Human Kallikrein-2 and Free Prostate Specific Antigen as Biomarkers for Early Detection of Prostate Cancer, Sudan: A Case-Control Study

Background: Prostate cancer (PCa) is considered one of the major health threats facing males in Sudan. Prostate-specific antigen (PSA) test is the most important laboratory test used in the diagnosis of prostate cancer, the main disadvantage of PSA is its limited specificity, which triggered a lot of interest in development, more research on other markers such as serum human kallikrein-2 (KLK-2) and free prostate specific antigen (fPSA). Objectives: To evaluate the validity of serum kallikrein-2 (KLK-2) and free prostate specific antigen (fPSA) in the early detection of prostate cancer among Sudanese patients. Method: In this study seventy men were considered as a case subject, who were diagnosed as cancer prostate at Gezira Hospital for Renal Disease and Surgery (GHRDS), Sudan during the period February 2018 to July 2019. Randomly selected sixty patients of BPH patients and forty-five apparently healthy men as control subject. KLK-2, fPSA and PSA estimations were performed from serum samples using the principle of Enzyme Linked Immunosorbent Assay (ELISA). Results: The results revealed a highly significant difference between the serum levels individual biomarkers (KLK-2, fPSA, PSA) and multiple biomarkers (fPSA/PSA, KLK-2/fPSA, KLK-2/PSA) for patients with prostate cancer when compared with the control groups. Furthermore, the fPSA/PSA ratio was lower in the patients with prostate cancer (P value = 0.00) than in the control group, the fPSA/PSA ratio showed that best accuracy to differentiate prostate cancer from control group, fPSA cut-off value was found to be more than (18 ng/ml), with sensitivity (93%), specificity (80%), and odds ratio (55). Conclusions: The use of multiple biomarkers rather than individual biomarkers especially fPSA/PSA ratio improves the specificity as well as maintenance of higher sensitivity for early diagnosis of the prostate cancer.

Development and evaluation of an ELISA method for the measurement of kallikrein-related peptidase 5 (KLK5) in human serum

Kallikrein-related peptidases (KLKs) have been proposed as potential cancer biomarkers. Kallikrein-related peptidase 5 (KLK5) is a secreted trypsin-like protease of the KLKs. Until now, detection of KLK5 in both biological fluids and tissues has been described frequently due to the potential of being a new cancer biomarker. Our objective was to prepare KLK5 antibodies and establish an ELISA method for KLK5 to study the possible clinical application of KLK5 as a biomarker for malignancies. In this study, recombinant KLK5 protein was produced and purified using a prokaryotic expression system, and then used as immunogen to generate antibodies. High titers of specific antibodies were measured in serum of rabbits after the forth booster injection. And the titer of the antiserum reached 1:106. We have also generated monoclonal antibodies using hybridoma technology and the titer reached 1:105. The activity of KLK5 antibodies was characterized by Western blot and immunohistochemistry. To quantitatively examine KLK5 in serum samples, we established double antibody sandwich ELISA method using mouse mAb as capture and rabbit pAb as tracer antibody. We have detected KLK5 levels in ovarian cancer serum to ensure that our sandwich ELISA measurement to have high sensitivity and specificity. The ranges of linearity reached by the standard curves of the newly developed ELISA were 0.45 ng/mL to 125 ng/mL. The detection limit of the method, defined as the concentration of KLK5 can be distinguished, was 0.20 ng/mL. Median serum KLK5 levels were 3.77 ng/mL and 0.86 ng/mL in ovarian cancer patients and normal female, respectively (P ELISA assay for KLK5. Our preliminary findings prompt that KLK5 may be a new potential biomarker for the diagnosis and prognosis in patients with ovarian.

人类Kallikrein基因家族与肿瘤

人组织型激肽释放酶基因家族（Kallikrein，简称KLK）是丝氨酸蛋白酶家族的一个亚群，近年新的研究发现位于人染色体19q13．3-13．4区域的基因克隆与经典的KLK基因（KLK1-编码hK1或肾／胰腺Kallikrein；KLK2-编码hK2／人腺型Kallikrein；KLK3-编码hK3／前列腺特异性抗原PSA）具有高度的同源性，使得该家族成员扩大到15个，并在GENBANK中首次注册命名。研究发现，除KLK3编码的hK3／PSA外，多种KLK基因家族成员在肿瘤尤其激素依赖性肿瘤中表达异常，并参与肿瘤发生发展的多个环节。本文旨在对近年来KLK基因家族与肿瘤的研究进展作一总结。

Kallikreins基因结构及功能的研究前景

最近关于Kallikrein基因结构及其表达的研究发现,Kallikrein是一类新的肿瘤标记家族.过去认为人Kallikrein基因位点仅含有3个基因--KLK1、KLK2和KLK3[1].目前,几个独立的研究小组证明并报告Kallikrein基因位于人染色体的19q13.4,已克隆出15种Kallikrein基因家族成员,在GenBank首次注册并命名.该方面的研究取得了引人注目的进展,现作一介绍.

