Colchicine reduces procollagen Ⅲ and increases pseudocholinesterase in chronic liver disease

AIM:To test whether colchicine would be an effective antif ibrotic agent for treatment of chronic liver diseases in patients who could not be treated with α-interferon.METHODS:Seventy-four patients(46 males,28 females) aged 40-66 years(mean 53±13 years) participated in the study.The patients were affected by chronic liver diseases with cirrhosis which was proven histologically(n=58);by chronic active hepatitis C(n=4),chronic active hepatitis B(n=2),and chronic persistent hepatitis C(n=6).In the four patients lacking histology,cirrhosis was diagnosed from anamnesis,serum laboratory tests,esophageal varices and ascites.Patients were assigned to colchicine(1 mg/d) or standard treatment as control in a randomized,double-blind trial,and followed for 4.4 years with clinical and laboratory evaluation.RESULTS:Survival at the end of the study was 94.6% in the colchicine group and 78.4% in the control group(P=0.001).Serum N-terminal peptide of type Ⅲ procollagen levels fell from 34.0 to 18.3 ng/mL(P=0.0001),and pseudocholinesterase levels rose from 4.900 to 5.610 mU/mL(P=0.0001) in the colchicine group,while no signif icant change was seen in controls.Best results were obtained in patients with chronic hepatitis C and in alcoholic cirrhotics.CONCLUSION:Colchicine is an effective and safe antifibrotic drug for long-term treatment of chronic liver disease in which fi brosis progresses towards cirrhosis.

Serum hyaluronic acid, procollagen type Ⅲ and Ⅳ in histological diagnosis of liver fibrosis

OBJECTIVE: To assess the significance of serum hyaluronic acid (HA), proeollagen type Ⅲ (PCⅢ), collagen type Ⅳ (CⅣ) in the histological diagnosis of liver fibrosis. METHODS: The concentrations of serum HA, PCⅢ, CⅣ in 253 patients with chronic liver diseases were measured by radioimmunoassay. Liver biopsies were performed in all patients at the same time. The liver was pathologically evaluated by a pathologist according to a scoring system. Combined with the results of liver pathological diagnosis, the accuracy of serum HA, PCⅢ, CⅣ in diagnosing patients with hepatic fibrosis (staging≥S_2) or cirrhosis (S_4) was assessed using the receiver operating curve (ROC). RESULTS: The cutoff values of serum HA, PCⅢ and CⅣ for identifying patients with hepatic fibrosis (≥S_2) or cirrhosis (S_4) were determined. The cutoff values of serum HA, PCⅢ and CⅣ for detecting patients with fibrosis (stage≥S_2) were 90μg/L, 90μg/L, 75μg/L, respectively; their sensitivity (Se) was 80.4%, 82%, 63.1%; their specificity (Spe) was 70.2%, 60.8%, 83.8%; their positive predictive values (PPV) were 86.7%, 83.5%, 90.4%; their negative predictive values (NPV) were 59.8%, 58.4%, 48.4%, respectively. The cutoff values for detecting patients with liver cirrhosis were 210μg/L for HA, 96.2% for Se, 85.3% for Spe, 65.4% for PPV, 98.8% for NPV; 150μg/L for PCⅢ, 76.4% for Se, 68.7% for Spe, 40.4% for PPV, 91.3% for NPV; 90μg/L for CⅣ, 80% for Se, 75.8% for Spe, 47.8% for PPV, 93.2% for NPV, respectively. CONCLUSIONS: Serum HA, PCⅢ and CⅣ can be determined for an accurate diagnosis of hepatic fibrosis in various stages. HA is the best for screening liver cirrhosis.

SERUM CONCENTRATIONS OF HYALURONIC ACID, PROCOLLAGEN TYPE III NH_2-TERMINAL PEPTIDE, AND LAMININ IN PATIENTS WITH CHRONIC CONGESTIVE HEART FAILURE

