Preparation and in vivo Distribution of Essential Oil from Alpinia zerumbet Fructus Encapsulated Nanoemulsions / 中国实验方剂学杂志

ObjectiveTo prepare oral nanoemulsions encapsulating essential oil from Alpinia zerumbet fructus（EOFAZ） and to investigate its pro-absorption effect in vitro and distribution in vivo. MethodThe proteoglycan conjugate polysaccharides of vinegar-processed Bupleuri Radix-bovine serum albumin（VBCP-BSA） was prepared by Maillard reaction of VBCP and BSA. Taking VBCP-BSA as emulsifier， vitamin B12（VB12） as absorption enhancer， and medium chain triglycerides mixed with EOFAZ as oil phase， the nanoemulsions loaded with EOFAZ was prepared by high energy emulsification method. The particle size， particle size distribution， surface Zeta potential， EOFAZ content and appearance and morphology of the nanoemulsions were characterized， and fluorescein tracer method was used to investigate the absorption effect of fluorescein-labeled EOFAZ nanoemulsions in vitro and their distribution in vivo. ResultVBCP-BSA was formed by Maillard reaction for 48 h with high grafting rate. Using VBCP-BSA as emulsifier， the homogeneous pink nanoemulsions was prepared and denoted as EOFAZ@VBCP-BSA/VB12. The particle size of the nanoemulsions was less than 100 nm and the particle size distribution was uniform. The surface of the nanoemulsions was a weak negative charge， and the shape was spherical. The encapsulation rate of the nanoemulsions for EOFAZ was greater than 80%， which had a good absorption effect in vitro and could enhance liver accumulation after oral administration. ConclusionThe designed proteoglycan nanoemulsions can effectively load EOFAZ， promote oral absorption and enhance liver distribution， which can provide experimental basis for the development of oral EOFAZ liver protection preparations.

Assessment of serum endocan levels in patients with beta-thalassemia minor

SUMMARY OBJECTIVE: Beta-thalassemia minor is a blood disease caused by a hereditary decrease in beta-globin synthesis, frequently leading to hypochromic microcytic anemia. Formerly called endothelial cell-specific molecule 1, endocan is a proteoglycan released by vascular endothelial cells in many organs. Our aim was to investigate the relationship between the beta-thalassemia minor patients and the healthy control group in terms of serum endocan level. METHODS: The study was performed in a total of 80 subjects. They were divided into two groups, the beta-thalassemia minor group (n=40) and the healthy control group (n=40). Serum endocan levels, age, sex, body mass index value, and tobacco use data of these groups were compared. RESULTS: No statistically significant difference was detected between the two groups in terms of age, sex, and body mass index values (p>0.05). Endocan levels were measured to be 206.85±88.1 pg/mL in the beta-thalassemia minor group and 236.1±162.8 pg/mL in the control group with no significant difference between the groups in terms of serum endocan levels (p>0.05). CONCLUSIONS: In our study, there was no change in endocan level in beta-thalassemia minor. This might be because serum endocan levels are affected by multi-factorial reasons. Serum endocan levels may be altered secondarily to decreased beta-globin chain, increased sympathetic activity due to anemia, or platelet dysfunction induced by oxidative stress in beta-thalassemia minor. Further multicenter studies involving more patients are necessary to demonstrate this.

Correlation between proteoglycan and alveolar bone resorption in a mouse periodontitis model / 口腔疾病防治