Overexpression of kallikrein gene 10 is a biomarker for predicting poor prognosis in gastric cancer

AIM:To analyze the expression of kallikrein gene 10(KLK10)in gastric cancer and to determine whether KLK10 has independent prognostic value in gastric cancer.METHODS:We studied KLK10 expression in 80 histologically confirmed gastric cancer samples using realtime quantitative reverse transcription-PCR and hK10expression using immunohistochemistry.Correlations with clinicopathological variables(lymph node metastasis,depth of invasion and histology)and with outcomes(disease-free survival and overall survival)during a median follow-up period of 31 mo were assessed.Gastric cancer tissues were then classified as KLK10 positive or negative.RESULTS:KLK10 was found to be highly expressed in 57/80(70%)of gastric cancer samples,while its expression was very low in normal gastric tissues.Positive relationships between KLK10 expression and lymph node metastasis(P=0.048),depth of invasion(P=0.034)and histology(P=0.015)were observed.Univariate survival analysis revealed that gastric cancer patients with positive KLK10 expression had an increased risk for relapse/metastasis and death(P=0.005 and0.002,respectively).Cox multivariate analysis indicated that KLK10 was an independent prognostic indicator of disease-free survival and overall survival in patients with gastric cancer.CONCLUSION:KLK10 expression is an independent biomarker of unfavorable prognosis in patients with gastric cancer.

Clinical significance of human kallikrein 12 gene expression in gastric cancer

AIM:To investigate whether the expression of kallikrein 12(KLK12) is related to the development of gastric cancer(GC) and to determine the role of KLK12 in gastric cancer cells growth,invasion and migration.METHODS:Between September 2007 and March 2008,133 patients with histologically confirmed GC were recruited for the study.Expression of KLK12 was detected in samples from GC patients by quantitative real-time reverse transcription polymerase chain reaction and immunohistochemistry.The relationship between KLK12 protein expression and clinicopathological features of GC was analyzed.The difference in 5-year survival rates between the high KLK12 protein expression group and the low KLK12 expression group was compared.Additionally,the expression of KLK12 was examined in various human GC cell lines,including MKN-28,SGC-7901 and MKN-45.Small interfering RNA(siRNA) was used to inhibit KLK12 expression in MKN-45 cells.Cell clones stably transfected with KLK12 siRNA were tested for KLK12 expression by quantitative real-time reverse transcription-polymerase chain reaction and Western blotting.Furthermore,a series of functional assays were performed in this study to assess the biological features of transfected cells.Cell proliferation was assessed using the methylthiazolyltetrazoliumassay.Finally,cell migration and invasion were assessed using transwell chamber assays.RESULTS:Of the 133 GC patients included in the study,126(94.7%) showed a higher expression level of KLK12 mRNA when compared to noncancerous tissue specimens.Expression of KLK12 mRNA was significantly higher in GC tissues than in normal tissue(P < 0.001).KLK12 protein expression was detected in 96 of 133(72.2%) GC samples with moderate or strong staining primarily in the cytoplasm.In contrast,negative immunostaining for KLK12 protein was observed in the corresponding normal gastric mucosal tissue.Overexpression of KLK12 protein was significantly associated with lymph node metastasis(P = 0.001),histological type(P < 0.001) and tumor-node-metastasis stage(P = 0.005),while no significant correlation was observed between expression of KLK12 protein and sex,age,depth of invasion,tumor size or lymphatic invasion.Furthermore,patients with high KLK12 expression had a significantly poorer 5-year survival rate than those with low KLK12 expression(P = 0.002).Expression of KLK12 mRNA was significantly higher in MKN-45 GC cells compared to normal mucosal cells or two other GC cell lines(P < 0.01).Expression of KLK12 in MKN-45 cells was downregulated after transfection with siRNA.Knockdown of KLK12 markedly decreased the proliferation of MKN-45 cells when compared with parent or mock-transfected cells(P = 0.001),especially from the 3rd to the 5th day of the assay.In migration assays,fewer KLK12 siRNA cells migrated through the chambers(22.00 ± 1.81) when compared to the parent(46.47 ± 2.42) or mock-transfected cells(45.40 ± 1.99);these differences were statistically significant(P < 0.001).However,in the invasion assay,the number of KLK12 siRNA cells that invaded the chambers was 18.40 ± 1.12,closely similar to both the parent(18.67 ± 0.98) and mock-transfected cells(18.53 ± 0.92).There was no significantly difference between the three groups in the invasion assay(P = 0.054).CONCLUSION:The KLK12 gene is markedly overexpressed in GC tissue,and its expression status may be a powerful prognostic indicator for patients with GC.KLK12 might serve as a novel diagnosis and prognosis biomarker in GC.

