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

Objective To evaluate the accuracy and feasibility of time-resolved immunofluorometric assay (TRI FA) for detection of HBsAg based on Abbott automated chemiluminescence immunoassay(CMIA),so as to carry out this project in primary hospitals,and provide reference for individual antiviral strategy and prediction of therapeutic effect.Methods Serum of 157 patients infected with hepatitis B virus were detected with CMIA and TRIFA,specimens with HBsAg titers exceeding the detection limit were firstly diluted,then performed quantitative analysis.HBsAg levels were divided into 4 groups:≤100 IU/mL,101-1 000 IU/mL,1 001-20 000 IU/mL,and ＞ 20 000 IU/mL,quantitative correlation between two methods was analyzed.Results The linear regression equation of two methods was Y=2.323X-896.3,correlation coefficent r=0.943,P＜0.001.CMIA was as a reference,4 groups were divided for analysis,results showed that when detected specimens was at low concentration of HBsAg,TRIFA value was low compared with CMIA method,while detected specimens was at high concentration of HB sAg,CMIA value was high,two reagents had good consistency in the detection of different concentrations of HBsAg(both P＜0.05),when concentration was at 1 001-20 000 IU/mL,consistency was the best.Conclusion The accuracy of two reagents in the quantitative detection of HBsAg is similar,and the best correlation of detection value is 1 000-20 000 IU/mL.TRIFA assay has wide application for its low-cost and easy to be operated,which is especially suitable for primary hospitals.

Objective: To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 μg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control. Methods: This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (CI) were calculated, and non-inferiority was demonstrated if the lower limit of 95% CI was > -10%. The t-test, chi-square test, or rank sum test was used according to the types and features of data. Results: A total of 855 HBeAg-positive CHB patients were enrolled and 820 of them received treatment (538 in the trial group and 282 in the control group). The data of the full analysis set showed that HBeAg seroconversion rate at week 72 was 27.32% in the trial group and 22.70% in the control group with a rate difference of 4.63% (95% CI -1.54% to 10.80%, P = 0.1493). The data of the per-protocol set showed that HBeAg seroconversion rate at week 72 was 30.75% in the trial group and 27.14% in the control group with a rate difference of 3.61% (95% CI -3.87% to 11.09%, P = 0.3436). 95% CI met the non-inferiority criteria, and the trial group was non-inferior to the control group. The two groups had similar incidence rates of adverse events, serious adverse events, and common adverse events. Conclusion In Peg-IFN-α regimen for HBeAg-positive CHB patients, the new drug Peg-IFN-α-2b (Y shape, 40 kD) has comparable effect and safety to the control drug Peg-IFN-α-2a.

Numerical simulations of volume conduction can be used to analyze the process of energy transfer and explore the effects of some physical factors on energy transfer efficiency. We analyzed the 3D quasi-static electric field by the finite element method, and developed A 3D coupled field-circuit model of volume conduction basing on the coupling between the circuit and the electric field. The model includes a circuit simulation of the volume conduction to provide direct theoretical guidance for energy transfer optimization design. A field-circuit coupling model with circular cylinder electrodes was established on the platform of the software FEM3.5. Based on this, the effects of electrode cross section area, electrode distance and circuit parameters on the performance of volume conduction system were obtained, which provided a basis for optimized design of energy transfer efficiency.
Algorithms ; Electric Conductivity ; Electrodes ; Electromagnetic Fields ; Equipment Design ; Finite Element Analysis ; Models, Theoretical ; Prostheses and Implants ; Skin Physiological Phenomena

