In this study, insulin (insulin, INS)/Ca₃PO₄ complex and glucose oxidase (glucose oxidase, GOx)/Cu₃(PO₄)₂ complex were prepared by coprecipitation method. The mineralized insulin (mineralized insulin, m-INS) showed irregular crystalline clusters, and the mineralized glucose oxidase (m-GOx) showed flower spherical morphology, with a diameter of about 1-2 μm. In vitro simulated release experiment showed that m-INS released INS as the pH value of the medium decreased. When the pH value was 4.5, the release amount reached 96.68%. The enzyme activity detection experiment showed that the enzyme activity stability of m-GOx was higher than that of free GOx. It still maintained high activity after 10 days at room temperature, while the activity of GOx was less than 60%. The glucose solution was prepared to simulate the state of normal blood glucose (5.6 mmol/L) and hyperglycemia (22.2 mmol/L). When m-INS and m-GOx were added to the glucose solution, the release amount of INS showed a significant glucose concentration dependence. The higher the glucose concentration, the greater the release amount and release rate of INS. Finally, m-INS, m-GOx and hyaluronic acid (HA) solution were mixed to prepare HA microneedle arrays loaded with m-INS and m-GOx. Type 1 diabetes mice were constructed to evaluate the effect of drug-loaded HA microarray on blood glucose control in diabetic rats. The results show that the HA microneedles loaded with m-INS/m-GOx could deliver drugs effectively. The average blood glucose concentration in diabetic rats dropped to about 7 mmol/L within 1 h, normal blood glucose concentration could be maintained for 10 h, and the overall blood glucose concentration was lower than the level before administration for 36 hours. Compared with HA microneedles loaded with INS only, m-ins microneedles showed better glucose tolerance, longer-lasting glucose control effect and less risk of hypoglycemia. Compared with other sustained-release systems, the preparation process of the core components in this study is simple, efficient, safe and effective, and has great commercial potential.
Animals ; Blood Glucose ; Delayed-Action Preparations/therapeutic use* ; Diabetes Mellitus, Experimental/drug therapy* ; Drug Delivery Systems/methods* ; Glucose Oxidase/chemistry* ; Hyaluronic Acid ; I Blood-Group System ; Insulin/therapeutic use* ; Mice ; P Blood-Group System ; Rats

BACKGROUND: The fabrication of microchannels in hydrogel can facilitate the perfusion of nutrients and oxygen, which leads to guidance cues for vasculogenesis. Microchannel patterning in biomimetic hydrogels is a challenging issue for tissue regeneration because of the inherent low formability of hydrogels in a complex configuration. We fabricated microchannels using wire network molding and immobilized the angiogenic factors in the hydrogel and evaluated the vasculogenesis in vitro and in vivo. METHODS: Microchannels were fabricated in a hyaluronic acid-based biomimetic hydrogel by using “wire network molding” technology. Substance P was immobilized in acrylated hyaluronic acid for angiogenic cues using Michael type addition reaction. In vitro and in vivo angiogenic activities of hydrogel with microchannels were evaluated. RESULTS: In vitro cell culture experiment shows that cell viability in two experimental biomimetic hydrogels (with microchannels and microchannels + SP) was higher than that of a biomimetic hydrogel without microchannels (bulk group). Evaluation on differentiation of human mesenchymal stem cells (hMSCs) in biomimetic hydrogels with fabricated microchannels shows that the differentiation of hMSC into endothelial cells was significantly increased compared with that of the bulk group. In vivo angiogenesis analysis shows that thin blood vessels of approximately 25–30 µm in diameter were observed in the microchannel group and microchannel + SP group, whereas not seen in the bulk group. CONCLUSION: The strategy of fabricating microchannels in a biomimetic hydrogel and simultaneously providing a chemical cue for angiogenesis is a promising formula for large-scale tissue regeneration.
Angiogenesis Inducing Agents ; Biomimetics* ; Blood Vessels ; Cell Culture Techniques ; Cell Survival ; Cues ; Endothelial Cells ; Fungi ; Humans ; Hyaluronic Acid ; Hydrogel* ; Hydrogels* ; In Vitro Techniques ; Mesenchymal Stromal Cells ; Oxygen ; Perfusion ; Regeneration ; Substance P

OBJECTIVETo study the inhibitory effect of flavonoids from Glycyrrhiza uralensis on thioacetamide-induced chonic hepatic fibrosis in rats and the effect on the protein expressions of transforming growth factor-β1 (TGF-β1) and Caspase-3 in livers.