Objective@#To analyze changes in proteoglycan and its correlation with alveolar bone resorption in periodontitis. @*Methods @#Twelve eight-week-old C57BL/6J male mice were selected, and the periodontitis model was established by ligating the right maxillary second molar with 6-0 silk thread. The nonligated part of the left maxilla was used as the control. The mice were killed 14 days after the operation. Micro-CT was used to assess alveolar bone resorption. HE staining was used to observe the alveolar bone profile, and TRAP staining was conducted to examine the positive rate of osteoclasts. The expression of proteoglycan-related genes, such as aggrecan (ACAN), biglycan (BGN), versican (VCAN), decorin (DCN), osteoclast-related genes, such as cathepsin K (CTSK), matrix metalloprotein-9 (MMP-9), and receptor activator of nuclear factor kappa-B ligand (RANKL), and inflammation-related genes, such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), was detected by real-time quantitative PCR. Additionally, the correlation of the expression of proteoglycans with osteoclast-related genes and inflammation-related genes was evaluated by Pearson correlation analysis.@* Results@#The resorption of alveolar bone on the periodontitis side increased. TRAP staining showed that the number of osteoclasts was substantially increased in the maxilla with periodontitis. Real-time quantitative PCR demonstrated that compared with the control side, the expression of proteoglycan-related genes, such as ACAN, BGN, and DCN, was decreased, whereas the expression of the VCAN gene was significantly increased in the periodontitis side. Meanwhile, the expression of osteoclast-related genes, such as CTSK, MMP-9, and RANKL, and inflammation-related genes, such as IL-1β, IL-6, and TNF-α, was markedly increased in the periodontitis side (P<0.05). Pearson correlation analysis indicated a negative correlation between the expression of proteoglycans and the mRNA levels of osteoclast-related genes and inflammation-related genes (P<0.05). @*Conclusion @#The expression of proteoglycan was closely related to alveolar bone resorption in a periodontitis model.

Mechanism of Proteoglycan TPG-1 from Trametes robiniophila Inhibiting Growth of Human Hepatoma SK-HEP-1 Cells / 中国实验方剂学杂志

ObjectiveProteoglycan TPG-1 isolated from Trametes robiniophila（Huaier） has proved to have anti-hepatoma activity， and this paper aims to explore the molecular mechanism. MethodHuman hepatoma SK-HEP-1 cells were treated with TPG-1 （0， 0.05， 0.1， 0.25， 0.5， 1 g·L-1）. Then cell survival was detected by methyl thiazolyl tetrazolium （MTT） and apoptosis by flow cytometry. In addition， expression of genes in SK-HEP-1 cells treated with or without TPG-1 was examined by DNA microarray to preliminarily explore the anti-hepatoma molecular mechanism of TPG-1. ResultTPG-1 inhibited the proliferation of SK-HEP-1 cells as compared with the blank group （P<0.01）. After treatment with 1 g·L-1 TPG-1 for 48 h， the apoptosis rate of SK-HEP-1 cells increased （P<0.01）， and TPG-1 promoted the cleavage of cysteinyl aspartate specific proteinase （Caspase）-3 and Caspase-7， the key mediators of apoptosis （P<0.01）. Additionally， TPG-1 （1 g·L-1） suppressed the migration of SK-HEP-1 cells （P<0.05）. A total of 971 differentially expressed genes （DEGs） were identified in SK-HEP-1 cells after treatment with TPG-1， with 486 up-regulated and 485 down-regulated. These DEGs were mainly involved in the Gene Ontology （GO） terms of interleukin-6 （IL-6） biosynthesis， antigen processing and presentation， superoxide dismutase activity， positive regulation of mitogen-activated protein kinase kinase kinase （MAPKKK） cascade， nature killer （NK） cell chemotaxis， and chemokine biosynthesis， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathways of nucleotide-binding oligomerization domain （NOD）-like receptor signaling pathway， apoptosis， Toll-like receptor signaling pathway， retinoic acid-inducible gene-Ⅰ （RIG-Ⅰ）-like receptor signaling pathway， T-cell receptor signaling pathway， and chemokine signaling pathway. Western blot results showed that TPG-1 （1 g·L-1） activated mitogen-activated protein kinase （MAPK） signaling pathway in SK-HEP-1 cells （P<0.01）. ConclusionProteoglycan TPG-1 inhibited the proliferation and migration， and induced apoptosis of human hepatoma SK-HEP-1 cells. Up-regulation of MAPK signaling pathway may be responsible for the growth inhibition of human hepatoma SK-HEP-1 cells by TPG-1.