PTD-kallikrein对大鼠MCAO模型缺血再灌注损伤的保护作用

目的:观察PTD-kallikrein对大鼠MCAO模型缺血再灌注损伤的保护作用,并探讨其可能的作用机制。方法:以化学交联法构建PTD-kallikrein,鉴定PTD-kallikrein对血脑屏障的通透性,并测定PTD肽段对原代神经元、星形胶质细胞及PC12细胞的毒性作用,制备大鼠MCAO模型,梗死90min后再灌注,并分为生理盐水组、Kallikrein组、PTD-kallikrein组和PTD-kallikrein+B2抑制剂组。再灌注后24h,分别用NSS量表测定神经功能,TTC染色测定梗死灶体积,ELISA测定梗死灶细胞因子IL-1β、TNF-α及PGE2的含量。结果:PTD-kallikrein能通过血脑屏障,1μmol/LPTD肽段对上述3种细胞无毒性,10μmol/LPTD肽段对原代神经元和PC12细胞有轻度毒性;PTD-kallikrein能显著改善脑缺血导致的神经功能障碍、减小梗死体积、抑制细胞因子IL-1β、TNF-α及PGE2的释放,B2受体抑制剂能明显抑制PTD-kallikrein的神经保护作用。结论:PTD-kallikrein对大鼠MCAO模型缺血再灌注损伤有显著保护作用,并且很可能通过B2受体介导而发挥该功效。

What have we learned about the kallikrein-kinin and renin-angiotensin systems in neurological disorders?

The kallikrein-kinin system(KKS) is an intricate endogenous pathway involved in several physiological and pathological cascades in the brain. Due to the pathological effects of kinins in blood vessels and tissues, their formation and degradation are tightly controlled. Their components have been related to several central nervous system diseases such as stroke, Alzheimer's disease, Parkinson's disease, multiple sclerosis, epilepsy and others. Bradykinin and its receptors(B1R and B2R) may have a role in the pathophysiology of certain central nervous system diseases. It has been suggested that kinin B1R is up-regulated in pathological conditions and has a neurodegenerative pattern, while kinin B2R is constitutive and can act as a neuroprotective factor in many neurological conditions. The renin angiotensin system(RAS) is an important blood pressure regulator and controls both sodium and water intake. AngⅡ is a potent vasoconstrictor molecule and angiotensin converting enzyme is the major enzyme responsible for its release. AngⅡ acts mainly on the AT1 receptor, with involvement in several systemic and neurological disorders. Brain RAS has been associated with physiological pathways, but is also associated with brain disorders. This review describes topics relating to the involvement of both systems in several forms of brain dysfunction and indicates components of the KKS and RAS that have been used as targets in several pharmacological approaches.

Prostate-specific antigen kallikrein and the heart

Currently,there is growing interest regarding prostatespecific antigen(PSA) and the cardiovascular system.Increased PSA serum levels have been reported after prolonged cardiopulmonary resuscitation,cardiac surgery,extracorporeal cardiopulmonary bypass,acute myocardial infarction(AMI) and coronary artery stenting.The possible role of PSA in cardiac events has been questioned due to the finding of PSA decrease during AMI and by the correlation of variation in PSA levels with coronary lesions and occurrence of major adverse cardiac events.Complexed PSA forms and uncomplexed PSA forms are observed in the bloodstream but the increasing formation of irreversible bound PSA seems to be a crucial finding during AMI.Large studies need to be carried out to confirm these preliminary results and to elucidate unclear aspects.These findings present many potential directions for future research including the role of uncomplexed forms of PSA,the possible distribution of PSA in the heart,the relative expression levels in heart disease states,the mode of expression regulation and other potential specific substrates.The journey of PSA investigation could be longer than initially expected.