Objective To explore the role of serum fibrotic indices including hyaluronic acid （HA）, procollagen type Ⅲ NH2-terminal peptide （PCIIIP）, and laminin （LN） in assessing the severity of myocardial fibrosis in chronic congestive heart failure （CHF）. Methods Serum levels of HA, PCIIIP, and LN in 39 patients with CHF E [14 with New York Heart Association （NYHA） functional class II, 21 with class Ⅲ, 4 with class Ⅳ] and in 46 patients with NYHA functional class I were assessed by radioimmunoassay. Results The serum concentrations of HA, PCMP, and LN were 359.75 ± 84.59 μg/L, 77.88 ± 24. 67 μg/L, 86. 73 ± 23.90 μg/L in CHF group, and 211.60 ±54. 80 μg/L, 64.82 ±23.99 μg/L, 82. 26 ±23.98 μg/L in NYHA functional class Ⅰ group, respectively. The HA level was significantly higher in CHF patients as compared with NYHA functional class Ⅰ group （ P 〈 0.05 ）. However, no difference was found in the levels of PCIIIP and LN between CHF group and NYHA functional class Ⅰ group. The serum HA concentration was negatively correlated with left ventricular ejection fraction （ r = - 0.71, P 〈 0.05 ）. Conclusion Serum HA level may act as an indicator for myocardial fibrosis.

Procollagen C-proteinase enhancer 1 promotes physiologic retinal angiogenesis via regulating the process of collagen

AIM:To investigate the role of procollagen C-proteinase enhancer 1(PCPE1)in retinal angiogenesis and relevant mechanisms.METHODS:The Pcolce1-knockout(KO)mice were used to explore the effect of PCPE1 on retinal angiogenesis in vivo.Pcolce1 si RNA were designed,cell count kit 8(CCK8)assays and tube formation assays were performed to investigate the cell proliferation and tube formation abilities of retinal microvascular endothelial cells(h RMECs)in vitro.Mouse embryo fibroblasts(MEF)cells were isolated and cultured to analyze the effect of PCPE1 on enhancing procollagen cleavage.RESULTS:In vivo studies showed that the retinal vascular density of Pcolce1-/-mice was significantly lower than that of the control group.Furthermore,silencing of Pcolce1 inhibited cell proliferation and tube formation abilities of h RMECs in vitro.Additionally,much more procollagen was found in Pcolce1-/-MEF cells,compared to wild type MEF cells.CONCLUSION:PCPE1 may promote physiological retinal angiogenesis by regulating the processing of collagen,which may provide a potential therapeutic target of retinal vascular disease.

Lentinula edodes extract inhibits matrix metalloproteinase expression and increases typeⅠprocollagen expression via the p38 MAPK/c-Fos signaling pathway in ultraviolet A and B-irradiated HaCaT keratinocytes

Objective:To determine the effect of Lentinula edodes extract on ultraviolet(UV)A and UVB-induced changes in matrix metalloproteinase(MMP)and type I procollagen expression using human immortalized HaCaT keratinocytes.Methods:Lentinula edodes ethanol extract(LEE)was obtained by extraction with 80%ethanol for 4 h at 80℃.Effect of LEE on UVinduced alteration on the expression and production of MMPs and type I procollagen in keratinocytes was investigated using ELISA,RT-PCR,and Western blotting assay.To determine the underlying mechanism of LEE-mediated effects,mitogen-activated protein kinase(MAPK)and activator protein 1 signaling pathways were analysed by Western blotting assay.Results:LEE significantly inhibited the expression of MMP-1 and MMP-9 and increased the expression of type I procollagen in UVA and UVB-irradiated HaCaT keratinocytes.The phosphorylation levels of p38 were significantly inhibited by LEE whereas it did not affect c-Jun N-terminal kinase and extracellular signal-regulated kinase phosphorylation.Suppression of p38 phosphorylation was also accompanied by downregulation of UVA and UVB-induced increase in c-Fos.Conclusions:LEE effectively inhibits the expression of MMP-1 and MMP-9 and increases type I procollagen production through the p38 MAPK/c-Fos signaling pathway in UVA and UVB-irradiated HaCaT keratinocytes.This findings suggest that Lentinula edodes may be developed as a cosmetic material to suppress UV exposuremediated skin aging.

Effect of cytokines on promoter activity in mouse α2(Ⅰ) procollagen gene

Objective:To investigate the effect of TGFβ, IFNs and TNFα on promoter activities in mouse α2(I )procollagen gene. Methods: Recombinant constructions PCOL 0. 4, pCOL0. 8, pAZ1009 containing CAT reportergene fused respectively with 400, 800 and 2 000 hp length of the 5'--flanking region in mouse α2(Ⅰ) procollagengene were used for transfection. Promoter activities were determined by CAT reporter gene assay(CAT--ELISA) intransfected cells treated with different cytokines. Results: TGFβ(2 ng/ml ) increased the activities of -- 2kb-- +54hp, -- 780~ + 54hp and -- 348~ +54 hp fragments as promoter respectively, with the increase of activity of 2kb -- + 54hp being most evident. TNFα (100 u/ml ) decreased the activity of -- 2kb ~ + 54hp sequence aspromoter. A slight increase, but not decrease of CAT activities was found in cells treated with IFNα(1000 u/ml)or IFNγ(1 000 u/ml) when pAZ1009 was trans feeted. But CAT expression was decreased more greatly if IFNα orIFNγ was used in combination with TNFα compared with TNFα treatment alone. The same change was seen ifPCOLO. 4 or PCOLO. 8 was used for transfection. Conclusion: Positive TGFβ--responsive elements exist within Zkbflanking region in mouse α2 (Ⅰ ) procollagen gene mediating the fibrogenic effect, while some negative elementsmediate the antifibrotic effect of TNFα. The target antifibrotic effect of IFNs might lie outside this 2kb-- +54bpregion or posttranscriptionally.

