Effects of gentiana scabra bage on expression of hepatic type Ⅰ,Ⅲ collagen proteins in Paragonimus skrjabini rats with liver fibrosis

Objective:To exploit- the effects of gentiana scabra bage on the expression of hepatic collagen proteins in Paragonimus skrjabini rats with liver fibrosis.Methods:Immunohistochemical technique was used to observe the changes of content of hepatic type Ⅰ,Ⅲ collagen proteins in Paragonimus skrjabini rats with Liver fibrosis before and after the gentiana scabra bage treatmeat.Results:Comparing with the model group,changes of hepatic tvpe Ⅰ and type Ⅲ collagen proteins in gentiana scabra bage treated group were significantly weakened.Conclusions:Gentiana scabra bage treatment can reduce the content of hepatic type Ⅲ and type Ⅰ collagen protein significantly in Paragonimus skrjabini rats with liver fibrosis,thereby,playing the role against hepatic fibrosis.

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。

微针导入重组Ⅲ型人源化胶原蛋白在皮肤屏障功能修复中的应用效果

目的探究微针导入重组Ⅲ型人源化胶原蛋白治疗因皮肤炎性衰老导致的皮肤屏障功能下降的有效性及安全性。方法选取2020年6月至2021年5月在本院接受面部皮肤治疗的30例患者作为研究对象,采用随机数字表法将其分为对照组(重组Ⅲ型人源化胶原蛋白)与治疗组(微针导入重组Ⅲ型人源化胶原蛋白),各15例。治疗后定期对患者进行随访,分别进行主观疗效评价及客观疗效评价。结果治疗后4、8、12周,治疗组的满意度显著高于对照组(P<0.05)。治疗后,治疗组的皮肤皱纹、纹理、红区、毛孔、弹性、皮肤经皮失水及皮肤含水量改善明显(P<0.05);对照组治疗后油脂改善明显(P<0.05)。治疗后12周,治疗组的总症状评分显著低于对照组(P<0.05)。结论微针导入重组Ⅲ型人源化胶原蛋白对皮肤修复有较好的疗效,可增强皮肤屏障功能,为改善皮肤屏障功能、治疗因皮肤屏障受损引起的炎性衰老提供了新选择。

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.

An enriched environment increases the expression of fibronectin type Ⅲ domain-containing protein 5 and brain-derived neurotrophic factor in the cerebral cortex of the ischemic mouse brain

Many studies have shown that fibronectin type III domain-containing protein 5(FDNC5) and brain-derived neurotrophic factor(BDNF) play vital roles in plasticity after brain injury. An enriched environment refers to an environment that provides animals with multi-sensory stimulation and movement opportunities. An enriched environment has been shown to promote the regeneration of nerve cells, synapses, and blood vessels in the animal brain after cerebral ischemia;however, the exact mechanisms have not been clarified. This study aimed to determine whether an enriched environment could improve neurobehavioral functions after the experimental inducement of cerebral ischemia and whether neurobehavioral outcomes were associated with the expression of FDNC5 and BDNF. This study established ischemic mouse models using permanent middle cerebral artery occlusion(pMCAO) on the left side. On postoperative day 1, the mice were randomly assigned to either enriched environment or standard housing condition groups. Mice in the standard housing condition group were housed and fed under standard conditions. Mice in the enriched environment group were housed in a large cage, containing various toys, and fed with a standard diet. Sham-operated mice received the same procedure, but without artery occlusion, and were housed and fed under standard conditions. On postoperative days 7 and 14, a beam-walking test was used to assess coordination, balance, and spatial learning. On postoperative days 16–20, a Morris water maze test was used to assess spatial learning and memory. On postoperative day 15, the expression levels of FDNC5 and BDNF proteins in the ipsilateral cerebral cortex were analyzed by western blot assay. The results showed that compared with the standard housing condition group, the motor balance and coordination functions(based on beam-walking test scores 7 and 14 days after operation), spatial learning abilities(based on the spatial learning scores from the Morris water maze test 16–19 days after operation), and memory abilities(based on the memory scores of the Morris water maze test 20 days after operation) of the enriched environment group improved significantly. In addition, the expression levels of FDNC5 and BDNF proteins in the ipsilateral cerebral cortex increased in the enriched environment group compared with those in the standard housing condition group. Furthermore, the Pearson correlation coefficient showed that neurobehavioral functions were positively associated with the expression levels of FDNC5 and BDNF(r = 0.587 and r = 0.840, respectively). These findings suggest that an enriched environment upregulates FDNC5 protein expression in the ipsilateral cerebral cortex after cerebral ischemia, which then activates BDNF protein expression, improving neurological function. BDNF protein expression was positively correlated with improved neurological function. The experimental protocols were approved by the Institutional Animal Care and Use Committee of Fudan University, China(approval Nos. 20160858 A232, 20160860 A234) on February 24, 2016.

