Collagen type Ⅳ对周围神经中再生轴突及非神经元细胞的作用和影响

本文用抗ｃｏｌｌａｇｅｎｔｙｐｅⅣ对抗体阻断ｃｏｌｌａｇｅｎｔｙｎｅⅣ的方法，研究了ｃｏｌｌａｇｅｎｔｙｐｅⅣ失活的移植神经段（长１０ｍｍ）植入大鼠坐骨神经后对再生轴突和非神经元细胞的作用和影响．实验结果显示：在移植神经段近端距近侧吻合口１ｍｍ处，术后１０ｄ抗ｃｏｌｌａｇｅｎｔｙｐｅⅣ组再生轴突数为对照组的５４％，术后１５ｄ增加到６６％，术后３０ｄ高达９４％．在移植神经段远侧距近侧吻合口９ｍｍ处，术后３０ｄ抗ｃｏｌｌａｇｅｎｔｙｐｅⅣ组再生轴突数为对照组的５８％。表明抗ｃｏｌｌａｇｅｎｔｙｐｅⅣ组再生轴突的生长启动和生长速度明显慢于对照组．巨噬细胞在移植神经段内的滞留数量抗ｃｏｌｌａｇｅｎｔｙｐｅⅣ组明显多于对照组．这些结果揭示ｃｏｌｌａｇｅｎｔｙｐｅⅣ在神经损伤和再生中对促进轴突的生长和维持神经微环境的平衡起着积极的作用．本文对ｃｏｌｌａｇｅｎｔｙｐｅⅣ在神经再生中的作用机制作了初步的分析和探讨。

基于上皮细胞-间充质细胞转分化(EMT)理论的艾灸配合化纤Ⅳ号方对实验大鼠Collagen Type Ⅲ和PDGF干预作用实验研究

目的:基于上皮细胞-间充质细胞转分化(EMT)学说观察化纤Ⅳ号方、艾灸以及二者相配合治疗肺纤维化大鼠Collagen TypeⅢ(Ⅲ-C)和PDGF的变化,探讨其治疗效应及生物学机制。方法:将鼠龄约为6周的SD大鼠随机分为空白组、模型组、化纤Ⅳ号方组、艾灸组、化纤Ⅳ号方与艾灸配合治疗组(简称为"灸药组"),治疗30 d后处死观察其肺组织病理改变,并检测其Collagen TypeⅢ、PDGF的基因和蛋白表达情况。结果:实时荧光定量结果显示:与空白组相比,各组Ⅲ-C和PDGF m RNA表达增高(P<0.05)。与模型组相比,各组的Ⅲ-C和PDGF m RNA表达有明显降低(P<0.01)。而各组中,灸药组疗效最明显,Ⅲ-C和PDGF的表达最低。蛋白免疫印迹法检测结果显示:与模型组相比各组的Ⅲ-C蛋白表达有差异。结论:1艾灸、化纤Ⅳ号方均可减轻博莱霉素诱导肺纤维化大鼠的肺纤维化程度。2艾灸配合化纤Ⅳ号方可减轻博莱霉素诱导肺纤维化大鼠的肺纤维化程度,且其效果优于单用艾灸或单用化纤Ⅳ号方。3艾灸、化纤Ⅳ号方及其二者配合使用不同程度阻抑博莱霉素诱导肺纤维化大鼠肺纤维化进程的效应机制,可能与通过调控其EMT过程中的Ⅲ-C和PDGF表达环节紧密相关。

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 type Ⅳ collagen level is predictive for esophageal varices in patients with severe alcoholic disease

