantly been developed. Biological agents represent an important addition to the conventional treatments for immuno-inflammatory conditions, acting as antagonists of adhesion molecules or various inflammatory cytokines. The interleukin 12(IL-12)/IL-23 common pathway has been found to play a determinant role in the induction of inflammation in adaptive immune responses. In particular, IL-23 promotes the differentiation of na?ve T helper cells into Th17 phenotype with the concomitant secretion of several inflammatory cytokines such as IL-17 and IL-22, whereas IL-12 induces the Th1 polarization and production of critical cytokines such as interferon-γ and tumor necrosis factor. Nowadays, there is increased interest regarding the role of IL-23 as a therapeutic target of CD through the blockage of IL-23 mediated pathways. In this editorial, we focus on the role of IL-12/IL-23 pathway in the regulation of mucosal immunity and in the induction and maintenance of chronic inflammation. In parallel, we critically discuss the available data regarding the therapeutic effect of the IL-12/IL-23 inhibitors and especially of ustekinumab, a human monoclonal antibody which has been recently approved by the United States Food and Drug Administration for the management of moderateto-severe CD and its potential to be used as first-line therapy in everyday clinical practice.

Relationship of Serum Interleukin-18 and Interleukin-12 Levels with Clinicopathology in Renal Cell Carcinoma

Objective： To investigate the relationship between serum interleukin-18 and interleukin-12 levels and clinicopathology of renal cell carcinoma. Methods： Peripheral blood samples were obtained from 20 healthy volunteers and 60 patients with renal cell carcinoma before curative surgery. IL-12 and IL-18 levels were determined by enzyme-linked immunosorbent assay. Results： Mean serum IL-12 and IL-18 levels were significantly higher in patients with renal cell carcinoma compared with healthy volunteers （P〈0.05） and mean serum IL-12 and IL-18 levels increased in patients as the pathologic stage progressed. A positive correlation was observed between serum IL-12 and IL-18 levels （P〈0.05）. In patients with renal cell carcinoma, increasing serum IL-12 and IL-18 levels correlated with pathological stage and Fuhrman grade. Conclusion： Serum IL-12 and IL-18 might be useful tumor markers in patients with renal cell carcinoma.

Immune Killing Activity of Lymphocytes on Hela Cells Expressing Interleukin-12 In Vitro

The killing effects of lymphocytes on Hela cells expressing interleukin-12 （IL-12） in vitro were explored. By using gene transfection technique, full length IL-12 gene was transfected into Hela cells. The expression of IL-12 in Hela cells was detected quantitatively by ELISA; Changes in killing effects of lymphocytes on Hela cells expressing IL-12 were observed by MTT. It was found that Hela cells could express IL- 12 between 24 h and 72 h after transfection. Killing activity of lymphocytes on Hela cells expressing IL-12 was significantly enhanced. It was concluded by cell transfection technique, Hela cells could express 1L-12 and were more easily killed by lymphocytes.

CONSTRUCTION OF THE DICISTRONIC ADENOVIRUS VECTOR EXPRESSING BIOACTIVE HUMAN INTERLEUKIN-12

The full-length cDNA encoding the subunits p40 and p35 of human interleukin12(hIL12) were cloned separately by RTPCR, linked together by internal ribosomal entry site (IRES) of encephalomyocarditis virus which initiates capindependent translation to form a dicistronic gene fragment. The dicistronic fragment was placed between the cytomegalovirus (CMV) promoter and SV40 polyA signal to form a dicistronic expression cassette. Subsequently, the dicistronic expression cassette was inserted into E1 region of Ad5 genome in cosmid vector pAx1cw of E1substitution type. By homologous recombination with EcoT22Idigested Ad5 DNATPC in 293 cells, the replicationdeficient recombinant adenoviruses of hIL12 were generated efficiently. After infected with hIL12 recombinant adenoviruses in vitro, 293 cells, human hepatocellular carcinoma cells HepG2, and primary human skin fibroblasts expressed and secreted hIL12 at comparable levels (30～60ng/ 106cells/24hr), which could stimulate the proliferation and IFNγ production of human lymphoblasts. These suggest that the dicistronic adenovirus vector of hIL12 could effectively mediate the expression of bioactive hIL12 and might be used in cancer gene therapy.

Modulation of cellular and humoral immune responses to anHIV-1 DNA vaccine by interleukin-12 and interleukin-18 DNA immunization

Objective: To investigate the effect of interleukin-12 (IL-12) and interleukin-18 (IL-18)DNA immunization on immune response induced by HIV-1 DNA vaccine and to explore new strategies for therapeutic HIV DNA vaccine. Methods: The recombinant expression vector pCI-neoGAG was constructed by inserting HIV Gag gene into the eukaryotic expression vector pCI-neo. Balb/c mice were immunized with pCI-neoGAG alone or co-immunized with the DNA encoding for IL-12 or IL-18.Anti-HIV antibody and IFN-γ were tested by ELISA,and splenocytes were isolated for detecting antigen-specific lymphoproliferative responses and specific CTL response by MTT assay and LDH assay respectively. Results: The anti-HIV antibody titers of mice co-immunized with pCI-neoGAG and the DNA encoding for IL-12 or IL-18 were lower than that of mice immunized with pCI-neoGAG alone(P<0.01). In contrast, the IFN-γ level of mice co-immunized with pCI-neoGAG and the DNA encoding for IL-12 or IL-18 was higher than that of mice immunized with pCI-neoGAG alone (P<0.01).Furthermore, compared with mice injected with pCI-neoGAG alone, the specific CTL cytotoxity activity and antigen-specific lymphoproliferative responses of mice immunized with pCI-neoGAG and the DNA encoding for IL-12 or IL-18 were significantly enhanced respectively (P<0.01). Conclusion: The DNA encoding for IL-12 or IL-18 together with HIV DNA vaccine may enhance specific Th-1 responses and cellular immune response elicited in mice. Hence, the DNA encoding for IL-12 or IL-18 are promising immune adjuvants for HIV-1 DNA vaccine.