BACKGROUND: Studies have demonstrated that intermittent high glucose can have a more severe impact on vascular endothelial function in comparison with persistent hyperglycemia.OBJECTIVE: To investigate the effect of intermittent high glucose on the proliferation and apoptosis of endothelial progenitor cells (EPCs) from human peripheral blood in vitro as well as the production of malondialdehyde (MDA) and antioxidant. METHODS: Total mononuclear cells were isolated from human peripheral blood by Ficoll density gradient centrifugation and then the cells were placed on fibronectin-coated culture dishes. After 7 days of culture, the adherent cells were identified as EPCs by laser scanning confocal microscope. The cells were synchronized and then stimulated with glucose 5.5 mmol/L (normal control group), 20 mmol/L (constant high glucose group), and 5.5/20 mmol/L (intermittent high glucose group, 5.5 and 20 mmol/L glucose culture solution was changed every 8 hours) for 72 hours. EPCs proliferation and apoptosis was measured by MTT assay and flow cytometry, respectively. The content of MDA and the activity of superoxide dismutase (SOD) in culture solution were detected with colorimetry.RESULTS AND CONCLUSION: After EPCs were exposed to constant high glucose (20 mmol/L) and intermittent high glucose (5.5/20 mmol/L) for 72 hours, proliferated cells were significantly reduced and the apoptosis rate was significantly increased compared with those exposed to normal glucose (P < 0.01). Furthermore, there was a significant increase in MDA contents as well as a significant reduce in SOD activities in the constant high glucose and intermittent high glucose group (P < 0.01), especially in the latter group. These findings indicated that both intermittent high glucose and constant glucose could inhibit the proliferation and promote the apoptosis of EPCs; however, intermittent high glucose appears to worsen the effects on EPCs. This is maybe due to the increased oxidative stress.

OBJECTIVE: To investigate the impact of coagulative parameters on different anticoagulation systems in molecular adsorbent recirculating system (MARS) in subjects with liver failure, and to evaluate the safety of different anticoagulation methods . METHODS: A prospective experimental observation was designed. According to anticoagulation Methods , 174 MARS treatment sessions for 146 patients with liver failure and prothrombin time activity percentage (PTA) ≤ 40% were randomly divided into 2 groups: 92 MARS treatment sessions in the heparin-free group and 82 in the low-dose heparin group. Time points of 0, 0.5, 1, 2, 3, 4, 5 and 6 h were selected to observe the coagulation changes of prothrombin time (PT), PTA, thrombin time (TT), activated partial thromboplastin time (APTT) and international normalized ratio (INR) dynamically. Adverse events such as line / filter coagulation, rupture and bleeding were also investigated and compared due to frequency and severity between the 2 groups. RESULTS: There was no difference in PT, PTA, INR between the 2 groups, but significant differences were observed in APTT and TT and fibrinogen (Fbg). APTT and TT levels in the low-dose heparin group was increased rapidly after the first given dose of anticoagulant heparin and reached the peak within 30 min.The levels at each time point was statistically different between the 2 groups (P<0.05). A significant difference in the Fbg level was obtained between the 2 groups. In the low-dose heparin group it was stabilized and increased slightly at the end of the treatment. While in the heparin-free group it was decreased gradually and reached a ravine at the end of the treatment. A curve was observed after 2.5 h treatment between the 2 groups (P=0.001). There were 2 cases of severe bleeding after MARS was finished in the heparin group, and 1 was terminated because of degree III clotting in the heparin-free group. CONCLUSION Fibrinogen should be adsorbed while the blood touches the MARS circuit path and anticoagulants can prevent it. Comprehensive analysis of blood platelet count (BPC), fibrin degradation products (FDP), D-dimer and clinical symptoms is critical and required to determine the coagulation status to select an anticoagulation system before MARS. The use of low dose heparin in MARS improves the disorder of hypercoagulable state during the high coaguation period, while heparin-free during low coagulation period can effectively prevent the occurrence of bleeding and improve the mechanism of blood coagulation by reducing heparin-like substance in the blood.
Adolescent ; Adsorption ; Adult ; Anticoagulants ; administration & dosage ; Disseminated Intravascular Coagulation ; prevention & control ; Female ; Heparin ; administration & dosage ; Humans ; Liver Failure ; therapy ; Liver, Artificial ; Male ; Middle Aged ; Prothrombin Time ; Sorption Detoxification ; methods ; Young Adult