AIM： To determine factors predictive for esophagea varices in severe alcoholic disease （SAD）. METHODS： Abdominal ultrasonography （US） was performed on 444 patients suffering from alcoholism. Forty-four patients found to have splenomegaly and/ or withering of the right liver lobe were defined as those with SAD. SAD patients were examined by upper gastrointestinal （UGI） endoscopy for the presence of esophageal varices. The existence of esophageal varices was then related to clinical variables. RESULTS： Twenty-five patients （56.8%） had esophageal varices. A univariate analysis revealed a significant difference in age and type Ⅳ collagen levels between patients with and without esophageal varices. A logistic regression analysis identified type Ⅳ collagen as the only independent variable predictive for esophageal varices （P = 0.017）. The area under the curve （AUC） for type Ⅳ collagen as determined by the receiver operating characteristic （ROC） for predicting esophageal varices was 0.78. CONCLUSION： This study suggests that the level of type Ⅳ collagen has a high diagnostic accuracy for the detection of esophageal varices in SAD.

Pre-hepatectomy type Ⅳ collagen 7S predicts post-hepatectomy liver failure and recovery

BACKGROUND Liver resection is an effective treatment for benign and malignant liver tumors.However,a method for preoperative evaluation of hepatic reserve has not yet been established.Previously reported assessments of preoperative hepatic reserve focused only on liver failure in the early postoperative period and did not consider the long-term recovery of hepatic reserve.When determining eligibility for hepatectomy,the underlying pathophysiology needs to be considered to determine if the functional hepatic reserve can withstand both surgery and any postoperative therapy.AIM To identify pre-hepatectomy factors associated with both early postoperative liver failure and long-term postoperative liver function recovery.METHODS This study was a retrospective cohort study.We retrospectively investigated 215 patients who underwent hepatectomy at our hospital between May 2013 and December 2016.Early post-hepatectomy liver failure(PHLF)was defined using the International Study Group of Liver Surgery’s definition of PHLF.Long-term postoperative recovery of liver function was defined as the time taken for serum total bilirubin and albumin levels to return to levels of<2 mg/dL and>2.8 g/dL,respectively,and the time taken for Child-Pugh score to return to Child-Pugh class A.RESULTS Preoperative type IV collagen 7S was identified as a significant independent factor associated with both PHLF and postoperative long-term recovery of liver function.Further analysis revealed that the time taken for the recovery of Child-Pugh scores and serum total bilirubin and albumin levels was significantly shorter in patients with type IV collagen 7S≤6 ng/mL than in those with type IV collagen 7S>6 ng/mL.In additional analyses,similar results were observed in patients without chronic viral hepatitis associated with fibrosis.CONCLUSION Preoperative type IV collagen 7S is a preoperative predictor of PHLF and longterm postoperative liver function recovery.It can also be used in patients without chronic hepatitis virus.

Clinical efficacy of intradermal type Ⅰ collagen injections in treating skin photoaging in patients from high-altitude areas

BACKGROUND Photoaging,a result of chronic sun exposure,leads to skin damage and pigmentation changes.Traditional treatments may have limitations in high-altitude areas like Yunnan Province.Intradermal Col Ⅰ injections stimulate collagen production,potentially improving skin quality.This study aims to assess the efficacy and safety of this treatment for photoaging.AIM To evaluate the efficacy and safety of intradermal typeΙcollagen(ColΙ)injection for treating photoaging.METHODS This prospective,self-controlled study investigated the impact of intradermal injections of ColΙon skin photodamage in 20 patients from the Yunnan Province.Total six treatment sessions were conducted every 4 wk±3 d.Before and after each treatment,facial skin characteristics were quantified using a VISIA skin detector.Skin thickness data were assessed using the ultrasound probes of the Dermalab skin detector.The Face-Q scale was used for subjective evaluation of the treatment effect by the patients.RESULTS The skin thickness of the right cheek consistently increased after each treatment session compared with baseline.The skin thickness of the left cheek significantly increased after the third through sixth treatment sessions compared with baseline.The skin thickness of the right zygomatic region increased after the second to sixth treatment sessions,whereas that of the left zygomatic region showed a significant increase after the fourth through sixth treatment sessions.The skin thickness of both temporal regions significantly increased after the fifth and sixth treatment sessions compared with baseline(P<0.05).These findings were also supported by skin ultrasound images.The feature count for the red areas and wrinkle feature count decreased following the treatment(P<0.05).VISIA assessments also revealed a decrease in the red areas after treatment.The Face-QSatisfaction with Facial Appearance Overall and Face-Q-Satisfaction with Skin scores significantly increased after each treatment session.The overall appearance of the patients improved after treatment.CONCLUSION Intradermal ColΙinjection improves photoaging,with higher patient satisfaction and fewer adverse reactions,and could be an effective treatment method for populations residing in high-altitude areas.

