Objective To investigate the relationship between PrfA-dependent promoters and PrfA regulation. Methods LacZ reporter gene fusions used to investigate the inhibitory elements for PrfA-dependent transcription were carried on two promoters of Listeria monocytogenes: a PrfA-dependent promoter of the phospholipase gene pica (PplcA) and a putative promoter of the aroA gene (ParoA2) which contains a similar PrfA-binding site (PrfA-box) and a similar-10 box as PplcA but does not function as PrfA-dependent promoter. A series of hybrid plcA-aroA promoters by exchanging corresponding sequence elements of these two "promoters" were constructed and incorporated into upstream of a promoterless lacZ gene. The variant promoter-lacZ transcriptional fusions were then electroporated into L. monocytogenes wild-type strain P14, prfA mutant P14a and prfA deletion mutant A42, respectively. The expression level of PrfA is the highest in the P14a and the lowest in A42. The corresponding transcription activities of hybrid promoters were measured by the β-galactosidase assay. Results The two critical elements of PrfA-dependent promoters, the PrfA-box and the-10 box, can be functionally exchanged as long as the distance in between is maintained 22 or 23 bp. However, the interspace sequence and the sequence downstream of the -10 box of ParoA2 were strongly inhibitory for PrfA-dependent transcription. Conclusion Downstream sequence together -10 box of ParoA2 might fold into a hairpin structure when present in a single stranded DNA and possibly block the formation of the transcriptional initiation open complex, hence, inhibit the PrfA-dependent transcription from ParoA2.

Sufficient embryos are needed for the preservation of Beagle dogs germplasm resources and the prepara-tion of gene?modified human disease animal models. Up to now, the induced ovulation technique has no effect on dogs,it is hard to obtain mature oocytes in vivo, although the scientists try a lot in many aspects, but still could not make a break?through. The in vitro maturation rate is too low to support the preservation of germplasm resources, application in gene?modified disease models and biomedical research. Aiming to provide useful information on breakthrough in dog oocytes mat?uration, this review will summarize the effect of different age and reproductive stage,different morphology and size of the oo?cytes and lipid droplet on the in vitro maturation of dog oocytes.

After a description of the demand for medical information in grass-root PLA health units, the experiences of Medical Library of Chinese PLA in providing medical information service for grass-root PLA health units were summarized, and suggestions were proposed for medical library and information institutions to provide information service for them.

Objective To study the effects of 8-methoxypsoralen (8-MOP) on a ctivation, proliferation and melanin synthesis of hair-follicle melanocytes. Me thods The cultured human hair follicle melanocytes were treated with various con centrations of 8-MOP(10～500 ?mol/L) in vitro. The effects of 8-MOP were obse rved on cellular morphologic changes, proliferation, tyrosinase activity and mel anin contents of cultured melanocytes. Results Seven days after treatment with 8-MOP(50 ?mol/L), striking changes were found in cellular morphology with many elongated dendrites, and deposition of brown granules in cytoplasm. Tyrosinase a ctivity and melanin contents also remarkably increased in treated cells compared to those in untreated cells(P

