Chitosan has drawn more and more attention for their particular biological effect. Nanometer materials displayed great perspective in medical application. In the present paper, we reviewed the biological characteristic of chitosan, the preparation methods of chitosar nanoparticles, as well as the applications of chitosan and chitosan nanoparticles in tissue engineering.

Objective To perform gene diagnosis of Duchenne muscular dystrophy (DMD) in 3 family members who were negative for DMD gene detected by multiplex PCR and to provide genetic counseling for their family members accordingly .Methods The clinical data and genomic DNA of patients and their family members were collected ,DMD gene mutation were detected by multiplex ligation dependent probe amplification (MLPA) or the 2nd generation of high‐throughput sequencing .Results In the first family ,3 male patients were detected deletion of Exon 7 and 2 female were heterozygous carriers .In the second family ,it was found in the proband that point mutation of c .3127C> T in the Exon 23 of chrX‐32486626 and c .3127C> T heterozygous mutations was confirmed in his mother ,the mother was pregnant with a girl .In the third family ,point mutation of c .2411G>A was detected in the Exon 20 of chrX‐32509581 in the proband and his mother had c .2411G> A heterozygous mutation .Conclusion MLPA or combining with the 2nd generation of high‐throughput sequencing can offer effective gene diagnosis for the patients of DMD and their family members ,and provide the basis for genetic counseling and prenatal diagnosis .

Acupuncture Points ; Animals ; Brain ; pathology ; physiopathology ; Brain Edema ; etiology ; pathology ; physiopathology ; Cerebral Hemorrhage ; chemically induced ; pathology ; physiopathology ; Collagenases ; Electroacupuncture ; Heparin ; Male ; Rats ; Rats, Sprague-Dawley

Objective To find an ideal liver cirrhosis portal hypertension model in big animals suitable for surgical study.Methods Twenty canines were treated by subcutaneous injection of 60% CCl4 colza oil emulsion into the back,at a dose of 1.0-1.3 mL/kg every 10 days for a total of 6-8 times.At the same time,it was combined with the control of food intake.All canines were fed with rice mixed with 10% hog fat.The amount of rice was 15 g/kg per day from the first day to the forth day,but the amount of food was not controlled from the fifth day to the tenth day.During the process,the canines′ general condition was observed and the changes of hepatic functions such as ALT,TP,ALB,G and A/G were measured.The changes of liver morphology,the diameter and blood flow of portal vein were monitored with color Doppler.Every two weeks,a piece of hepatic tissue to observe the pathological change of liver was excised operatively.Results During the process,the body weight of the canines decreased continually;ALT increased during early and middle stages and decreased during advanced stage;TP and ALB always decreased,and A/G continuously decreased,but G did not change significamtly.Eight weeks after injection,liver became progressively smaller.its density was not well-distributed,and liver particles gradually became coarse,and liver capsule became rough.The diameter of portal veins became larger and the velocity of portal vein became slower.Liver cirrhosis with psudolobules was found on light microscopy from the tenth week to the twelfth week.Three canines died,with mortality of 15.0%.Conclusions It took 10-12 weeks to establish this liver cirrhosis portal hypertension models in canines by subcutaneous injection of CCl4.This method is convenient,has high success rate,and is suitable for use in surgieal research.

Objective To evaluate whether the comparability of 3 automatic blood cell analyzers meet the clinical requirements by conducting the comparative study on the detection results of these instruments.Methods With the Sysmex 2100 automatic blood cell analyzer as the reference instrument,Sysmex 1000i and Abbott 1800 as the experimental instrument,the original quality control provided by the instrument factory and the patient′s fresh anticoagulant blood samples in the laboratory were adopted to monitor for continuous 40 d by these three instruments and the detection results of WBC,RBC,HGB,HCT and PLT were analyzed.Results The detection results of these 3 instruments were statistically tested by the F test,the differences showed no statistical significance (P >0.05)and the bias was in 1/2 of the maximum permissible error range in America department clinical test revised regulations (CLIA′88).Conclusion The detection results by these 3 instruments are comparable and can meet the clinical requirements.

Objective To evaluate the role of prostaglandin E2 (EP) receptors in H9c2 cardiomyocyte hypertrophy induced by prostaglandin E2 (PGE2).Methods Primary cultured H9c2 cardiomyocytes were seeded in culture flasks (3 ml/flask) or in 24-well plate (1 ml/hole) or 6-well plate (2 ml/hole) with density of 4 × 104/ml.The cells were randomly divided into 4 groups (n=24 each): control group (group C),PGE2 group,AH6809 (EP1 and EP2 receptor antagonist) group (group A) and GW627368X (EP4 receptor antagonist) group (group G).The cells were continuously cultured for 48 h.PGE2 (final concentration 1 μmol/L) was added to the culture medium in PGE2 group.PGE2 (final concentration 1 μmol/L) and A H6809 (final concentration 10 μmol/L) were added to the culture medium in group A.PGE2 (final concentration 1 μmol/L) and GW627368X (final concentration 10 μmol/L) were added to the culture medium.The cells were then cultured for 48 h in groups PGE2,A and G.Then the cell morphology was observed by using fluorescent microscope.The cell diameter was measured by using the Image J medical image analysis system.Total protein content in the cells was measured with BCA method.The expression of atrial natriuretic peptide (ANP) mRNA and brain natriuretic peptide (BNP) mRNA in the cytoplasm was determined using RT-PCR.Results Compared with group C,the total protein in the cells and cell diameter were significantly increased,and the expression of ANP mRNA and BNP mRNA in the cytoplasm was up-regulated in groups PGE2,A and G (P ＜ 0.05).Compared with group PGE2,the total protein in the cells and cell diameter were significantly decreased,and the expression of ANP mRNA and BNP mRNA in the cytoplasm was downregulated in group G (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group A (P ＞ 0.05).Conclusion EP4 receptor mediates H9c2 cardiomyocyte hypertrophy induced by PGE2 and the effect is not related to EP1 and EP2.

