OBJECTIVE To establish a method for determining the content of 11 antioxidants and their degradation products in recombinant Exendin-4-FC fusion protein injection by HPLC. METHODS The protein was precipitated with saturated ammonium sulfate. After centrifugation, the supernatant was transferred to a C18 solid phase extraction cartridge activated by methanol. Then the cartridge was eluted with 4 mL of methanol and 5 mL of ethyl acetate respectively, and the eluent was diluted with methanol-ethyl acetate(2∶3) mixed solvent and passed through a 0.22 µm PTFE hydrophobic filter. It was analyzed by HPLC and quantified by external standard method. Chromatographic conditions: Kinetex® XB-C18 100Å (100 mm×4.6 mm, 2.6 µm)column, the detection wavelength was 230 nm, the column oven was 30 ℃, the injection volume was 5 µL and the flow rate was 0.4 mL·min–1, mobile phase was 0.1% formic acid-methanol(A)-0.1% formic acid aqueous solution(B), the running time was 45 min. RESULTS The 11 target substances showed a good linear relationship in the range of 2.5−35 μg·mL–1 with R2 ≥0.99. At three different concentration(25, 10, 5 μg·mL–1) of spiked samples, the average recovery rates of 11 antioxidants ranged from 88.1% to 106.5%, with RSDs in the range of 0.10%–9.05%. The RSDs of 6 repeatable samples was 2.01%–4.77%, which of 12 intermediate precision samples was 2.58%–9.75%. The positive/inverted samples of three batches of recombinant Exendin-4-FC fusion protein injection were detected at 0 month, 3 months and 6 months(25 ℃), and the results showed that there was no antioxidant and its degradation leaching in all batches of samples at different detection points. CONCLUSION The method has good specificity, high accuracy and precision, good solution stability, high durability and can be used for the content detection of antioxidants in drugs.

Objective: To assess the safety and immunogenicity of freeze-dried rabies vaccine (Vero-cells) for human use on different immunization procedures in healthy people aged 9-65 years. Methods: A randomized, blind, positive-controlled clinical study was conducted in March 2015. The eligible residents aged 9-65 were recruited in Dengfeng city and Biyang County, Henan Province. A total of 1 956 subjects were enrolled. The subjects were randomly (1∶1∶1) assigned to 5-dose control group, 4-dose trial group and 5-dose trial group, with 652 subjects in each group. The subjects of 5-dose control group were immunized with control vaccine on days 0, 3, 7, 14 and 28. The subjects of 4-dose trial group were immunized with trial vaccine on days 0, 7 and 21 (2-1-1 phases) and the subjects of 5-dose trial group were immunized with trial vaccine on days 0, 3, 7, 14 and 28. A combination of regular follow-up and active reporting was used to observe local and systemic adverse reactions till 30 days after the first and full immunization, and the incidence rate of adverse reactions in three groups was analyzed and compared. The venous blood was collected before the first immunization, 7 days after the first immunization, 14 days after the first immunization and 14 days after the full immunization. The neutralizing antibody of rabies virus was detected by rapid fluorescent focus inhibition test (RFFIT), and the seropositive conversion rate and geometric mean concentration (GMC) of antibody were calculated. Results: The adverse reaction rates in 5-dose control group, 4-dose trial group and 5-dose trial group were 41.87% (273/652), 35.43% (231/652) and 34.97% (228/652), respectively. The adverse reaction rates of 4-dose trial group and 5-dose trial group were lower than those of the 5-dose control group (P<0.05). The local reactions were mainly pain, itching, swelling and redness in injection site, while the systemic reactions were mainly fever, fatigue, headache and muscle pain. The severity of adverse reactions was mainly mild (level 1), accounting for 85.33% (518/607), 89.02% (373/419) and 88.96% (427/480) of the total number of adverse reactions in each group. At 14 days after the first immunization and 14 days after the full immunization, the antibody positive conversion rates of three groups were all 100%. At 7 days, 14 days after the first immunization and 14 days after the full immunization, the GMCs of three groups were 0.60, 0.72, 0.59 IU/ml, 20.42, 23.99, 24.38 IU/ml and 22.95, 23.52, 24.72 IU/ml, respectively, with no significant difference (P>0.05). Conclusion: The freeze-dried rabies vaccine (Vero-cells) for human use has good safety and immunogenicity when inoculated according to 5-dose and 4-dose immunization procedures.
Humans ; Rabies Vaccines ; Antibodies, Viral ; Antibodies, Neutralizing ; Rabies virus ; Vaccination ; Rabies/prevention & control*