Association of htra1 and hapln1 gene polymorphism with intervertebral disc degeneration / 中国组织工程研究

BACKGROUND: Studies have found that single nucleotide polymorphism genotypes in the HTRA1 gene promoter region are associated with intervertebral disc degeneration, while HAPLN1 is associated with osteoarthritis caused by intervertebral disc degeneration. OBJECTIVE: To explore the role of human secretory serine protease HTRA1 and the key group of extracellular matrix HAPLN1 in the pathogenesis of intervertebral disc degeneration. METHODS: This study included 498 postmenopausal female subjects who underwent a physical examination at Dingzhou People’s Hospital from April 2015 to December 2018. TaqMan PCR was used to detect HTRA1 gene promoter rs11200638 single nucleotide polymorphism and HAPLN1 gene 5' flanking rs975563, intron 1 rs10942332, intron 2 rs179851 and intron 4 rs4703570 single nucleotide polymorphism in 498 postmenopausal Chinese women. The correlation between the HTRA1orHAPLN1 gene polymorphisms and the radiographic features of spinal disc degeneration was analyzed. The trial has been approved by the Ethics Committee of Dingzhou People’s Hospital. RESULTS AND CONCLUSION: Among the 498 subjects with the HTRA1 gene rs11200638 single nucleotide polymorphism, 178 were GG homozygotes, 222 were GA heterozygotes, and 98 were AA homozygotes. We compared the parameters of intervertebral disc degeneration in subjects with at least one G allele (GG+GA, n=400) and without G allele (AA, n=98). In HTRA1 gene rs11200638 single nucleotide polymorphism, the score on intervertebral space stenosis in the subjects with GG+GA allele genome was lower than that in the subjects with AA allele (P < 0.001). With the increase of the score on intervertebral space stenosis, the proportion of the subjects with AA alleles increased (P ≤ 0.001). Among the 498 subjects with single nucleotide polymorphisms of the HAPLN1 gene, 137 were homozygous for TT, 230 were heterozygous for CT, and 131 were homozygous for CC. Intervertebral disc degeneration parameters of CC+TT allele (n=361) and TT allele (n=137) were compared. In the HAPLN1 gene, there was a significant difference between the CC+TT and TT alleles of the rs179851 single nucleotide polymorphism in osteophyte formation and intervertebral space stenosis (P < 0.01). Among the HAPLN1 gene rs179851 single nucleotide polymorphisms, the proportion of subjects with TT alleles and intervertebral space stenosis ≥ 6 points increased (P < 0.05). With an increase in osteophyte formation score, the proportion of subjects with TT allele increased (P < 0.001). These results reveal that HTRA1 and HAPLN1 genetic variations at specific genetic loci are associated with intervertebral disc degeneration.

Research Progress on Mechanical Characteristics of Articular Cartilage / 医用生物力学

Osteoarthritis (OA) is a chronic degenerative joint disease characterized by joint pain and stiffness, which predisposes to the elderly. The onset of OA is slow, the course of disease is long, and the early clinical manifestations and histological changes are not obvious, which limits the early diagnosis and treatment of the disease. The micro-structure of articular cartilage determines the macro-mechanical properties of cartilage. The micro-structure of articular cartilage changes in a depth-dependent manner, which makes the mechanical properties of cartilage also depth-dependent. From superficial to deep areas of cartilage, the anti-load and anti-deformation ability of cartilage increases gradually. However, with development of the disease, the change of cartilage micro-structure leads to the decrease in load resistance and deformation resistance of OA cartilage. Therefore, the mechanical properties of articular cartilage can be inferred by detecting the micro-structure of articular cartilage. On the other hand, the mechanical properties of articular cartilage can be used to understand the micro-changes of cartilage, which is helpful to understand OA development and facilitate early diagnosis of the disease. This paper reviewed the recent research literatures on mechanical properties of articular cartilage under normal and acute or chronic injuries, and elaborated the relationship between the structure and mechanical properties of articular cartilage, which further provided the theoretical basis for the OA development, early diagnosis and treatment.

Matrix Production in Chondrocytes Transfected with Sex Determining Region Y-Box 9 and Telomerase Reverse Transcriptase Genes: An In Vitro Evaluation from Monolayer Culture to Three-Dimensional Culture

BACKGROUND: This study aimed to observe the cartilaginous matrix production in SRY (sex determining region Y)-box 9 (SOX9)- and/or telomerase reverse transcriptase (TERT)-transfected chondrocytes from monolayer to three-dimensional (3D) culture.