“Hepatic hilum area priority,liver posterior first”:An optimized strategy in laparoscopic resection for type Ⅲ-Ⅳ hilar cholangiocarcinoma

BACKGROUND In recent years,pure laparoscopic radical surgery for Bismuth-Corlette type Ⅲ and Ⅳ hilar cholangiocarcinoma(HCCA)has been preliminarily explored and applied,but the surgical strategy and safety are still worthy of further improvement and attention.AIM To summarize and share the application experience of the emerging strategy of“hepatic hilum area dissection priority,liver posterior separation first”in pure laparoscopic radical resection for patients with HCCA of Bismuth-Corlette types Ⅲ and IV.METHODS The clinical data and surgical videos of 6 patients with HCCA of Bismuth-Corlette types Ⅲ and Ⅳ who underwent pure laparoscopic radical resection in our department from December 2021 to December 2023 were retrospectively analyzed.RESULTS Among the 6 patients,4 were males and 2 were females.The average age was 62.2±11.0 years,and the median body mass index was 20.7(19.2-24.1)kg/m^(2).The preoperative median total bilirubin was 57.7(16.0-155.7)μmol/L.One patient had Bismuth-Corlette type Ⅲa,4 patients had Bismuth-Corlette type Ⅲb,and 1 patient had Bismuth-Corlette type Ⅳ.All patients successfully underwent pure laparoscopic radical resection following the strategy of“hepatic hilum area dissection priority,liver posterior separation first”.The operation time was 358.3±85.0 minutes,and the intraoperative blood loss volume was 195.0±108.4 mL.None of the patients received blood transfusions during the perioperative period.The median length of stay was 8.3(7.0-10.0)days.Mild bile leakage occurred in 2 patients,and all patients were discharged without serious surgery-related complications.CONCLUSION The emerging strategy of“hepatic hilum area dissection priority,liver posterior separation first”is safe and feasible in pure laparoscopic radical surgery for patients with HCCA of Bismuth-Corlette types Ⅲ and Ⅳ.This strategy is helpful for promoting the modularization and process of pure laparoscopic radical surgery for complicated HCCA,shortens the learning curve,and is worthy of further clinical application.

Combination of type Ⅳ collagen 7S, albumin concentrations, and platelet count predicts prognosis of non-alcoholic fatty liver disease