Crystal Structures of the Serine Protease Domain of Murine Plasma Kallikrein

Plasma kallikrein（PK）, a serine protease in the trypsin clan（SA）, plays critical roles in many physiological and pathological pathways. Regulating the abnormal activity of PK has been successfully used in the clinical therapy of hereditary angioedema. In this study, the serine protease domain of murine plasma kallikrein（m PK） was expressed in the pichia pastoris system. The recombinant protein was a glycosylated active enzyme after purification by the cation exchange and size-exclusion chromatography, and was crystallized at the precipitant condition of 25% PEG 3350, 0.1 M Tris-HCl pH 8.5 and 0.1 M Na Cl. The crystal structure of m PK was determined at 2.6 . This is the first published crystal structure of m PK, showing some distinctive features at S2＇ and S3＇ pockets when compared to its human analogue（human plasma kallikrein, h PK）. In addition, m PK show unique structural features in the non-conservative 67-72 and 76-81 loops when compared to other serine proteases. These results provide insights for the design of potent and selective PK inhibitors.

Kallikrein-kinin in stem cell therapy

The tissue kallikrein-kinin system exerts a wide spectrum of biological activities in the cardiovascular, renal and central nervous systems. Tissue kallikrein-kinin modulates the proliferation, viability, mobility and functional activity of certain stem cell populations, namely mesenchymal stem cells(MSCs), endothelial progenitor cells(EPCs), mononuclear cell subsets and neural stem cells. Stimulation of these stem cells by tissue kallikrein-kinin may lead to protection against renal, cardiovascular and neural damage by inhibiting apoptosis, inflammation, fibrosis and oxidative stress and promoting neovascularization. Moreover, MSCs and EPCs genetically modified with tissue kallikrein are resistant to hypoxia- and oxidative stress-induced apoptosis, and offer enhanced protective actions in animal models of heart and kidney injury and hindlimb ischemia. In addition, activation of the plasma kallikrein-kinin system promotes EPC recruitment to the inflamed synovium of arthritic rats. Conversely, cleaved high molecular weight kininogen, a product of plasma kallikrein, reduces the viability and vasculogenic activity of EPCs. Therefore, kallikrein-kinin provides a new approach in enhancing the efficacy of stem cell therapy for human diseases.

Stereotaxic microinjection of adenovirus-mediated human tissue Kallikrein gene reduces apoptosis in a rat model of middle cerebral artery occlusion

BACKGROUND：Several studies have demonstrated that Kallikrein gene transfer provides neuroprotection. Whether the neuroprotective effects of human tissue Kallikrein （HTK） are associated with apoptosis remains unclear. OBJECTIVE： To investigate the effects of HTK on apoptosis in the peripheral cerebral infarct region. DESIGN, TIME AND SETTING： The completely randomized grouping, gene engineered, controlled experiment was performed at the Lin Baixin Laboratory Center, the Second Affiliated Hospital of Sun Yat-sun University between September 2007 and April 2008. MATERIALS： Ninety clean, healthy, male, Sprague Dawley rats were included. pUC19-HTK plasmid was constructed in the Laboratory for Neurology, the Second Affiliated Hospital of Sun Yat-sen University, China. bcl-2, bax, caspase-3, and β-actin were designed and purified by Shanghai Shuiyuan Company, China. METHODS： Middle cerebral artery occlusion （MCAO） model was established in all rats. At 72 hours after MCAO model establishment, rats were randomly divided into 3 groups, with 30 rats per group： blank control, saline, and pAdCMV-HTK. The saline and pAdCMV-HTK groups were respectively stereotactically micro-injected with 5 μL physiological saline or pAdCMV-HTK at the area surrounding the cerebral infarction region. Only puncture was performed, without any injection, in the blank control group. MAIN OUTCOME MEASURES： At 72 hours after MCAO establishment, as well as at 24 hours, 72 hours, and 7 days subsequent to treatment, exogenous HTK expression was detected by immunohistochemistry. Apoptosis was examined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL）, while mRNA levels of bcl-2, bax, and caspase-3 were detected by reverse transcription-polymerase chain reaction. In addition, neurological severity scores were evaluated prior to and after treatments. RESULTS： Ninety rats were included in the final analysis. HTK was primarily detected in the cytoplasm at 24 hours after pAdCMV-HTK injection. Thereafter, HTK expression gradually increased and reached a peak level at 72 hours after injection, which was significantly different from the blank control and saline groups （P 〈 0.05）. At 7 days, HTK expression began to decrease, but remained higher than the saline and blank control groups （P 〈 0.05）. Apoptotic cells aggregated around the cerebral infarction region. Compared with the saline and blank control groups, the mean number of TUNEL-positive cells was notably decreased in the pAdCMV-HTK group at each time point after treatment （P 〈 0.05）. mRNA levels of bcl-2, bax, and caspase-3 were elevated in all groups at 24 hours after treatment, peaked at 72 hours, and then gradually decreased again at 7 days. Compared with the saline and blank control groups, bcl-2 slightly increased, but was not significantly different from the pAdCMV-HTK group （P 〉 0.05）. bax and caspase-3 mRNA levels were significantly reduced at 24 and 72 hours after treatment （P 〈 0.05）. At 72 hours and, in particular, at 7 days after treatment, neurological severity scores were significantly less in the pAdCMV-HTK group compared with the saline and blank control groups （P 〈 0.05–0.01）. CONCLUSION： HTK could protect neural cells in the peripheral cerebral infarction region from apoptosis, which resulted in a better outcome. This may be related to modulated bcl-2 expression and reduced bax and caspase-3 expression.