Glycosylation of dentin matrix protein 1 is critical for fracture healing via promoting chondrogenesis / 医学前沿

Fractures are frequently occurring diseases that endanger human health. Crucial to fracture healing is cartilage formation, which provides a bone-regeneration environment. Cartilage consists of both chondrocytes and extracellular matrix (ECM). The ECM of cartilage includes collagens and various types of proteoglycans (PGs), which play important roles in maintaining primary stability in fracture healing. The PG form of dentin matrix protein 1 (DMP1-PG) is involved in maintaining the health of articular cartilage and bone. Our previous data have shown that DMP1-PG is richly expressed in the cartilaginous calluses of fracture sites. However, the possible significant role of DMP1-PG in chondrogenesis and fracture healing is unknown. To further detect the potential role of DMP1-PG in fracture repair, we established a mouse fracture model by using a glycosylation site mutant DMP1 mouse (S89G-DMP1 mouse). Upon inspection, fewer cartilaginous calluses and down-regulated expression levels of chondrogenesis genes were observed in the fracture sites of S89G-DMP1 mice. Given the deficiency of DMP1-PG, the impaired IL-6/JAK/STAT signaling pathway was observed to affect the chondrogenesis of fracture healing. Overall, these results suggest that DMP1-PG is an indispensable proteoglycan in chondrogenesis during fracture healing.

Mechanism of thermosensitive moxibustion on knee osteoarthritis in rabbit models / 中国针灸

<p><b>OBJECTIVE</b>To observe the impacts of thermosensitive moxibustion (TSM) on the expressions of nitric oxide (NO), typeⅠdisintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), typeⅡcollagen and proteoglycan (PG) in the rabbit models of knee osteoarthritis (KOA) and explore the mechanism of TSM on KOA.</p><p><b>METHODS</b>A total of 42 Japanese long-eared male rabbits were divided into a blank group (6 rabbits), a model group (6 rabbits), a moxibustion group (24 rabbits) and a sham-operation group (6 rabbits) according to the random number table. In the blank group, the rabbits were fed normally. In the model and moxibustion groups, the papain injection was given to establish KOA models. The rabbits in the sham-operation group were treated with the intracavity injection of 0.9% NaCl solution. The rabbits were forced to move for 30 min every day, continuously for 15 days during modeling. At the end of modeling, in the moxibustion group, moxibusiton was applied at "Dubi" (ST 35), once a day, 40 min each time, for 14 days totally. According to the temperature changes during moxibustion, the rabbits were divided into a TSM group and a non-TSM group. 6 rabbits were collected randomly from the two groups. The usual feeding was given in the blank group, the model group and the sham-operation group, without any intervention. The body mass and behavioristics changes were observed in each group. At the end of treatment, the nitrate reduction method was adopted to determine NO expression in the serum. The real-time PCR was adopted to determine the expressions of ADAMTS-4, typeⅡcollagen and PG in the cartilage.</p><p><b>RESULTS</b>① After modeling, compared with the blank group, the body mass was all reduced and the Lequesne MG score was increased in the model group, TSM group, non-TSM group and sham-operation group (<0.05, <0.01). After intervention, compared with the blank group, the body mass was decreased and the Lequesne MG score was increased in the model and sham-operation groups (<0.05, <0.01). Compared with the model group, the body mass was increased and the lequesne MG score was decreased in the TSM, non-TSM, and sham-operation groups (<0.05, <0.01). Compared with the non-TSM group, the body mass in the TSM group was increased remarkably (<0.05), but the difference in Lequesne MG score was not statistically significant (>0.05). ② After intervention, compared with the blank group, the expressions of NO and ADAMTS-4 were all increased and the expressions of typeⅡcollagen and PG were decreased in the model group, TSM group, non-TSM group and sham-operation group (<0.05, <0.01). Compared with the model group, the expressions of NO and ADAMTS-4 were all remarkably lower and the expressions of typeⅡcollagen and PG were increased in the TSM group, non-TSM group and sham-operation group (<0.05, <0.01). Compared with the non-TSM group, the expressions of NO and ADAMTS-4 were all remarkably lower and the expressions of typeⅡcollagen and PG were increased in the TSM group after intervention (all <0.05).</p><p><b>CONCLUSION</b>The thermosensitive moxibustion alleviates the inflammatory reactions and protects the joint cartilage through inhibiting the expressions of NO and ADAMTS-4 to achieve the effects in the treatment of KOA.</p>