妊娠高血压孕妇血清25(OH)D、Hcy、IL-12水平变化及其临床意义

目的观察妊娠高血压孕妇的血清25羟维生素D[25(OH)D]、同型半胱氨酸(Hcy)、白细胞介素-12(IL-12)水平变化,分析其临床意义,以减少不良妊娠结局。方法回顾性收集40例妊娠高血压孕妇的临床资料为研究组,另选取同期体检的健康孕妇40例为对照组,比较两组孕妇的血清25(OH)D、Hcy、IL-12的水平,并采用受试者工作特征曲线(ROC)分析上述指标诊断妊娠高血压的效能以及对预后的判断价值。结果研究组孕妇的收缩压、舒张压、血清Hcy及IL-12水平均高于对照组,研究组的新生儿体重、孕妇的血清25(OH)D水平低于对照组,差异均有统计学意义(P<0.05)。ROC结果显示:血清25(OH)D、Hcy、IL-12水平诊断妊娠高血压的AUC分别为0.880、0.921、0.914,95%CI分别为0.808~0.952、0.865~0.976、0.853~0.975,早期诊断的Cutoff值分别为18.581 mg/mL、14.847μmol/L、56.846 pg/mL。研究组孕妇的不良妊娠结局发生率(27.50%)高于对照组(7.50%),差异有统计学意义(χ^(2)=5.541,P=0.019)。结论妊娠高血压孕妇的血清25(OH)D水平降低、血清Hcy和IL-12水平升高,25(OH)D、Hcy、IL-12对妊娠高血压有一定诊断价值。

基于通量测序的CXCL12、IL-6和IDO基因在去势抵抗性前列腺癌细胞中的表达及意义

目的结合生物信息学分析,探讨趋化因子CXc配体L12(CXCL12)、白细胞介素6(IL-6)和吲哚胺-2,3-双加氧酶(IDO)在去势抵抗性前列腺癌细胞中的表达水平。方法利用激素敏感性前列腺癌细胞株LNCaP和去势抵抗性前列腺癌细胞株C4-2细胞的基因表达谱,采用R3.6.1软件对2组的表达数据进行差异基因分析,并根据差异基因进行基因本体(GO)功能富集分析,筛选差异基因富集的京都基因和基因组百科全书(KEGG)信号通路,最终选定CXCL12、IL-6和IDO三个差异基因;将LNCaP和C4-2细胞分为control组(常规处理)、si-NC组(将si-NC转染入细胞内)、si-IL-6组(将si-IL-6转染入细胞内)和si-CXCL12组(将si-CXCL12转染乳细胞内);采用实时荧光定量PCR(RT-PCR)及Western blot法检测LNCaP和C4-2细胞株中CXCL12、IL-6和IDO各自mRNA及蛋白表达水平;使用si-IL-6、si-CXCL12敲低IL-6、CXCL12在LNCaP、C4-2细胞中的表达,采用qRT-PCR法检测细胞中IDO、CXCL12和IL-6基因表达,WB检测相关蛋白表达,MTT法检测LNCaP或C 4-2细胞增殖、流式凋亡术检测细胞凋亡、Transwell实验检测细胞侵袭能力、划痕恢复实验检测细胞迁移能力。结果生物信息学分析结果显示,细胞株LNCaP和细胞株C4-2之间共有3505个差异基因,其中上调基因1695个、下调的基因1810个;RT-PCR法和WB检测显示,与C4-2细胞比较,LNCaP细胞中IL-6和IDOmRNA及蛋白表达量均明显增高,CXCL12 mRNA及蛋白量显著降低,差异有统计学意义(P<0.05);将si-IL-6转染入LNCaP和C4-2细胞后,与si-NC组比较,LNCaP细胞的si-IL-6组中细胞增殖、侵袭和迁移能力受到显著抑制,细胞凋亡率显著增加,差异有统计学意义(P<0.01);与si-NC组比较,si-IL-6组中IL-6和IDO蛋白和mRNA表达水平显著降低,差异有统计学意义(P<0.01);将si-ICXCL12转染入LNCaP或C4-2细胞后,与正常C4-2细胞比较,si-CXCL12组中C4-2细胞增殖、侵袭和迁移能力受到显著抑制,同时细胞凋亡率显著增加,差异有统计学意义(P<0.01);与si-NC组比较,si-CXCL12组中CXCL12蛋白和mRNA表达水平显著降低,差异有统计学意义(P<0.01)。结论CXCL12、IL-6和IDO在不同前列腺癌中表达异常,可能是导致前列腺癌治疗方案差异性较大的原因。