Effects of kallikrein gene transfer on penumbral microvascular proliferation and on regional cerebral blood flow following cerebral ischemia/reperfusion injury

BACKGROUND： Recent findings have demonstrated that the kallikrein-kinin system （KKS） participates in the pathological process of cerebral ischemia/reperfusion injury. Kallikrein gene transfer exhibits neural protective effects following cerebral infarction. OBJECTIVE： To observe the effects of kallikrein gene transfer on vascular proliferation in the peripheral infarct focus and on regional cerebral blood flow （rCBF） following cerebral ischemia/reperfusion injury. DESIGN, TIME AND SETTING： The completely randomized, controlled experiment was performed at the Lin Baixin Laboratory Center, the Second Affiliated Hospital of Sun Yat-sun University between September 2007 and April 2008. MATERIALS： pUCI9-HTK plasmid was constructed and maintained in the Laboratory for Neurology, the Second Affiliated Hospital of Sun Yat-sen University, China. Mouse anti-human kallikrein 1 monoclonal antibody was purchased from R＆D Systems, USA. METHODS Ninety healthy, male, Sprague Dawley rats were used. Middle cerebral artery occlusion （MCAO） was established in all rats to induce cerebral ischemia/reperfusion injury. Following MCAO establishment, all rats were randomly divided into three groups （n = 30）： blank control, saline, and pAdCMV-HTK. The saline and pAdCMV-HTK groups were stereotactically micro-injected with 5μL of physiological saline or with pAdCMV-HTK [multiplicity of infection （MOI） = 20], respectively, into the ischemic penumbra. In the blank control group, only sham injection was performed. MAIN OUTCOME MEASURES： At 12, 24, and 72 hours after treatment, cerebral infarction volume was measured by 2, 3, 5-triphenyltetrazolium chloride （TTC） staining. Exogenous HTK expression, as well as regional vascular endothelial growth factor （VEGF） expression, was detected by immunohistochemistry. rCBF was examined by 14C-iodoantipyrine micro tracing. In addition, neurological severity score （NSS） was performed. Higher scores indicated more severe neurological deficits. RESULTS： NSS results demonstrated that compared with the saline and the blank control groups, the pAdCMV-HTK group exhibited lower NSSs 24 hours after pAdCMV-HTK injection （P 〈 0.05）. The NSSs were further decreased after 72 hours （P 〈 0.01）. Cerebral infarction volume at 24 hours, and in particular at 72 hours after treatment, was significantly reduced in the pAdCMV-HTK group compared with the blank control and saline groups （P 〈 0.05）. The rCBF in the area surrounding the infarction lesion was slightly decreased in all groups compared with the contralateral area. At 24 and 72 hours following treatment, the rCBF in the peripheral infarction lesion was significantly elevated in the pAdCMV-HTK group compared with the blank control and saline groups （P 〈 0.05）. Immunohistochemistry results revealed that VEGF-positive cells were primarily found in the cortex and in some white matter surrounding the cerebral infarction lesion. In addition, the expression of VEGF in the pAdCMV-HTK group was significantly higher compared with that in the blank control and saline groups at 12, 24, and 72 hours following treatment （P 〈 0.05）. CONCLUSION： Following cerebral ischemia/reperfusion, kallikrein gene transfer can promote vascular proliferation in the brain tissue surrounding the infarction lesion, improve rCBF, and reduce infarction volume, thereby exhibiting protective effects to attenuate neurological deficits.

