ObjectiveTo explore whether the biocompatibility of phosphorylcholine (PC) modified alginate-chitosan microcapsules could be improved. MethodsPC modified alginate-chitosan microcapsules were obtained by high-voltage electrostatic system.Bradford method was adopted to determine the adsorption amounts of bovine serum albumin by chitosan alone and PC modified chitosan.Alginate-chitosan-PC microcapsules (experimental group) and alginate-chitosan microcapsules ( control group) were respectively implanted into the peritoneal cavity of mice and retrieved 4 weeks after transplantation.Fibrosis of the capsules was evaluated by HE staining.Glucose stimulated insulin secretion (GSIS) assay was used to assess the insulin secretion response of encapsulated and nonencapsulated rat islets. Results The adsorption amount of protein was 189.4 μg/mg and 90.5 μg/mg respectively by chitosan alone and PC modified chitosan.The difference had statistical significance ( t =5.549, P ＜ 0.05 ).In contrast to the control group, the cellular reaction on the surface of the modified microcapsules was weaker, with no obvious fibrosis found.The insulin secreted by encapsulated islets and nonencapsulated islets was( 3.298 ± 1.680 ) μIU/ml VS (4.299 ± 1.159 ) μIU/ml ( t =1.096, P ＞ 0.05 ) in response to low-glucose stimulus and( 11.783 ± 4.175 ) μIU/ml VS ( 12.875 ± 2.268 ) μIU/ml ( t =0.514, P ＞ 0.05 ) in response to high-glucose stimulus.Conclusions PC can improve the biocompatibility of alginate-chitosan microcapsules, with no effect on the biological function of encapsulated islets.It may be more appropriate to use modified microcapsules encapsulating islets for transplantation.

Objective: To explore the mechanism of An-pressing manipulation in improving post-stroke muscle spasticity, by observing the changes of γ-aminobutyric acid (GABA) and glycine (Gly) in plasma and gray matter of L1-L3 spinal cord anterior horn in post-stroke rats with muscle spasticity after An-pressing manipulation intervention. Methods: Ten of 80 adult male Sprague-Dawley (SD) rats were randomly selected as the blank group, and the remaining 70 were used for modeling. The middle cerebral artery occlusion (MCAO) rat model was established by insertion suture occlusion method in the left external carotid artery. Thirty rats with a Longa neurological score of 2-3 points and a modified Ashworth spasticity scale score of 1-, 1+, or 2 were included in the experiment. Using the random number table method, the 30 successfully modeled rats were randomly divided into a model group, an An-pressing tendon group and an An-pressing muscle belly group. Two days after modeling, rats in the An-pressing tendon group and An-pressing muscle belly group received An-pressing manipulation on the tendon and belly of quadriceps femoris muscle respectively, with the pressure of (350±50) g and the frequency of 5 s/time, 15 min per session, once a day for 5 continuous days. After the 5th treatment, the tension of the rat quadriceps femoris muscle was evaluated using the modified Ashworth spasticity scale. The Gly levels in rat plasma and L1-L3 segments of spinal cord were determined by enzyme-linked immunosorbent assay (ELISA). The GABA levels in rat plasma and L1-L3 segments of spinal cord were measured by high performance liquid chromatography (HPLC). Results: The decrease in rat muscle tension scored by the modified Ashworth spasticity scale in the An-pressing tendon group was more significant than that in the An-pressing muscle belly group (P<0.01); the increases in Gly and GABA levels in the rat plasma and L1-L3 segments of spinal cord were more significant in the An-pressing tendon group than those in the An-pressing muscle belly group (all P<0.01). Conclusion: Based on the theory of 'anti-stretch reflex' of tendon organs, the use of An-pressing manipulation to induce the 'anti-stretch reflex' by stimulating the tendon organs can improve the muscle spasticity of rats, which is better than An-pressing the muscle belly. Increased levels of Gly and GABA in rat plasma and L1-L3 segments of spinalcord may be one mechanism of An-pressing manipulation to improve muscle spasticity by stimulating tendon organs.

OBJECTIVETo evaluate the immunohistochemical detection of epidermal growth factor receptor(EGFR) mutations using two EGFR mutation-specific monoclonal antibodies: delE746-A750 and L858R.

METHODSA total of 175 paraffin-embedded lung adenocarcinoma tissue samples previously genotyped by directive DNA sequencing were subject to immunostaining using delE746-A750 and L858R antibodies.

RESULTSThere was no significant difference of mutation detection between DNA sequence analysis and delE746-A750 and/or L858R immunostaining (33.7% vs 30.9%, P > 0.05). The overall sensitivity, specificity, positive predictive value and negative predictive value of immunostaining using these two EGFR mutation-specific antibodies were 83.1%, 95.7%, 90.7% and 90.9%, respectively.