BACKGROUND Non-alcoholic fatty liver disease(NAFLD)is the most common cause of chronic liver disease and affects approximately 25%of the general global adult population.The prognosis of NAFLD patients with advanced liver fibrosis is known to be poor.It is difficult to assess disease progression in all patients with NAFLD;thus,it is necessary to identify patients who will show poor prognosis.AIM To investigate the efficacy of non-invasive biomarkers for predicting disease progression in patients with NAFLD.METHODS We investigated biomarkers associated with mortality in patients with NAFLD who visited the Kawasaki Medical School General Medical Center from 1996 to 2018 and underwent liver biopsy and had been followed-up for>1 year.Cumulative overall mortality and liver-related events during follow-up were calculated using the Kaplan-Meier analysis and compared using log-rank testing.We calculated the odds ratio and performed receiver operating characteristic curve analysis with logistic regression analysis to determine the optimal cut-off value with the highest prognostic ability.RESULTS We enrolled 489 patients who were followed-up for a period of 1-22.2 years.In total,13 patients died(2.7%of total patients enrolled);7 patients died due to liverrelated causes.Poor prognosis was associated with liver fibrosis on histological examination but not with inflammation or steatosis.Blood biomarkers associated with mortality were platelet counts,albumin levels,and type IV collagen 7S levels.The optimal cutoff index for predicting total mortality was a platelet count of 15×10^(4)/μL,albumin level of 3.5 g/dL,and type IV collagen 7S level of 5 mg/dL.In particular,only one-factor patients with NAFLD presenting with platelet counts≤15×10^(4)/μL,albumin levels≤3.5 g/dL,or type IV collagen 7S≥5 mg/dL showed 5-year,10-year,and 15-year survival rates of 99.7%,98.3%,and 94%,respectively.However,patients with two factors had lower 5-year and 10-year survival rates of 98%and 43%,respectively.Similarly,patients with all three factors showed the lowest 5-year and 10-year survival rates of 53%and 26%,respectively.CONCLUSION A combination of the three non-invasive biomarkers is a useful predictor of NAFLD prognosis and can help identify patients with NAFLD who are at a high risk of all-cause mortality.

Enrichment of putative human epidermal stem cells based on cell size and collagen type IV adhesiveness

The enrichment and identification of human epidermal stem cells （EpSCs） are of paramount importance for both basic research and clinical application. Although several approaches for the enrichment of EpSCs have been established, enriching a pure population of viable EpSCs is still a challenging task. An improved approach is worth developing to enhance the purity and viability of EpSCs. Here we report that cell size combined with collagen type IV adhesiveness can be used in an improved approach to enrich pure and viable human EpSCs. We separated the rap- idly adherent keratinocytes into three populations that range in size from 5-7 μm （population A）, to 7-9 μm （population B）, to ≥9μm （population C） in diameter, and found that human putative EpSCs could be further enriched in population A with the smallest size. Among the three populations, population A displayed the highest density of plintegrin receptor, contained the highest percentage of cells in G0/G1 phase, showed the highest nucleus to cytoplasm ratio, and possessed the highest colony formation efficiency （CFE）. When injected into murine blastocysts, these cells participated in multi-tissue formation. More significantly, compared with a previous approach that sorted putative EpSCs according to pl-integrin antibody staining, the viability of the EpSCs enriched by the improved approach was significantly enhanced. Our results provide a putative strategy for the enrichment of human EpSCs, and encourage further study into the role of cell size in stem cell biology.

1型糖尿病大鼠肾脏病变与ADPN、TGF-β1、collagen Ⅳ、ICAM-1的关系

目的探讨脂联素、TGF-β1、collagenⅣ、ICAM-1在1型糖尿病肾脏病变发生及发展中的变化及作用。方法大鼠随机分为正常对照组和糖尿病组,每组分别于4周和8周,测24h尿白蛋白排泄量(UPro/24h)、空腹血浆葡萄糖、血肌酐、尿素氮、血清脂联素(ADPN)、肾重/体重,观察肾脏TGF-β1、Ⅳ型胶原蛋白(collagenⅣ)、ICAM-1的表达。结果糖尿病大鼠UPro/24h增高、TGF-β1、collagenⅣ、ICAM-1的表达增加、脂联素水平降低。结论 TGF-β1、collagenⅣ、ICAM-1在肾脏组织中表达水平与血清ADPN呈负相关。随着ADPN水平降低,TGF-β1、collagenⅣ、ICAM-1在肾脏组织的表达增加。