Molecular Cloning and Bioinformatics Analysis of TypeⅢSecretion System Effector Protein Va1686 Gene of Vibrio alginolyticus

In this study, Va 1686 gene was cloned from Vibrio alginolyticus . The total length of the gene is 1 164 bp, and it could encode 387 amino acids. The physicochemical properties, protein structure, genetic evolutionary relationship and antigenic characteristics of the effect protein Va1686 of V. alginolyticus HY9901 type Ⅲ secretion system were studied and analyzed by bioinformatics methods and tools. The results showed that Va1686 is a stable hydrophilic and acidic protein without a transmembrane region and a signal peptide, and secondary structure to α-helix. The evolutionary analysis showed that V. alginolyticus HY9901 and V. harveyi were clustered together, which indicated that the genetic relationship between the two species was the closest. Va1686 contains a Fic superfamily conserved domain associated with cell division. Bioinformatics analysis showed that the B-cell preponderant epitopes of Va1686 might be localized in the regions of 48-49, 82-85, 125-126, 150-153, 185-186, 236-237 and so on. The 3D structure model of Va1686 subunit was simulated by SWISS-MODEL software and it was found that the vopS of V. parahaemolyticus was similar and the similarity was 89.46%. In this study, the feasibility of Va1686 as a common antigen of Vibrio was verified from the perspective of bioinformatics, which laid the foundation for the next step in vaccine development.

Collagen type Ⅲ is an important linking molecule between generated renal tubules and an artificial polyester interstitium--Reticulin as a linker between generated tubules and the interstitium

在再生医学领域,已经有很多人考虑在将来用干/祖细胞治疗急慢性肾功能衰竭。但是,要使这种治疗方案行之有效,需要了解关于患病肾脏肾小管发育的细胞生物学信息。与干/祖细胞治疗急慢性肾功能衰竭相关尚待明晰的细胞生物学问题包括:①干/祖细胞的整合。②干/祖细胞的定向分化类型。③新形成肾小管的空间构成。为了更好的了解这项技术的相关机制,文章应用了先进的培养技术在人工非细胞外基质材料界面条件下构建肾小管。将新生兔肾来源的干/祖细胞用多层聚酯纤维网覆盖,放置在灌注培养器中,用含1×10-7mol/L醛固酮的新鲜合成的在DMEM基础上改良的IMDM培养基诱导培养13d。扫描电镜下,在大豆凝集素、银染和抗Ⅲ型胶原或层粘连蛋白γ1单克隆抗体标记的全标本包埋爬片或冰冻切片上观察肾小管的生长情况。扫描电镜观察结果显示,新生成的肾小管完全被基底膜覆盖,纤维网状板有大量的纤维与生成的肾小管基底面和周围人工聚酯材料相互交联。抗Ⅲ型胶原染色和银染结果表明,在新生成的肾小管的基底膜和邻近的人工聚酯材料间有大量的纤维交织,在发育成熟的肾小管中有与新生成肾小管相同排列形式的Ⅲ型胶原纤维。以上实验结果说明,Ⅲ胶原是联系新生成的肾小管基底面和人工基质聚酯纤维的分子纽带。

EFFECTS OF TGF-β_1 ON CELLULAR PROLIFERATION ANDEXPRESSION OF TYPE Ⅲ COLLAGEN BY CULTURED RATRENAL INTERSTITIAL FIBROBLASTS

Objective To investigate the effects of TGF- β1 on proliferation and type N collagen mRNAexpression of rat renal interstitial libroblasts in vitro for elucidating the role of fibroblasts in renal interstitialfibrosis. Methods The cells were cultured in media containing various concentrations of TGF- β1. Theproliferation and the type Ⅲ collagen mRNA expression of the cells were assayed with MTT method andRT- PCR respectively. Results TGF- β1, has a stimulating proliferation effect with dose- dependence and aincreasing expression of type Ⅲ collagen mRNA with both dose- and time- dependence to the renal fibroblasts. Conclusion It is suggested that TGF- β, is involved in proliferation of renal fibroblasts and their type Ⅲcollagen mRNA expression, and may play an important role in renal interstitial fibrosis.

