BACKGROUND: Although cultured technique and purified method of Schwann cells (SCs) are changed successively,purity is still limited. Whether double negative selections can culture high-purified SCs still needs a further study.OBJECTIVE: To investigate the effects of double negative selections on growth and purity of SCs.DESIGN: Controlled animal study.SETTING: Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University,and the Beijing Institution of Otolaryngology.MATERIALS: The experiment was carried out in the Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University and the Beijing Institution of Otolaryngology between May 2006 and July 2006. Twenty new-born Wistar rats of 3 days (without gender limitation) were provided by the Experimental Animal Center of the Chinese Medical Academy.METHODS: The sciatic nerves of rats were digested by 0.2% collagenase. ① Grouping: All the rats were divided into 4 groups with 5 each. The experimental group was depurated by differential adhesion and Ara-c for 30 minutes. SCs in the control group 1 and 2 were managed by differential adhesion and Ara-c for 30 minutes, respectively. Cells in the control group 3 were not purified. ② Evaluation: The cell growth was observed under invert microscope; growth curves were measured with MTT method in the experimental group and control group 3 to observe cell proliferation. In addition, the SCs were identified by immunocytochemistry; the culture purity was calculated (ratio of S100 positive cells in each sight).MAIN OUTCOME MEASURES: ① Growth of SCs; ② immunocytochemistry stain of SCs; ③ growth curve of SCs; ④ purity of SCs.RESULTS: ① Growth of SCs: At 5 days after incubation, the SCs proliferated well. SCs were spindle, bipolar and sometimes tripolar, with obvious nuclei and a little cytoplasm. There was more fibroblast cell in the group 3 than that in experimental group. ② Growth curve: The SCs in experimental group entered into the logarithm multiplication period at the third day. ③ Immunocytochemistry: The positive-stained SCs were brown in its body and process, and arranged like swirl. The FB with spread and anomaly shape was slight blue. ④ The ratio of anti-S100 positive cells was higher in experimental group than that in group 3 (P ＜ 0.01).CONCLUSION: With the double negative selection, high-purified SCs are obtained effectively.

The status of anesthesia and psychotropic substances use for cancer pain control was analyzed in 355 cancer patients in Huanggu District of Shenyang City.The results revealed that patients aged 60 and above accounted for 62.8％ of the total; the 5 leading diseases were lung cancer ( 132 cases,37.2％ ),colorectal cancer (34 cases,9.6％ ),liver cancer (33 cases,9.3％ ),stomach cancer ( 31cases,8.7％ ) and pancreatic cancer (20 cases,5.6％ ).There was no significant difference in constituent ratio of disease categories in the last 4 years ( x2 =18.75,P ＞ 0.05 ).The daily oral dose of morphine sulphate was 60 - 200 mg with an effective rate of 91.5％ (325/355). The side effects including constipation,nausea,vomiting and itching can be effectively reduced by prophylactic medication.

Objective To investigate the role of IL-13 in the patients with acute and chronic urticaria. Methods In 22 patients with acute urticaria, 20 patients with chronic urticaria and 19 normal controls, the levels of IL-13, IL-4 and IFN-? of peripheral T lymphocytes were measured by flow cytometry. The serum concentrations of IL-13 and total IgE were tested by ELISA. Results The results of flow cytometry showed that the level of IL-13 of the patients with acute urticaria was significantly higher than that of the normal controls (P

Objective To explore the role of Annexin Ⅱ and urokinase-type plasminogen activator(uPA) proteins in the development and lymph node metastasis of thyroid cancer.Methods Immunohistochemistry was used to detect the expression of Annexin Ⅱ and uPA proteins in 35 cases of papillary thyroid cancer tissues,10 cases of thyroid adenoma tissues,16 cases of nodular goiter tissues and 7 cases of normal thyroid tissues.Results The positive expression rate of Annexin Ⅱ and uPA protein was significantly higher in papillary thyroid cancer than in normal thyroid tissues,nodular goiter tissues and thyroid adenoma tissues( P ＜0.05 ).The expression of Annexin Ⅱ and uPA protein was positively correlated in papillary thyroid cancer tissues(P ＜0.05).The expression of Annexin Ⅱ protein in thyroid cancer tissues was associated with tumor size and lymph node metastasis while the expression of uPA protein was only related to lymph node metastasis.Conclusions Annexin Ⅱ and uPA proteins are involved in the occurrence and development of papillary thyroid cancer.They also play a facilitating role in lymph node metastasis of papillary thyroid cancer.

