Objective: To investigate the effects of bone marrow stromal cells (BMSCs) isolated from leukemia patients on the invasion capacity leukemia SHI-1 cells in vitro and the underlying mechanism. Methods: BMSCs were isolated from leukemia patients and their conditional culture medium were collected. The expressions of extracellular matrix metalloproteinase inducer (EMMPRIN) in SHI-1 cells and BMSC were detected by RT-PCR. The BMSC and SHI-1 cells were mixed at 1: 10 and inoculated in Matrigel-coated transwells. Then CXC chemokine receptor 4 (CXCR4, final concentration of 2 μg/mL) or functional antibody of EMMPRIN were added. The BMSC cultured medium was used as control on cell invasion test. The alteration of matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinase 2 (TIMP-2), and CXCR4 mRNA were determined by real-time PCR before and after co-culture of SHI-1 cells and BMSC. The content of stromal cell-derived factor 1 (SDF-1) in serum-free supernatent was measured by ELISA. Results: Both SHI-1 and BMSC expressed EMMPRIN. The invasion capacity of SHI-1 cells increased significantly after co-culture with BMSC, which could be blocked by CXCR4 and functional antibody of EMMPRIN. The cultured medium of BMSC did not increase the invasion capacity of SHI-1 cells. The mRNA expression levels of MMP-2, MMP-9, TIMP-2, and CXCR4 as well as SDF-1 contents in serum-free supernatent increased significantly after the SHI-1 cells were co-cultured with BMSC. Conclusion: When BMSC islated from leukemia patients contacted with leukemia SHI-1 cells, they increases the invasion capacity of SHI-1 cells through multiple molecule pathways on the surface of them on cell invasion test. It may be an important mechanism responsible for the invasion of leukemia cells to the outer space of bone marrow.

Objective To observe the cervical regeneration outcome after different loop electrosurgical excision procedures(LEEP) for the management of cervical lesions.Methods A total of 209 patients with cervical lesions including cervical epithelial neoplasia,cervical HPV infection,cervical polyp and condyloma,and severe cervicitis were performed LEEP in our hospital.The types of LEEP included shallow ring excision,deep ring excision,LEEP conization,and that similar to cold-knife conization.The width and depth of removed cervical tissues were recorded,and the cervical regeneration was observed during the follow-up. Results Among the 209 cases,179(85.6%) were satisfactory cervical regeneration,24(11.5%) were little satisfactory cervical regeneration,and 6(2.9%) were unsatisfactory cervical regeneration.Shallow ring excision,deep ring excision and cone excision had higher satisfactory situation.Extroversion of cervical columnar epithelium was observed in 15 cases(7.2%) in little satisfactory cervical regeneration.Severe extroversion of cervical columnar epithelium of 3 cases occurred in unsatisfactory cervical regeneration.Besides,there were 2 case of cervical shortening and 1 case of severe erythema in unsatisfactory cervical regenerations. Conclusion Although having such complications as extroversion of cervical columnar epithelium,cervical shortening and erythema,LEEP performs a high satisfactory cervical regeneration after management of cervical lesions.It is in great need to analyze the condition of different patients in managing cervical lesions by LEEP.