A rapid, sensitive and convenient LC-MS/MS method was developed for the determination of γ-aminobutyric acid (GABA) in human plasma. d2-γ-Aminobutyric acid (d2-GABA) was synthesized as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all analytes were separated on a Luna HILIC column (100 mm x 3.0 mm, 3 μm) using an isocratic mobile phase of water: acetonitrile: formic acid (20 : 80 : 0.12) with a flow rate of 0.5 mL x min(-1). Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode (MRM) in positive electrospray ionization using the transitions of m/z 104 --> 69 for GABA and m/z 106 --> 71 for d2-GABA. The method was linear in the concentration range of 5.00 to 1 000 ng x mL(-1). The intra- and inter-day precisions were within 9.9%, and accuracy ranged from 99.1% to 104%, within the acceptable limit across all concentrations. The method was successfully applied to a pharmacokinetic study of GABA tablets in healthy Chinese volunteers.
Chromatography, Liquid ; Humans ; Tandem Mass Spectrometry ; gamma-Aminobutyric Acid ; blood

A rapid, sensitive and convenient LC-MS/MS method was developed for the determination of γ-aminobutyric acid (GABA) in human plasma. d2-γ-Aminobutyric acid (d2-GABA) was synthesized as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all analytes were separated on a Luna HILIC column (100 mm x 3.0 mm, 3 μm) using an isocratic mobile phase of water: acetonitrile: formic acid (20 : 80 : 0.12) with a flow rate of 0.5 mL x min(-1). Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode (MRM) in positive electrospray ionization using the transitions of m/z 104 --> 69 for GABA and m/z 106 --> 71 for d2-GABA. The method was linear in the concentration range of 5.00 to 1 000 ng x mL(-1). The intra- and inter-day precisions were within 9.9%, and accuracy ranged from 99.1% to 104%, within the acceptable limit across all concentrations. The method was successfully applied to a pharmacokinetic study of GABA tablets in healthy Chinese volunteers.

Objective Analysis the risk factors which maybe caused the cerebral hemorrhage after the liver transplantation, and then make out the counter nursing measures. Methods Selected 12 patients with cerebral hemorrhage after the liver transplantation as the experimental group, and then matching 48 patients without cerebral hemorrhage after the liver transplantation as the control group. Analysis the factor which related the cerebral hemorrhage. Results The factors which related the cerebral hemorrhage after the liver transplantation included in the Intensive Care Unite, heme level, Child level, APTT, albumin, blooding volume during the operation, operation time and the postoperative blood platelet level. The factors of Child level and the blooding volume during the operation were the independence risk factors of cerebral hemorrhage. Conclusion The factors of Child level and the blooding volume during the operation were the independence risk factors of cerebral hemorrhage, we should carry out counter nursing measures to prevent them.

Background The construction of tissue-engineered corneal endothelium needs the functional seeding cells,so how to culture a large amount of functional corneal endothelial cells (CECs) is an urgent problem to be solved.Objective The aim of this study was to evaluate the role of aqueous humor on bovine CECs in vitro.Methods Aqueous humor of 1.2 ml was collected from the anterior chamber of bovine and sterilized,and the liquid supernatant was obtained.The bovine CECs were isolated from bovine cornea and then cultured in low glucose Dulbecco Modified Eagle Medium with 10％ fetal bovine serum (FBS) in vitro.Aqueous humor was added into the medium with the final concentration of 2.5％,5.0％,l0.0％,15.0％ and 20.0％,respectively,and no aqueous humor was added in the control group.Cell counting kit-8 (CCK-8) assay was used to detect the absorbency value of CECs for the evaluation of cell proliferation.Progression of the cell cycle was analyzed by flow cytometry (FCM).After confluence of the cells was reached,1 ml plastic spear tip was used to scratch the cell single layer,and the cells were incubated consequently in medium with 10％ FBS and with or without aqueous humor for 24 hours.Healing area of the cell single layer was measured.The cells were incubated at a density of 6 × 105 cells/ml and cultured using medium with or without 10.0％ aqueous human for 5 days,and the number of the cells was analyzed by DAPI fluorescence technique.Results Under the phase-contrast microscopy,the confluent CECs showed a slabstone-like and hexagonal appearance.CCK-8 assay revealed that the absorbance values of CECs was significantly different among the various culture groups (F=4.051,P =0.007),and the absorbance value in different concentrations of aqueous human culture groups was significantly higher than that in the control group (P ＜ 0.01).FCM showed that the percentage of the cells in S-G2 phases was (34.80-±3.13)％ in the 10.0％ aqueous humors group and (23.06±1.13)％ in the control group,showing a significant difference (t =-5.729,P=0.005).The scratch test showed that the healing area of the cell signal layer was (0.116±0.019) mm2 in the 10.0％ aqueous humors group and (0.358 ±0.049) mm2 in the control group,showing a significant difference (t =13.842,P =0.000).The density of cells in the 10.0％ aqueous humor group was (1439± 1 10)/field,which was more than (1162±45)/field in the control group (t =-11.020,P=0.000).Conclusions Aqueous humor at the concentration of 10.0％ promote the growth and proliferation of bovine CECs.The result suggests that 10.0％ aqueous humor can be used as a promoting agent during the culture of CECs.