【Objective】 To study the removal effect of fibronectin(Fn) from von willebrand factor(vWF) by ion-exchange chromatography through processing human coagulation factor Ⅷ chromatographic washing products, in order to select a method that can effectively reduce Fn without compromising the activity yield. 【Methods】 In a multi-batch process development experiment, Fractogel^® EMD TMAE(M) strong anion filler produced by Merck(Germany) was used to conduct chromatography to investigate vWF ristomycins titer (vWF: RCof), vWF recovery, protein content and Fn content. 【Results】 During the development of vWF pilot purification process, the content of Fn in the samples can be effectively reduced by ion-exchange chromatography, with removal rate more than 87%, titer recovery of vWF more than 80%, and no significant change in other quality indexes. 【Conclusion】 The use of ion-exchange chromatography to purify vWF can effectively reduce the content of Fn, which has positive significance for developing new product process and improving the product quality of blood products manufacturers.

Tripartite motif (TRIM) family proteins are important effectors of innate immunity against viral infections. Here we identified TRIM35 as a regulator of TRAF3 activation. Deficiency in or inhibition of TRIM35 suppressed the production of type I interferon (IFN) in response to viral infection. Trim35-deficient mice were more susceptible to influenza A virus (IAV) infection than were wild-type mice. TRIM35 promoted the RIG-I-mediated signaling by catalyzing Lys63-linked polyubiquitination of TRAF3 and the subsequent formation of a signaling complex with VISA and TBK1. IAV PB2 polymerase countered the innate antiviral immune response by impeding the Lys63-linked polyubiquitination and activation of TRAF3. TRIM35 mediated Lys48-linked polyubiquitination and proteasomal degradation of IAV PB2, thereby antagonizing its suppression of TRAF3 activation. Our in vitro and in vivo findings thus reveal novel roles of TRIM35, through catalyzing Lys63- or Lys48-linked polyubiquitination, in RIG-I antiviral immunity and mechanism of defense against IAV infection.
A549 Cells ; Animals ; Apoptosis Regulatory Proteins/immunology* ; DEAD Box Protein 58/immunology* ; Dogs ; HEK293 Cells ; Humans ; Influenza A Virus, H1N1 Subtype/immunology* ; Madin Darby Canine Kidney Cells ; Mice ; Mice, Knockout ; Orthomyxoviridae Infections/pathology* ; Proteolysis ; RAW 264.7 Cells ; Signal Transduction/immunology* ; THP-1 Cells ; TNF Receptor-Associated Factor 3/immunology* ; Ubiquitination/immunology* ; Viral Proteins/immunology*