Objective To investigate clinical features and antifungal therapeutic effect of chronic severe hepatitis (CSH) patients with invasive fungal infection (IFI), and to improve the diagnosis and treatment.Methods Clinical manifestation, blood routine, imageology and mycetology characteristic, antifungal treatment perscription and therapeutic effect of 79 CSH patients with IFI were retrospectively analyzed. Antifungal therapeutic effect was compared between fluconazole and voriconazole. Results Thirteen (16.5%) patients received glucocorticoid or other immunodepressants for a relatively long time, 40 (50.6%) patients had invasive operation, and 61 (77.2 %) patients were administered 1-6 kinds of broad-spectrum antibiotics. Seventy-three patients had fever. Leucocytes and neutrophilic granulocyte increased in 96.2% of the patients. Lung (31.6%), intestinal tract (26.2%) and oral cavity (14%) infections were common. Fungus was found in 70.9% of the patients. Candida albicans (40.9%) and aspergillus (21.1%) were often seen. Halo signs and crescent signs on lung CT were relatively specific in 40% of the patients with fungal pneumonia. Voriconazole was more effective than fluconazole(71.4% vs. 39.0%, P<0.05). Twelve patients with lung aspergillus infection were administered voriconazole, 8 (66.7%) patients of whom was effective, and the other 4 (33.3%) patients died. Conclusion There are high risk factors in major CSH patients with IFI. The most common clinical manifestations of CSH patients with IFI are fever, leukocytosis, lung and intestinal tract infection. Candida albicans and aspergillus infection are common. Voriconazole is more effective than fluconazole, and can increase the survival rate of CSH patients with IFI.

Objective To investigate the potential complications of renal transplant patients, to develop appropriate treatment programs to reduce the incidence. Methods Clinical data of 42 patients with renal transplantation were retrospectively analyzed. Blood type of the donors and recipients were same in 32 cases; Blood type of the donors was 0 and recipients blood type was A,3 cases;Donors blood type was 0 and recipients blood type was B,2 cases; Donors blood type was O and recipients blood type was AB,4 cases; Donors blood type was B and recipients blood type was AB,lease. Results In alll cases, there were acute rejection in 2 cases,pulmonary infection in 2 cases, wound infection in 1 case,urinary leakage in 1 case,urinary tract infection in 2 cases.42 cases were all cured up. One patient lost kidney function and required dialysis therapy 4 years after kidney transplantation, the remaining 40 patients were good. Conclusion Early detection and timely management of complications of renal transplantation played a very important role in improving the survival rate.

BACKGROUND: The key of tissue engineered blood vessel research depends on the appropriate scaffolds. The porcine vascular tubes have frequently been used as candidates for tissue engineering vessel construction, but high immunogenicity and poor mechanical strength limit its application for tissue engineering scaffolds.OBJECTIVE: To prepare a novel tissue engineered vascular scaffold with good mechanical properties and biocompatibility using acellular porcine aorta matrix.METHODS: Porcine aorta was decellularized and modified by thermal cross-linking to improve the mechanical strength and biodegradation properties and to prepare acellular porcine aorta matrix scaffolds. Hematoxylin-eosin staining of histological sections and biomechanical tests were performed to assess the decellularization effects and the mechanical strength of the vascular matrix respectively. Vascular endothelial cells from human umbilical cord veins were isolated and seeded on the acellular matrix scaffolds and cultured in vitro. The biocompatibility was evaluated. RESULTS AND CONCLUSION: After the porcine aorta was treated by 1% Triton X-100 solution for 84 hours, the vessel was fully decellularized, and the architecture of matrix was well preserved. The acellular vascular matrix demonstrated improved biomechanical properties after modification by thermal cross-linking under vacuum at 120 °C for 12 hours. Its tensile strength reached 1.70 MPa. After 7 days of in vitro culture, the seeded endothelial cells formed a typical vascular endothelial layer structure on the surface of acellular matrix, as observed by scanning electron microscopy. Results proved that acellular vascular matrix of porcine aorta could maintain the morphology and structure of natural vessels, its mechanical strength could be greatly improved after successive freeze-drying and thermal cross-linking. A good compatibility between the acellular matrix and endothelial cells of umbilical vein is also achieved.

A total of 189 stool samples from swine with diarrhea, collected in various porcine farms in the central region of China were tested for porcine enteric caliciviruses (PEC) member porcine sapoviruses (SaV) by reverse transcription polymerase chain reaction (RT-PCR) amplification using primers designed to detect porcine SaV. Selected amplicons were sequenced to establish phylogenetic relationships with reference strains. Porcine SaV were detected in 12.70% (24/189) of the samples. Phylogenetic studies based on partial RNA polymerase gene sequences indicated that the field strains of viruses isolated in China were closely related (75.6 88.3% identity) to the porcine SaV Cowden reference strain. These results provide evidence that caliciviruses of the genus sapovirus circulate in piglets in China, but further studies are needed to clarify their importance as cause of diarrhea. This is the first report of PEC in China.