Objective:To explore the anti-fatigue effect of Tibet maca in mice. Methods: The mice were respectively given the powder or the alcohol extract of Tibet maca. The lactic acid concentration in blood, serum lactate dehydrogenase ( LDH) , the time of weight loading swimming and serum urea ammonia level after the exercise in the mice were detected, and the anti-fatigue effect of the powder and the alcohol extract of Tibet maca was compared. Results: After the 30-day feeding, the serum LDH activity of the mice taking the powder or alcohol extract of Tibet maca was obviously higher than that of the mice in the control group(P<0. 05), the time of weight loading swimming was significantly longer than that in the control group (P<0. 05), and the blood lactic acid concentration after the exercise was obviously lower than that in the control group (P<0. 05). Conclusion: Tibet maca can improve the time of weight loading swimming of mice, and reduce the level of serum urea ammonia after exercise and blood lactic acid concentration, sug-gesting the powder and alcohol extract of Tibet maca have obvious anti-fatigue effect.

BACKGROUND:The in vitro construction, maturation and differentiation of cellscaffold complexes into tissue-engineered bone is the necessary process of bone tissue engineering construction, but there are no uniform
methods and standards.
OBJECTIVE:To summarize the basic method and technology to build bone tissue engineering at present and to discuss the related development.
METHODS:A compute-based online search was conducted on the PubMed database and CNKI database for the articles related to the in vitro construction and culture of bone tissue engineering bone from January 1997 to
January 2013 with the key words of“bone tissue engineering, cellbiological scaffold, cellinoculation, seeding density, culture in vitro, bioreactor”in English and Chinese. Final y, 44 articles were included according to the inclusion and exclusion criteria.
RESULTS AND CONCLUSION:As the carrier of bone tissue engineering seed cells, the primary prerequisite of biological scaffold is sterile, because the sterile biological scaffold can be able to survive. Sterilization of biological scaffolds includes ultraviolet sterilization, 60Coγ-ray sterilization, soaking in ethanol with the volume fraction of 75%, autoclave method, and ethylene oxide sterilization. 60Coγ-ray sterilization is the common method in the biological scaffold sterilization. The inoculation density of seed cells is the key factors that influence the adhesion growth and proliferation of seed cells on the scaffolds. The adhesion between cells and scaffold materials wil be affected by the affinity of scaffolds, celladhesion and gravity, and other factors. The method for the inoculation of bone tissue engineering seed cells includes static inoculation and dynamic inoculation. Each construction method has its advantages and disadvantages. Overcomeing these disadvantages, forming a uniform construction method and ful y clinical application are the direction of future development.

Objective: To evaluate the quality of gentiana macrophylla from different origins in Gansu by determination the content of gentiopicroside.Methods: HPLC method was used.Results: The calibration curve of gentiopicroside was linear within 0.52-5.2?g(r=0.9991),the average recovery rate was 99.2%(RSD=1.23%,n=5).Conclusion: The content of gentiopicroside among gentiana macrophylla from different origins in Gansu are different,and all of them are higher than the standard of China Pharmacopoeia.Gentiopicroside content in planting gentiana macrophylla is higher than that of wild gentiana macrophylla.It provides a scientific basis of cultivating gentiana macrophylla and protecting wild gentiana macrophylla.

Objective:To assess the genetic relationship of clinical isolates of carbapenem-resistant A.baumannii (resistant to both imipenem and meropenem) from January 2007 to March 2008 in Peking University Third Hospital for measures to decrease the isolates; to investigate the characteristics of patients with carbapenem-resistant A. baumannii colonization or infection and to evaluate antibiotic treatment for health care-associated infections caused by carbapenem-resistant A. baumannii. Methods: The medical records of patients with carbapenem-resistant A. baumannii colonization or infection were reviewed. Antibiotic susceptibilities of the isolates were determined by the standardized disk-diffusion method and the clonal relationship of the isolates was analyzed by pulsed-field gel electrophoresis. Results: A total of 49 carbapenem-resistant A. baumannii strains were isolated from the 49 patients hospitalized during the study period and pulsed-field gel electrophoresis typing yielded 7 different patterns. A total of 45 (91.8%)genotyped strains showed clonal relationship. The most frequently identified predisposing factors were intensive care unit stay, invasive procedures, and hypoalbuminemia. Chronic obstructive pulmonary disease (12 cases) and cerebrovascular disease (10 cases) were the most common comorbid conditions.The mortality of patients with carbapenem-resistant A. baumannii infection was 38. 1% (8 of 21 patients), and the acute physiology and chronic health evaluation Ⅱ score, initial antibiotic therapy failure rate and the presence of hypoalbuminemia were significantly increased in the death group. Combination therapy regimens had higher success rates than monotherapy regimens (11/13, 84. 6% vs. 3/17,17.6%). Conclusion: There has been clonal spread of carbapenem-resistant A. baumannii strains among patients in our hospital since 2007. Intensive care unit stay, invasive procedures, hypoalbuminemia, chronic obstructive pulmonary disease and cerebrovascular disease were common in patients with carbapenem-resistant A. baumannii colonization or infection. Antibiotic combination therapy may be effective for carbapenem-resistant A. baumannii infection.