Construction and Identification of Human Tissue Kallikrein Gene Eukaryotic Expressing Vector

To clone and sequence the human tissue kallikrein gene of Chinese, and to construct eukaryotic expression recombinant of KK, total RNA was extracted from human pancreas and human tissue kallikrein gene cDNA was amplified by PCR after reverse-transcription by using Oligo（dT） primer. The original kallikrein cDNA was recovered and filled with Klenow enzyme and inserted into KS plasmid. After restriction endonuclease digestion, KK cDNA was sequenced by ABI 377 analyzer. Then the KK gene was amplified from pBluescript KSKK and inserted into pcDNA3. A sequence comparison showed that the cloned kallikrein gene was only one nucleotide different from that reported in the Genbank. The coding amino acid was Asp in the Genbank gene, while the coding amino acid of Chinese kallikrein gene was Asn. The KK cDNA fragment was inserted into the eukaryotic expression vector pcDNA3. The cloned kallikrein gene and the pcDNA3KK can be used for further study in gene therapy.

A STUDY OF THE RELATIONSHIP BETWEEN URINARY KALLIKREIN AND HYPERTENSION IN CHILDREN

hour urinary kallikrein excreation was measured(by fluorometry)in 300 children aged 10￣ 15 in Hanzhong. By comparison and retrospective study,the results showed:(1)in children with higher blood pressure(HBP)and positive family history of hypertension (FH+ ),the urinary kallikrein excreation was significantly lower than that in controls(P< 0.01).(2) the 12-hour urinary kallikrein excreation was negatively correlated with the SBP and DBP in the following up 3 times.(3)6-year retrospective review of blood pressure evolution showed that the blood pressure increased degree(△SBP) in children with lower urinary kallikrein excreation was much greater than that in those with higher one(P<0.01),and the percentiles or systolic blood pressure(△SBP)for children with lower urinary kallikrein mostly kept rising or kept up higher percentiles during the period. It is indicated that low urinary kallokrein may be a genetic marker to identify the children who are in danger or developing essential hypertension in future.

Kallikrein 10基因在前列腺癌组织中的表达及意义

目的:探讨Kallikrein 10(KLK10)基因在不同前列腺组织中的表达情况。方法:采用荧光定量PCR方法检测3例正常前列腺组织、5例BPH组织、6例前列腺癌细胞株、35例前列腺癌组织中KLK10 mRNA的表达水平。结果:正常前列腺组织、良性前列腺增生组织、前列腺癌细胞株、前列腺癌组织KLK10 mRNA表达相对值分别为0.521、0.487、0.021、0.018,组间比较差异有统计学意义(P<0.01)。骨转移前列腺癌与未转移前列腺癌KLK10 mRNA表达相对值分别为0.003、0.023,两组比较差异有统计学意义(P<0.05)。结论:KLK10在前列腺癌组织中表达下调,在转移性前列腺癌中表达更低。KLK10表达下调可促进肿瘤的发生与进展。

卵巢癌血清CCL18、HK10水平变化及与肿瘤恶性生物学行为的关系

目的探讨卵巢癌血清趋化因子18(CCL18)、人激肽释放酶10(HK10)水平变化及与肿瘤恶性生物学行为的关系。方法选取2018年1月至2023年4月河北省沧州中西医结合医院收治的100例卵巢癌患者作为研究组,50例良性卵巢肿瘤患者作为对照组,50名健康志愿者作为健康组。比较三组血清CCL18、HK10水平,研究组不同肿瘤恶性生物学行为患者血清CCL18、HK10水平,研究组不同化疗灵敏度患者化疗前和化疗3、6个周期后血清CCL18、HK10水平变化值,分析化疗前后血清CCL18、HK10水平变化值与化疗灵敏度的关系及对化疗灵敏度的预测价值。结果研究组血清CCL18、HK10水平均高于对照组、健康组(P<0.05)。研究组不同国际妇产科联盟(FIGO)分期、淋巴结转移、肌层浸润程度血清CCL18和HK10水平、不同分化程度血清HK10水平比较,差异具有统计学意义(P<0.05)。研究组化疗不灵敏患者CCL18△1(△1为化疗3个周期后与化疗前变化值的绝对值)、HK10△1均低于化疗灵敏患者(P<0.05)。CCL18△1、HK10△1联合检测的曲线下面积(AUC)显著高于单一指标(P<0.05)。结论卵巢癌血清CCL18、HK10水平升高与肿瘤恶性生物学行为关系密切,且其变化值能够辅助临床预测化疗灵敏度,两者联合检测具有较高的化疗灵敏度预测价值。

