Objective To simply the method of how to obtain cultures rich in astrocytes from SD rat cerebral cortex which could be utilized in vitro experiments.Methods The neonatal rat cerebral cortex was made into suspension by mechanical dissociation,and then reduced other cells by differential velocity adherent technique,shaking in orbital shaker and passage of cultured cells.After purification,the cultured cells were identified by double immunofluorescence staining and SABC.Results We successfully obtain cultures rich in astrocytes and the proportion of astrocytes were more than 95%.Conclusion The method described above was reliable in obtaining the high purity astrocytes from neonatal rat cerebral cortex and double immunofluorescence staining was more vivid and direct.

0.05).Conclusion The peptide growth factors can activate the Akt kinase by phosphorylation.This process depends on the production of H2O2 and the presence of PTEN.

OBJECTIVE:To study clinical efficacy and adverse drug reaction(ADR) of Cyclopentolate hydrochloride eye drops,Tropicamide eye drops and Atropine eye drops.METHODS:600 asthenopic children were divided into 3 groups according to visiting time sequence(n=200).3 groups were treated with 1% Cyclopentolate hydrochloride eye drops,1% Tropicamide eye drops and 1% Atropine eye drops respectively.Mydriasis effects of 3 groups were observed and results of retinoscopy were recorded.Ocular ADR and systemic ADR were surveyed.RESULTS:The level of ocular ADR and systemic ADR of tropicamide group were lower than other 2 groups(P

Objective To investigate the clinical application of VCS parameters of leukocyte in the detection of blood bacterial infection, Methods The subjects consisted of 120 patients with blood bacterial infection,69 non-infectious fever patients and 67 health controls.The VCS parameters of neutrophil and lymphocyte were examined with Coulter LH 750 hematology analyzer.The parameters examined including mean channel of neutrophil volume(MNV),neutrophil volume distribution width (NDW) ,mean channel of neutrophil conductivity (MNC),mean channel of neutrophil scatter (MNS),mean channel of lymphocyte volme(MLV),lymphocyte volume distfibufion width (LDW),mean channel of lymphocyte conductivity (MLC) and mean channel of lymphocyte scatter (MLS).Additionally,120 blood bacterial infection patients were grouped according to WBC count(WBC≤10×109/L group and WBC＞10×109/L group),neutrophii rate(≥85%group and＜85%group)and bacterial stain(Gram positive bacteria group and Gram negative bacteria group).VCS parameters among these groups were compared.Results The results of blood infection group were as follows:MNV 156±15,NDW 23.31±3.72,MNS 137±7,MLV 87±12,LDW 17.50±3.38.MLC 110±5 and MLS 69±12.The results of non-infectious fever group were as follows:MNV 151±8,NDW 21.33 ±2.62,MNS 132±10,MLV 91±4.LDW 15.78±1.96.MLC 117±4 and MLS 62±6.The results of control group were as follows:MNV145 ±5.NDW 18.43±0.93.MNS 143 ±4,MLV 84±2,LDW 13.30±0.76.MLC 108±1 and MLS 62±2.There were significant diffierences among these three groups (F value were 19.295,26.272,32.767,6.226,31.016,23.739 and 12.662 respectively,P＜0.05 or P＜0.01).In the infection group.the MNV and NDW were 152 ±16 and 22.19±3.45 respectively for WBC≤10×109/L group.159±12 and 25.29±3.43 respectively for WBC＞10×109/L group.They were both significantly different compared with control group (F valRe were 21.575 and 40.856 respectively,P＜0.01).Also in the infection group.the MNV and NDW were 159±12 and 24.88 ±3.74 respectively for neutrophil rate≥85%group.151±16 and 21.68±2.29 respectively for neutrophil rate＜85%group.They were both significantly different compared with control group(F value were 23.76 and 43.22 respectively,P＜0.01).The MNV and NDW were 157±15 and 24.25±3.39 respectively in those cases with gram-negative bacteremia,153±14 and 21.51±3.78 respectively in those cases with gram-positive bacteremia.They were both signifieanfly difierent compared with control group (F value were 18.74 and 37.47 respectively,P＜0.01).With a cut-off value of 20.50 for the NDW,a sensitivity of 76.7%and specificity of 98.3% were achieyed in diagnosing blood hacterial infection.Conclusion The VCS parameters can reflect the morphologic change of leukocyte in blood bacterial infection.Additionally.the NDW can detect blood bacterial infection more sensitively and specifically.