Quantitative Study on Articular Cartilage By Fourier Transform Infrared Spectroscopic Imaging and Support Vector Machine / 分析化学

Fourier transform infrared spectroscopic imaging (FTIRSI) technology can simultaneously obtain microstructure information and infrared spectral information of the samples. The method of FTIRSI combined with chemometric algorithms can be used for quantitative analysis of sample spectral information and tissue discrimination research. Based on this, FTIRSI and support vector machine classification (SVC) for the first time were used in this work to discriminate healthy and degenerated articular cartilage, with high accuracies of 100％ and 95. 4％ , respectively, and sum accuracy of 97. 7％ . The support vector regression (SVR) model was used to quantitatively study the contents and distribution of two biomacromolecules, collagen and proteoglycan, in articular cartilage. The proteoglycan loss occurred in the degenerated articular cartilage, especially in the superficial area. This study indicates that the combination of FTIRSI and support vector machine (SVM) is expected to become a new diagnostic tool for osteoarthritis, which is of great significance for the early diagnosis and research of osteoarthritis.

Glycosylation of dentin matrix protein 1 is a novel key element for astrocyte maturation and BBB integrity

The blood-brain barrier (BBB) is a tight boundary formed between endothelial cells and astrocytes, which separates and protects brain from most pathogens as well as neural toxins in circulation. However, detailed molecular players involved in formation of BBB are not completely known. Dentin matrix protein 1 (DMP1)-proteoglycan (PG), which is known to be involved in mineralization of bones and dentin, is also expressed in soft tissues including brain with unknown functions. In the present study, we reported that DMP1-PG was expressed in brain astrocytes and enriched in BBB units. The only glycosylation site of DMP1 is serine89 (S89) in the N-terminal domain of the protein in mouse. Mutant mice with DMP1 point mutations changing S89 to glycine (S89G), which completely eradicated glycosylation of the protein, demonstrated severe BBB disruption. Another breed of DMP1 mutant mice, which lacked the C-terminal domain of DMP1, manifested normal BBB function. The polarity of S89G-DMP1 astrocytes was disrupted and cell-cell adhesion was loosened. Through a battery of analyses, we found that DMP1 glycosylation was critically required for astrocyte maturation both in vitro and in vivo. S89G-DMP1 mutant astrocytes failed to express aquaporin 4 and had reduced laminin and ZO1 expression, which resulted in disruption of BBB. Interestingly, overexpression of wild-type DMP1-PG in mouse brain driven by the nestin promoter elevated laminin and ZO1 expression beyond wild type levels and could effectively resisted intravenous mannitol-induced BBB reversible opening. Taken together, our study not only revealed a novel element, i.e., DMP1-PG, that regulated BBB formation, but also assigned a new function to DMP1-PG.

MRS in quantitative assessment of lumbar discs degeneration / 中国医学影像技术

Objective To evaluate the value of MRS in quantitative assessment of degeneration of lumbar discs.Methods Totally 82 patients with lumbago underwent lumbar MR scanning.All the discs were classified with Pfirrmann grades in the sequences of sagittal T2WI.The area under N-acetyl peak,under water peak and the ratio of N-acetyl/Water were measured by MRS.Correlation between MRS values and Pfirrmann grade,age were analyzed.Results In 82 patients,204 lumbar discs were measured by MRS.There were 89,73,39,3 discs in Pfirrmann Ⅱ,Ⅲ,Ⅳ,V respectively.The areas of N-acetyl,water peak and N-acetyl/Water ratio of nucleus region were positively correlated with Pfirrmann grading,respectively (rs =-0.460,-0.204,-0.526,all P＜0.05).There were 62,25,37,51,29 discs in patients aged ＜30,30-39,40-49,50-59,＞59 years respectively.The ares of N-acetyl peak,N-acetyl/Water ratio of nucleus region was negatively correlated with the age (rs=-0.247,-0.385,both P＜0.05).Conclusion MRS can be used for quantitative assessment of lumbar discs degeneration.