CONCLUSIONWith high sensitivity and good specificity, immunohistochemistry using EGFR mutation-specific monoclonal antibodies is an adequate, easy and cost-effective prescreening method to detect EGFR mutations using paraffin-embedded tissue specimens of lung adenocarcinomas.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Monoclonal ; DNA Mutational Analysis ; Female ; Gene Deletion ; Humans ; Immunohistochemistry ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Paraffin Embedding ; Point Mutation ; Receptor, Epidermal Growth Factor ; genetics ; immunology ; metabolism ; Sensitivity and Specificity

OBJECTIVETo investigate the osteogenesis mechanism by analysis of the expression of insulin-like growth factor-I (IGF-1) and alkaline phosphatase (ALP) in the reconstruction of cleft palate (CP) with distraction osteogenesis (DO) in rhesus.

METHODSThe CP animal models were established surgically. 21 rhesus in experimental group underwent DO to close the soft and bony defect, followed by consolidations. Every 3 animals were killed and the specimen were taken out after consolidation of 1, 2, 4, 6, 8, 12, 24 weeks. The mRNA of IGF-1 and ALP were detected with Real-time RT-PCR technique. The expression of IGF-1 and ALP was quantitatively analyzed by ELISA. The results were compared with those in control and sham groups (each of 2 animals), respectively.

RESULTSSince consolidation, the mRNA of IGF-1 and ALP increased significantly at one week and reached the peak at two weeks, but decrease to control level after 12 weeks of consolidation. The expression of IGF-1 also increased to peak level after two weeks of consolidation. The expression of ALT increased significantly since consolidation and reach the peak value after six weeks. They all decreased to nearly control level after 8-12 weeks.

CONCLUSIONSThe palate cleft can be successfully closed with new formed bone after DO. The mechanism of bone consolidation is intramembranous bone formation.

Alkaline Phosphatase ; metabolism ; Animals ; Bone Regeneration ; Cleft Palate ; metabolism ; surgery ; Insulin-Like Growth Factor I ; metabolism ; Macaca ; Osteogenesis, Distraction ; Postoperative Period

OBJECTIVETo construct an eukaryotic expression vector of open reading frame of unknown KH gene (KH-ORF), and investigate its effect on cell proliferation.

METHODSThe pCI-neo-KH-ORF expression vector was constructed by DNA recombinant technique and was introduced into COS-7 cells and K562 cells by lipofectactin-mediated DNA transfection. Expression of KH-ORF mRNA was detected by RT-PCR. The effect of KH-ORF on cell cycle of COS-7 cells and K562 cells was evaluated by flow cytometry (FCM). Effect on cell proliferation of COS-7 cells was tested by MTT assay and that on K562 cells was analyzed by growth curves and LDH activity measurement.

RESULTS(1) KH-ORF mRNA was expressed both in COS-7 cells and K562 cells. (2) The cell cycle and cell proliferation of COS-7 cells were unaffected significantly. (3) The proportion of cells in S phase was increased in pCI-neo-KH-ORF-transfected K562 cells; and growth curves and LDH activity indicated enhanced cell proliferation.

CONCLUSIONKH gene may be a leukemia gene related to proliferation of K562 cells.

Animals ; COS Cells ; Cell Proliferation ; Cercopithecus aethiops ; Gene Expression Regulation, Neoplastic ; Genes, Neoplasm ; genetics ; physiology ; Genetic Vectors ; Humans ; K562 Cells ; L-Lactate Dehydrogenase ; metabolism ; Open Reading Frames ; genetics ; physiology ; Plasmids ; RNA, Messenger ; biosynthesis ; genetics ; RNA-Binding Proteins ; genetics ; S Phase ; Transfection

With the wide application of electronic data management (EDC), the data management is shifting to a new mode. In order to recognize the advantages of EDC, we choose 20 representative registered clinical trials, which involve 5 404 subjects and 321 sites. We found that EDC has many beneficial impacts on the course of clinical trial data management, including the process of data collection, data cleaning, data quality control and clinical trial decision-making. The result also provides a reference for the adoption of EDC in clinical trials.
Clinical Trials as Topic ; Data Collection ; standards ; Information Storage and Retrieval ; standards ; Quality Control

To observe the serum samples and the anti-inflammatory effect of Tripterygium wilfordii in treating RA by using the pharmacokinetic-pharmacodynamic model, make a correlation analysis on concentration-time and effect-time curves, and explore RORγt, IL-17, STAT3, IL-6 mRNA transcriptional levels in rats by PCR. Methotrexate, tripterine and high-dose T. wilfordii could down-regulate RORγt, IL-17, STAT3, IL-6 mRNA transcriptional levels in AA rat lymph nodes. The study on PK-PD model showed correlations between inflammatory factors and blood concentration of T. wilfordii. T. wilfordii and its main active constituent tripterine could show the inflammatory effect and treat RA by inhibiting IL-17 cytokine.
Animals ; Arthritis, Rheumatoid ; drug therapy ; immunology ; Biomarkers ; Female ; Interleukin-17 ; antagonists & inhibitors ; genetics ; Interleukin-6 ; genetics ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Tripterygium ; Triterpenes ; pharmacokinetics ; pharmacology

OBJECTIVETo develop a fast, high-throughput screening method with suspension array technique for simultaneous detection of biothreat bacteria.