血管球瘤中FLi-1、h-caldesmon及collagen Ⅳ的表达及临床病理分析

目的探讨FLi-1、h-caldesmon及collagenⅣ在血管球瘤中的表达及不同组织学类型与临床病理指标的关系。方法采用免疫组化En Vision法检测35例血管球瘤中FLi-1、h-caldesmon及collagenⅣ的表达。分析35例血管球瘤不同组织学分型与临床病理参数间的关系。结果镜下见肿瘤细胞圆形或多边形,呈片状分布在血管之间或呈环状围绕在血管周围,瘤细胞边界清晰,外形规则,有时可见瘤细胞与梭形平滑肌细胞移行过渡。免疫表型:血管球瘤可表达FLi-1、h-caldesmon及collagenⅣ,阳性率分别为58.6%、97.1%和100%。血管球瘤的不同类型与患者性别和肿瘤体积无关,与患者年龄相关(P=0.01)。35例血管球瘤中vimentin和SMA均呈(+),2例CD34呈局灶(+),1例desmin呈(+),EMA、S-100、Cg A、CD68及CD99均呈(-)。结论 h-caldesmon和collagenⅣ可作为血管球瘤诊断的标志物,FLi-1可作为一种辅助参考标志物,有助于血管球瘤的诊断。

维药买朱尼含药血清对IL-1β作用下大鼠软骨细胞MMP-13、Type-ⅡCollagen表达的影响

目的:观察维药买朱尼含药血清对IL-1β作用下体外培养的大鼠关节软骨细胞Type-ⅡCollagen、MMP-1、MMP-13表达的影响,并进一步探讨维药买朱尼防治OA的作用机制。方法:从1周龄SD大鼠关节软骨中分离培养原代软骨细胞,经鉴定后选用第二代细胞随机分为空白对照组、模型组、维药买朱尼组,培养第3d时模型组、维药买朱尼组采用细胞因子IL-1β(10ng/ml)继续培养,分别在培养后第24h、36h、48h采用RealTimePCR方法检测并分析各组软骨细胞中MMP-13、Type-ⅡCollagen的表达情况。结果:在第24h时各组MMP-13表达增加,Type-ⅡCollagen有一定降低,但无统计学差异;而在第36h和48h时空白对照组与模型组MMP-13、Type-ⅡCollagen表达差异均有统计学意义(P<0.05),维药买朱尼组较模型组MMP-13与Type-ⅡCollagen的表达在48h、72h时间点有显著性差异(P<0.05)。结论:维药买朱尼可抑制IL-1β对Type-ⅡCollagen的降解和破坏作用,并能抑制IL-1β对MMP-13的诱导和激活作用。

Sublytic C5b-9诱导肾小球系膜细胞产生的TSP-1和TGF-β1对其合成FN和collagenⅣ的影响

目的:探讨亚溶解型(sublytic)C5b-9复合物刺激大鼠肾小球系膜细胞(glomerular mesangial cell,GMC)后,诱生的血小板反应蛋白-1(thrombospondin-1,TSP-1)与转化生长因子-β1(transforming growth factor-β1,TGF-β1)对其促进细胞外基质(ex-tracellular matrix,ECM),如纤维连接蛋白(fibronectin,FN)和Ⅳ型胶原蛋白(collagenⅣ)合成的影响。方法:体外培养大鼠GMC,进行不同分组处理并给予sublytic C5b-9刺激,然后分别检查受刺激的GMC合成TSP-1和TGF-β1因子水平,同时检测GMC分泌FN和collagenⅣ的水平。此外,应用人工合成的TSP-1封闭肽段(GGWSHW)和TGF-β1中和抗体处理培养的大鼠GMC,研究TSP-1和TGF-β1在sublytic C5b-9诱导的GMC分泌上述ECM中的作用及其相互关系。结果:培养的大鼠GMC在sublytic C5b-9刺激后18 h,其FN和collagenⅣ表达水平均明显升高。同时,TSP-1蛋白表达和TGF-β1分泌(包括活化的TGF-β1含量)也显著增多。用TSP-1封闭肽段(GGWSHW)处理大鼠GMC后亦能显著抑制由sublytic C5b-9诱导的TGF-β1活化,并减少FN、collagenⅣ的产生。同样用TGF-β1中和抗体处理GMC后也能明显抑制由sublytic C5b-9导致的FN、collagenⅣ的分泌。结论:体外用sublytic C5b-9刺激大鼠GMC,能诱导其ECM分泌与TSP-1和TGF-β1的合成及活化,而sublytic C5b-9促进GMC分泌ECM的机制可能与其诱导TSP-1合成及活化的TGF-β1存在一定的关系。