Prediction of miR-199a-3p targets gene and its bioinformatics analysis / 西安交通大学学报(医学版)

Objective To provide theoretical guidance for further research on the role of miR-1 99a-3p in formation and development of bladder cancer.Methods Mature sequence of miR-1 99a-3p was analyzed;target genes and transcription factors of miRNA-1 99a-3p were predicted,and the target genes were analyzed for gene ontology (GO)enrichment and Kyoto Encyclopedia of Genes and Genome (KEGG)pathway.Then TF-miRNA-mRNA network diagram was constructed.Results Sequences of miR-1 99a-3p were highly conserved in various species.In GO analysis,the target genes of miR-1 99a-3p were enriched in many biological processes,such as regulation of cellular process,regulation of macromolecule metabolic process,and regulation of biological process (P <0.01 ).In KEGG pathway,the target genes were mainly located in bacterial invasion pathway of epithelial cells,ECM-receptor interaction pathway,PI3K-Akt signaling pathway,MAPK signaling pathway,small cell lung cancer pathway,and proteoglycans pathway in the cancer (P <0.05).According to the TF-miRNA-mRNA network diagram,the important genes that might be regulated by miR-1 99a-3p were MYC,SP1,mTOR,NFκB,and NFκB1.Conclusion miR-1 99a-3p may directly target mTOR and participate in the formation and development of bladder cancer through regulating PI3K-Akt-mTOR signaling pathway.

The research progression on SPOCK1 and tumor / 实用肿瘤学杂志

Testicular proteoglycan 1 ( SPOCK1 ) , a kind of extracellular matrix glycoprotein, can inhibit the activity of cathepsin and promotes the combination of low affinity for calcium.SPOCK1 is essential to the de-velopment of mammals.It not only regulates cell,and the interactions between cells and matrix are also related to cell migration and proliferation.Currently,SPOCK1is confirmed that expression can promote the occurrence and development of tumor,suggesting it could be a new anti-tumor target.This article is to review SPOCK1 molecular structure,biological function and its relationship with tumor.

Recent advances in corneal stroma protein studies / 中华实验眼科杂志

Cornea stroma mostly consists of proteins in extracellular matrix secreted by corneal epithelial cells,stroma and endothelial cells.A total of 1 679 unique Swiss-Prot annotated proteins are identified by LC-MS/MS analyses in human corneal stroma based on SDS-PAGE separation followed by in-gel trypsin digestion.As the principal component of corneal stroma, corneal stroma proteins play an important role in the physiological and pathological activities of eye.Studies towards their functions are helpful for revealing the pathogenesis of eye diseases and providing new ideas of prevention and therapy.Mainly through their manifestations,proteins in corneal stroma play a key role in forming gel-like organic material and network like structure of filaments.The gel-like organic material is composed of proteoglycan,enzymes,haemocyanin and binding proteins,while the network of filaments are constituted by collagen, elastin,keratin,vimentin as well as interconnected filaments made from fibronectin and laminin.The categories and functions of corneal stroma proteins were summarized.

Progress in technology for glycomics / 中国药理学与毒理学杂志

Technologies for glycomics usually involve methods for separation and purification of poly-saccharides, and separation, structure resolution, quantification, property investigation and function comment of glycan chains. Because of the different biochemical properties of glycoproteins, proteogly-cans and glycolipids, the separation and purification of polysaccharides involve corresponding fractional precipitation, boric acid affinity, titanium dioxide, affinity chromatography, size exclusion method, and gel filtration chromatography column chromatography methods. The lectins, water affinity chromatogra-phy , solid phase extraction and other technologies could be applied to the oil enrichment of high pure and specific glycan chains. The structure of glycan chains can be analyzed using lectin microarray technolo-gy, mass spectrometry, and derivatization markers of glycan chains. lsotope labelling and metabolic labeling can be used to quantify glycan chains. The glycan biological function can be better understood using glycan chain structure analysis software and database of glycan chains by bioinformatics.

