BACKGROUND: Studies have shown that 160 mg/L tramethylpyrazine (TMP) can inhibit the proliferation of endothelial cells cultured in vitro,but whether TMP also inhibits vascular endothelial growth factor (VEGF)induced proliferation of vascular endothelial cells remains unkown. OBJECTIVE: To study the effect of rat serum containing TMP on vascular endothelial cell proliferation and its proliferation induced by VEGF. DESIGN: Randomized controlled experiment. SETTING: College of Traditional Chinese Medicine, Chongqing University of Medical Sciences. MATERIALS: The experiment was carried out in the Institute of Pediatrics, Chongqing University of Medical Sciences from March, 2002 to March, 2003 using human umbilical vein endothelial cell (HUVEC) line ECV304 and 30 female Wistar rats. METHODS: Thirty rats were randomly divided into 3 equal groups to receive intraperitoneal TMP injection at 143.0 mg/kg and 71.5 mg/kg and 0.8 mL normal saline (blank control group). All the injections were performed once a day for consecutive 7 days, then blood was collected from the intraperitoneal artery and diluted to 20％, 10％, and 5％ in triplicate.Effect of TMP-containing serum on the proliferation of ECV304 cells was observed by means of in vitro culture and 3H-TdR incorporation as well as methyl thiazolyl tetrazolium (MTT) methods. MAIN OUTCOME MEASURES: ① Effect of TMP-containing serum on proliferation of ECV304 cells in in vitro culture and VEGF-induced proliferation of the cells.RESULTS:All the 30 rats survived the experiment without losses. The absorbance (A) and scintillation counting (minute-1) of the cells were significantly lower in 143.0 mg/kg TMP group than those in the control group treated with rat serum at the dilution of 5％ (0.720±0.024 vs 0.816±0.068,3340.45±567.7 vs 5120.84±301.49), 10％ (0.630±0.017 vs 0.798±0.015,2430.06±265.98 vs 5225.83±100.10), and 20％ (0.765±0.027 vs 0.823±0.031,3570.45±130.52 vs 5256.82±183.18), and those in 71.5 mg/kg TMP group were also significantly lower than those in the control group after treatment with the serum of 10％ (0.775±0.023, 4571.14±275.39) and 20％(0.749±0.012, 3287.25±144.82). In the experiment evaluating the effect of TMP serum on VEGF-induced proliferation of ECV304 cells, the A value and scintillation counting in the 143.0 mg/kg TMP group were significantly lower than those in the control group after the cells were treated with the serum of 5％ (0.726 ±0.004 vs 0.964 ±0.004, 5760.46 ±49.64 vs 9821.82±128.05), 10％ (0.712±0.004 vs 0.933±0.014, 5024.48±100.57 vs 9052.76±65.19), and 20％ (0.717±0.003 vs 0.924±0.004, 5405.45±140.90vs 9197.07±169.92], and those of the cells treated with 71.5 mg/kg TMP serum of 10％ and 20％ were also significantly lower than those in the control group (0.703±0.005 and 7526.47±169.21 for 10％ serum, and 0.693±0.006 and 5720.09±279.03 for 20％ serum). CONCLUSION: The sera at the concentrations of 5％,10％, and 20％from rats treated with 143 mg/kg TMP and at higher concentrations from 71.5 mg/kg TMP-treated rats can inhibit the proliferation of ECV 304 in in vitro culture as well as VEGF-induced proliferation of the cells.

Objective: To improve the diagnosis and treatment level of secretory otitis media (SOM) of children. Method :Clinical information of 80 children with SOM were colletcted and analysed. All of the 80 cases (144 ears)were treated by myringotomy with Grommet insertion. Among them, 30 cases by adenotonsillectomy. 10 cases by palatoplasty. Secretions of 54 ears were carded out by bacterial culture. Result:80 cases were followed up for 6 to 12 mothes. 120 ears of 144 were cured (83％) ,with hearing improved. The tympanograms and hearing threshold of pure tone were found to be normal. The Gram＇s positive and negative germs of bactevial culture were 20.7％ (6/29 samples ) and 24％ (6/25 samples ) respectively. Conclusion: Children with secretory otitis media should be treated as early as posible to improve the patients＇ hearing and develope their speech.

Thirty-one children suffered from speech retardation were found to have normal hearing level through ABR and/or acoustic immittance tests. Some of them were further checked by psychiatrists and pediatricians. The results demonstrate that among reasons for speech retardation in children,autism and mental retardation are important factors that can not be ignored. A-mong children with unclear diagnosis .some harmful factors were found on themselves and in surroundings. We emphasize that for children with speech retardation,not only hearing disorders,but also other factors should be considered.