Molecular Cloning and Bioinformatics Analysis of TypeⅢSecretion System Effector Protein HY 322 Gene of Vibrio alginolyticus

In this study,Hy322 gene was cloned from Vibrio alginolyticus.The total length of its gene was 969 bp,and it could encode 322 amino acids.The physicochemical properties,protein structure,genetic evolutionary relationship and antigenic characteristics of the effector protein Hy322 of V.alginolyticus HY9901 type Ⅲ secretion system were studied and analyzed by bioinformatics methods and tools.The results showed that Hy322 is an unstable hydrophilic and acidic protein without a transmembrane region and a signal peptide,and secondary structure to α-helix.The evolutionary analysis showed that V.alginolyticus HY9901 and V.harveyi were clustered together,which indicated that the genetic relationship between the two species was closest.HY322 contains a FliN super family conserved domain associated with Flagellar motor switch.Bioinformatics analysis showed that the B-cell preponderant epitopes of Hy322 might be localized in the regions of 32-33,100-102,138-140,215-216,235-238 and 246-249.The 3D structure model of Hy322 subunit was simulated by SWISS-MODEL software and itwas found that the yscQ of Yersinia were similar and the similarity was 42.25%.In this study,the feasibility of Hy322 as a common antigen of Vibrio was verified from the perspective of bioinformatics,which laid the foundation for the next step in vaccine development.

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。

Vitamin D deficiency: Correlation to interleukin-17, interleukin-23 and PⅢNP in hepatitis C virus genotype 4

AIM: To assess vitamin D (Vit D) abnormalities in hepatitis C infected patients and their relationship with interleukin (IL)-17, IL-23 and N-terminal propeptide of type Ⅲ pro-collagen (PⅢNP) as immune response mediators. METHODS: The study was conducted on 50 Egyptian patients (36 male, 14 female) with hepatitis C virus (HCV) infection, who visited the Hepatology Outpatient Clinic in the Endemic Disease Hospital at Cairo University. Patients were compared with 25 ageand sexmatched healthy individuals. Inclusion criteria were based on a history of liver disease with HCV genotype 4 (HCV-4) infection (as new patients or under followup). Based on ultrasonography, patients were classified into four subgroups; 14 with bright hepatomegaly; 11 with perihepatic fibrosis; 11 with hepatic cirrhosis; and 14 with cirrhosis and hepatocellular carcinoma (HCC).Total Vit D (i.e., 25-OH-Vit D) and active Vit D [i.e., 1,25-(OH) 2 -Vit D] assays were carried out using commercial kits. IL-17, IL-23 and PⅢNP levels were assayed using enzyme linked immunosorbent assay kits, while HCV virus was measured by quantitative and qualitative polymerase chain reaction. RESULTS: Levels of Vit D and its active form were significantly lower in advanced liver disease (hepatic cirrhosis and/or carcinoma) patients, compared to those with bright hepatomegaly and perihepatic fibrosis. IL-17, IL-23 and PⅢNP levels were markedly increased in HCV patients and correlated with the progression of hepatic damage. The decrease in Vit D and active Vit D was concomitant with an increase in viral load, as well as levels of IL-17, IL-23 and PⅢNP among all subgroups of HCV-infected patients, compared to normal healthy controls. A significant negative correlation was evident between active Vit D and each of IL-17, IL-23 and PⅢNP (r = -0.679, -0.801 and -0.920 at P < 0.001, respectively). HCV-infected men and women showed no differences with respect to Vit D levels. The viral load was negatively correlated with Vit D and active Vit D (r = -0.084 and -0.846 at P < 0.001, respectively), and positively correlated with IL-17, IL-23 and PⅢNP (r = 0.951, 0.922 and 0.94 at P < 0.001, respectively). Whether the deficiency in Vit D was related to HCVinduced chronic liver disease or was a predisposing factor for a higher viral load remains to be elucidated. CONCLUSION: The negative correlations between Vit D and IL-17, IL-23 and PⅢNP highlight their involvement in the immune response in patients with HCV-4related liver diseases in Egypt.