EFFECTS OF ALVEOLAR MACROPHAGE CONDITIONED MEDIA FROM INTERSTITIAL LUNG DISEASE PATIENTS ON THE PROCOLLAGEN mRNA EXPRESSION IN HUMAN LUNG FIBROBLASTS

Progressive inflammation and fibrosis are the central processes in the pathogenesis of pulmonary fibrosis. It is believed that macrophages in areas of chronically inflamed lung play a key role in fibrotic response. Therefore, we investigated the effects of alveolar macrophage (AmΦ) conditioned media from interstitial lung disease (ILD) patients on lung fibroblast proliferation and procollagen mRNA expression. After stimulating with AmΦ conditioned media from ILD patients, the fibroblast proliferation increased 71. 4 % compared with the control, but for media from bronchial carcinoma (BC) patients, it just increased 14. 3%. There is a significant difference between the two groups (P<0. 05). The procollagen a1 (Ⅰ) mRNA in fibroblasts stimulated with AmΦ conditioned media from ILD patients was increased 21. 3 %, and a1 ( Ⅲ) was 37. 2% higher than control (P<0.05). It increased 6. 8% and 12. 8% for media from BC patients respectively, but there was no difference when compared to the control. We considered that AmΦ from ILD patients might be in an activated state and could release some growth factors to stimulate fibroblast proliferation and promote collagen DNA expression.

Age-Related Changes of Procollagen Alpha Polypeptide in Vascular Remodeling in Rat Vascular Smooth Muscle Cell

The current study revealed that increased synthesis and secretion of collagen types I and III play major roles in arterial wall remodeling, aneurysm formation, and atherosclerotic cap stability. The aim is to investigate the age-related changes of the procollagen alpha polypeptide gene mRNA and protein expression in the vascular smooth muscle cells (VSMCs) in rats, as well as the possible underlying mechanisms. We tested in vitro culture of VSMC from the thoracoabdominal aorta in neonate and 9-month-old healthy male Wistar rats;procollagen alpha polypeptide mRNA and procollagen alpha polypeptide protein expression were detected, using RT-PCR, VG staining, Western blot and ELISA methods. Semi-quantitative analysis displayed that, in the real-time reverse transcription polymerase chain reaction (RT-PCR), the type I collagen α polypeptide chain mRNA increased in the adult group, but not significantly (P = 0.05). Further, there was no significant difference between the two groups of type III collagen α polypeptide chain mRNA (P > 0.05). Both the type I and type III procollagen alpha polypeptide protein expression were increased significantly in the older group as compared with the young group (P < 0.05). This phenomenon mainly lies in the fact that the regulatory pathway on age-related changes of procollagen alpha polypeptides may be one of the molecular mechanisms in vascular remodeling during aging.

The Level of Serum Intact Terminal Peptide of Procollagen and Its Clinical Significance in Patients with Chronic Keshan Disease

Objectives To evaluate the changes of serum intact terminal peptide of procollagen in patients with chronic Keshan disease (KD) and investigate their clinical significance. Methods The concentrations of serum intact N-terminal peptide of type procollagen (P NP) and intact N-terminal peptide of type procollagen were measured by radioimmunoassay in 35 patients with chronic KD and 31 normal control. Doppler ultrasounds was used to determine several parameters of left ventricular systole and diastole functions. Results The concentration of serum P NP (74.07±16.74)μg/L and the ratio of P NP/ P NP (18.02 ±4.60) in chronic KD were significantly increased as compared to the control (39.63±12.07 μg/L, 12.12±4.24; P< 0.001). Serum P NP (4.19±0.64)μg/L in chronic KD was higher than that in the control (3.36±0.65 μg/L,P < 0.001) too. The higher of serum concentration of P NP and the ratio of P NP/ P NP, the worse of cardiac function in patients with chronic KD. A negative correlation was found between serum P NP/ P NP, P NP and VE/VA, LVEF (γ=-0.4502, -0.4608, P< 0.01 and γ=-0.3936, -0.3904, respectively; P<0.05). Conclusions These findings suggested that tissue synthesis of collagen type and type was abnormally increased in chronic KD. On the other hand, our results indicated that P NP and P NP were related to several functional alterations of the left ventricle. Serum procollagen peptide measurements might therefore provide indirectly diagnostic information on myocardial fibrosis associated with chronic KD.

