AIMTo study the cellular uptake of chitosan oligosaccharide nanoparticles by A549 cells and evaluate the possibility of chitosan oligosaccharide nanoparticles used as a potential drug carrier.

METHODSChitosan oligosaccharide (CSO) was obtained by ultrafiltration separation after regulation of the condition of chitosanase degradation. The molecular weight of CSO was determined by gel permeation chromatography (GPC). Chitosan oligosaccharide nanoparticles (CSO-NPs) were prepared by a novel solvent diffusion method in an oil system after the carrier material grafted fluorescein isothiocyanate (FITC) and the particle size distribution and zeta potential were determined by light scattering and electrophoretic mobility. The cytotoxicity and uptake of FITC-labeled CSO-NPs in A549 cells following various incubation periods were studied by the MTT method and fluorescence microscopy, flow cytometric analysis, respectively.

RESULTSThe molecular weight (MW) of CSO was 18,678 u and the particles sizes of CSO-NPs were 133.3 nm (number average) and 368.2 nm (volume average), respectively. The IC50 of CSO and CSO-NPs were 944.36 and 643.16 mg x L(-1), respectively, and the result showed low cytotoxicity. Cellular uptake of CSO and CSO-NPs were relative to the concentration and the incubation time. Internalization of CSO-NPs increased 0.49 - 13.9 times more than that of the CSO with the same incubation time.

CONCLUSIONCSO and CSO-NPs have low cytotoxicity. CSO-NPs can significantly improved the uptake of CSO-NPs by A549 cells compared to the same molecular weight of CSO.

Adenocarcinoma ; pathology ; Chitin ; analogs & derivatives ; metabolism ; toxicity ; Chitosan ; Dose-Response Relationship, Drug ; Drug Carriers ; metabolism ; toxicity ; Epithelial Cells ; drug effects ; metabolism ; Humans ; Lung Neoplasms ; pathology ; Nanotechnology ; Oligosaccharides ; metabolism ; toxicity ; Particle Size ; Time Factors ; Tumor Cells, Cultured

Objective To investigate the relationship of stress hyperglycemia,cystatin C and estimated glomerular filtration rate (eGFR) with short-term prognosis in elderly patients with acute myocardial infarction.Methods 242 consecutive patients with acute myocardial infarction were divided into two groups according to age:the elderly group (n=182),and the non elderly group (n=60).The clinical data including cystatin C (Cys C),eGFR and stress hyperglycemia levels were collected.The major adverse cardiovascular events (MACE) were observed during hospitalization and 30 days after discharge.Results The incidences of stress hyperglycemia,the levels of creatinine,Cys C and brain natriuretic peptide (BNP),as well as the total MACE were higher and eGFR was lower in elderly group than in non-elderly group (all P＜0.05).Cys C level was positively correlated with age,body mass index and levels of creatinine and BNP (all P＜0.05),and negatively correlated with fasting glucose and eGFR in elderly group (both P＜0.05).The eGFR was positively correlated with body mass index (P＜0.05),and negatively correlated with age,creatinine and BNP levels in elderly group (all P＜0.05).Logistic regression analysis indicated that stress hyperglycemia [OR=1.871,95％CI:1.071-3.269,P=0.03],Cys C [OR=7.093,95％CI:2.261-22.249,P=0.00] were the independent risk factors for MACE.Conclusions Cys C level and eGFR can predict the early renal dysfunction and its prognosis in elderly patients with acute myocardial infarction.The incidence of stress hyperglycemia is higher in the elderly,and stress hyperglycemia and Cys C level are the independent risk factors for MACE.

Objective To discuss how to protect the intracranial vertebral artery and posterior inferior cerebellar artery by observing and measuring the intracranial vertebral artery in the surgery adopt far lateral approach. Methods Mimicking far lateral approach, 20 adult cadaveric heads connected to neck fixed with 10% formalin were dissected. Intracranial segment of the vertebral arteries and their main branches were exposed and measured under operating microscope. Results The intracranial vertebral artery joined with the contralateral one into the basilar artery after traveling through the atlanto-occipital sulcus. The relationship between the vertebral artery and the hypoglossal nerve is close. Thirty sides (75%) of the vertebral arteries traveled to pons medulla sulcus in front of the hypoglossal nerve roots and 2 sides (5%) behind the hypoglossal nerve roots, while 8 sides (20%) traveled among the hypoglossal nerve roots; 70% of the vertebral arteries were contacted to the hypoglossal nerve roots, 30% of which compressed the hypoglossal nerve. The main branches of intracranial segment of the vertebral arteries were the posterior inferior cerebellar arteries, the anterior spinal arteries, the posterior meningeal arteries,and some perforating arteries. Posterior inferior cerebellar arteries all originated from the intracranial vertebral artery were the largest vertebral artery's branches; their trip was mostly loop-shaped and they had close relationship with Ⅸ, Ⅹ, Ⅺ cranial nerves. The starting points of the posterior inferior cerebellar arteries were different, even in the same specimen, but most of them originated from the upper 1/3intracranial vertebral artery. No anterior inferior cerebellar artery was noted originated from the vertebral artery in our specimen. Anterior spinal arteries originated from the vertebral arteries joined with the branches of the bilateral vertebral arteries and traveled down through the tortuous anterior median fissure to supply the spinal cord. Conclusion Being familiar with the characteristics and anatomic vertebral arteries variations of the intracranial vertebral artery and its branches can contribute to identify and protect the intracranial segment of the vertebral artery and its main branches in the surgery adopt far-lateral approach.

OBJECTIVETo explore in vivo effects of Qizheng-xiaotong plaster on soft tissue injury in rabbit ears at different periods and to offer theoretical bases for clinical application.

METHODSThe experimental models of soft tissue injury in ears were produced in 10 New Zealand white rabbits, and the ears were divided into three groups at the 1st, 2nd, 3rd week. The normal group and treatment group were given the Qizheng-xiaotong plaster extract, and the model group with normal saline. Microscopic analysis, digital collection system, infrared temperature tester and thickness tester were applied to determine the changes of soft tissue injury in local microcirculation and the temperature change after 0, 0.5, 3 and 5 hours, and swelling change at 1 to 5 days respectively.

RESULTSAt the 3rd hour, blood velocity speeded up in normal group and model group, and it lasted for two hours in model group. As compared with model group, it slowed down to original level in treatment group at the 5th hour and the soft tissue swelling decreased from the 3rd to the 5th day as well.

CONCLUSIONThe application of Qizheng-xiaotong plaster is effective in preventing further soft tissue oedema and haematoma. It can make the soft tissue swelling decreased at chronic stage compared with that at acute stage.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Ear, External ; drug effects ; injuries ; physiopathology ; Humans ; Microcirculation ; drug effects ; Rabbits ; Soft Tissue Injuries ; drug therapy ; physiopathology

OBJECTIVETo develop a transparent, non-toxic, non-irritating anti-fogging agent with long-lasting effect for nasal endoscopy.

METHODSThe anti-fogging agent was prepared by mixing ethanol, propylene glycol, polyoxyethylene lauryl ether, sodium dodecyl sulfate, polyethylene glycol 400 and deionized water at different proportions based on an orthogonal test design. Twenty-seven test samples of the anti-fogging agents were obtained, which were colorless, transparent, and non-irritating, with a pH value of 7-8. Storz00 nasal endoscopy and its imaging system were used to test the anti-fogging time of the 27 samples, and each agent was tested for 3 times with medical Seoul iodine and 95% ethanol as control.

RESULTSThe optimal composition of the anti-fogging agent was 20% ethanol, 10% propylene glycol, 20% polyoxyethylene lauryl ether, 4% sodium dodecyl sulfate, 4% polyethylene glycol, 42% deionized water. The anti-fogging time of this agent reached 15 min, significantly longer than that of medical Seoul iodine (4 min) and 95% ethanol (18 s).

CONCLUSIONThis anti-fogging agent for nasal endoscopes is colorless and safe and has a long anti-fogging time by forming a homogenous transparent membrane over the endoscopic lens.

Endoscopes ; Endoscopy ; methods ; Ethanol ; Nose ; surgery ; Polyethylene Glycols ; Sodium Dodecyl Sulfate ; Solutions ; chemistry

ObjectiveTo study the mRNA expression of rat Insulin-like growth factors- Ⅰ (IGF- Ⅰ ) and Transforming growth factor-β1 (TGF-β1) in thyroid and placenta with different iodine intakes during pregnancy.MethodsOne hundred and fifty female Wistar rats,weighting 80 - 100 g,were randomly divided into five groups according to body weight,30 rats in each group.Each group was given deionized water containing different concentrations of iodine,50 μg/L(control group,NI),0 μg/L(iodine deficiency 1 group,LI1 ),5 μg/L(iodine deficiency 2 group,LI2),3000 μg/L(iodine excess 1 group,HI1 ),and 10 000 μg/L(iodine excess 2 group,HI2),respectively.After feeding for 12 weeks,the female rats were mated with male rats.The female rats were sacrificed at first(6,7 days),trimester( 12,13 days),and third trimesters( 19,20 days),respectively,then their thyroid and placenta were collected.The mRNA expressions of IGF- Ⅰ and TGF-1 in thyroid and placenta were detected by real-time quantitative PCR.Results①The actual thyroid weights of LI1 and LI2 groups[ (12.17 ± 5.41 ) × 10-2 g,(3.54 ± 1.21) × 10-2 g] were significantly higher than that of NI group[ (2.05 ± 0.50) × 10-2 g,all P ＜ 0.05] ;actual weights of HI1 and HI 2 groups[ (1.64 ± 0.27) × 10-2 g,(1.66 ± 0.29) × 10-2 g] were compared with that of NI group,the difference was not statistically significant(all P ＞ 0.05).②The mRNA expression of IGF- Ⅰ: at the first trimester,LI1 and LI2 groups(l.98 ± 0.35,1.47 ± 0.22) were all higher than that of NI group(1.01 ± 0.18,all P＜ 0.01 ),HI1 and HI2 groups(0.68 ± 0.16,0.75 ± 0.09) were lower than that of NI group(all P ＜ 0.01 );at the second trimester,HI2 group( 1.14 ± 0.17) was lower than that of NI group( 1.58 ± 0.33,P ＜ 0.01 ) ; at the third trimester,LI2 and HI2 groups(1.47 ± 0.20,1.45 ± 0.35) were lower than that of NI group(2.20 ± 0.37,all P＜0.01).The mRNA expression of IGF- I level in NI group at the first,second,and third trimesters(1.01 ±0.18,1.58 ±0.33,2.20 ± 0.37) was up regulated gradually,pairwise comparisons were statistically significant(all P ＜ 0,01 ).③The mRNA expression of TGF-β1: at the first trimester,LI1 group (1.37 ± 0.13) was higher than NI group (1.05 ±0.18,P ＜ 0.01 ),HI1 and HI2 groups(0.50 ± 0.09,0.44 ± 0.11) were lower than NI group(all P＜ 0.01); at the second trimester,LI1 and HI2 groups(1.39 ± 0.28,1.17 ± 0.12) were higher than NI group(0.63 ± 0.22,all P ＜0.01 ) ; at the third trimester,LI1 and LI2 groups ( 1.57 ± 0.30,1.23 ± 0.20) were higher than NI group ( 0.68 ± 0.17,all P＜ 0.01).TGF-β1 mRNA expressions of NI group at the second (0.63 ± 0.22) and third trimesters(0.68 ± 0.17) were lower than that of the first trimester (1.05 ± 0.18,all P ＜ 0.01).④ Rats' IGF-Ⅰ mRNA expression in placental: at the second trimester HI1 group,HI2 group( 1.48 ± 0.16,1.45 ± 0.25) were all higher than the NI group ( 1.00 ± 0.10,all P ＜ 0.01 ) ; at third trimester,HI1 group ( 1.75 ± 0.15 ) were higher than the NI group ( 1.54 ± 0.29,P＜ 0.05),HI2 group(l.94 ± 0.31) were higher than the NI group(P ＜ 0.01 ).IGF- Ⅰ mRNA expression in placental of NI group at the third trimester was higher than the second trimester(P＜ 0.01).⑤ Rats' TGF-β1 mRNA expression in the placenta: at the second trimester and the third trimester of pregnancy there were no significant difference between the five groups(all P ＞ 0.05) ; NI group at the third trimester(0.83 ± 0.16) was lower than the second trimester(0.98 ± 0.20,P ＜ 0.05).Conclusions During pregnancy,IGF- I mRNA expression increases in thyroid under the conditions of iodine deficiency,and this effect is particularly significant in the first trimester; at the same time,TGF-β1 mRNA expression is increased,and this inhibition becomes clear with the deepening of iodine deficiency.Under the condition of iodine excess,the functions of IGF- Ⅰ and TGF-β1 in thyroid above-mentioned were relatively weak.With the development of gestational period,promoting tissues growth and differentiation effect of placenta's IGF- Ⅰ was more significant gradually,but,inhibited effect of TGF-β1 was weaken.

Objective To observe the effect of different levels of iodine nutrition on rat maternal thyroid function during pregnancy.Methods A total of 225 Wistar rats one month after weaning were involved in the study(female 165,male 60,body mass 80 to 100 g).Female rats were randomly divided into six groups by body mass:control group(NI group),iodine deficiency 1 and 2 groups(LI1,LI2 groups),iodine excess 1 and 2 groups (HI1,HI2 groups),and the control of not pregnant group(NNI group).There were 30 rats in 1-5 groups and 15 rats in group 6.LI1,LI2 groups:low iodine diet + deionized water of no iodine or iodine-containing 5 μg/L; HI1,HI2 groups:normal diet + deionized water of iodine 3000,10 000 μg/L; NI,NNI groups:normal diet + deionized water of iodine-containing 50 μg/L.After 12 weeks,the females(except group 6) mated the male by 2 ∶ 1,and then each pregnant female rat was fed in a single cage.The female mice were sacrificed in the first(5 ± 2)d,the second (12 ± 2)d and the third trimesters of pregnancy (17 ± 2)d,respectively,and there blood samples and thyroid were obtained.Serum total thyroxine(TT4),free thyroxine(FT4),total triiodothyronine (TT3),free triiodothyronine (FT3) and thyroid stimulating hormone(TSH) were determined by radioimmunoassay and serum thyroglobulin(TG) and thyroid-binding globulin (TBG) were determined by enzyme-linked immunosorbent assay.Results ①Thyroid absolute quality and relative quality was compared among groups,and the differences were statistically significant (F =16.55,24.25,F ＜ 0.01 or ＜ 0.05).②At the first,the second and the third trimesters of pregnancy,the differences of maternal serum TT4 and FT4 between groups were statistically significant(F =5.02,13.41,17.39,41.89,23.72,48.64,P ＜ 0.01 or ＜ 0.05).Female rats in NI,HI1 and HI2 groups in different pregnant periods among inner groups were compared,and the differences of serum TT4 and FT4 were statistically significant(F=3.27,6.98,8.22,8.65,29.68,7.90,P ＜ 0.01 or ＜ 0.05).③ In the first and the third trimesters of pregnancy,maternal serum TT3 was compared among groups,and the differences were statistically significant(F=3.59,8.22,P ＜ 0.05 or ＜ 0.01) ; in the second and the third trimesters of pregnancy,maternal serum FT3 was compared among groups,and the difference was statistically significant(F =3.86,4.26,P ＜ 0.05 or ＜ 0.01).Female rats in NI,LI1 and HI1 groups in different pregnant periods among inner groups were compared,and the differences of maternal serum TT3 were statistically significant(F =8.77,7.11,6.28,P ＜ 0.01 or ＜ 0.05).④At the first,the second and the third trimesters of pregnancy,the differences of maternal serum TG and TBG were compared in groups,and the differences were statistically significant(F =5.47,3.62,9.35,4.15,13.16,22.78,P ＜ 0.01 or ＜ 0.05).The differences of maternal serum TG of HI1 group and of serum TBG of NI group in different pregnant periods among inner groups were statistically significant (F =3.18,7.94,P ＜ 0.05).⑤At the first,the second and the third trimesters of pregnancy,the differences of maternal serum TSH in groups were statistically significant(F =4.83,7.08,6.52,P ＜ 0.01); the differences of maternal serum TSH of all the 5 groups in different pregnant periods among inner groups were statistically significant (F =3.26,8.89,11.45,4.04,3.78,P ＜ 0.05).Conclusions Different levels of iodine nutrition can cause changes in thyroid function in rats maternal thyroid function during pregnancy; serum TT4,FT4 level decreases when iodine deficiency,and increase with iodine excess.Serum TT3,FT3 level of does not changed significantly due to compensatory regulation of the body.

Objective To study the effect of different levels of iodine nutrition on secretion of placental hormone in pregnant rats.Methods Two hundred and twenty five Wistar rats (165 female,60 male),weighing about 80 - 100 g were used in the study.Female rats were randomly divided into five groups according to their body weights:low iodine group Ⅰ(LⅠ),low iodine group Ⅱ (LⅡ),adequate iodine(control) group(Al),high iodine group Ⅰ ( HⅠ ),and high iodine group Ⅱ (H Ⅱ ),and 33 rats in each group.Animals in the low iodine groups were fed low-iodine diet,the iodine content was 13.46 μg/kg,in addition,these rats drank deionized water which containing potassium iodated,the dose was 0 and 5 μg/L,respectively.The rats of adequate and the two high iodine groups were fed normal diet,the iodine content was 22.00 μg/kg,they also drank deionized water,containing potassium iodated 50,3000,and 10000 μg/L,respectively.The rats mated after 3 months of feeding,and were respectively sacrificed at early pregnancy(5 ± 2)d,second trimester( 12 ± 2)d,and third trimester of pregnancy(17 ± 2)d,and then their serum was taken.Serum human chorionic gonadotropin(HCG),human chorionic thyrotropin(HCT),and progesterone were measured by enzyme-linked immunosorbent assay (ELISA).Results In the third trimester,the serum levels of rat HCG were significantly different between groups(F =4.16,P ＜ 0.05).The means of rats serum HCG of the two low iodine groups [ (16.08 ± 4.45),(17.43 ± 2.70)U/L] were significantly higher compared with that of AI group[ (13.68 ± 3.52)U/L] in the third trimester(all P ＜ 0.01 ).In the second and third trimester,the levels of rats serum HCT were significantly different between groups(F =3.59,3.40,all P ＜ 0.05).The means of rats serum HCT of HI group [(70.11 ± 10.97)μU/L] in the second trimester and HII group[(74.93 ± 13.22)μU/L] in the third trimester were higher than those of AI group[ (57.14 ± 12.56),(58.17 ± 8.54)μU/L] significantly(all P ＜ 0.01 ).There were statistical differences of the means of serum progesterone among trimester of pregnancy(F =4.06,4.43,all P ＜ 0.05).The level of serum progesterone of the third trimester[ ( 1462.80 ± 286.48 )pmoL/L] compared to those of the first[ (1929.93 ± 158.37) pmol/L] and the second trimester[ (1856.44 ± 542.08)pmol/L] was decreased significantly(all P ＜ 0.05) in LI group.In the control group,the level of serum progesterone of the second trimester [ (2046.45 ± 475.67)pmol/L ] was significantly higher than the first trimester[ (1714.39 ± 461.71 )pmol/L,P ＜ 0.05 ].Conclusions During pregnancy,placenta could promote HCG secretion under iodine-deficient conditions.In addition,the placenta increases the secretion of HCT under conditions of excess iodine.In the condition of severe iodine deficiency,the secretion of serum progesterone decreases,and further decreases with prolongation of pregnancy,but it is opposite to the change of HCG during pregnancy.This phenomenon could lead to harmful pregnant outcomes easily.

Objective To explore the effect ofmiR-137 on proliferation and apoptosis of U87 glioma cells.Methods The miR-137 mimics were transfected to human glioma cell line U87 as miR-137 transfection group; blank control group and negative control group were also employed in our study.The expression ofmiR-137 was identified by real-time polymerase chain reaction (PCR) after transfection.Methylthiazol tetrazoliu (MTT) assay,flow cytometry-Annexin V/PI assay and fluorescence microscope were employed to detect the cell proliferation and apoptosis.MiR-137 and its relative targeting protein expressions were detected by Westem blotting.Results MTT assay showed that the relative cell survival rates in the blank control group,negative control group and miR-137 transfection group were (105.16±8.57)％,(98.57±8.21)％ and (45.84±6.93)％,with significant differences (F=82.157,P=0.000).Annexin V/PI assay showed that the cell apoptosis rates in the blank control group,negative control group and miR-137 transfection group were (4.3±0.63)％,(4.7±0.77)％ and (16.6±1.98)％,with significant differences (F=63.837,P=0.000); and apoptotic body was detected by fluorescence microscope.Moreover,CDC42,pERK1/2,AKT and pAKT protein expression levels were inhibited after transfected by miR-137 mimics.Conclusion MiR-137 inhibits proliferation and induces apoptosis of U87 glioma cells,suggesting that miR-137 could be tumor suppressor gene in glioma.

Objective:To study the optimal nutritional support strategy in critical patients with acute ischemic stroke.Methods:50 critical patients with acute ischemic stroke were divided into two groups.One group were started with enteral nutrition alone within 24h after admission,while the other group received sequential parenteral and enteral nutrition.The incidence of pulmonary infection,gastric retention,upper gastrointestinal bleeding,hypoglycemia and hyperglycemia were compared between the two groups.Another endpoint was discharge from ICU at day 20,and it was compared between the two groups using Kaplan-Meier methods.Results:There was no difference between the two groups in the rate of hypoglycemia or hyperglycemia (P ＞ 0.05).The incidences of pulmonary infection,gastric retention,upper gastrointestinal bleeding and diarrhea were lower in the PN+EN group than EN group (P ＜ 0.01).The length of ICU stay was also shorter in the PN+EN group.Conclusion:The strategy of sequential parenteral and enteral nutrition decreased the complication rate related to nutrition support,such as pulmonary infection,gastric retention,upper gastrointestinal bleeding.Also,it shortened the ICU stay in critical patients with acute ischemic stroke.