METHODMale Sprague-Dawley rats were randomly divided into totally seven groups: the normal control group, the model group, LF groups s (400, 200, 100, 50 mg · kg(-1) · d(-1)) and the silymarin positive control group (30 mg · kg(-1) · d(-1)). The hepatic fibrosis model was induced in the rats through intraperitoneal injection with 3% thioacetamide (TAA) at a dose of 150 mg · kg(-1) body weight twice a week for 12 weeks. During the course, the control group and the model group were orally administered with saline (1 mL · kg(-1) · d(-1)). After the modeling and drug intervention, the pathologic changes and fibrosis in liver tissues were observed by HE staining and Masson's Trichrome staining. The serum alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and liver hydroxyproline (HYP) contents were assayed by biochemical process. The serum hyaluronic acid (HA) was assessed by radioimmunoassay. In addition, the protein expressions of liver TGF-β1 and Caspase-3 were examined by immunohistochemical method. The mRNA expression of TGF-β1 in hepatic tissues was examined by quantitative Real-time PCR analysis.

RESULTCompared with the model group, flavonoids can protect the integrity of the structure of liver tissues, significantly reduce the hepatic cell degeneration and necrosis and the proliferation of fibrous tissues, notably reduce the serum AST, ALT, ALP and HA and HYP in hepatic tissues and down-regulate the protein expressions of liver TGF-β1 and Caspase-3 and the mRNA expression of TGF-β1 in hepatic tissues.

CONCLUSIONThe licorice flavonoids can resist the thioacetamide-induced hepatic fibrosis in rats. Its mechanism may be related to the down-regulation of the protein expressions of TGF-β1 and Caspase-3.

Animals ; Caspase 3 ; analysis ; Flavonoids ; pharmacology ; Glycyrrhiza uralensis ; chemistry ; Hyaluronic Acid ; blood ; Liver ; pathology ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; prevention & control ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Thioacetamide ; Transforming Growth Factor beta1 ; analysis ; genetics

OBJECTIVETo investigate the correlation between four serum fibrosis markers and liver function in patients with infantile hepatitis syndrome (IHS), and to explore the clinical significance of these markers in the diagnosis of IHS and the assessment of disease severity.

METHODSA retrospective study was performed on 60 patients with IHS who were divided into hepatic fibrosis and normal groups based on ultrasound diagnosis. Levels of four liver fibrosis markers, i.e., hyaluronic acid (HA), type III procollagen (PC-III), type IV collagen (IV.C), and laminin (LN), were compared between the two groups, and the correlation between these markers and liver function was analyzed.

RESULTSLevels of liver function markers (alanine aminotransferase (ALT), glutamyl transpeptidase (GGT), total bilirubin (TBil), direct bilirubin (DBil), indirect bilirubin (IBil), and total bile acid (TBA)) in the hepatic fibrosis group were significantly higher than those in the normal group (P<0.05). Levels of HA and IV.C in the hepatic fibrosis group were significantly higher compared with those in the normal group (P<0.05). Furthermore, HA, IV.C, and PC-III levels were positively correlated with those of ALT, TBil, GGT, DBil, IBil, and TBA (r=0.25-0.49), and the strongest correlation existed between HA/IV.C and ALT/jaundice markers.

CONCLUSIONSAssay measuring serum fibrosis markers (HA, IV.C, and PC-III) in combination with liver function tests and ultrasound examination has an important clinical value in the early diagnosis of IHS and evaluation of disease severity.