生、醋莪术对大鼠免疫性肝纤维化及HSC-T6增殖和α-SMA,Procollagen Ⅰ表达的影响

比较生、醋莪术对猪血清所致大鼠免疫性肝纤维化及对活化的肝星状细胞(HSC-T6)增殖和α-SMA,ProcollagenⅠ表达的影响。采用腹腔注射猪血清0.5 mL/只,每周2次,共14周制备大鼠免疫性肝纤维化模型。观察生、醋莪术(0.95,1.90 g·kg^(-1))对大鼠血清ALT,AST,PC-Ⅲ,IV-C,LN,HA和肝组织HYP,MDA的表达水平;采用HE染色观察大鼠肝组织病理改变情况,Masson染色和天狼猩红染色观察大鼠肝组织中胶原表达情况;体外培养HSC-T6细胞,MTT法测定不同浓度生、醋莪术含药血清作用12,24,36,48 h对HSC-T6增殖的影响;采用Real-time PCR法检测各组α-SMA和ProcollagenⅠ表达情况。结果显示,给药生、醋莪术后,大鼠血清ALT,AST,PC-Ⅲ,IV-C,LN,HA表达水平均下降,与生莪术组相比,醋莪术组作用效果优于生莪术组;生、醋莪术含药血清各剂量组均能抑制HSC-T6的增殖,呈现一定的量效和时效关系;各给药组作用24 h后,HSCT6中α-SMA和ProcollagenⅠ表达水平降低,且醋莪术组降低更显著。生、醋莪术均能不同程度减轻肝纤维化,其抗肝纤维化机制可能与抑制HSC-T6增殖,减少细胞外基质的生成并促进其降解有关。

基于病因和国际骨微循环研究协会分期的股骨头坏死患者骨代谢水平研究

背景:目前缺乏大样本研究对不同病因、分期的股骨头坏死患者的骨代谢水平进行分析,这不利于制定更优的促坏死修复策略。目的:研究不同病因、国际骨微循环研究协会(ARCO)分期股骨头坏死患者的骨代谢水平。方法:回顾性纳入股骨头坏死的患者401例作为试验组,81例健康体检者作为对照组。试验组按照病因不同分为激素性股骨头坏死、酒精性股骨头坏死和创伤性股骨头坏死3组,并按ARCO分期的不同分为Ⅱ期/Ⅲ期/Ⅳ期。收集所有受试者的7项骨代谢相关指标,包括骨代谢调控激素25-羟基维生素D,骨转换标志物Ⅰ型前胶原N末端前肽、β-胶原降解产物、骨钙素N端中分子片段,骨代谢一般生化标记血清Ca、P、碱性磷酸酶。比较各组受试者的骨代谢水平,并应用二元Logistic回归分析与股骨头坏死发病相关的独立因素。结果与结论:①与对照组比较,试验组患者的β-胶原降解产物、Ⅰ型前胶原N末端前肽、骨钙素N端中分子片段、血清P、碱性磷酸酶水平均显著升高(均P<0.05);②以是否发病为结局,应用二元Logistic回归分析得出β-胶原降解产物、Ⅰ型前胶原N末端前肽、骨钙素N端中分子片段是与股骨头坏死发病相关的独立因素;③不同病因3组患者的β-胶原降解产物、Ⅰ型前胶原N末端前肽水平均高于正常参考值;酒精性股骨头坏死组患者的25-羟基维生素D、血清Ca水平均高于其他2组(P<0.05);激素性股骨头坏死、创伤性股骨头坏死组患者的25-羟基维生素D水平均低于正常值;④ARCOⅡ、Ⅲ、Ⅳ期股骨头坏死患者的7项骨代谢指标水平比较差异均无显著性意义(均P>0.05),但3组患者的β-胶原降解产物、Ⅰ型前胶原N末端前肽均高于正常参考值;ARCOⅡ期、Ⅳ期患者的25-羟基维生素D低于正常参考值;⑤结论:股骨头坏死患者的骨代谢水平异常,不同病因、ARCO分期的股骨头坏死患者的β-胶原降解产物、Ⅰ型前胶原N末端前肽均高于正常参考值,处于高骨转换状态;β-胶原降解产物、Ⅰ型前胶原N末端前肽、骨钙素N端中分子片段可能是股骨头坏死发病的危险因素。