Expression of kallikrein-related peptidase 7 is decreased in prostate cancer

Recent evidence suggests that the human kallikrein 7 （KLK7） is differentially regulated in a variety of tumors. The aim of this study was to determine the expression of kallikrein-related peptidase 7 and KLK7 in our large collection of prostate samples. Between August 2000 and December 2012, 116 patients with histologically confirmed prostate cancer （PCa） and 92 with benign prostate hyperplasia （BPH） were recruited into the study. Using immunohistochemistry, quantitative reverse transcription polymerase chain reaction （RT-PCR） and western blot, kallikrein-related peptidase 7 expression in BPH and PCa tissues was determined at the mRNA and protein levels. The relationships between kallikrein-related peptidase 7 mRNA expression and clinicopathological features were analyzed. A total of 64 of 92 （69.57%） benign cases showed positive staining for KLK7 and 23 of 116 （19.83%） malignant cases showed positive, the difference of KLK7 expression between PCa and BPH was statistically significant （P〈 0.001）. The expression level of kallikrein-related peptidase 7 mRNA was significantly decreased in PCa tissues compared with that in BPH tissues and normal prostate tissue. Kallikrein-related peptidase 7 mRNA exhibited different expression patterns in terms of localization depending on pathological category of PCa. Similarly, our western immunoblot analyses demonstrated that the protein expression levels of KLK7 was lower in PCa than in BPH tissues and normal prostate tissue. Kallikrein-related peptidase 7 and KLK7 expression are down-regulated in PCa and lower expression of kallikrein-related peptidase 7 closely correlates with higher Gleason score and higher prostate-specific antigen level.

血清Kal、TREM-1对高血压性脑出血患者病情严重程度及预后的影响

目的探讨血清人源性激肽释放酶结合蛋白(Kal)、髓系细胞触发受体-1(TREM-1)对高血压性脑出血(HICH)患者病情严重程度及预后的影响。方法选取该院2020年1月至2023年3月该院收治的180例HICH患者作为HICH组,另选取同期在该院体检的180例健康体检者作为对照组。根据HICH患者的病情严重程度分为轻度组、中度组、重度组,根据患者的预后情况分为预后良好组和预后不良组。收集HICH患者基本资料并检测所有研究对象的Kal、TREM-1水平。采用多因素Logistic回归分析HICH患者预后不良的危险因素,绘制受试者工作特征(ROC)曲线分析血清Kal、TREM-1对HICH患者预后不良的预测价值。结果HICH组血清Kal水平低于对照组,TREM-1水平高于对照组(P<0.05)。轻度组有56例患者,中度组有82例患者,重度组有42例患者。重度组血清Kal水平低于轻度组和中度组,且中度组低于轻度组(P<0.05)。重度组血清TREM-1水平高于轻度组和中度组,且中度组高于轻度组(P<0.05)。预后良好组有122例患者,预后不良组有58例患者。预后不良组有高血压史患者的比例、TREM-1水平、出血量均高于预后良好组,Kal水平低于预后良好组(P<0.05);预后良好组和预后不良组年龄、男性比例、体质量指数、糖尿病史情况比较,差异均无统计学意义(P>0.05)。多因素Logistic回归分析结果显示,有高血压史、TREM-1水平升高、出血量大、Kal水平降低是HICH患者预后不良的独立危险因素(P<0.05)。ROC曲线分析结果显示,2项指标联合检测预测HICH患者预后不良的曲线下面积为0.920,高于Kal、TREM-1单独预测的0.857和0.860(Z=2.765、2.324,P=0.006、0.020)。结论HICH患者血清Kal水平降低,TREM-1水平升高,与患者病情严重程度及预后密切相关,2项指标联合检测对HICH患者预后不良具有较高预测价值。