Biomarkers ; blood ; Collagen Type III ; blood ; Collagen Type IV ; blood ; Female ; Hepatitis ; blood ; diagnosis ; physiopathology ; Humans ; Hyaluronic Acid ; blood ; Infant ; Laminin ; blood ; Liver ; physiopathology ; Liver Cirrhosis ; blood ; diagnosis ; Male ; Retrospective Studies ; Syndrome

INTRODUCTIONTo prevent long-term unfavourable consequences to the articular cartilage of weight-bearing joints, serum biomarkers can be used to identify optimum loading of activities. This study aimed to investigate the circulation pattern of serum cartilage biomarkers in healthy adults in response to an uphill walk.

METHODSThis study recruited 58 healthy participants for the experimental group and 24 matched participants for the control group. Participants in the experimental group walked continuously for 14 km on a pathway with a 5.97° incline, while participants from the control group walked on a horizontal pathway. Serum was collected from both groups preactivity (i.e. T1), immediately after activity (i.e. T2) and 24 hours after T1 (i.e. T3). The serum cartilage oligomeric matrix protein (COMP), chondroitin sulfate-WF6 (WF6) and hyaluronic acid (HA) levels at each time point were quantified using enzyme-linked immunosorbent assays, and the results analysed.

RESULTSBoth groups shared similar demographic characteristics and activity duration. At T2, the serum COMP level of the experimental group was significantly higher than that of the control group, but the serum HA level of the experimental group was significantly lower than that of the control group. No significant difference between the serum WF6 levels of the experimental and control groups was observed at T2.

CONCLUSIONIncreasing levels of serum COMP demonstrate articular cartilage susceptibility to the increasing load. An unsustainable, high serum COMP level and an undetectable change in WF6 level were considered to be a reversible physiological change of the cartilage. A change in ser um HA level could be related to intensive physical activity and dynamic clearance rather than a change in cartilage structure.

Adolescent ; Adult ; Biomarkers ; blood ; Cartilage Oligomeric Matrix Protein ; blood ; Cartilage, Articular ; metabolism ; Chondroitin Sulfates ; blood ; Female ; Healthy Volunteers ; Humans ; Hyaluronic Acid ; blood ; Male ; Time Factors ; Walking ; Young Adult

Thirty-one dogs with patellar luxation (grades 2 and 3) were categorized into three groups. Group 1 (G.1; n = 12) had sodium hyaluronate (SHA) intra-articularly injected into the stifle joint that received surgery. Group 2 (G.2; n = 10) received SHA twice: first after surgery and then 1 week later. Group 3 (G.3; n = 9) served as a control, without injection. Blood was collected before injection and then once a week for 4 weeks after injection for evaluation of chondroitin sulfate (CS-WF6) and hyaluronan (HA). The results revealed significantly (p < 0.05) improved clinical scores by the end of week 4 in G.1 and G.2 relative to G.3; however, there was no significant difference between G.1 and G.2. There was a significant decrease (p < 0.05) in serum CS-WF6 levels beginning at week 2 in G.1 and G.2. At weeks 3 and 4, serum HA in G.1 and G.2 differed from that in G.3 (p < 0.05). No significant difference (p > 0.05) was observed in serum biomarkers between G.1 and G.2. In conclusion, intra-articular injection with SHA after joint surgery may improve homeostasis of the joint, retarding the process of OA.
Animals ; Blood Chemical Analysis/veterinary ; Chondroitin Sulfates/metabolism ; *Dogs ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Hyaluronic Acid/*administration & dosage/metabolism ; Injections, Intra-Articular/veterinary ; Male ; Osteoarthritis, Knee/drug therapy/prevention & control/*veterinary ; Stifle/*surgery ; Viscosupplements/*administration & dosage

BACKGROUNDChronic liver disease causes aberrant formation of fibrous tissue that impedes normal liver function, ultimately resulting in liver cirrhosis. Iron uptake can occur within the hepatic parenchyma or within the various nodules that form in a cirrhotic liver, termed siderotic nodules (SN). We aimed to investigate the diagnostic performance of susceptibility weighted imaging (SWI) for detection of SN in patients with liver cirrhosis, and to evaluate the potential of SN numbers for assessing the degree of hepatic iron deposition, liver function, and liver fibrosis stage.