骨化三醇对老年骨质疏松性脊柱骨折病人PKP术后骨折愈合的影响

目的研究骨化三醇用于老年骨质疏松性椎体压缩性骨折(OVCF)病人经皮椎体后凸成形术(PKP)后治疗对骨折愈合和血清骨碱性磷酸酶(BALP)、Ⅰ型前胶原N-端前肽(PINP)、胶原羧基端肽β特殊序列(β-CTX)表达水平的影响。方法纳入2019—2020年我院接受PKP手术的106例OVCF病人为研究对象,并采用随机数表法均分为观察组和对照组,每组53例。对照组PKP术后给予钙剂+阿仑膦酸钠片进行治疗,观察组在此基础上加用骨化三醇进行治疗,2组疗程均为6个月,比较2组临床疗效、Cobb角、Oswestry功能障碍指数(ODI)、骨密度和骨代谢指标的差异。结果2组治疗6个月有效率分别为90.57%和83.02%,差异无统计学意义(P>0.05),优良率分别为73.58%和52.83%,差异有统计学意义(P<0.05)。2组治疗1、3、6个月时Cobb角和ODI均明显低于治疗前(P<0.05),且观察组治疗6个月时ODI低于对照组,差异有统计学意义(P<0.05)。2组治疗3、6个月时骨密度均明显高于治疗前(P<0.05),且观察组治疗6个月时骨密度高于对照组,差异有统计学意义(P<0.05)。与治疗前相比,2组治疗1、3、6个月时BALP和PINP水平明显升高(P<0.05),β-CTX水平均明显降低(P<0.05),且观察组治疗1、3、6个月时BALP和PINP水平均高于对照组,差异有统计学意义(P<0.05)。结论OVCF病人PKP术后应用骨化三醇进行治疗具有良好效果,有利于改善骨代谢并增加骨密度,对促进骨折愈合、改善脊柱功能和提升日常生活能力具有积极作用。

唑来膦酸联合PVP治疗对骨质疏松性椎体压缩骨折患者血清PⅠNP、β-CTX及骨密度的影响

目的:探讨骨质疏松性椎体压缩骨折(OVCF)患者采用唑来膦酸联合经皮椎体成形术(PVP)治疗对血清Ⅰ型胶原氨基端延长肽(PⅠNP)、β胶原降解产物(β-CTX)及骨密度的影响。方法:按随机数字表法将2021年6月—2022年6月于南昌市洪都中医院治疗的86例OVCF患者分为两组,各43例。两组均给予碳酸钙D_(3)片(Ⅰ)、阿法骨化醇软胶囊治疗,对照组采用PVP治疗,观察组采用PVP联合唑来膦酸治疗。比较两组PⅠNP、β-CTX、骨密度、腰痛程度、功能状况、不良反应及再次骨折发生情况。结果:术后两组PⅠNP、β-CTX及视觉模拟评分法(VAS)、Oswestry功能障碍指数(ODI)评分均低于术前,骨密度均高于术前,且观察组PⅠNP、β-CTX及VAS、ODI评分均低于对照组,骨密度高于对照组,差异均有统计学意义(P<0.05);观察组再次骨折发生率低于对照组,不良反应发生率高于对照组(P<0.05)。结论:OVCF患者采用唑来膦酸联合PVP治疗可降低血清PⅠNP、β-CTX水平,改善患者骨密度、腰痛程度及功能状态,降低再次骨折发生率。

血清P1NP、β-CTX水平联合骨髓细胞形态学检查对多发性骨髓瘤的诊断价值

目的 探讨骨髓细胞形态学检查联合血清Ⅰ型前胶原氨基端前肽(P1NP)、Ⅰ型胶原羧基端肽β特殊序列(β-CTX)检测对多发性骨髓瘤(MM)及其分期的诊断价值。方法 选取2020年1月至2023年2月在该院就诊的104例MM患者作为研究对象,根据国际分期标准(ISS)分为Ⅰ期、Ⅱ期和Ⅲ期,分别为32例、36例和36例。同时选取同期因为缺铁性贫血、再生障碍性贫血、骨髓增生异常综合征等原因接受骨髓穿刺的40例作为对照组。对所有受试者进行骨髓细胞形态学检查,测定血清P1NP和β-CTX水平,并分析其与MM及其分期的关系。结果 MM组骨髓浆细胞比[(23.4±8.6)%]、血清P1NP[(120.5±35.6)ng/mL]和β-CTX水平[(820.4±210.3)pg/mL]均显著高于对照组[(3.2±1.4)%、(52.3±12.4)ng/mL、(320.6±80.2)pg/mL],差异均有统计学意义(P<0.05),且随着MM分期的升高而增高。以骨髓浆细胞比例≥10%、血清P1NP≥80 ng/mL和β-CTX≥500 pg/mL为诊断标准,对MM的灵敏度和特异度分别为92.3%和95.0%,联合检测的灵敏度和特异度分别为98.1%和100.0%。以血清P1NP≥100 ng/mL和β-CTX≥700 pg/mL为分期标准,对MMⅢ期的灵敏度和特异度分别为86.1%和88.2%,联合检测的灵敏度和特异度分别为94.4%和97.1%。结论 骨髓细胞形态学检查联合血清P1NP、β-CTX检测对MM及其分期具有较高的诊断价值,有助于评估MM患者的病情和预后。

血清透明质酸、层粘连蛋白、Ⅲ型前胶原N端肽和Ⅳ型胶原辅助诊断原发性肝癌的临床意义分析

目的 分析血清透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原N端肽(PⅢNP)和Ⅳ型胶原(CⅣ)辅助诊断原发性肝癌的可行性和临床价值。方法 选取82例原发性肝癌患者、33例肝硬化患者及127例健康对照者作为研究对象,分别作为原发性肝癌组、肝硬化组及健康对照组。检测并比较三组血清HA、LN、PⅢNP、CⅣ水平;分析血清HA、LN、PⅢNP、CⅣ的受试者工作特征曲线(ROC曲线)及曲线下面积(AUC);分析血清HA、LN、PⅢNP和CⅣ的诊断界值及诊断效能。结果 原发性肝癌组血清HA 80.32(42.18, 165.50)ng/ml、LN 42.91(31.47, 74.91)ng/ml、PⅢNP 16.15(10.93, 31.33)ng/ml和CⅣ126.00(73.16, 236.60)ng/ml均显著高于健康对照组的26.83(17.25, 40.02)、30.46(25.02, 40.95)、6.38(4.93, 7.86)、48.49(40.24, 57.57)ng/ml(P<0.05);原发性肝癌组血清HA、LN、PⅢNP水平显著低于肝硬化组(P<0.05),原发性肝癌组血清CⅣ水平与肝硬化组比较无统计学差异(P>0.05);肝硬化组血清HA、LN、PⅢNP和CⅣ水平均显著高于健康对照组(P<0.05)。ROC曲线及AUC分析血清HA、LN、PⅢNP和CⅣ的诊断效能显示:血清HA、LN、PⅢNP和CⅣ诊断原发性肝癌的AUC分别为0.8453、0.7195、0.9310和0.8842;血清HA、LN和PⅢNP鉴别原发性肝癌和肝硬化的AUC分别为0.7749、0.6208和0.6384。根据ROC曲线计算原发性肝癌和肝硬化的诊断界值,血清HA+LN+PⅢNP+CⅣ诊断原发性肝癌的灵敏度为92.7%。结论 血清HA、LN、PⅢNP和CⅣ可用于原发性肝癌的辅助诊断,具有重要的临床价值。