Objective To discuss the effect of melatonin on sleep and agitation in patients who had undergone tracheotomy in ICU.Methods Sixty-six tracheotomisd patients of ASA Ⅱ or Ⅲ in ICU were included,age 18-60 years,weaning from mechanical ventilation not receiving continuous sedation.We conducted a double-blind, placebo-controlled randomized pilot study comparing melatonin with placebo in such patients.They were allocated into two groups by compute randomiza-tion-melatonin group(n =32)and control group(n = 34).We administered either oral melatonin (3 mg)or placebo at 20∶00 for 7 days.We collected pre-and post-dosage blood samples on days 1 and 3 to confirm drug delivery.The hours of sleep at day and night were assessed by the bedside nurse as the primary outcome.And the secondary outcome included comparison of the incidence of agitation, assessed by score on the Riker Sedation Agitation Scale,and requirement for sedatives or haloperidol to settle agitation.Results Pre-treatment melatonin levels in the two groups were similarly low.Post-treatment,melatonin levels increased significantly in the melatonin group compared with the placebo group (3 233±1 961 )pg/ml versus (3.8 ± 2.2 )pg/ml(P < 0.05 ).And subsequently the observed sleep time in the two groups was (1 1.9±2.0)hours for melatonin vs (7.8±1.6)hours for placebo, respectively (P < 0.01 ).The incidence of agitation was significantly lower in the melatonin group (0% vs 8.8%).Conclusion Melatonin is well absorbed,and a standard dose increases blood levels approximately 1000-fold.Furthermore it would help the tracheotomisd patients sleep and ease agitation in ICU.

The clinical significance of a myeloperoxidase (MPO) gene polymorphism and inducible nitric oxide synthase (iNOS) expression in cirrhotic patients with hepatopulmonary syndrome (HPS) was explored. Enrolled subjects were divided into three groups according to their disease/health conditions: the HPS group (cirrhotic patients with HPS; n=63), the non-HPS group (cirrhotic patients without HPS; n=182), and the control group (healthy subjects without liver disease; n=35). The distribution of the MPO -463 G/A genotype and its relationship with iNOS expression in a typical cell block from ascitic fluid were detected by immunohistochemistry and polymerase chain reaction-restricted fragment length polymorphism analysis (PCR-RFLP). In the HPS group, the partial pressure of oxygen in blood and ascitic fluid was significantly decreased (8.95+/-1.58 kPa and 6.81+/-0.95 kPa, respectively; both P<0.01), while the partial pressure of carbon dioxide significantly increased (4.62+/-0.20 kPa and 5.92+/-0.45 kPa, respectively; P<0.01). MPO and iNOS levels were significantly increased in the HPS group as compared with the non-HPS group. These increases were even more remarkable in ascitic fluid (41.36+/-11.62 and 13.23+/-4.81 mug/L; 10.27+/- 3.20 and 4.95+/-1.12 mug/L) than in blood (16.66+/-5.24 and 4.87+/-1.73 mug/L; 5.79+/-2.31 and 2.35+/-0.84 mug/L). The distribution of the MPO genotypes GG, GA, and AA were 76.2%, 22.2% and 1.6% in the HPS group, and 57.7%, 37.9% and 4.4% in the non-HPS group (P<0.05). The expression of iNOS was significantly higher in patients with the G alleles (G/G and G/A) (61.54%, 48/78) than in patients with A alleles (G/A and A/A) (38.46%, 30/78) (P<0.01). It was suggested that the expression levels of iNOS and MPO were correlated with HPS-induced hypoxemia. The MPO-463 G/A mutation might be a protective factor that prevents the development of HPS. The MPO might be involved in the regulation of iNOS expression. In humans, MPO pathways, the iNOS/NO system, and their interaction might have an impact on the occurrence and development of HPS.

Objective:To investigate the effectiveness of heart rate deceleration capacity (DC) measurement in predicting the car-diotoxicity of malignant tumor patients treated with epirubicin-based chemotherapy. Methods:The clinical medical records, including CK-MB and cTnI levels and dynamic electrocardiogram (ECG) parameters before and after each chemotherapy cycle, of 140 patients treated with epirubicin-based chemotherapy were analyzed. Patients were divided into the DC>4.5 ms group and the DC≤4.5 ms group based on the calculated DC values. The CK-MB and cTnI levels and the dynamic ECG parameters of the two groups were compared af-ter two and four cycles of chemotherapy. Results:Patients in the two groups exhibited no statistically significant difference in their rele-vant clinical and pathological data before receiving chemotherapy (P>0.05). However, after four cycles of chemotherapy, the DC≤4.5 ms group showed a significantly greater increase in serum CK-MB and cTnI concentrations over the pre-chemotherapy levels compared with the DC>4.5 ms group. After two and four cycles of chemotherapy, the DC≤4.5 ms group also exhibited a significantly greater in-crease in mean heart rate (beats/min) and supraventricular and ventricular arrhythmia counts (times/24 h) over the pre-chemotherapy values compared with the DC>4.5 ms group (P<0.05). After four cycles of chemotherapy, 23 cases showed abnormally elevated cTnI levels in the DC≤4.5 ms group. In this group, patients with elevated cTnI level exhibited no statistically significant difference in CK-MB and cTnI concentrations, mean heart rates, and supraventricular and ventricular arrhythmia counts compared with those with nor-mal cTnI level before chemotherapy (P>0.05). However, the DC values of patients with elevated cTnI were significantly lower than those with normal cTnI level (P<0.05). Conclusion:The risk of epirubicin-induced cardiotoxicity increased with decrease in DC value. The DC test was shown to be an effective predictor of the risk of epirubicin-induced cardiotoxicity.

Objective:To elucidate the characterization of antigen-specific memory T cells from PPD~+ individuals after stimulation with BCG in vitro.Methods:PBMCs were isolated from PPD~ -/+ normal human peripheral blood and stimulated with BCG. The level of IFN-? in the culture supernatants was assayed by ELISA and the frequency of IFN-?-producinging cells was detected by ELISPOT. The subsets and frequency of cytokine-producing cells were determined at a single cell level by flow cytometry.Results:After stimulation with BCG, PBMCs from PPD~+ but not PPD~- individuals produced significantly high levels of IFN-? in culture supernatants detected by ELISA(P

OBJECTIVE:To observe the clinical efficacy and safety of Bromfenac sodium eyed drops in the treatment of xe-rophthalmia. METHODS:80 patients with xerophthalmia were randomly divided into observation group and control group,with 40 cases in each group. Control group was given 0.1% Sodium hyaluronate,one drop,qd;observation group was given 0.1% Sodi-um hyaluronate eye drops,one drop,bid. Both groups received 14 d of treatment. The subjective symptom and sign,the time of lacrimal film break-up,fluorescent staining score and schemer test were observed in 2 groups before treatment,3,7,14 d after treatment. Clinical efficacy and the occurrence of ADR were observed in 2 groups. RESULTS:There was no statistical significance in subjective symptom and sign,the time of lacrimal film break-up,fluorescent staining score and tear between 2 groups before treatment,3,7 d after treatment(P>0.05). 14 d after treatment,subjective symptom and sign and fluorescent staining score of 2 groups were decreased significantly,and the time of lacrimal film break-up was prolonged and secretion was increased significant-ly;the observation group was significantly better than the control group,with statistical significance (P<0.05). The cure rate of observation group was 60.0%,which was significantly higher than that of control group(35.0%),with statistical significance(P<0.05). There were no statistical significance in ADR between 2 groups(P>0.05). CONCLUSIONS:Sodium hyaluronate eye drops is effective for xerophthalmia,and can relieve the symptoms with good safety.