METHODSNinety-one patients with chronic liver cirrhosis, who underwent megnetic resonance imagine (MRI) scanning in our department between November 2010 and April 2011, were included in the study. A 3.0T MRI scanner was used to acquire T1WI, T2WI, T2WI, and SWI images. The number of nodules, signal intensity ratio (SIR), and contrast noise ratio (CNR) were recorded and analyzed by chi-square and ANOVA statistical tests. Correlation analysis was performed to evaluate the correlations between the number of SN and Child-Pugh classification, ferritin and hyaluronic acid levels.

RESULTSThe sensitivity of SWI, T1WI, T2WI, and T2 WI for detecting SN was 62.5%, 12.1%, 24.2% and 41.8%, respectively. SWI detected significantly more nodules than routine T1WI, T2WI, and T2 WI procedures (P < 0.05). The SIR was the lowest in SWI (0.361 ± 0.209), as compared to T1WI (0.852 ± 0.163), T2WI (0.584 ± 0.172), and T2 WI (0.497 ± 0.196). The CNR was the highest in SWI (13.932 ± 5.637), as compared to T1WI (9.147 ± 5.785), T2WI (9.771 ± 5.490), and T2 WI (11.491 ± 4.573). The correlation coefficients of the number of SN with ferritin, Child-Pugh classification, and hyaluronic acid levels were 0.672, -0.055, and 0.163, respectively.

CONCLUSIONSThe sensitivity and contrast of SWI for detecting SN in patients with liver cirrhosis are higher than conventional MRI. The number of SN can help to assess the degree of iron deposition in patients with liver cirrhosis.

Adult ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Ferritins ; blood ; Humans ; Hyaluronic Acid ; blood ; Liver ; pathology ; Liver Cirrhosis ; blood ; pathology ; Male ; Middle Aged ; Sensitivity and Specificity

PURPOSE: Aplasia Cutis Congenita(ACC) is a rare disease characterized by the focal defect of the skin at birth, frequently involving scalp, but it may affect any region of the body. There are no etiology known but some conditions such as intrauterine vascular ischemia, amniotic adherences and viral infections are associated. The ideal treatment for the ACC is not known. Superficial and relatively small sized defects(<3x5cm) may heal spontaneously and large defects related with risks of infection and bleeding may require aggressive surgical treatment. Hyalomatrix(R) is a bilayer of an esterified hyaluronan scaffold beneath a silicone membrane. It has been used as a temporary dermal substitute to cover deep thickness skin defect and has physiological functions derive from the structural role in extracellular matrix and interaction with cell surface receptor. This material has been used for the wound bed pre-treatment for skin graft to follow and especially in uncooperative patient, like a newborn, this could be a efficient and aseptic way of promoting granulation without daily irritative wound care. For this reason, using Hyalomatrix(R) for the treatment of ACC was preferred in this paper. METHODS: We report a case of a newborn with ACC of the vertex scalp and non-ossified partial skull defect. The large sized skin and skull defect(6x6cm) was found with intact dura mater. No other complications such as bleeding or abnormal neurologic sign were accompanied. Escharectomy was performed and Hyalomatrix(R) was applied for the protection and the induction of acute wound healing for 3 months before the split-thickness skin graft. During the 3 months period, the dressing was renewed in aseptic technique for every 3 weeks. The skin graft was achieved on the healthy granulation bed. RESULTS: The operative procedure was uneventful without necessity of blood transfusion. Postoperative physical examination revealed no additional abnormalities. Regular wound management was performed in out-patient clinic and the grafted skin was taken completely. No other problems developed during follow-up. CONCLUSION: Hyalomatrix(R) provides protective and favorable environment for wound healing. The combination of the use of Hyalomatrix(R) and the skin graft will be a good alternative for the ACC patients with relatively large defect on vertex.
Bandages ; Blood Transfusion ; Dura Mater ; Ectodermal Dysplasia ; Extracellular Matrix ; Follow-Up Studies ; Hemorrhage ; Humans ; Hyaluronic Acid ; Infant, Newborn ; Ischemia ; Membranes ; Neurologic Manifestations ; Outpatients ; Parturition ; Physical Examination ; Rare Diseases ; Scalp ; Silicones ; Skin ; Skull ; Surgical Procedures, Operative ; Transplants ; Wound Healing

OBJECTIVETo explore therapeutic effect of Haobieyangyinruanjianfang (HBYYRJ) on mouse liver fibrosis by schistosomiasis.

