Objective: To investigate the effect of leukemia inhibitory factor (LIF) on the of proliferation and differentiation of ectomesenchymal cells of mandibular process in Balb/c fetal mice . Methods: Ectomesenchymal cells from the E12.5 mice mandibular process were cultured in DMEM/F12 with 10 6u/L LIF (experimental group) or without LIF (control). The proliferation effect was detected by MTT assay, Brdu test and flow cytometry. Immunohistochemistry were used to identify the differentiation state. Results: By day 7 the A value of the experimental group was 0.38?0.03,that of the control 0.30?0.02 (P

To establish the online two-dimensional liquid chromatography by using double gradient liquid chromatography system and UV detector, in order to simultaneously determine the content of paeoniflorin, paenol, amygdaloside and cinnamic acid. A pump of the two-dimensional liquid chromatography was adopted as the one-dimensional separation pump. C18 (3.0 mm x 150 mm, 3 microm) was used as the analytical column, with acetonitrile as the organic phase and 0.08% phosphoric acid + 0.08% triethylamine as the aqueous phase for gradient elution at the flow rate of 0.5 mL x min(-1). Another pump of the two-dimensional liquid chromatography was adopted as the two-dimensional separation pump. PAII C18 was used as the analytical column, with acetonitrile as the organic phase and 20 mmol, pH 3.0 monopotassium phosphate as the aqueous phase for gradient elution at the flow rate of 0.8 mL x min(-1). The detection wavelengths were set at 218, 230, 275 nm by using wavelength time-switching program. The linearity range of paeoniflorin, amygdaloside, paeonol and cinnamic acid were 5.55-222 (r = 0.999 7), 6.6-264 (r = 0.999 8), 3.3-132 (r = 0.999 5) and 0.315-12.6 mg x L(-1) (r = 0.999 7), respectively. The average recoveries of the four components were between 96.12% and 103.9%. The experiment proved that this method was so rapid and accurate in determination results that it could be used for evaluating drug quality.
Capsules ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Online Systems ; Time Factors

Objective To explore the effects of the autoantibody against the second extracellular loop of ?3-adrenoceptor(?3AR)from children with heart failure on isolated cardiomyocytes of adult rats.Methods The synthetic peptide corresponding to the sequence of the se-cond extracellular loop(176-202 amino acid residues)of the human ?3AR was used as the antigen to screen the anti-?3AR autoantibodies from the sera of the 50 children with heart failure and 50 healthy children by enzyme linked immunosorbent assay(ELISA);IgG in the children with heart failure of positive autoantibodies sera was purified.The effects of purified IgG per each group on contractile response of isolated cardiomyocytes of adult rats were observed using motion-edge detection system and double-provocation fluorescence photomultiplier system.Results Among the 50 children with heart failure,the anti-?3AR autoantibodies positive rate was 30%(15/50 cases),and ob-viously higher than that of healthy children[12%(6/50)cases,P

Based on the Miller Pyramid for assessing clinical competence, this article analyzed the existing problems in the practice of teaching physical examination and provided suggestions for possible reforms.

Objective To assess the effectiveness of donepezil versus huperzine in the treatment of elderly patients with mild cognitive impairment (MCI).Methods Total 122 elderly patients with MCI were divided into two groups:donepezil treatment (5.0 mg once daily) (n=71) and huperzine treatment group (0.1 mg twice daily) (n=51).All the patients were followed up for 24 weeks.Before and 12 weeks,24 weeks after drug treatment,the cognitive functions were evaluated,including MMSE,MOCA,ADAS-cog,CDR,GDS,ADL,HIS and HAMD.Results There was no significant difference in age,sex,education time and neuropsychology rating scales between the groups before drug use.As compared with the score before drug use,the donepezil group showed a significant increase in MMSE after 12-weeks (t=4.47) or 24-weeks (t=6.16) (P＜0.01),a decrease in the score of ADAS-cog after 12-weeks (t=2.33,P＜0.05) or 24-weeks( t=3.68,P＜0.05),and an increase in the score of MOCA after 24-weeks drug use (t=2.56,P＜0.05).The huperzine group showed significant improvement in MMSE after 24-weeks drug use (t=2.80,P＜0.05),but there was no difference in other time points or in the score of MOCA and ADAS-cog as compared with the score before drug use.After 24 weeks' treatment,the donepezil group had higher MMSE (t=2.01,P＜0.05) and lower ADAS-cog (t=2.09,P＜0.05) scores than the huperzine group.30 patients (total effective rate was 42.3 ％) and 9 patients (total effective rate was 17.6 ％ ) became improved in donepezil and huperzine group,respectively,with significant difference (x2 =8.26,P＜0.01 ).There were 5 cases in the donepezil group and 3 cases in the huperzine group getting slight side-effects which disappeared by continuing to take drugs or by adjusting drug taking time.Conclusions Donepezil and huperzine as the cholinesterase inhibitors are effective and safe,and the efficacy of donepezil is faster and better in treating elderly patients with MCI.

AIM:To compare the reliability and plaque area between using high-cholesterol diet and high-cho-lesterol diet with corn oil to establish a rabbit atherosclerotic model .METHODS:Eighteen New Zealand rabbits were ran-domly divided into 3 groups (6 rabbits each):normal diet group (group C), high-cholesterol diet group (group H1) and high-cholesterol diet containing 6%corn oil group (group H2).All rabbits were fed for 12 weeks, and their body mea-sured was weighed at the end of every weeks .The serum levels of high-density lipoprotein cholesterol ( HDL-C) , low-den-sity lipoprotein cholesterol ( LDL-C) , total cholesterol ( TC) and triglyceride ( TG) were measured by automatic chemistry analyzer at 0 week and 12 weeks.At the end of 12 weeks, the thoracic aorta of 8-cm length since aortic root was isolated from the rabbit after anesthesia , and stained with Sudan IV or oil red O to verify the existence of plaque .The percentage of plaque area (PA/IA) in the intima area was further calculated by ImageJ 2x software.RESULTS:At the end of 12-week feeding, the serum levels of HDL-C, LDL-C and TC in both group H1 and group H2 were significantly higher than those in group C, and serum TG in group H2 was significantly higher than that in group C .Serum HDL-C in group H2 was signifi-cantly higher than that in group H1, but no significant difference of serum LDL-C, TC and TG between group H1 and group H2 was found .There was no plaque in the intima in group C , and plaques were observed in the intima of all rabbits in group H1 and group H2.Rabbit atherosclerotic models in both group H 1 and group H2 were established with a success rate of 100%.The values of PA/IA in group H1 [(49.74 ±18.78)%] and group H2 [(56.95 ±26.74)%] were both sig-nificantly higher than that in group C (0%), and no significant difference of PA/IA between group H1 and group H2 was observed.CONCLUSION:High-cholesterol diet with or without corn oil can establish a rabbit atherosclerotic model with a success rate of 100%after 12-week feeding , and the percentage of plaque area in the total aortic intimal area is not differ-ent in the 2 feeding methods .

AIM: To study the immunological protection of H. pylori vaccine with chitosa as adjuvant. METHODS: One-grade female BALB/c mice were randomly divided into nine groups and immunized by ①PBS alone; ②chitosan solution alone; ③chitosan particles alone; ④H. pylori antigen alone; ⑤H. pylori antigen plus chitosan solution; ⑥H. pylori antigen plus chitosan particles; ⑦H. pylori antigen plus CT; ⑧H. pylori antigen plus chitosan solution and CT; ⑨H. pylori antigen plus chitosan particles and CT. At 4 weeks after the last immunization, these mice were challenged by alive H. pylori(1?1012CFU/L) twice at two-day intervals. At 4 weeks after the last challenge, these mice were all killed and gastric mucosa were embedded in paraffin, sectioned and assayed with Giemsa staining. The other gastric mucosa were used to quantitatively culture with H. pylori. ELISA was used to detect H.pylori IgA in saliva and gastric mucosa and anti-H.pylori IgG, IgG1, IgG2a in serum, and immunohistochemical method was used to examine sIgA in gastric mucosa. RESULTS: ①In the groups with chitosan as adjuvant, 60% mice achieved immunological protection, which was according to that with CT as adjuvant (58.33%), and was significantly higher than H. pylori antigen alone and other groups without H. pylori antigen(P0.05)and were significantly higher than those in non-adjuvant groups, while those in the groups with chitosan plus CT were significantly higher than those in the group with CT as an adjuvant(P

AIM: To establish an experimental model of high intraocular pressure in mice by laser photocoagulation and to prepare for future research.
METHODS: Experimental model of high intraocular pressure was induced unilaterally in 44 C57BL/6 mice. The fellow eye served as a control. TONO-PEN AVIA Tonomter was used to measure intraocular pressure (IOP) to guarantee IOP value at 1, 2, 4, 8wk. Slit-lamp biomicroscopy was performed throughout the period and the structural changes were assessed histologically. And then, their eyes were enucleated, postfixed, cryoprotected, and embedded in optimal cutting temperature medium. After hematoxylin and eosin stain ( HE stain ) , cryosections of the retina were observed under light microscope. TdT-mediated biotin-dUTP nick end labeling ( TUNEL ) was performed on the retinal sections to determine apoptosis rate.
RESULTS: IOP of laser-treated eyes was significantly higher than that of control eyes from 1-8wk (P<0. 05). The highest IOP was 31mmHg, but only one eye. The IOP was mainly around 20mmHg. In laser-treated eyes, the angle of anterior chamber were narrow. Number of cells in the inner nuclear layer and retial gangllion cell layer was slightly lower than that in control eyes at 2wk, but by 4 and 8wk the number of cells was significantly lower than that in the control contralateral eyes.
CONCLUSION: The laser photocoagulation of limbus causes chronic elevation of IOP and this method may be a promising experimental model for the investigation of biological mechanisms of glaucomatous retinal ganglion cell damage.

Aim To investigate the effect of hypercholesterolemia on L-type Ca2+(ICa-L) current and intracellular calcium concentration ([Ca2+]i) in single ventricular myocytes of hypercholesterolemic rats.Methods 12 male wistar rats were randomly divided into two dietary groups:a group fed a normal diet(n=6)and a group fed high-cholesterol diet(n=6) for 4 weeks,respectively. The level of serum lipid was examined.Zymolytic method was used to isolate single ventricular myocytes from hypercholesterolemic and normal rats,which were loaded with Ca2+-sensitive fluorescent indicator Fluo-3/AM.[Ca2+]i represented by fluorescent intensity(FI)was measured by laser scanning confocal microscope.Whole cell patch clamp technique was used to record ICa-L.Results There was no significant influence exhibited on TG level.However, the serum total cholesterol(TC)level of hypercholesterolemic rats was much higher than that of model control group; at the test potential of 0 mV, ICa-L decreased from(-8.56?1.29)pA/pF(Control)to(-5.24?0.90) pA/pF(HC)(n=6 in each group,P

OBJECTIVETo investigate the clinical indications of asthma control test (ACT).

METHODSA total of 120 asthmatic patients with a diagnosis in line with the American Thoracic Society criteria and treated for over a month were enrolled in this study. The patients were asked to complete a survey to assess their symptoms and asthma attacks, and ACT evaluation was conducted by physicians familiar with ACT evaluation. The patients were classified into two groups based on the pulmonary function test (positive for bronchodilator test and provocation test) or based on disease severity (mild and moderate-to-severe asthma groups). The effect of ACT evaluation was graded as good (no less than 4 item available for evaluation), fair (2-3 items available) and poor (no more than 1 item). To further analyze the ACT sensitivity in relation to different disease severity, 29 asthmatic patients with an initial diagnosis and BDT positivity were included, and the ACT score of the patients with mild, moderate and severe asthma based on FEV1% were compared.

RESULTSIn patients positive for bronchodilator test, good, fair and poor evaluation effects were found in 48, 15, and 5 cases, as compared to 10, 15, and 27 in those positive for provocation test, respectively, showing significant differences between the two groups (P < 0.001). In mild asthma group, good, fair and poor evaluation effects were found in 12, 15, and 18 cases, respectively, significantly different from those in moderate-to- severe asthma group (50, 21, and 4 cases, P < 0.001). ACT scores showed a positive correlation to FEV1% in 29 patients with positive BDT (r = 0.55, P = 0.003). ACT scores had no significant difference between mild and moderate asthma groups (P > 0.05), but showed significant differences between mild and severe groups (P = 0.009) and between moderate and severe groups (P = 0.008).

CONCLUSIONACT is more suitable for evaluating patients positive for bronchodilator test or with moderate to severe asthma.

Adolescent ; Adult ; Aged ; Asthma ; diagnosis ; physiopathology ; Female ; Humans ; Male ; Mass Screening ; Middle Aged ; Predictive Value of Tests ; Respiratory Function Tests ; Sensitivity and Specificity ; Severity of Illness Index ; Surveys and Questionnaires ; Young Adult