The resting cells of Citrobacter freundii 48003 3 expressing high tyrosine phenol lyase activity under the inducing of L tyrosine were used for L DOPA synthesis from catechol, pyruvate and ammonia In this paper, the effects of temperature, pH and substrate concentrations on the synthesis of L DOPA were studied At the optimal conditions of reaction, 9 5g/L of L DOPA was obtained in 12h

OBJECTIVE:To investigate the satisfaction degree of outpatients on policy of separation of clinical pharmacy from 4 third grade hospitals in Shanghai,and provide reference for promoting medical reform policy. METHODS:Parts of outpatients in the 4 new suburb third grade hospitals were investigated by the questionnaire and interview in Shanghai city,and the data was sta-tistically analyzed by Excel software and SPSS 19.0 software. RESULTS:Totally 400 questionnaires were sent out and 368 were valid with effective recovery of 94.3%. The awareness rate of medical separation policy was 81.3% in the outpatients;55.5% of outpatients thought that medicine cost savings effect was not obvious;the overall satisfaction of patients was 91.6%,and the high-est of satisfaction was the service attitude and the lowest was medical expenses. CONCLUSIONS:Policy of separation of clinical pharmacy has reduced the medical expenses,guided the triage treatment of patients partly and improved the satisfaction of patients. It is suggested to shorten the waiting time by optimizing examination process,strengthen price regulation in drug,standardize pro-duction and distribution process,improve use rate of essential medicines and promote clinical rational drug use for further improve-ment of satisfaction.

OBJECTIVE:To systematically review the efficacy and safety of Magnesium isoglycyrrhizinate injection versus 4 comnon medicines in the treatment of drug-induced liver damage,and to provide evidence-based reference for clinic treatment. METHODS:Retrieved from PubMed,EMBase,Cochrane Library,CBM,CJFD,Wanfang Database and VIP Database,random-ized controlled trials (RCT) about Magnesium isoglycyrrhizinate injection versus other medicines in the treatment of drug-induced liver damage were enrolled. Meta-analysis was performed by using Rev Man 5.3 software after literature selection,data extract and quality assessment. RESULTS:A total of 13 RCTs were included,involving 1 093 patients. Results of Meta-analysis showed clini-cal effective in magnesium isoglycyrrhizinate group was significantly higher than tiopronin group[RD=0.29,95%CI(0.17,0.42), P<0.001] and diammonium glycyrrhizinate group [RD=0.07,95%CI(0.01,0.12),P=0.02],compared with glutathione group and compound ammonium glycyrrhetate group,there were no significant differences ;incidence of adverse reactions in magnesium iso-glycyrrhizinate group was significantly lower than diammonium glycyrrhizinate group [RD=-0.07,95%CI(-0.11,-0.03),P<0.001] and compound ammonium glycyrrhetate group[RD=-0.21,95%CI(-0.38,-0.04),P=0.02],compared with triopro-nin group and glutathione group,there were no significant differences among 3 groups. CONCLUSIONS:Magnesium isoglycyrrhiz-inate injection has better efficacy and safety than other 4 commons hepatoprotective medicines in the treatment of drug-induced liver damage. Due to the limit of methodological quality,more large-scale and long-term follow-up studies with strict designed are need-ed for the further verification of the conclusion.