补肾运脾方促矮小大鼠生长的效用机制研究

目的:观察补肾运脾方对矮小大鼠生长的影响,探讨其作用机制。方法:将雌性30只、雄性15只大鼠合笼过夜,雌鼠受孕后按随机数字表随机分为低蛋白饲料组与标准饲料组,所有孕鼠分娩后采用标准饲料喂养。仔鼠2周龄去除雌仔鼠,3周龄离乳。标准饲料组孕鼠分娩的新生仔鼠中按随机数字表随机方法选取10只雄鼠仔鼠标记为正常组,选矮小模型仔鼠50只分为模型组、西药组、低剂量组、中剂量组、高剂量组,每组10只。正常组、模型组给予无菌生理盐水灌胃,1 mL/100 g;低、中、高剂量组分别给予补肾运脾方药液0.76 g/mL、1.52 g/mL、3.04 g/mL灌胃,1 mL/100 g;西药组给予重组人生长激素注射液腹部皮下注射(0.84 IU/kg)。每天上午8:00给药,1次/d,连续3周。对比各组身长、体质量、骨密度(BMD)、骨矿盐(BMC)及血清Ⅰ型前胶原羧基端肽(PICP)、Ⅰ型胶原羧基吡啶交联肽(ICTP)、生长激素(GH)、胰岛素样生长因子-1(IGF-1)。结果:各组大鼠治疗后身长、体质量高于治疗前(P<0.05),低剂量组、高剂量组BMD高于模型组(P<0.05),正常组、西药组、中剂量组、高剂量组血清PICP含量低于模型组(P<0.05),中药3组血清PICP含量大于正常组(P<0.05),正常组、中药3组、西药组血清ICTP含量小于模型组(P<0.01),中药3组血清ICTP含量大于正常组(P<0.05);正常组、中药3组、西药组血清GH含量小于模型组(P<0.01),低剂量组、中剂量组血清GH含量大于西药组(P<0.05),低剂量组血清GH含量大于正常组(P<0.05);低剂量组、中剂量组血清IGF-1水平小于西药组(P<0.05)。结论:补肾运脾方可促进矮小大鼠身长、体质量增长,促进骨代谢,增加BMD,其效用机制为通过促进血清GH利用、提高血清IGF-1水平发挥促生长作用。

消化性溃疡患者HMGB1、PINP和CD_(4)^(+)T细胞变化及与Hp感染的关系

目的探讨消化性溃疡患者高迁移率族蛋白B1(HMGB1)、I型原胶原N端前肽(PINP)和CD_(4)^(+)T细胞变化及与幽门螺杆菌(Hp)感染的关系。方法选择2019年9月—2021年9月我院收治的消化性溃疡患者97例作为研究对象,根据是否Hp感染分为Hp阳性组(n=63)与Hp阴性组(n=34);另选择同期我院健康体检者60例作为对照组。采用酶联免疫吸附法测定HMGB1和PINP水平;采用流式细胞仪测定CD_(4)^(+)T细胞水平。比较消化性溃疡组与对照组HMGB1、PINP和CD_(4)^(+)T细胞水平;不同Hp感染HMGB1、PINP和CD_(4)^(+)T细胞水平;采用Pearson分析Hp感染与HMGB1、PINP和CD_(4)^(+)T细胞相关性;采用多因素Logistic回归分析HMGB1、PINP和CD_(4)^(+)T细胞与Hp感染的关系。结果消化性溃疡组HMGB1水平高于对照组,PINP和CD_(4)^(+)T细胞水平低于对照组(P<0.05)。Hp阳性组HMGB1水平高于Hp阴性组,PINP和CD_(4)^(+)T细胞水平低于Hp阴性组(P<0.05)。经Pearson分析,HMGB1与Hp感染呈线性正相关,PINP和CD_(4)^(+)T细胞与Hp感染呈线性负相关(P<0.05)。经多因素Logistic回归分析HMGB1、PINP和CD_(4)^(+)T细胞为影响Hp感染危险因素。结论消化性溃疡患者HMGB1水平升高,PINP和CD_(4)^(+)T细胞水平下降,且与Hp感染密切相关,为影响Hp感染的危险因素。

安石榴苷促进成骨治疗绝经后骨质疏松

背景:安石榴苷作用广泛,安全性高,但其对成骨细胞和绝经后骨质疏松症的作用尚不清楚。目的:探讨安石榴苷对成骨细胞和绝经后骨质疏松症的作用。方法:安石榴苷作用于MC3T3-E1细胞,检测其促成骨细胞增殖作用。在成骨诱导液中添加安石榴苷,检测其促成骨分化作用。卵巢切除大鼠术后灌胃安石榴苷,3个月后进行MicroCT扫描和血清1型前胶原氨基末端肽检测,观察治疗效果。结果与结论:①CCK-8检测发现安石榴苷能促进成骨细胞增殖(P<0.05);②qRT-PCR和Western blot检测发现安石榴苷能促进碱性磷酸酶、Runx2 mRNA和蛋白的表达(P<0.05);③Micro CT扫描和血清学检测结果显示,安石榴苷能改善卵巢切除大鼠的骨密度、骨体积分数、骨小梁厚度、骨小梁数目和1型前胶原氨基末端肽水平;④结果表明,安石榴苷能促进成骨细胞增殖和分化,且对绝经后骨质疏松大鼠有治疗作用。