[Abstract] Objective To investigate the neuroprotection of progesterone on neonatal rats after sevoflurane inhalation anesthesia and its mechanisms. Methods A total of 120 newborn Sprague-Dawley rats were randomly divided into three groups (n=40): blank control group (group C), sevoflurane group (group S) and progesterone plus sevoflurane group (group S+P), half male and half female in each group. The rats in group S were exposed to 3% sevoflurane for two hours on postnatal days (P) seven, eight and nine, which was used to establish the developmental sevoflurane neurotoxicity model. The rats in group C were exposed to mixture of gases (2 L/min, 2 hours a day). The rats in group S+P received a daily injection of progesterone (8 mg/kg) from P4 to P9 and then were exposed to 3% sevoflurane (2 L/min, 2 hours a day) for 3 consecutive days between P7 to P9. The apoptosis of nerve cells in the CA1 area of the hippocampus evaluated by TUNEL assays in neonatal rats. The relative expression of apoptosis protein (caspase-3) in the hippocampus determined by Western blotting. Rats in each group evaluated for the space orientation ability and the learning and memory ability by Y maze, Morris water maze and platform test 6 weeks after birth. Results Sevoflurane significantly increased the neuronal apoptosis in CA1 area of the hippocampus in the central nervous system of newborn rats and increased the expression of Caspase-3 in the hippocampus (P<0.01); progesterone significantly reduces neuronal apoptosis which induced by sevoflurane and the expression of Caspase-3 in the hippocampus (P<0.01). The results of the Y maze, Morris water maze and platform test showed that sevoflurane reduced the alternating scoring rate of rats (P<0.05), prolonged the time required to find the platform in the water maze (P<0.05) and increased the number of errors in the platform test (P<0.01). Progesterone significantly increased the alternating scoring rate of rats (P<0.05), shortened the time required to find the platform in the water maze (P<0.05) and significantly reduced the number of errors in the platform test (P<0.01). Conclusion Repeated inhalation of 3% sevoflurane in neonatal rats can cause neurotoxic damage and induce cognitive dysfunction. Progesterone may have a neuroprotective effect on the neurotoxic damage of neonatal rats induced by sevoflurane.

Objective To evaluate the migrating regulation and ultrastructure of the corneal epithelial stem cells in human fetuses. Methods We examined the corneal cryosections of 14-38 weeks of gestation. Hematoxylin-eosin staining showed the stratified corneal epithelium and the corneal epithelial stem cells were localized by mouse monocolonal antibody against human 64-kilodalton keratin (mAE5), and the ultrastructure of the corneal epithelial stem cells was observed. Results At 14 weeks of gestation, the corneal epithelium was composed of a single basal cells layer and 1-2 superficial squamous cells layers. Some superficial squamous cells were mAE5 positive in the limbus as well as the central and peripheral cornea. At 17-29 weeks of gestation, the limbus epithelium developed from 3 to 5 cells layers and the central region from 2 to 3 cells layers. mAE5 positive cells were found in the suprabasal layers of all 3 regions examined but not in the basal layer. At 33-38 weeks of gestation, the corneal epithelium consisting of 4-6 cells layers was morphologically mature. mAE5 immunoreaction showed the negative cells were confined to limbus basal layer. The ultrastructure of basal layer cells showed they had more heterochromatin in the nucleus, less organells in the cytoplasm and less desmosomes among them. Conclusion The migration of corneal epithelial stem cells in the human fetuses was from the whole layers to basal layer and confined to limbus region finally, and their ultrastructure was immature.

Objective To study the production of ?-lactamase in multidrug-resistant Pseudomonas aeruginosa and to guide the proper use of antibiotics in clinical practice. Methods The modified three-dimensional extract test was employed to detect ?-lactamase in 30 multidrug-resistant Pseudomonas aeruginosa strains screened from antimicrobial susceptibility test in our hospital,and isoelectric focusing electrophoresis was performed on the enzyme-producing strains. Results No metalloenzyme was detected in all the 30 strains.Twenty-six strains produced ?-lactamase,among which 25 continuously yielded large amount of AmpC enzyme and the other one both AmpC enzyme and extended-spectrum ?-lactamases(ESBLs).86.7% of multidrug-resistant Pseudomonas aeruginosa produced enzyme. ConclusionThe majority of the multidrug-resistant Pseudomonas aeruginosa in our hospital yielded large amount of AmpC enzyme in a continuous way.The modified three-dimensional extract test can eliminate some interference such as the decrease of outer membrane permeability and overexpression of efflux pump,facilitating the effective and accurate detection of ESBLs in Pseudomonas aeruginosa.