Objective To explore the effects of nursing intervention based on Information-motivation-behavioral model (IMB) on self-efficacy and quality of life of patients with primary liver cancer. Methods A total of 84 patients with primary liver cancer who were hospitalized in the First Affiliated Hospital of Ji'nan University from January to December in 2016 were selected by convenience sampling method and divided into control group (40 cases) and observation group (44 cases) according to the time of admission. The patients in the control group received routine care, while the patients in the observation group received nursing intervention based on IMB. Chinese Strategies Used by People to Promote Health (C-SUPPH) and Quality of Life-Liver Cancer (QOL-LC) were applied to investigate patients' self-efficacy and quality of life before and after intervention, so as to evaluate the effects of intervention. Results The C-SUPPH and QOL-LC scores of the observation group and the control group were (87.18±14.83), (105.01±18.56) and (78.65±15.75),(94.75±18.15) respectively on the day of discharge, and the differencs were statistically significant (t=-2.557,-2.068; P< 0.05). The C-SUPPH and QOL-LC scores of the observation group and the control group were (92.34±16.04), (123.11±20.86) and (80.50±15.86), (101.90±19.95) respectively 1 month after discharge, and the differences were statistically significant (t=-3.397,-4.753; P< 0.05). Conclusions Nursing intervention based on IMB can improve the self-efficacy and quality of life of patients with primary liver cancer, which is worthy of clinical promotion.

Objective:To establish a method for the determination of asperosaponinⅥin Dieda Cuyu tablets by HPLC. Methods:A Hypersil C18(250 mm ×4.6 mm,5 μm)column was used. The mobile phase was acetonitrile-water(30∶70) with a flow rate of 1.0 ml·min-1 . The detection wavelength was 212 nm, the column temperature was room temperature,and the injection volume was 10μl. Results:AsperosaponinⅥ showed a good linear relationship within the range of 0. 04-0. 32 μg(r=0. 999 6). The average recovery was 97. 84%(RSD=1. 70%, n=6). Conclusion:The method is simple,accurate and reproducible, which can be used in the deter-mination of asperosaponinⅥ in Dieda Cuyu tablets.

Objective To review the basic clinical characteristics and the pathogens and their antimicrobial susceptibilities to neonatal sepsis in very low birth weight infants (VLBWI) and extremely low birth weight infants ( ELBWI) for selection of appropriate antibiotics. Methods A retrospective chart review of 56 cases with neonatal sepsis(early onset neonatal sepsis 3 cases, late onset 53 cases) in VLBWI and ELBWI admitted to the neonatal intensive care unit of Yuying Children's Hospital of Wenzhou Medical College from January 1, 1999 to December 31, 2008 was conducted. The basic clinical characteristics and the results of blood culture and antimicrobial susceptibilities were analyzed. Results Among the 56 cases, the clinical presentations were non-specific. A total of 43 strains of bacteria were isolated, and the most important pathogens responsible for neonatal sepsis in VLBWI and ELBWI were opportunistic pathogenic bacteria. In early onset neonatal sepsis, there was only one culture-proven sepsis that was Chryseobacterium meningosepticum. In the late onset neonatal sepsis cases, the main pathogens of Gram-negative organisms were Klebsiella pneumoniae (33. 3%, 14/42), and the most common Gram-positive organisms were coagulase-negative Staphylococci (26. 2%, 11/42), followed by Enterococcus species (11. 9%,5/42). Furthermore, there were 2 fungal sepsis(4. 8%, 2/42), which were infected by Candida albicans. All of the coagulase-negative Staphylococci were methicillin-resistant coagulase-negative Staphylococci, and they were resistant to common antibiotics and sensitive to vancotnycin and rifampicin. And all of the Klebsiella pneumoniae produced extended-spectrum (Hactamases, which were sensitive only to a few antibiotics such as carbopenems, aminoglycosides and quinolones. Among those 56 cases, 43 patients were cured, 13 died, including six patients who refused any treatments, the mortality rate of neonatal sepsis in VLBWI and ELBWI was 23. 2%. Conclusions The clinical presentations of neonatal sepsis in VLBWI and ELBWI were non-specific, and the most important pathogens were opportunistic pathogenic bacteria, which were multi-drug resistant. Routine blood culture should be taken from infants who are suspected of neonatal sepsis and empirical use of appropriate antibiotics should be initiated as soon as the blood specimen for culture has been drawn. To reduce the occurrence of multi-drug resistant bacteria, we should restrict the use of antibiotics especially the third generation of cephalosporins in neonates as much as possible.