METHODS16 S rDNA universal primers for Bacillus anthracis, Francisella tularensis, Yersinia pestis, Brucella spp.and Burkholderia pseudomallei were selected to amplify corresponding regions and the genus-specific or species-specific probes were designed. After amplification of chromosomal DNA by 16 S rDNA primers 341A and 519B, the PCR products were detected by suspension array technique. The sensitivity, specificity, reproducibility and detection power were also analyzed.

RESULTSAfter PCR amplification by 16 S rDNA primers and specific probe hybridization, the target microorganisms could be identified at genus level, cross reaction was recognized in the same genus. The detection sensitivity of the assay was 1.5 pg/microl (Burkholderia pseudomallei), 20 pg/microl (Brucella spp.), 7 pg/microl (Bacillus anthracis), 0.1 pg/microl (Francisella tularensis), and 1.1 pg/microl (Yersinia pestis), respectively. The coefficient of variation for 15 test of different probes was ranged from 5.18% to 17.88%, it showed good reproducibility. The assay could correctly identify Bacillus anthracis and Yersinia pestis strains in simulated white powder samples.

CONCLUSIONThe suspension array technique could be served as an opening screening method for biothreat bacteria rapid detection.

Bacillus anthracis ; isolation & purification ; Bioterrorism ; prevention & control ; DNA Primers ; DNA, Bacterial ; analysis ; Francisella tularensis ; isolation & purification ; Oligonucleotide Array Sequence Analysis ; methods ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 16S ; genetics ; Yersinia pestis ; isolation & purification

OBJECTIVETo investigate the relationship between the single nucleotide polymorphisms(SNPs) in the redox domain of aprimidinic/apurinic endonuclease/redox factor-1(APEX) gene and the development of sporadic colorectal cancer.

METHODSOne hundred and fifty cases of sporadic colorectal cancers and 143 peripheral blood samples from healthy population were screened for genetic polymorphisms or mutations in the redox domain by denaturing gradient gel electrophoresis followed by DNA sequencing.

RESULTSThere were two SNPs identified in the redox domain of APEX gene, namely, 453G to T and 1247A to G. The gene frequencies of 453T and 1247G were 1.3% and 5.7%, respectively, in patient group, while 1.05% and 4.55%, respectively, in healthy population. The genotype distribution at the two sites in healthy population was consistent with Hardy-Weinberg equilibrium. There was no difference in gene frequencies at the two sites between cancer patients and healthy population.

CONCLUSIONThe polymorphisms in the redox domain of APEX gene are irrelevant to the development of sporadic colorectal cancer, but their distribution may vary greatly among tribes.

Aged ; Alleles ; Base Sequence ; Binding Sites ; genetics ; China ; Colorectal Neoplasms ; enzymology ; genetics ; pathology ; DNA Mutational Analysis ; DNA, Neoplasm ; chemistry ; genetics ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; genetics ; metabolism ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Oxidation-Reduction ; Point Mutation ; Polymorphism, Single Nucleotide

Objective To evaluate the clinical efficacy and safety of Shenfu injection in the treatment of leukopenia induced by chemotherapy docetaxel, epirubicin and cyclophosphamide adjuvant chemotherapy ( TEC chemotherapy) after surgery of breast cancer.Methods A total of 106 patients with breast cancer were randomly divided into control group ( n=53 ) and treatment group ( n =53 ) .Control group was re-ceived 75 mg? m-2 docetaxel, intravenous infusion on day 1 +50 mg? m-2 epirubicin, intravenous infusion on day 1 +500 mg? m-2 cy-clophosphamide, intravenous infusion on day 1, 3 weeks for a course. Treatment group was given 50 mL Shenfu injection added 500 mL of 5%glucose intravenous infusion and plus the treatment of control group, 1 to 10 days for a course.Patients of two groups were received 2 courses of treatment.The clinical efficacy, the incidence of leukopenia, white blood cell count to improve the rate and quality of life before and after treatment, and adverse drug reactions in two groups were compared.Results After treatment, total effective rate in treatment group was significantly higher than that in control group (84.91%vs 69.81%, P<0.05).After treatment 7,10 d, leukopenia rates in treatment group were 9.43%and 11.32%, significantly lower than 37.74%and 30.19%in control group (P<0.05).After treatment, the white blood cell count in the two groups all decreased than those before treatment, and the degree of decline in treatment group was higher than that in control group (P<0.05).After treatment, the rate of improvement of quality of life in treatment group was significantly higher than that in control group (88.68%vs 54.72%, P<0.05).The incidence of adverse drug reactions in treatment group was significantly lower than that in control group ( P<0.05).Conclusion Shenfu injection has a definitive clinical efficacy for the treatment of chemotherapy-induced leukopenia, which may improve immune function and the quality of life, worthy of further promotion in clinical practice.