Objective To explore the underlying mechanism of potential effect of sildenafil on porcine pulmonary artery smooth muscle cells (SMCs) proliferation induced by serotonin. Methods Porcine pulmonary artery SMCs at 3-5 passages were randomly divided into 4 groups: control group (CON), 5-HT group (HT), sildenafil group (SIL) and sildenfil-5HT combined group (S-HT). Pulmonary artery SMCs at exponential growth phase were serum starved with 0.2% FBS for 72 h, followed by sterile PBS, serotonin (10 μmol/L) and sildenafil (1 μmol/L) incubation in CON group, HT group and SIL group, respectively. In combined group (S-HT): pulmonary artery SMCs were serum starved and then exposed to sildenafil for 20 min, followed by serotonin incubation for indicated time. The phosphorylation of extracellular signal regulated kinase (ERK1/ERK2) was measured by Western blot at indicated time points. Flow active cell sorting (FACS) was used to evaluate the ratio of S phase cells in the cell cycle after 24 h of treatment. MTT color metric method was used to analyze SMCs proliferative index after 72 h of treatment. Results Compared with CON group, the phosphorylation of ERK1/ERK2, the percentage of cells in S phase, and the cell proliferation index increased remarkably after incubation with 5-HT (P<0.05). Preincubation with sildenafil followed by serotonin enhanced the phosphorylation of ERK1/ERK2 (p-ERK1/ERK2) and further elevated the percentage of cells in S phase and the cell proliferation index compared with that of HT group. While the total ERK1/ERK2 (t-ERK1/ERK2) was not statistically different among these groups. Conclusions Sildenafil potentiates the proliferative effect of serotonin on pulmonary arterial SMCs via upregulating phosphorylation of ERK1/ERK2.

Objective　To monitor KBD prevalence rate in Changdu of Tibet.Methods　The aged 7～12 year children are tested with X- ray and epidemiological investigation.Results　Xizang are still high yet,the 3 of 4 porints X-rates are more than 20% and the highest one is 55.34%.Conclusions　The Changdu is the severest point of KBD in our country.

Objective To construct the recombinant plasmid pCMV-Myc-PIASX? and express the fusion protein in mammalian cells.Methods PIASx? fragment was digested from the original vector pGADT7 with SalⅠand NotⅠ,and then was inserted into the targeted pCMV-Myc vector by the recombinant DNA technique.After identification,the recombinant plasmid was transfected into CHO cells.The expression of recombinant Myc-PIASx? fusion protein was detected by Western blotting.Results By the restriction enzyme digestion,fragment purification,ligation and transformation,the recombinant plasmid was obtained.The right recombinant plasmid pCMV-Myc-PIAS3 was identified with enzyme digestion and sequencing.By EcoRⅠ digestion analysis,pCMV-Myc-PIASx? showed a 5641 bp band.By XbaⅠdigestion analysis,pCMV-Myc-PIASx? showed two expected band of 3291 bp and 2349 bp.A specific protein expression band at 68 000(PIASx? fusion protein) was showed in Western blotting,which matched recombinant plasmid.Conclusion The recombinant plasmid of pCMV-Myc-PIASx? is sucssesefully constructed,which provids a good tool for further function study on PIAS family.

OBJECTIVE:To establish the quality standard of Fructus arctii concentrated granules. METHODS:TLC was con-ducted for the qualitative identification and HPLC was used for the content determination of arctiin in F. arctii concentrated gran-ules. The determination was performed on Dionex C18 column with mobile phase consisted of methanol-water(40∶60,V/V) at the flow rate of 1.0 ml/min. The detection wavelength was set at 280 nm,and the column temperature was 30 ℃. The samples size was 10 μl. RESULTS:TLC spectrum showed that the spots of F. arctii concentrated granules and arctiin reference medicinal material had the same color;the linear range of arctiin was 0.5-12.5 μg(r=0.999 8)with the average recovery of 98.41%(RSD=0.81%, n=9). The RSDs of precision,stability,repeatability tests were no more than 1.0%. CONCLUSIONS:The method is feasible and reproducible,and can effectively control the quality of F. arctii concentrated granules.