According to 45 hemagglutinin (HA) gene sequences of H7 subtype of avian influenza virus (AIV), a pair of specific oligonucleotide primers was designed. We developed one step RT-PCR for detecting AIV subtype H7. Sensitivity to detection of allantoic fluid by one step RT-PCR reached 10(5.5) EID50/mL and detection of swab samples reached 10(3) EID50/mL. We simultaneity detected the tissue and swab samples infected with H7 subtypes AIV by one step RT-PCR and virus isolation method. The results showed that the sensitivity of the assay gave an excellent correlation with the conventional virus isolation method. H1-H15 subtypes of avian influenza and other avian diseases were detected by the one step RT-PCR. The results showed the assays were high specific, without cross-reaction with other subtypes or other avian diseases. Development of one step RT-PCR will provide effective technical support for the rapid diagnosis and surveillance of molecular epidemiology of AIV subtype H7.
Animals ; Birds ; Chick Embryo ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Influenza A Virus, H7N1 Subtype ; classification ; genetics ; isolation & purification ; Influenza in Birds ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective To estimate time of gauze swabs left in abdomen with comparison of the number of foreign body giant cells,theirs nuclei and the proportion of type Ⅰ collagenous fibers to type Ⅲ and argentaffin fibers in different times.Methods F344 rat models dependent 2,8,30,120 days were established by gauze swab fixed in the abdomen,and were studied on gauze swab wrapped by greater omentum,the number of foreign body giant cells and theirs nuclei by HE staining,and the proportion of type Ⅰ collagenous fibers to type Ⅲ and argentaffin fibers by sirius red and silver staining respectively.The results were analyzed by image analysis system.Results The results showed that number of foreign body giant cells,theirs nuclei and the proportion of type Ⅰ collagenous fibers to type Ⅲ and argentaffin fibers increased gradually(P<0.01)followed the time delayed.The Proportion of type Ⅰ collagenous fibers to argentaffin fibersis Power Function of Days of guaze swab left in rat abdomen(r=0.972).Conclusion The number of foreign body giant cells,theirs nuclei and the proportion of type Ⅰ collagenous fibers to type Ⅲ and argentaffin fibers contribute to the estimation of foreign body(eg.gauze swab)left in abdomen.

BACKGROUND:Operation is the main therapy for protrusion of lumbar intervertebral disc.The researches have indicated that non-operation therapy can improve effectively its clinical symptoms.OBJECTIVE:To observe the effect of "three points and five methods"therapy on analgesia of radiculalgia in protrusion of lumbar intervertebral disc and on recovery of motor function in lumbar region and lower limbs.DESIGN:Non-randomized concurrent control study based on the patients.SETTING:Orthopedic department and college of acupuncture and massage of one university-affiliated Hospital.PARTICIPANTS:Totally 210 inpatients of lumbar vertebral disorders were selected from Orthopedic Department of First Affiliated Hospital of Henan College of Traditional Chinese Medicine and Department of Acupuncture and Massage of Third Affiliated Hospital from March 1999 to June 2003,which were randomized into experimental group (n=108) and control group(n=102).METHODS:"Three points and five methods" therapy was adopted in experimental group and the traditional manipulation was for the control.The evaluation on pain and motor function was carried on at minutes 5,10,30,and hours 1,2,...,24 successively for statistical analysis.RESULTS:Total effective rate was 94.44% and the average time of analgesia was(18.40±2.11) hours in the experimental group.Total effective rate was 82.35% and the average time of analgesia was (10.60±2.63) hours in the control group. The analgesia effect and evaluation results on comprehensive function in the experimental group were superior to that in the control group(t=4.86,4.42,P ＜ 0.01).CONCLUSION: "Three points and five methods" therapy releases lumbar pain, improves lumbar symptoms and recovers motor function of lower limbs for the patients with radiculalgia in protrusion of lumbar intervertebral disc.