Application of combined detection of Golgi glycoprotein 73,phosphatidylinositol proteoglycan 3 and percentage of AFP heteroplasmon in diagnosis of primary hepatocellular carcinoma / 国际检验医学杂志

Objective To study the application of the combined detection of Golgi glycoprotein 73(GP-73),phosphatidylinositol proteoglycan 3(GPC3 )and percentage of AFP heteroplasmon(AFP-L3%)in the diagnosis of primary hepatocellular carcinoma (PHC).Methods The concentrations of GP-73,GPC3 and AFP-L3 were detected by enzyme-linked immunosorbent assay(ELISA) in 154 patients with PHC(PHC group),78 patients with cirrhosis(cirrhosis group)and 56 healthy subjects(control group).Then the detection results were statistically analyzed.Results The levels of GP-73,GPC3 and AFP-L3% in the PHC group were signifi-cantly higher than those in the liver cirrhosis group and the control group(P <0.05).The positive rates of GP-73,GPC3 and AFP-L3% in the PHC group were 66.2%,72.1% and 53.2% respectively.The positive rate in the combined detection of these three in-dices could reach 97.9%,which was higher than the sensitivity and accuracy in any single index detection and the combination de-tection.In the PHC group,the comparison between different levels of GP-73 and AFP-L3% with the AFP levels showed the statis-tically significant difference(P <0.05 ).Conclusion The combination detection of GP-73,GPC3 and AFP-L3% can improve the sensitivity and accuracy for diagnosing PHC and has reference significance in the differential diagnosis of early PHC.

Proteoglycan and collagen type Ⅱ in the adjacent intervertebral disc of the cervical instability models / 中国组织工程研究

BACKGROUND:Cervical decompression and fusion internal fixation wil accelerate adjacent segment disc degeneration, and it is not clear whether single segment instaibility can increase the adjacent segment disc degeneration. OBJECTIVE:To study the changes of morphology, proteoglycan and col agen type Ⅱ in the adjacent intervertebral disc of the cervical instability models. METHODS:Sixteen New Zealand white rabbits were divided into two groups randomly, with eight rabbits in the control group and eight rabbits in the model group. The animal cervical instability models were made by destroyed partly annulus fibrosus and absorbed C 5/6 nucleus pulposus through anterior cervical puncture. After 12 weeks, the animal models were tested by X-ray film. Al rabbits were sacrificed and 10 mg nucleus pulposus of the intervertebral discs of C 4/5 cut from sagittal plane were harvested and stored under 0 ℃. The content of proteoglycan in nucleus pulposus was tested with phloroglucinol method. Then, the paraffin sections of intervertebral disc tissues were taken for hematoxylin-eosin staining and SABC immunohistochemical staining. RESULTS AND CONCLUSION:The notochord cells of C4/5 intervertebral discs in the experimental group was decreased, and being replaced by fibroblast-like cells. Round chondrocytes could be seen occasional y and intervertebral discs annulus fibrosus became rough and arranged disorderly, the hyaline degeneration and pigmentation were observed as wel as the fibrochondrocytes, and there was a gap between inner and outer annulus fibrosus. The content of proteoglycan was decreased in the nucleus pulposus, and there was significant difference between two groups. The col agen type Ⅱ in the degenerative disc nucleus pulposus and annulus fibrosus of the experimental group was lower than that of the control group. Cervical instability can lead to adjacent intervertebral disc degeneration with the morphological changes and decreased content of proteoglycan and col agen type Ⅱ.

Demyelination and changes in chondrotin sulfate proteoglycan expression after spinal cord compression injury / 中华物理医学与康复杂志