METHODSMice except for normal control were infected with Japanese schistosome cercarias, after 12 weeks, infected mice were divided into 7 groups: low HBYYRJ group, middle HBYYRJ group, high HBYYRJ group, Fufangbiejiaruangan tablet (FFBJRG) group, colchicine group, 3 months infection group and 6 months infection group. Hepatic fibrosis was found in 3 months infection group. Liver hydroxyproline (Hyp) was determined, matrix metalloproteinase-2 and 9 (MMP-2, MMP-9) were detected with gelatin zymography, serum hyaluronic acid (HA) and precollagen III (PC-III) were detected using RIA.

RESULTSHBYYRJ obviously reduced hepatic fibrosis (probability value less than 0.01). Collagen and HA in 3 months infection group and 6 months infection group were higher than that in normal group (probability value less than 0.01), collagen in high and middle HBYYRJ groups and HA in middle and low HBYYRJ groups were lower than that in 6 months infection group (P less than 0.01, probability value less than 0.05). The expression of MMP-9 and MMP-2 in 3 months infection group and 6 months infection group was higher than that in normal group (probability value less than 0.01), The expression of MMP-9 in three HBYYRJ groups and the expression of MMP-2 in high HBYYRJ group were lower than that in 6 months infection group (probability value less than 0.05).

CONCLUSIONHBYYRJ can reduce liver fibrosis caused by schistosomiasis.

Animals ; Collagen Type III ; blood ; Disease Models, Animal ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; therapeutic use ; Female ; Hyaluronic Acid ; blood ; Hydroxyproline ; metabolism ; Liver ; drug effects ; metabolism ; pathology ; Liver Cirrhosis, Experimental ; drug therapy ; etiology ; metabolism ; pathology ; Male ; Materia Medica ; isolation & purification ; pharmacology ; therapeutic use ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Schistosoma japonicum ; Schistosomiasis japonica ; complications ; Severity of Illness Index ; Sex Factors ; Treatment Outcome

OBJECTIVETo investigate the effects of anluohuaqianwan on experimental hepatic fibrosis induced by dimethyl nitrosamine (DMN) in rats.

METHODS36 male SD rats were randomly dividied into three groups: model group, normal group, anluohuaqianwan group. The rats in the three groups were treated with DMN daily for 4 weeks. The liver function was detected using auto biochemistry analyzer, the serum HA, LN, IV-C, PIIIP were detected by immunoradiometry, the histopathology was observed in the left liver lobe after HE staining, the expression of matrix metalloproteinase-2 (MMP-2) in liver tissue were detected by immunohistochemistry.

RESULTSThe serum levels of ALT, AST, ALP, TP, ALB and the contents of HA, LN, IV-C in model group were significantly increased compared to these in the normal group (P less than 0.01). The serum levels of ALT, AST and the contents of HA in anluohuaqianwan group were significantly lower than those in the model group (P less than 0.01). The liver MMP-2 in the model group was significantly increased compared to that in the normal group (P less than 0.05). The expression of MMP-2 in liver tissue of model group was lower than that in the anluohuaqianwan group (P less than 0.05).

CONCLUSIONAnluohuaqianwan can inhibit liver fibrosis in rats induced by DMN.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Dimethylnitrosamine ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hyaluronic Acid ; blood ; Hydroxyproline ; metabolism ; Immunohistochemistry ; Liver ; drug effects ; metabolism ; pathology ; Liver Cirrhosis, Experimental ; chemically induced ; drug therapy ; metabolism ; pathology ; Liver Function Tests ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley