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.

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。

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.

Cell therapy of a patient with type Ⅲ Osteogenesis imperfecta caused by mutation in COL1A2 gene and unstable collagen type I

The allogenic bone marrow derived mesenchymal stem cells transplantation was given to the newborn girl diagnosed with osteogenesis imperfecta type III, with multiple bone fractures, extreme shortness and limbs deformities. The treatment was performed at the age of 4 and 6 weeks. The clinical diagnosis was supported by biochemical analysis of collagen type I recovered from culture medium of cultivated patient’s skin fibroblast, which revealed its triple helix instability at temperature about 2?C lower than normal. Sequencing of both genes encoding procollagen type I revealed heterozygous substitution G23569Ain COL1A2 gene causing change of glycine at position 517 to aspartate. The donor of mesenchymal stem cells was the girl’s father. She received two intravenous infusions of suspended cultured mesenchymal cells in 16 days apart without any side effects. An analysis of procollagen type I secreted to the culture medium by bone marrow-derived mesenchymal stem cells obtained from the patient, 3 months following transplantation revealed its normal triple helix stability. During the subsequent two years of follow up two new bone fractures were noted. Currently a two-year-old girl’s presents extreme growth and weight deficiency. The motoric development is also retarded, but the patient constantly improves and makes progresses.

Effects of connective tissue growth factor and collagen type Ⅰ scleroderma

Objective： To investigate the effects of connective tissue growth factor（CTGF） and collagen type I（COL-I） on the pathogenesis of scleroderma and explore the relationship between the level of COL-I and CTGF. Methods： 12 mice model of scleroderma was established by the injection of Bleomycin. The level of CTGF and COL-I were detected by immunohistochemical method. The relationship was analyzed between CTGF and COL-I level. As control group, 12 healthy mice were selected. Results： The levels of CTGF and COL-I in sclerotic models were higher than in normal controls （P 〈 0.05）. It was found that there was a correlation between the level of CTGF and COL-I. Conclusion： CTGF and COL-I played an important role in the hardening process of the skin lesions of the mice model, which may be involved in the pathogenesis of scleroderma.

Effect of Collagen Type I or Human Fibronectin on Imatinib Cytotoxicity in Oral Squamous Cell Carcinoma

Extracellular matrix (ECM) components are critical for all aspects of cell proliferation, adhesion, and morphological alteration. Recent progress has yielded multiple molecular drugs that specifically target gene products which are expressed at high levels in tumor cells. We investigated whether the sensitivity of tumor cells to molecular target drugs could be altered when cells were cultured on surfaces with various coating conditions such as lysine, laminin, Matrigel, collagen type I, and human fibronectin (HFN). This study evaluates the IC50 values of imatinib in oral squamous cell carcinoma (OSCC) cell lines when cells are cultured on plates coated with ECM components such as collagen type I and HFN. Four OSCC cell lines—SQUU-A, SQUU-B, SAS, and NA— are used. Cell proliferation was assessed using WST-8 reagent. Collagen type I and HFN significantly enhanced OSCC cell proliferation compared with control. Imatinib cytotoxicity was demonstrated following culture of OSCCs in culture plates coated with collagen type I or HFN. However, there were no significant changes in imatinib IC50 values between collagen type I and HFN. These results indicate that some molecular target drugs exhibit cancer cell cytotoxicity without being influenced by cell environment factors such as the ECM. These results may aid in the search for molecular target drugs to apply in the clinical chemotherapy of OSCC.

The Use of Bone Morphogenetic Protein-7 and Resveratrol in Collagen Type II of Articular Cartilage

This study aimed to investigate the effects of resveratrol and bone morphogenetic protein 7 on type II collagen from superficial and middle zone of porcine articular chondrocytes. Articular cartilage was isolated from dissected porcine knee joint n = 12. Isolated cells were plated as monolayers at a density of 1 × 105 cells/well in 12-well culture plates and incubated at 37℃ in a humid atmosphere of 5% carbon dioxide and 95% air. Cell cultures were treated for four days with various concentrations of bone morphogenetic protein-7 and resveratroL Cells were then collected and analysed for collagen type II expression by real time polymerase chain reaction and protein level quantification by enzyme-linked immunosorbent assay. Cartilage tissue sections were localised for collagen type II by immunohistochemistry. Moreover, resveratrol and bone morphogenetic protein-7 effects on cartilage matrix contents were analysed by histology. Resveratrol and bone morphogenetic protein-7 stimulates expression of collagen type II mRNA and protein level accumulation in the surface zone and middle zone at 50μM ＋ 300 ng/ml （RSV ＋ BMP-7）. Immunohistochemistry results confirmed the presence of collagen type II on articular cartilage. Histological tissue sections confirmed that chondrocytes were obtained from different zones of articular cartilage. The study suggests that a combination of bone morphogenetic protein-7 and resveratrol up-regulate the expression and synthesis of collagen type II.

Thermal,infrared spectroscopy and molecular modeling characterization of bone:An insight in the apatite-collagen type I interaction

An insight into the interaction of collagen type I with apatite in bone tissue was performed by using differential scanning calorimetry, Fourier transform infrared spectroscopy, and molecular modeling. Scanning electron microscopy shows that bone organic content incinerate gradually through the different temperatures studied. We suggest that the amide regions of the type I collagen molecule (mainly C=O groups of the peptide bonds) will be important in the control of the interactions with the apatite from bone. The amide I infrared bands of the collagen type I change when interacting to apatite, what might confirm our assumption. Bone tissue results in a loss of thermal stability compared to the collagen studied apart, as a consequence of the degradation and further combustion of the collagen in contact with the apatite microcrystals in bone. The thermal behavior of bone is very distinctive. Its main typical combustion temperature is at 360°C with a shoulder at 550°C compared to the thermal behavior of collagen, with the mean combustion peak at ca. 500°C. Our studies with molecular mechanics (MM+ force field) showed different interaction energies of the collagen-like molecule and different models of the apatite crystal planes. We used models of the apatite (100) and (001) planes;additional two planes (001) were explored with phosphate-rich and calcium-rich faces;an energetic preference was found in the latter case. We preliminary conclude that the peptide bond of collagen type I is modified when the molecule interacts with the apatite, producing a decrease in the main peak from ca. 500°C in collagen, up to 350°C in bone. The combustion might be related to collagen type I, as the ΔH energies present only small variations between mineralized and non-mineralized samples. The data obtained here give a molecular perspective into the structural properties of bone and the change in collagen properties caused by the interaction with the apatite. Our study can be useful to understand the biological synthesis of minerals as well as the organic-inorganic interaction and the synthesis of apatite implant materials.

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。

Collagen typeⅠ在胰腺癌发展中的作用

胰腺癌(PC)是高发病率、高死亡率的恶性肿瘤,尽管目前在胰腺癌的治疗策略中已取得了较大的进展,但胰腺癌的转移、复发及高耐药性仍是大部分患者预后不良的主要原因。Ⅰ型胶原蛋白(Col⁃I)是人体中主要的细胞外基质蛋白,在多种实体肿瘤中表达异常并参与肿瘤的形成。最近研究表明,Col⁃I在胰腺癌中高度表达与胰腺癌的生长、转移、侵袭、耐药以及治疗密切相关,是胰腺癌新的潜在治疗靶点。该文重点介绍了COl⁃I对胰腺癌发生发展的影响,为胰腺癌的诊断和靶向治疗提供新的思路。

Peptoids with aliphatic sidechains as helical structures without hydrogen bonds and collagen/ inverse-collagen type structures

Aliphatic homo-polypeptoids of NAla, NVal, NIle and NLeu both in the presence and absence of protecting groups adopt helical structures without hydrogen bonds with Φ, Ψ values of ~ 0, ± 90° with trans amide bonds. These structures are stabilized by carbonyl-carbonyl interactions and characterized by ~ 3.16 residues per turn with a pitch of ~ 6.13 ?. It has been shown that like polyvaline and polyleucine peptides, poly-peptoids can also be exploited for the construction of potential surfactant like molecules by incorporating charged amino acid residues at the N terminal. A single-handed template with Φ, Ψ values of ~ 0, 90° can be attained by incorporating L-leu or L-val at the C-terminal of poly-NIle. Analysis of the simulation results in water as a function of time reveals that the opening of helical structures without hydrogen bonds takes place at sub-picosecond time scale starting from the N-terminal. This leads to the formation of collagen or inverse-collagen type structures (Φ, Ψ ~ -60, 145° and 60, -145° respectively) stabilized by interactions of water molecules with the backbone carbonyl groups.

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.

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。

Combined expression of CTGF and tissue inhibitor of metalloprotease-1 promotes synthesis of proteoglycan and collagen type Ⅱ in rhesus monkey lumbar intervertebral disc cells in vitro

Background Low back pain has emerged as a widespread disease often caused by intervertebral disc degeneration.This study aimed to establish an in vitro cell culture model of rhesus monkey lumbar intervertebral discs and to investigate the effect of combined connective tissue growth factor （CTGF） and tissue inhibitor of metalloprotease-1（TIMP-1） expression mediated by adeno-associated virus （AAV） on collagen type Ⅱ and proteoglycan levels.The purpose of these investigations was to explore potential methods for relieving the degeneration of lumbar intervertebral disc cells.Methods Rhesus monkey lumbar intervertebral disc nucleus pulposus cells （NPCs） were isolated by enzyme digestion,cultured, and transduced with rAAV2-CTGF-IRES-TIMP-1, rAAV2-CTGF, or rAAV2-TIMP-1 at a multiplicity of infection （MOl） of 106.The expression of collagen type Ⅱ and proteoglycan was measured using RT-PCR and Western blotting.The synthetic rate of proteoglycan was measured using 35S incorporation.Results Rhesus monkey lumbar intervertebral disc NPCs were transduced with rAAV2-CTGF-IRES-TIMP-1,rAAV2-CTGF, and rAAV2-TIMP-1 and the transduced genes were expressed and detected.Compared to the control,CTGF promoted the synthesis of collagen type Ⅱ and proteoglycan.TIMP-1 showed an enhancing effect on the expression of proteoglycan but no effect on collagen type Ⅱ.Expression of both genes in rhesus monkey lumbar intervertebral disc NPCs significantly enhances the synthesis of proteoglycan and collagen type Ⅱ.Conclusions Single gene transduction of CTGF or TIMP-1 can enhanced synthesis of proteoglycan.CTGF expression can also enhance collagen type Ⅱ protein synthesis.Combined transduction of both CTGF and TIMP1 can significantly promote the expression of proteoglycan and collagen type Ⅱ to levels greater than transduction of a single gene alone.Our study provides a good basis for multi-gene therapy to treat lumbar intervertebral disc degeneration.

Interleukin-1 inhibits Sox9 and collagen type Ⅱ expression via nuclear factor-κB in the cultured human intervertebral disc cells

Background The most significant biological change in intervertebral disc degeneration is the decrease of chondrocyte specific gene and protein expression of Sox9 and collagen type Ⅱ. Interleukin-1 （IL-1） is not expressed in the normal intervertebral disc tissue but increases in the degenerated intervertebral disc tissue. This suggests that IL-1 may play a role in regulation of the expression of Sox9 and collagen type Ⅱ. Methods Human intervertebral disc cells were isolated and cultured. Sox9 and collagen type Ⅱ expression during treatment with IL-1, with or without the nuclear factor-κB （NF-κB） activity inhibitor curcumin, were detected by using reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting, and the activity of the NF-κB signaling pathway was detected by the electrophoretic mobility shift assay （EMSA）. Results IL-1 lowered the mRNA level and protein expression of Sox9 and collagen type Ⅱ in the cultured intervertebral disc cells in a dose dependent manner （P 〈0.05）, and this effect was attenuated by curcumin. Curcumin alone had no effect on Sox9 and collagen type Ⅱ expression （P 〉0.05）. IL-1 at concentrations of 0.1 ng/ml, 1 ng/ml and 10 ng/ml could stimulate the activity of NF-κB in the intervertebral disc cells in a dose dependent manner （P 〈0.05） that was inhibited by curcumin. Conclusions We demonstrated the previously unknown function of IL-1 in inhibiting Sox9 and collagen type Ⅱ via NF-κB in the intervertebral disc cells. This inhibition can be attenuated by curcumin, which is an effective NF-κB activity inhibitor.

Effects of adeno-associated virus (AAV) of transforming growth factors β_1 and β_3 (TGFβ_(1,3)) on promoting synthesis of glycosaminoglycan and collagen type II of dedifferentiated nucleus pulposus (NP) cells

The effects of AAV-TGF beta_1 and AAV-TGF beta_3 on promoting synthesis ofglycosaminoglycan and collagen type II of dedifferentiated rabbit lumbar disc NP cells were studiedin this work.The rabbit lumbar disc NP cells were isolated and cultured.The earlier and laterdedifferentiated NP cells were established by subculture.The AAV transfection efficiency todedifferentiated NP cells was analyzed with AAV-EGFP in vitro.After dedifferentiated NP cells weretransfected by AAV-TGFp,or AAV-TGF beta_3,their biological effects on promoting synthesis ofglycosaminoglycan or collagen type II were detected and compared by the methods of 35S incorporationor immunoblotting.The experimental results showed that AAV could transfect efficiently the earlierdedifferentiated NP cells,but its transfection rate was shown to be at a low level to the laterdedifferentiated NP cells.Both AAV-TGF beta_1,and AAV-TGF beta_3 could promote the earlierdedifferentiated NP cells to synthesize glycosaminoglycan and collagen type II,and the effect ofAAV-TGFp,was better than that of AAV-TGF beta_3.For the later dedifferentiated NP cells,the AAV-TGFbeta_3 could promote their synthesis,but AAV-TGFp,could slightly inhibit theirsynthesis.Therefore,AAV-TGFp,and AAV-TGF beta_3 could be used for the earlier dedifferentiated NPcells,and the TGF beta_3 could be used as the objective gene for the later dedifferentiated NPcells.

Marginal zone B cells are naturally reactive to collagen type II and are involved in the initiation of the immune response in collagen-induced arthritis

Antibodies against type II collagen(CII)are essential for development of collagen-induced arthritis(CIA),but how and where the B-cell response to CII is initiated is not fully known.We show here that naive DBA/1 mice display naturally reactive IgM and IgG anti-CII producing B cells prior to immunization.The CII-reactive B cells were observed in the spleen and recognized as marginal zone(MZ)B cells.After CII immunization,CII-specific B cells expanded rapidly in the spleen,in contrast to the lymph nodes,with the initial response derived from MZ B cells and later by follicular(FO)B cells.This was evident despite that the MZ B cells were subject to stringent tolerance mechanisms by having a greater Fc gamma receptor IIb expression than the FO B cells.Further,the MZ B cells migrated to the FO areas upon immunization,possibly providing antigen and activating FO T cells and subsequently FO B cells.Thus,around CIA onset increased numbers of IgG anti-CII producing FO B cells was seen in the spleen,which was dominated by IgG2a-and IgG2b-positive cells.These data demonstrate that CII-reactive MZ B cells are present before and expand after CII immunization,suggesting an initiating role of MZ B cells in the development of CIA.

Collagen Type I Alpha 1 Mutation Causes Osteogenesis Imperfecta from Mild to Perinatal Death in a Chinese Family

Osteogenesis imperfecta （01）, also known as brittle bone disease or Lobstein syndrome, is characterized by blue or gray sclerae, variable short stature, dentinogenesis imperfecta, hearing loss, and recurrent fractures. Based on clinical, genetic, and radiological features, Sillence et al. classified the OI into four subtypes including type I： Mild, common, with blue sclera; type Ⅱ： Perinatal lethal form; type Ⅲ： Severe and age-related progressive detbrmity, with normal sclera; and type Ⅳ： Moderate severity with normal sclera.

Extraction and characterization of bovine collagen Type V and its effects on cell behaviors

Collagen Type V(Col.V)plays an essential role in cell behaviors and has attracted increasing attention in recent years.Highpurity Col.V is needed for evaluating its biological properties.In this research,the enzymatic hydrolysis process was combined with ultrafiltration to purify Col.V from the bovine cornea.The purity of Col.V was determined to be above 90%by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and high-performance liquid chromatography methods.The effect of Col.V on cell behaviors was evaluated.The circular dichroism spectroscopy results demonstrated that the extracted Col.V exhibited a complete triple helix structure.SDS-PAGE suggested that the molecular weight of Col.V was 440 kDa.The self-assembly experiment revealed that the proportion of Col.V in the collagen mixture can affect the Col.I fiber diameter.The cell culture results implied that Col.V can inhibit fibroblasts(L929)proliferation.The L929 showed maximum mobility when the addition of Col.V was 30%.Thus,Col.V has the effect of inhibiting L929 proliferation and promoting migration.The high-purity Col.V provides useful information for further understanding its biological implications.