CaMKⅡ抑制剂抑制AngⅡ或电场刺激诱导的心肌成纤维细胞TNF-α、TGF-β_1及collagenⅠ、Ⅲ的表达

目的:观察钙-钙调素依赖性蛋白激酶(CaMK)Ⅱ在血管紧张素Ⅱ(AngⅡ)或电场刺激(EFS)诱导的大鼠心肌成纤维细胞增殖、分泌细胞因子及胶原酶表达中的作用及其机制。方法:培养新生1-3 d乳鼠心肌成纤维细胞(3代),分为正常对照组(control)、0.1μmol/L AngⅡ组、0.1μmol/L AngⅡ+0.5μmol/L CaMKⅡ抑制剂KN92组、0.1μmol/L AngⅡ+0.5μmol/L CaMKⅡ抑制剂KN93组、0.1μmol/L AngⅡ+0.5μmol/L CaMKⅡ抑制剂AIP组;10 V1.0 Hz EFS组、10 V 1.0 Hz EFS+0.5μmol/L KN92组、10 V 1.0 Hz EFS+0.5μmol/L KN93组、10 V1.0 Hz EFS+0.5μmol/L AIP组、10 V1.0 Hz EFS+0.1μmol/L AngⅡ组。MTT法测定心肌成纤维细胞增殖;ELISA法测定细胞因子(TGF-β1,TNF-α)分泌;RT-PCR检测TGF-β1、TNF-α、collagenⅠ、ⅢmRNA水平。结果:CaMKⅡ抑制剂(0.5μmol/L KN93,0.5μmol/L AIP)能预防AngⅡ或EFS诱导的心肌成纤维细胞增殖;CaMKⅡ抑制剂(0.5μmol/L KN93,0.5μmol/L AIP)可预防AngⅡ或EFS引起的细胞培养上清液TGF-β1、TNF-α含量及相应mRNA表达增加。CaMKⅡ抑制剂(0.5μmol/L AIP,1.0μmol/L AIP)预防0.1μmol/L AngⅡ引起的collagenⅠ、Ⅲ表达增加。结论:抑制CaMKⅡ对AngⅡ或EFS诱导的心肌成纤维细胞增殖具有预防作用,其机制可能与CaMKⅡ抑制剂抑制TGF-β1、TNF-α以及胶原的表达有关。

Urinary type IV collagen excretion predicts an increased urinary albumin-to-creatinine ratio in normoalbuminuric patients with diabetes

Aims: We evaluated whether urinary excretion of type IV collagen (U-COL) may predict an increase in the urinary albumin-to-creatinine ratio (ACR) and what factors regulate U-COL in 145 normoalbuminuric patients with type 2 diabetes. Methods: We measured HbA1c, systolic blood pressure (SBP), urinary 8-hydroxydeoxyguanosine (8-OHdG) and monocyte chemoattractant protein (MCP)-1 at start of this study (Baseline), ACR and U-COL in addition to these measurements at one year later (Evaluation-1), and ACR and SBP after two years of the Evaluation-1 (Evaluation-2). The relationships were investigated between the increase of ACR and the U-COL. The effect of angiotensin receptor blockers (ARB) treatment on the correlations between U-COL and ACR at Evaluation-2 on one hand, and between U-COL and percent change of ACR on the other, was also analyzed. Furthermore, we investigated whether the increase in 8-OHdG and in MCP-1 in a year prior to the Evaluation-1 were risk factors of the rise in U-COL levels. Results: Both U-COL and SBP at Evaluation-1, but not ARB treatment, were independent risk factors for an increased ACR after 2 years. ARB treatment significantly suppressed the increase in ACR after 2 years in patients with higher U-COL excretion. The percentage changes in 8-OHdG (%8-OHdG) and MCP-1 (%MCP-1) in one year prior to Evaluation-1 measurements are independent risk factors for U-COL. HbA1c and SBP values one year prior to Evaluation-1 are independent risk factors not only for %8-OHdG but also, for baseline U-COL. The %8-OHdG is an independent risk factor for %MCP-1. Conclusions: U-COL may predict an increase in the ACR. The U-COL seems to be increased with oxidative stress and inflammation induced by past hyperglycemia.