A novel high performance liquid chromatographic method was developed for the determination of 4-O-methylhonokiol in rabbit plasma and was applied to its pharmacokinetic investigation.Plasma samples were treated by one-fold volume of methanol and acetonitrile to remove the interference proteins.A reverse phase column of SHIMPACK VP-ODS (150 mm× 4.6 mm,5.0 μm) was used to separate 4-O-methylhonokiol in the plasma samples.The detection limit of 4-O-methylhonokiol was 0.2 μg/L and the linear range was 0.012- 1.536 mg/L.The good extraction recoveries were obtained for the spiked samples (84.7％,89.3％ and 87.7％ for low,middle and high concentrations of added standards,respectively).The relative standard deviation of intra-day and inter-day precisions was in the range from 0.6％ to 13.5％.The pharmacokinetic study of 4-O-methylhonokiol was made and the results from the plasmaconcentration curve of 4-O-methylhonokiol showed a two-apartment open model.This work developed a sensitive,stable and rapid HPLC method for the determination of 4-O-methylhonokiol and the developed method has been successfully applied to a pharmacokinetic study of 4-O-methylhonokiol.

BACKGROUND:Whether determination of tacrolimus blood concentration by different immunoassay methods can influence predictive ability to immunosuppressive effects and toxicity, and whether it can be more sensitive to reflect blood concentration in patients with renal dysfunction are worthy of studying.
OBJECTIVE:To analyze the correlation of tacrolimus (FK506) concentrations determined by enzyme-multiplied immunoassay technique (EMIT) and enzyme linked immunosorbent assay (ELISA) in combination with renal function parameters.
METHODS:133 clinical blood samples were col ected. EMIT and ELISA techniques were used to determine the FK506 concentration. The correlation of two determination methods were analyzed, combined with renal function. RESULTS AND CONCLUSION:In patients with renal dysfunction, the mean results and standard deviation mensurated by ELISA were higher than those by EMIT. For blood concentration in 5-20μg/L by ELISA, the incidence of renal dysfunction occurred less than by EMIT. The overal mean results of blood concentration for two methods appeared no significant difference (r=0.904 5, P>0.05). When the concentration was less than 2.0μg/L, the concentration results by EMIT were higher than those by ELISA (P<0.01). When the concentration was more than 2.0μg/L, there was no significant difference between two determination methods (P>0.05). These findings indicate that EMIT and ELISA has good correlation, which are both suitable for clinical routine determination of plasma concentration. It is not recommended for applying EMIT method to determine low blood concentrations (<2.0μg/L). The reference range of concentration should be compartmentalized depending on combination of determination methods and renal function.

Background and purpose: The metastasis-associated in colon cancer-1 (MACC1) is highly expressed in different cancers and has an effect on the proliferation and apoptosis of tumor cells through the regulation of extracellular signal-regulated kinase (ERK)1/2 pathway. However, the role of MACC1 in ovarian cancer has been rarely studied. The study was aimed to suppress MACC1 gene expression by siRNA and explore the relationship between MACC1 expression and chemosensitivity to cisplatin in ovarian cancer cell line SKOV3/DDP. Methods:Empty plasmid p-super-EGFP-1 (negative control group) and p-super-EGFP-MACC1 shRNA (experimental group) were transfected into ovarian cancer cell SKOV3/DDP respectively. SKOV3/DDP cells without transfection were used as blank group. Then, MACC1 mRNA and protein levels were measured by RT-PCR and Western blot, respectively. Cell proliferation and IC50 of cisplatin was determined by methyl thiazolyl tetrazolium test (MTT). Apoptosis rate was determined by lfow cytometer (FCM). ERK1/2 and p-ERK1/2 protein levels were determined by Western blot. Results:Compared with those in blank and negative control groups, MACC1 mRNA and protein levels deceased in experimental group. The IC50 of cisplatin in experimental group was lower than that in the other groups (26.094 vs 47.501/47.089μmol/L, P<0.05). There was a lower expression of p-ERK1/2 in experimental group (0.3979 vs 00.6712/0.6681, P<0.05). Apoptosis rate was significantly higher in the experimental group before and after treatment of cisplatin (1.32%vs 0.66%/0.48%, P<0.05;36.70%vs 18.53%/16.60%, P=0.000). Conclusion:MACC1 gene may be involved in cisplatin resistance phenomenon in SKOV3/DDP cells through ERK1/2 pathway.