经DCAG方案化疗后AML患者血清HOXA9、PCⅢ、SE-CAD水平的变化及其与预后的关系

目的:探讨急性髓系白血病(AML)患者采用DCAG方案化疗后血清同源盒基因A9(HOXA9)、可溶性E钙黏蛋白(SE-CAD)及Ⅲ型前胶原蛋白(PCⅢ)水平的变化及其与预后的关系。方法:回顾性分析2018年3月至2021年12月经本院确诊并收治的80例复发难治性AML患者的资料,按照治疗方案不同将其分为DCAG组(n=40)与CAG组(n=40),对比治疗前后各组的临床疗效及HOXA9、SE-CAD、PCⅢ水平的变化;另按照临床疗效将所有患者分为缓解组(n=58)与未缓解组(n=22),通过单因素和多因素分析影响AML患者预后的危险因素;采用ROC曲线分析HOXA9、SE-CAD、PCⅢ三项单一指标及三者联合对预后的预测效能。结果:相比于治疗前,DCAG组与CAG组患者在治疗后的HOXA9、SE-CAD、PCⅢ水平均有所下降,但DCAG组患者各指标的改善效果明显优于CAG组,且DCAG组患者在治疗后的临床疗效显著优于CAG组(均P<0.05);多因素分析结果显示,骨髓原始细胞比率、HOXA9 m RNA、SE-CAD及PCⅢ水平升高是影响AML患者化疗疗效的独立危险因素(均P<0.05);ROC曲线分析显示,HOXA9 m RNA、SE-CAD及PCⅢ联合能够有效预测AML预后情况,其敏感度为84.80%、特异度为88.20%。结论:应用DCAG的化疗方案能够显著改善AML患者的HOXA9 m RNA、SE-CAD及PCⅢ水平;且这三项指标作为影响AML患者预后的危险因素,通过联合检测能够对AML患者的预后情况进行有效预测。

PLOD1在口腔鳞状细胞癌组织和细胞中的表达及其意义

目的:探讨前胶原赖氨酸,2-酮戊二酸5-双加氧酶1(PLOD1)在口腔鳞状细胞癌(OSCC)组织和细胞中的表达及其对OSCC细胞生物学行为的影响,阐明PLOD1作为OSCC预后标志物的潜力。方法:采用肿瘤免疫评价资源(TIMER)数据库、基因表达谱交互分析(GEPIA)数据库和Kaplan-Meier Plotter数据库分析头颈部鳞状细胞癌(HNSC)组织中PLOD1 mRMA表达水平及其与HNSC患者生存期的关联。免疫组织化学法检测110例OSCC组织和64例癌旁组织中PLOD1蛋白表达情况,分析其与OSCC患者临床病理特征和预后的关系。采用受试者工作特征(ROC)曲线下面积(AUC)评估PLOD1在OSCC诊断中的价值。实时荧光定量PCR(RT-qPCR)法和Western blotting法检测人正常口腔上皮HOK细胞和OSCC SCC15和CAL27细胞中PLOD1 mRNA和蛋白表达水平。利用脂质体法将小干扰RNA(siRNA)片段转染至SCC15细胞,分为si-NC组(转染阴性对照si-NC)和si-PLOD1组(转染si-PLOD1),采用CCK-8法、细胞划痕愈合实验和Transwell小室实验分别检测2组细胞增殖活性、划痕愈合率和侵袭细胞数。结果:公共数据库分析,HNSC组织中PLOD1 mRNA表达水平高于癌旁组织(P<0.05);与PLOD1低表达组比较,PLOD1高表达组HNSC患者生存期较短(HR=1.41,P=0.018)。PLOD1蛋白主要表达于OSCC细胞的细胞质中,OSCC组织中PLOD1表达强度高于癌旁组织(P<0.01),并与OSCC患者T分期和TNM分期有关(P=0.021,P=0.004)。PLOD1诊断OSCC的AUC为0.811,特异度和灵敏度分别63.64%和90.63%。Kaplan-Meier生存分析,与PLOD1低表达组比较,PLOD1高表达组OSCC患者生存率降低(P<0.01);COX回归分析,PLOD1高表达是影响OSCC预后的独立危险因素(P=0.012)。OSCC细胞中PLOD1 mRNA和蛋白表达水平高于HOK细胞(P<0.05)。与si-NC组比较,si-PLOD1组细胞增殖活性和划痕愈合率降低(P<0.05),侵袭细胞数减少(P<0.01)。结论:PLOD1在OSCC组织和细胞中高表达,沉默PLOD1表达可抑制OSCC细胞增殖、迁移和侵袭。PLOD1可能是OSCC新的治疗靶点和预后标志物。