Objective To investigate the role of demyelination and the alteration of chondrotin sulfate proteoglycan (CSPG,NG2) expression after compression injury of the spinal cord (CSCI).Methods Seventy-five adult Sprague-Dawley rats were randomly divided into a normal group,a sham-operation group,a CSCI 1 day group,a CSCI 3 day group,and a CSCI 7 day group.There were 15 rats in each group.The injuries in the CSCI groups were inflicted using a technique devised in our laboratory.Basso-Beattie-Bresnahan (BBB) neurological function assessment was used to assess the rats' motor function,osmic acid staining and transmission electronic microscopy (TEM)were used to observe any pathological changes of myelinated nerve fibers in the white matter at 1,3 and 7 days after CSCI.The amount of myelinated nerve fibers in the posterior funiculus of the spinal cord and the ratio of myelin sheath thickness to axon diameter (the G-ratio) were calculated.Any alteration in NG2 expression was observed by Western blotting.Results The average neurological function assessment scores in the CSCI groups were (1.23 ±0.45),(0.65 ± 0.35) and (0.00 ± 0.00) respectively.Compared with the normal group (21.00 ± 0.00) and the sham operation group (21.00 ± 0.00),the differences were all statistically significant.The rats' motor function deteriorated gradually with time after the CSCI.Osmic acid staining showed that the white matter was intact in the normal and sham groups.After being compressed the myelinated nerve fibers became swollen,degenerated and broke down.The amount of myelinated nerve fibers in the normal group,the sham operation group and the three CSCI groups was (2771 ± 108),(2675 ± 199),(2403 ± 161),(1708 ± 70) and (8 10 ± 95) respectively.The amount of myelinated nerve fibers decreased in the CSCI groups and reached a minimum on the 7th day.The difference was statistically significant.The TEM quantity analysis showed that the G-ratios in the normal,sham operation,and CSCI 1 day,3 day and7 day groups were (18.10±0.4),(17.70±1.0),(6.69 ±0.8),(5.73 ±0.4) and (4.95 ±0.5) respectively.Compared with the normal and sham operation groups,the G-ratios in the 3 CSCI groups were lower and reached their minimum on the 7th day after injury.The difference was statistically significant.TEM observation showed that the axons and myelin sheaths were intact in the normal and sham groups.After CSCI the axons became swollen and cell organelles in the axoplasm degenerated and decreased.The layers of myelin sheath shrank,folded and even wrinkled,which had an onion-like appearance.The oligodendrocytes exhibited chromatin condensation.Macrophages showed infiltration.Western blotting showed that the expression of NG2 in the CSCI groups reached a maximum on the 1st day after injury and then decreased with time.The expression of NG2 in the CSCI groups was higher than in the normal and sham groups,and the difference was statistically significant.Conclusion Demyelination occurs after CSCI-the amount of myelinated nerve fibers decreases and neurological deficits increase with time.The expression of NG2 was associated with changes in the myelin sheaths after CSCI and contributed to recovery of the myelin sheath through proliferation and differentiation to oligodendrocytes and perhaps other kinds of cells.

A ROCK Inhibitor Blocks the Inhibitory Effect of Chondroitin Sulfate Proteoglycan on Morphological Changes of Mesenchymal Stromal/Stem Cells into Neuron-Like Cells

Chondroitin sulfate proteoglycan (CSPG) inhibits neurite outgrowth of various neuronal cell types, and CSPG-associated inhibition of neurite outgrowth is mediated by the Rho/ROCK pathway. Mesenchymal stromal/stem cells (MSCs) have the potential to differentiate into neuron-like cells under specific conditions and have been shown to differentiate into neuron-like cells by co-treatment with the ROCK inhibitor Y27632 and the hypoxia condition mimicking agent CoCl2. In this study, we addressed the hypothesis that a ROCK inhibitor might be beneficial to regenerate neurons during stem cell therapy by preventing transplanted MSCs from inhibition by CSPG in damaged tissues. Indeed, dose-dependent inhibition by CSPG pretreatment was observed during morphological changes of Wharton's jelly-derived MSCs (WJ-MSCs) induced by Y27632 alone. The formation of neurite-like structures was significantly inhibited when WJ-MSCs were pre-treated with CSPG before induction under Y27632 plus CoCl2 conditions, and pretreatment with a protein kinase C inhibitor reversed such inhibition. However, CSPG treatment resulted in no significant inhibition of the WJ-MSC morphological changes into neuron-like cells after initiating induction by Y27632 plus CoCl2. No marked changes were detected in expression levels of neuronal markers induced by Y27632 plus CoCl2 upon CSPG treatment. CSPG also blocked the morphological changes of human bone marrow-derived MSCs into neuron-like cells under other neuronal induction condition without the ROCK inhibitor, and Y27632 pre-treatment blocked the inhibitory effect of CSPG. These results suggest that a ROCK inhibitor can be efficiently used in stem cell therapy for neuronal induction by avoiding hindrance from CSPG.