Type XIX collagen: a promising biomarker from the basement membranes

Among collagen members in the collagen superfamily,type XIX collagen has raised increasing interest in relation to its structural and biological roles.Type XIX collagen is a Fibril-Associated Collagen with Interrupted Triple helices member,one main subclass of collagens in this superfamily.This collagen contains a triple helix composed of three polypeptide segments aligned in parallel and it is associated with the basement membrane zone in different tissues.The molecular structure of type XIX collagen consists of five collagenous domains,COL1 to COL5,interrupted by six non-collagenous domains,NCI to NC6.The most relevant domain by which this collagen exerts its biological roles is NCI domain that can be cleavage enzymatically to release matricryptins,exerting anti-tumor and anti-angiogenic effect in murine and human models of cancer.Under physiological conditions,type XIX collagen expression decreases after birth in different tissues although it is necessary to keep its basal levels,mainly in skeletal muscle and hippocampal and telencephalic interneurons in brain.Notwithstanding,in amyotrophic lateral sclerosis,altered transcript expression levels show a novel biological effect of this collagen beyond its structural role in basement membranes and its anti-tumor and anti-angiogenic properties.Type XIX collagen can exert a compensatory effect to ameliorate the disease progression under neurodegenerative conditions specific to amyotrophic lateral sclerosis in transgenic SOD1 G93 A mice and amyotrophic lateral sclerosis patients.This novel biological role highlights its nature as prognostic biomarker of disease progression in and as promising therapeutic target,paving the way to a more precise prognosis of amyotrophic lateral sclerosis.

重组Ⅲ型人源化胶原蛋白水光注射技术的操作规范

胶原蛋白是细胞外基质的主要组成成分,是动物体内分布最多、含量最广的蛋白质。在真皮层注射胶原蛋白后,即刻补充皮肤中的胶原蛋白含量,改善肤质、减少静态纹,改善皮肤松弛、增强皮肤弹性,同时刺激了新的胶原蛋白合成及组织的修复重塑。本文对重组Ⅲ型人源化胶原蛋白冻干纤维的产品特点、有效性和安全性、适应证、禁忌证、术中操作、注意事项、联合治疗等多个方面进行分析,为使用重组Ⅲ型人源化胶原蛋白的专业医师提供规范化的操作建议和参考资料,从而使治疗更加有效,同时尽可能降低治疗过程中不良反应的发生率。

实时荧光定量PCR法检测外源性DNA在重组Ⅲ型人源化胶原蛋白冻干纤维中的残留

目的对实时荧光定量PCR(qPCR)法检测重组Ⅲ型人源化胶原蛋白冻干纤维中的外源性DNA残留量进行探究和分析,确定该方法用于DNA质量控制的可行性。方法采用宿主细胞残留DNA样本前处理试剂盒(磁珠法)提取DNA,利用E.coli残留DNA检测试剂盒(PCR-荧光探针法)进行扩增反应,绘制标准曲线,建立重组Ⅲ型人源化胶原蛋白冻干纤维中外源性DNA残留量的qPCR检测方法。结果对8批重组Ⅲ型人源化胶原蛋白冻干纤维进行测定,外源性DNA残留量在0.03~300 pg/μl内,重复性良好,均远低于每剂10 ng,线性良好(R^(2)>0.99),回收率均在50%~150%之间。结论该方法可用于重组Ⅲ型人源化胶原蛋白冻干纤维中外源性DNA残留量的定量测定。