氧化苦参碱对皮肤创面愈合中PCNA、α-SMA及Type Ⅰ collagen的影响

目的探讨氧化苦参碱对小鼠皮肤创面愈合中血清增殖细胞核抗原(PCNA)、α-平滑肌肌动蛋白(α-SMA)及细胞Ⅰ型胶原蛋白(Type Ⅰ collagen)的影响。方法昆明小鼠背部手术制备1.5 cm×1.5 cm全层皮肤缺损创面模型。自第1 d始,除对照组给予等量生理盐水外,余各组分别按20、40、80 mg/kg给予氧化苦参碱。Van Gieson纤维胶原染色法观察小鼠背部皮肤创面组织胶原纤维的表达;免疫组学法评价创面组织中PCNA、α-SMA及Type Ⅰ collagen的表达。结果Van Gieson纤维胶原染色显示,氧化苦参碱可促进新生肉芽组织、毛细血管及新生纤维胶原生长。免疫组学研究显示,在第7 d时,氧化苦参碱20、40、80 mg/kg使小鼠皮肤创面组织中PCNA表达明显升高(P<0.05);在第9 d、11 d时,氧化苦参碱20 mg/kg使小鼠皮肤创面组织中α-SMA显著增加;氧化苦参碱20 mg/kg在第3 d、11 d,氧化苦参碱40 mg/kg在第3 d,氧化苦参碱80 mg/kg在第3 d、7 d引起Type Ⅰ collagen的表达显著升高(P<0.05)。结论氧化苦参碱能增加皮肤创面愈合中PCNA、α-SMA及Type Ⅰ collagen的表达。

TGF-β_1、BMP-2和TypeⅡ Collagen在黄韧带中的表达及意义

目的研究TGF-1β、BM P-2和typeⅡco llagen在退行性腰椎滑脱(degenerative lum bar spondy lo listhes is,DLS)和腰椎间盘突出症(lum bar d isc hern iation,LDH)黄韧带中的表达及其意义。方法37例手术切除的腰椎椎板间部黄韧带标本分为3组,第1组为退行性腰椎滑脱组(DLS)10例;第2组为腰椎间盘突出症组(LDH)17例,第3组为正常对照组10例,其中7例取自腰椎骨折手术病人,3例取自意外死亡者。应用EnV is ion二步免疫组化的方法检测其TGF-1β、BM P-2和typeⅡco llagen的表达情况,普通光镜观察,计算出各标本的表达阳性率和表达强度,数据以x-±s标准差及表达强度表示,结果分别用Spss统计软件和R id it进行分析。结果TGF-1β、BM P-2和typeⅡco llagen的阳性表达产物见于成纤维细胞、成软骨细胞和软骨细胞中,而Ⅱ型胶原染色还可同时见于基质。TGF-1β、BM P-2和typeⅡco llagen在DLS组中的表达明显高于LDH组和正常组(P<0.01或P<0.05),Ⅱ型胶原基质染色明显深于LDH组和对照组。LDH组的TGF-1β和typeⅡco llagen的表达阳性率和表达强度与正常组之间差异无显著性(P>0.05),而BM P-2的表达阳性率和表达强度在LDH组与正常组之间具有统计学意义(P<0.01)。结论黄韧带所受到的异常机械牵张力可以增加TGF-1β在黄韧带细胞中的合成,而TGF-1β则促进退行性腰椎滑脱黄韧带中的Ⅱ型胶原合成,导致黄韧带的退变和肥厚。BM P-2在退变黄韧带中的表达异常增高,可能与黄韧带的软骨化倾向有关。