ObjectiveTo evaluate the reliability and clinical value for detecting paraquat (PQ)concentration in serum by spectrometry. MethodsThe determinations of wave length for detecting serum PQ concentration by ordinary spectrometry and second-derivative spectrometry were carried out. When the second-derivative spectrometry was used for detecting PQ in serum, the linear range and precision for PQ concentration were well defined. The results of serum PQ concentration determined by second-derivative spectrometry and by HPLC (high performance liquid chromatography) were compared in 8 patient with PQ poisoning. A total of 21 patients with acute poisoning after PQ ingestion over 4 hours admitted from October 2008 through September 2010 were retrospectively studied. Patients were divided into two groups as per the serum concentrations more than 1.8 μg/mL or less than that by second-derivative spectrometry on the day of admission. The severity of clinical manifestations between two groups was analyzed with t-test or Fisher's exact probabilities analysis. Results ( 1 ) The absorption peak of 257 nm could not be found by using ordinary spectrometry to detect the PQ concentration in serum. (2) The calibration curve in the 0. 4 ～ 8.0μg/mL range for detecting PQ concentration by second-derivative spectrometry observed the Beer's law (r =0. 996) . The average retrieval rate of PQ was within the range of 95.0％～ 99. 5％ with relative standard deviation (RSD) within 1.35％～ 5.41％ ( n = 6), and the lowest detection limit was 0. 05μg/mL. (3) The results of PQ concentrations from 8 patients with PQ poisoning detected by second-derivative spectrometry were consistent with those of the quantitative determinations by HPLC ( r = 0. 995,P＜0. 01 ) . (4) The survival rate of patients with serum PQ concentration more than 1.8 μg/mL was 22. 2％ ,and the incidences of acidosis, oligouria and pneumomediastium in these patients were 55.6％,55. 6％ and 77.8％, respectively. These clinical manifestations were significantly different from those in patients with serum PQ concentration less than 1.8 μg/mL ( P ＜ 0. 05 ) . Conclusions ( 1 ) It was inappropriate to take 257 nm as the determination wave length for detecting serum PQ concentration by ordinary spectrometry. (2) The method of second-derivative spectrometry was reliable for detecting serum PQ concentration. (3) Serum PQ concentration detected by second derivative spectrometry could be used to predict the severity of clinical manifestations of patients with PQ poisoning and was an important predictive factor for poor prognosis if the serum PQ concentration more than 1.8 μg/mL after PQ ingestion over 4 hours.

ObjectiveTo explore the influences of age and duration of status convulsivus （SC） on mitochondrial membrane potential (△Ψm） in hippocampus. MethodsConvulsive seizures for 30 min or 3 h (30 min SC or 3 h SC) were induced in 80 infant (20 d after birth) and 80 adult Wistar rats (IRs ＆ ARs respectively) with lithium-pilocarpine ip. The rats were sacrificed at 6 different time points from the 3rd hour to 7th day after SC termination. The mitochondrial △Ψm in hippocampal cells was determined with flow cytometry. ResultsThe mitochondrial △Ψm in hippocampal cells started to decrease at the 3th hour after SC in both IRs and ARs. The bottom level was reached at the 6th hour after SC ［(6.08±0.43) in IRs and (5.70±0.63) in ARs ) ］. Both of them were significantly lower than that of control group (P<0.01） and began to increase at 12th hour after SC. On the 7th day after 30 minutes SC, the level of mitochondrial △Ψm in IRs increased to the level of control, while the level in ARs was still lower than that of control (P<0.05）. At the 3rd hour, the 3rd and the 7th day after SC, the levels of mitochondrial △Ψm in IRs were obviously higher than those in ARs. Compared with the same time point after 30 min SC, the levels of mitochondrial △Ψm at the 3rd and the 6th hour after 3 h SC were much lower in different age groups (P<0.05）. Except the effect of the age-related difference, there was a positive correlation between the duration of SC and the changes of mitochondrial △Ψm in partial correlation analysis （r=0.71，P<0.05）. ConclusionSevere seizure could induce the mitochondrial △Ψm decreased in hippocampus. Age and duration of SC were important factors associated with the mitochondrial △Ψm decrease. There may be an internal protective response against brain damage in premature brain.

Acoustic Stimulation ; Animals ; Corpus Striatum ; physiology ; Estrogens ; metabolism ; Estrus ; physiology ; Evoked Potentials, Auditory ; physiology ; Female ; Rats ; Rats, Sprague-Dawley

Anemia, Refractory ; Humans ; Male ; Middle Aged ; Thrombocytosis