骨质疏松症患者PINP、25-(OH)VitD_(3)、BMP-2与中医辨证分型的相关性研究

目的:分析骨质疏松症患者血清总I型胶原氨基末端前肽(PINP)、25-羟维生素D3[25-(OH)VitD_(3)]及骨形态发生蛋白2(BMP-2)与其中医辨证分型的相关性。方法:收集我院2020年8月~2023年8月医院收治的157例骨质疏松症患者的一般资料及中医四诊资料进行回顾性分析,并统计骨质疏松症患者中医辨证分型结果,对不同症型患者基本资料、PINP、25-(OH)VitD_(3)、BMP-2水平进行比较。结果:157例骨质疏松症患者中医辨证分型属肾阳虚证34例,脾肾阳虚证43例,肝肾阴虚证49例,血瘀气滞证31例。不同中医辨证分型的骨质疏松患者血清PINP、25-(OH)VitD_(3)及BMP-2水平整体相比,差异有统计学意义(P<0.05);其中,血瘀气滞组PINP水平显著高于肾阳虚组、脾肾阳虚组及肝肾阴虚组患者(P<0.05),而其余3组两两间相比,差异无统计学意义(P>0.05);脾肾阳虚证患者25-(OH)VitD_(3)水平明显低于肾阳虚组、肝肾阴虚组及血瘀气滞组(P<0.05),且肾阳虚组低于肝肾阴虚组及血瘀气滞组(P<0.05),而其余两组相比差异无统计学意义(P>0.05);血瘀气滞组BMP-2水平显著低于肾阳虚组、脾肾阳虚组及肝肾阴虚组患者(P<0.05),而其余3组两两间相比,差异无统计学意义(P>0.05);二分类logistics回归分析结果显示,血瘀气滞与PINP呈正相关(P<0.05),与BMP-2呈负相关(P<0.05);脾肾阳虚与25-(OH)VitD_(3)呈负相关(P<0.05)。结论:骨质疏松症患者的血清PINP、25-(OH)VitD_(3)、BMP-2水平与其中医辨证分型具有一定相关性,可作为评估患者中医辨证分型的参考指标。

生肌化瘀方及其拆方对大鼠创面修复早期肉芽组织中Ⅰ、Ⅲ型胶原的影响

目的:探讨生肌化瘀方及其拆方对创面修复早期Ⅰ、Ⅲ型胶原的影响。方法:将24 只清洁级雄性SD大鼠随机分为4组:生肌化瘀方组、生肌方组、化瘀方组和模型组。以大鼠皮肤全层缺损创面为模型,采用免疫组化和图像分析相结合的方法,观察各组大鼠创面修复早期肉芽组织中Ⅰ、Ⅲ型胶原的变化。结果:在创面修复的第3 天,生肌方组和生肌化瘀方组的Ⅰ型胶原水平均高于模型组,化瘀方组则低于模型组(P <0.05);生肌方组、化瘀方组和生肌化瘀方组的Ⅲ型胶原水平均低于模型组(P<0.05),以化瘀方组更为明显;在创面修复的第7 天,生肌方组、生肌化瘀方组的Ⅰ型胶原水平均低于模型组(P<0.05);生肌方组和化瘀方组的Ⅲ型胶原水平均高于模型组(P<0.05)。结论:祛瘀生肌法可以促进大鼠创面修复,减少瘢痕形成。

腹股沟疝患者腹直肌后鞘Ⅰ、Ⅲ型胶原蛋白含量的初步分析

目的分析腹股沟疝患者腹直肌后鞘Ⅰ、Ⅲ型胶原含量及Ⅰ型/Ⅲ型胶原比例的变化。方法切取104例开腹手术患者腹直肌后鞘组织,其中合并腹股沟疝者16例,应用免疫组织化学方法(SABC法)进行染色,比较Ⅰ、Ⅲ型胶原蛋白含量、胶原总量及Ⅰ型/Ⅲ型胶原蛋白比例在腹股沟疝组和非腹股沟疝组间的差异。结果腹股沟疝组Ⅰ型胶原含量降低,Ⅲ型胶原含量升高,胶原总量升高,Ⅰ型/Ⅲ胶原蛋白比例降低。结论腹股沟疝患者腹直肌后鞘的Ⅰ型/Ⅲ型胶原比例下降,Ⅰ型胶原含量降低,而Ⅲ型胶原含量增高。腹股沟疝可能是系统性胶原代谢紊乱在腹股沟区的局部表现。