芡实对糖尿病肾病大鼠肾组织MMP-9、TIMP-1及Collagen Ⅳ表达的影响

目的:探讨芡实对糖尿病肾病大鼠肾组织MMP-9、TIMP-1及CollagenⅣ表达的影响。方法:健康雄性Wistar大鼠65只,随机选取10只为正常对照组(N组);其余一次性腹腔注射链脲佐菌素(STZ)制作DN模型,造模成功后,随机分为DN组、芡实低、中、高剂量组及氯沙坦钾组,均采用灌胃给药,N组和DN组给予等体积蒸馏水。12周后检测大鼠生化指标;HE、Masson、PAS染色观察肾脏病理变化;免疫组化法测定MMP-9、TIMP-1及CollagenⅣ在肾组织表达情况;采用实时荧光定量PCR技术检测肾组织中MMP-9、TIMP-1及CollagenⅣ基因的表达。结果:实验12周末,与N组比较,DN组大鼠肾小球肥大,细胞外基质积聚,血糖、24 h尿蛋白定量、血尿素氮(BUN)、肌酐(Scr)、尿白蛋白排泄率(UAER)、尿蛋白/尿肌酐(Up/Ucr)比值显著升高(P<0.05),肾组织MMP-9蛋白和mRNA表达降低(P<0.05),TIMP-1、CollagenⅣ蛋白和mRNA表达升高(P<0.05);与DN组比较,EM组、EH组及LP组大鼠病理改变减轻,24 h尿蛋白定量、Scr、BUN、UAER、Up/Ucr显著降低(P<0.05),肾组织MMP-9蛋白和mRNA表达显著升高(P<0.05),TIMP-1、CollagenⅣ蛋白和mRNA表达显著减少(P<0.05)。结论:芡实可能通过调节DN大鼠肾脏MMP-9、TIMP-1的表达,减少细胞外基质积聚,从而减轻肾小球硬化,减少蛋白尿,保护肾功能,延缓DN病变的进展。

未知病变类型脑血管中β-amyloid、α-actin、collagen Ⅳ的含量

目的研究未知病变类型脑血管病变的结构特征。方法通过刚果红染色、免疫组织化学染色、计算机图像分析技术对未知病变类型脑血管病变的β-amyloid、α-actin、collagenⅣ的含量进行研究。结果未知病变类型脑血管壁α-actin、collagenⅣ呈少量阳性染色,与正常脑血管存在显著差异(P<0.05);β-amyloid染色呈阴性,与正常脑血管无差异(P>0.05)。病变血管壁中上述三种蛋白的表达特点与脑血管淀粉样变(cerebralamyloidangiopathy,CAA)及小动脉硬化玻璃样变不同。结论未知病变类型脑血管病变具有不同于CAA的病变特征。

Expressions of type I collagen, α2 integrin and β1 integrin in sclera of guinea pig with defocus myopia and inhibitory effects of bFGF on the formation of myopia

AIM:To investigate the expressions of type I collagen, α2 integrin and β1 integrin in the posterior sclera of guinea pigs with defocus myopia and whether basic fibroblast growth factor (bFGF) injection inhibits the formation and development of myopia by upregulating the expression of type I collagen, α2 integrin and β1 integrin. METHODS:After 14 days of treatment, the refractive state and axial length were measured and the levels of type I collagen, α2 integrin and β1 integrin were assayed in the posterior sclerae of groups of guinea pigs that wore a monocular-7D polymethylmethacrylate (PMMA) lens or had -7D lens wear followed by the peribulbar injection of Phosphate Buffer Solution (PBS) or bFGF. The untreated fellow eye served as a control. Guinea pigs with no treatment served as normal group. ·RESULTS:The results showed that 14 days of monocular defocus increased axial eye length and refraction, while bFGF delivery inhibited them markedly. Further, it was also found that the monocular-7D lens could decrease the levels of type I collagen, α2 integrin and β1 integrin expressions, while, unlike PBS, bFGF increased them significantly in comparison to contralateral control eyes and normal eyes. CONCLUSION:bFGF can prevent the formation anddevelopment of defocus myopia by upregulating the expressions of type I collagen, α2 integrin and β1 integrin. Taken together, our results demonstrate that bFGF promotes sclera remodeling to prevent myopia in guinea pigs.