Objective To study the relationship between cleft palate and secretory otitis media(SOM)and the change of hearing of cleft palate patients with SOM before and after tube insertion.Methods 203 cleft palate patients(406 ears)were investigated by pure tone audiometry,acoustic immitance audiometry and auditory brainstem response(ABR).21 cleft palate patients with SOM underwent palatoplasty and myringotomy with tube insertion at the same time.Results The incidence of middle ear disfunction was 68.5% in 203 cases with cleft palate,which in the cases under 6 years the incidence was 80%.After operation,21 cases were followed up one to three years,their average hearing improvement was 17dB by tube insertion( P

Objective To Clone,express,purify 38 ku protein of mycobacterium tuberculosis,to study its immunological characteristics,and to evaluate its potenial value for serodiagnosis of tuberculosis.Methods Extract DNA from standard strain of H37Rv as the template,amplify gene of 38 ku protein by PCR,insert to PET-30a(+)and construct the recombinant plasmid,express 38 ku protein in E.coli BL21(DE3),purify by Nickel affinity chromatography,at last get target proteins of higher purity,analyze its immol/Lunological characteristics by Western blotting and ELISA technology from Feb.2003 to Mar.2004. Results The clone was analyzed at the nucleotide lever and showed the same DNA sequence coding for natural 38 ku protein. The recombinant protein expressed in inclusion body in E.coli BL21(DE3). The purity of terget protein was 92.7% by Nickel affinity chromatography,Western blotting assays showed that the recombinant protein had satisfactory antigenicity . 38 ku protein detected TB postive and negative refference serum based on the mechanism of indirect ELISA,results showed that the sensitivity was 80.5%(33/41)and the specificity reached to 96%(25/26).Conclusion The recombinant protein expressed in inclusion body in E.coli BL21(DE3)and had satisfactory antigenicity,and might be selected as one of serodiagnostic antigen of tuberculosis.

OBJECTIVE:To study the effect of ligustrazine injection(LTZ)on DNA synthesis of vascular endothelial cell.METHODS:In vitro cultured cell line of human umbilical vein endothelial cells,ECV304,were adopted to measure the ef?fects of LTZ on DNA synthesis by 3 H-TdR incorporation.RESULTS:LTZ could inhibit DNA synthesis of ECV304in a dose-dependent manner.CONCLUSION:The mechanism of anti-angiogenesis of LTS might be associated with inhibiting DNA synthesis of vascular endothelial cell.

OBJECTIVE To investigate the antibiotic use during perioperative period in surgery after administration according to Antibiotic Drug Use Guideline.METHODS The antibiotics usage status among 462 outpatient cases in Mar 2005 and 483 cases in Mar 2006 during perioperative period in surgery were surveyed.RESULTS The preventive use rate of antibiotic medication in surgery in 2005 and 2006 were respectively 99.48% and 88.12%(P

0.05).The transfection efficiencies were improved significantly in the ultrasound plus contrast agent and VEGF plasmid group(MI 1.2,1.4,and 1.6) compared with pure plasmid group(P0.05);there were significant differences between ultrasound plus contrast agent and VEGF165 plasmid(MI 1.4,1.6) and ultrasound plus contrast agent and VEGF165 plasmid group(MI 1.2)(P

OBJECTIVE To offer scientific basis for reasonable usage and management of antibiotics through surveying the situation and problems of antibiotics usage in surgery during perioperative period.METHODS The antibiotics usage status among 462 cases of patients during perioperative period was retrospectively analyzed.RESULTS Antibiotics were used in 460 patients.The rate of prophylactic usage was 83.26%,the rate of therapeutic one was 16.74%;the duration of postoperative prophylaxis was:80.68% of the patients were treated with antibiotics for 4 days or more,the longest one was 28 days;43.26% of the patients received single antibiotic treatment,and 44.13%,11.96%,or 0.65% of patients received 2,3 or 4 kinds of antibiotics combined therapy respectively.CONCLUSIONS Improper application of antibiotics during perioperative period needs higher level administration.

Objective The mononuclear cells(MNCs) were cultivated and expanded into mesenchymal stem cells(MSCs) from human umbilical cord blood,and the purpose of this study was to explore the biological characteristics and induced differentiation ability in vitro.Methods Human umbilical cord blood samples were obtained and the mononuclear cells were isolated from it,then inoculated the MNCs into 25-mm culture flasks containing DMEM/F12 medium.The morphology was observed under microscope.Nissl body staining was used,The passage 2,4,7 of the expanded MSCs were induced to differentiate to neuron-like cells.The expressions of nestin and neuron-specific enolase (NSE) on the treated cells were detected by immunocytochemical method.Results Nissl body staining was positive;Nestin expression was found in(51.2?3.2)% of the second,(34.6?2.7)% of the fifth,(11.3?3.3)% of the seventh passage of MSCs;NSE expression was found in(11.4?2.3)% of the second,(21.78?3.1)% of the fifth,(40.7?3.4)% of the seventh passage of MSCs.Conclusion Cord bloodMSCs possess some features of neural stem cells,and have the capacity to differentiate into neuron-like cells under proper conditions.