Background The orbital fibroblasts (OFs) in thyroid associated ophthalmopathy (TAO) play important roles in the proliferative and inflammatory response.Seeking the drug which inhibit OFs growth is of a vital significance for the prevention and treatment of TAO.Research documented that colchicine has an anti-fibrosis effect.But its influence on OFs of TAO patient is few known.Objective This study was to investigate the effect of colchicine on growth and apoptosis of OFs in vitro.Methods The retroobital connective tissue was obtained form 3 TAO patients and cultured using explant method.OFs were passaged and identified by immunochemistry,and 3-8 genetaions of cells were used in the study.Colchicine at the concentrations of 1 × 10-8,1 × 10-7,1 × 10-6,1 × 10-5,1 ×10-4 mol/L was added into the RPMI 1640 with 10％ fetal bovine serum(FBS) to incubated the cells for 24,48 and 72 hours respectively,and only RPMI 1640 was used to culture the cells as the control group.Cell counting kit-8 (CCK-8)was used to detect the absorbance value (A450) of OFs for the evaluatuion of OFs and the inhibitory rate of colchicine to OFs.The colchicine of 1 ×10-6,1 ×10-5,1 × 10-4 mol/L was added into the culture medium for 48 hours,and then the apoptotic rate of the cells and the cell percentage in various cellular cycle was assayed by flow cytometry(FCM).The expression of transforming growth factor-β (TGF-β)in the cells was detected by immunochemistry to assess the influence of colchicine on the serection of the cells.Results Cultured cells showed the spindle-like in shape and the cell number was significantly increased with the incubation time.After incubated with 1 × 10-4,1 × 10 5,1 × 10-6,1 ×10-7,1 × 10-8 mol/L colchicines,the A450 values were gradually reduced with the increase of the concentrations of colchicine(F ion =62.004,P＜0.05),and significant differences were found between different contrations of colchicine groups(all P＜0.05).Aslo,gradually declined A450 values of the cells were seen with the lapse of culture time among the groups(Ftime =459.582,P＜0.05).The inbitory rate of colchicine to the cells was elevated with the increase of concentrations.The apoptotic rates of the cells were (1.73 ± 0.15) ％,(21.04 ± 4.56) ％,(31.84 ±6.21)％and(35.32±5.56)％ in the control group and 1 × 10-6,1 × 10-5,1 × 10 4 mol/L colchicine groups respectively,with statistically significant difference among the 4 groups (F =83.905,P＜0.05).With the increase of concentrations of colchicines,the cell percentage in G2 +M phase lessened gradually,showing significant difference among the control group and the 1 × 10-6,1 × 10-5,1 × 10-4 mol/L colchicine groups (F =20.443,P＜0.05).The expression of the TGF-β in the cells was (97.60± 2.09) ％ in the control group,and that in the 1 × 10-4 mol/L colchicine group was (44.43 ± 3.96) ％,presenting a significant difference between them (t =65.330,P ＜ 0.05).Conclusions Colchicine can induce apoptosis of OFs and inhibit the prolilferation of OFs in a time-and dose-dependent manner probably by decreasing the TGF-β secretion

Audiometry, Speech ; Humans ; Language

Objective To evaluate the effects of sevoflurane preconditioning and postconditioning on the activity of NF-κB during lung ischemia-reperfusion (I/R) in rats.Methods Fifty-four adult male Sprague-Dawley rats,weighing 300-350 g,were randomly divided into 3 groups (n =18 each) using a random number table:sham operation group (S group),I/R group and sevoflurane preconditioning and postconditioning group (SPP group).Lung I/R was induced by clamping the left pulmonary hilum for 45 min followed by 120 min reperfusion in I/R and SP groups.The left pulmonary hilum was only isolated but not ligated in group S.In group SP,2.1％ sevoflurane was inhaled for 30 min followed by 10 min washout,the left pulmonary hilum was then ligated for 45 min,and 2.1％ sevoflurane was inhaled for 30 min again starting from onset of reperfusion.Six rats were chosen at 30,60 and 120 min of reperfusion and sacrificed in each group,lungs were removed for determination of wet/dry lung weight (W/D) ratio and content of tumor necrosis factor-α (TNF-α) (by ELISA) and expression of nuclear transcription factor κB (NF-κB) p65 of nuclear protein (by Western blot) and for microscopic examination of the pathologic changes of lungs (with light microscope).Results Compared with S group,W/D ratio,TNF-α content and nuclear protein NF-κB p65 expression in lung tissues were significantly increased at each time point of reperfusion in I/R and SPP groups.Compared with I/R group,W/D ratio,TNF-α content and nuclear protein NF-κB p65 expression in lung tissues were significantly increased at each time point of reperfusion,and the pathological changes of lungs were attenuated in SPP group.Conclusion Sevoflurane preconditioning combined with postconditioning can reduce the lung I/R injury in rats through inhibiting NF-κB activity in lung tissues.