P92钢接头Ⅳ型开裂蠕变孔洞临界值的分析及应用

【目的】分析P92管道接头热影响区发生Ⅳ型开裂的蠕变孔洞临界值的影响因素,得到蠕变孔洞临界值计算方法,为评估接头开裂风险提供依据。【方法】收集服役后发生Ⅳ型开裂的P92接头,在未开裂处取样进行610℃下不同应力条件的高温持久蠕变试验,然后使用光学显微镜、电子显微镜观察服役开裂接头和持久断裂接头热影响区的蠕变孔洞分布形貌,并借助图像分析软件测量蠕变孔洞的面积分数,同时使用硬度仪测试热影响区软化情况,最后综合测试结果分析细晶区软化程度和接头应力水平对蠕变孔洞临界值的影响,并拟合应力、显微硬度、孔洞临界值三者之间的函数关系。【结果】研究发现,P92钢接头细晶区相比于接头其它区域软化速度更快,但软化作用并非其发生Ⅳ型开裂的必要条件,当应力水平较高时,细晶区没有发生明显软化时就可以触发蠕变开裂;得到接头轴向应力水平、蠕变孔洞面积分数临界值和显微硬度的函数关系式。【结论】引起Ⅳ型开裂的蠕变孔洞面积分数临界值与相对应力水平(轴向应力与硬度的比值)有关,相对应力水平越高,蠕变孔洞临界值越小,接头安全性就越低;通过测量接头细晶热影响区的蠕变孔洞面积分数和显微硬度,可以估算开裂接头的轴向应力水平,也可以评估在役未开裂接头的Ⅳ型开裂风险。

Adhesive properties of hepatoma cells to collagen Ⅳ coated surfaces

Objectives: To quantitatively study the adhesive pro- perties of hepatoma cells to collagen Ⅳ coated artifi- cial basement membrane and to investigate the rele- vance of cell adhesive forces to the concentration of collagen Ⅳ. Methods: Synchronous G1 and S phase cells were a- chieved using thymine-2-desoxyriboside and cochicine sequential blockage method and double thymine-2- desoxyriboside blockage method respectively. The adhesive forces of hepatoma cells were investigated by micropipette aspiration technique. Results: The adhesive forces of hepatoma cells to ar- tificial basement membrane were (107.78±65.44) ×10^(-10)N, (182.60±107.88)×10^(-10)N, (298.91± 144.13)×10^(-10)N when the concentration of the membrane coated by 1, 2, 5μg/ml collagen Ⅳ re- spectively (P<0.001). The adhesive forces of G1 and S phases hepatoma cells to artificial basement membrane were (275.86±232.80)×10^(-10)N and (161.16±120.40)×10^(-10)N respectively when the concentration of the membrane coated by 5μg/ml collagen Ⅳ (P<0.001). Conclusions: The adhesive forces of hepatoma cells to artifical basement membrane in direct proportion to the concentration of collagen Ⅳ suggests that the in- crease of basement membrane might be conducive to the chemotactic motion and adhesiveness of tumor cells. G1 phase cells are more capable of adhering to basement membrane than S phase cells. Hepatoma cells, especially G1 phase cells, may survive in blood circulation, and sequest and adhere in microcircula- tion, and get through basement membrane for re- mote metastasis.