BACKGROUND: Osthol is a simple coumarin from Cnidium monnier (L.)Cusson which has been long used of in China as a herbal medicine for arthritis. We have previously observed protective effects of osthol on Ca2+antagonism, oxidative stress and inflammation. And other researches reported that it could inhibit increase of serum xanthine oxidase induced by liver tumor.OBJECTIVE: To investigate the protective effect of osthol on carbon tetrachloride (CCl4)-induced liver injury of mice.DESIGN: Completely randomized controlled study.SETTING: Department of Pharmacology, Gannan Medical College; Physical Education, Gannan Normal College.MATERIALS: A total of 40 Kunming mice were of both genders and weighing (20±2) g. Osthol was provided by Chengdu Longquan High-Tech Natural Pharmaceutical Co. Ltd.METHODS: The experiment was performed at Department of Pharmacology, Gannan Medical College from March to July 2005. Forty mice were randomly divided into control, model, osthol (ip 50 g/kg) and osthol (ip 100 g/kg) groups with 10 in each. Separately once a day for 15 consecutive days, the control and model groups were equalized injected intraperitoneally with 10 mL/kg saline and osthol groups injected intraperitoneally with 50 and 100 g/kg osthol, respectively. On 15 day just after treatment,they, except the control, were challenged with CCl4 (ip 1 g/L peanut oil solution 10 mL/kg). Then all mice were free access to water but fast food.At specified time points 16 hours after the injection of CCl4, all mice were sacrificed and blood was collected in centrifuge tubes. The serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and liver malondialdehyde (MDA) were determined, and the pathological examination of liver was observed.MAIN OUTCOME MEASURES: Contents of serum ALT, AST, MDA and pathological examination.RESULTS: Forty mice were involved in the final analysis. ① At 16 hours after CCl4 induction, contents of ALT and AST were higher in model group than those in control group (P ＜ 0.001). The increase in the contents of ALT and AST was inhibited by osthol in a dose-dependent manner, especially in 100 mg/kg osthol group (P ＜ 0.05). Both 50 mg/kg and 100 mg/kg osthol could inhibit the increase of AST induced by CCl4 (P ＜ 0.01-0.001).② Content of MDA in model group was increased (P ＜ 0.05), and content of MDA in 100 mg/kg osthol group was similar to that in control group.100 mg/kg osthol could decrease content of MDA, 50 mg/kg osthol could increased the content; however, it still had the tendency of decrease. ③Effect of osthol on histopahtological changes, the livers of CCl4intoxicated mice showed massive fatty change, gross necrosis, broad infiltration of the lymphocytes, and Kupffer cells around the central vein, loss of cellular boundary. The histological pattern of the livers of the mice treated with 100 mg/kg and 50 mg/kg osthol showed a mild degree of fatty change,necrosis and lymphocyte infiltration. In contrast, the inhibitory potency of 100 mg/kg osthol on the histological changes significantly higher than those models.CONCLUSION: Osthol can protect against CCl4-induced hepatotoxicity in mice through decreasing activities ALT and AST and contents of MDA.

Aim To investigate the proliferative effect and the apoptosis of human hepatoma SMMC-7721 cells induced by gallic acid ( GA ) , and its underlying mechanism. Methods SMMC-7721 cells were cul-tured in vitro. MTT assay was used to observe the pro-liferation of SMMC-7721 cells induced on GA 24 , 48 , 72 h. The morphological and ultra structural changes of the SMMC-7721 cells were observed by inverted micro-scope and transmission electron microscope respective-ly. Annexin V-FITC/PI staining was used to quantify the percentages of apoptosis in the total cell popula-tion. The expression of p53 mRNA was investigated by RT-PCR. Western blot was used to determine the pro-tein expression of p53. Results GA(6. 25~50 μmol ·L-1 ) markedly inhibited the activity of proliferation and induced apoptosis of SMMC-7721 cells after 48 h in a dose-dependent manner. GA significantly induced cell nuclear condensation and fragmentation. RT-PCR and Western blot results showed that GA could improve the expression of p53 mRNA and protein. Conclusion GA can inhibit the proliferation of human hepatoma SMMC-7721 cells and induce cells apoptosis. The mechanism may be associated with improving tumor suppressor gene p53 expression.

Objective To evaluate the plasma levels of adrenomedullin (ADM) and urotensin Ⅱ ( U-Ⅱ ) in children with capillary bronchiolitis, and their clinical significance. Methods One hundred and fifty three children with capillary bronchiolitis and 36 healthy children were recruited. Plasma levels of ADM and U- Ⅱ were measured at acute stage ( <7days) and convalescent stage (>14days) of airway inflammation. The relationship of plasma ADM and U-Ⅱ levels with symptom scores was evaluated. Results Plasma levels of ADM and U-Ⅱ in acute stage of capillary bronchiolitis were significantly higher than those in convalescent stage and healthy controls ( ADM: t = 20. 57 and 26. 26, P < 0. 01; U-Ⅱ: t = 14. 27 and 7. 61, P < 0. 01 ) , and there were significant differences among mild, moderate and severe capillary bronohiolitis (F = 245. 94 and 304. 79, P <0. 01). Plasma level of U-Ⅱ in convalescent stage of capillary bronchiolitis was lower than that of healthy controls (t = 6. 99, P <0. 01 ) , but ADM was still in a higher level ( t = 8. 98, P < 0. 01 ). In the convalescent stage, there was significant difference on U-Ⅱ levels among mild, moderate and severe capillary bronchiolitis ( F = 25.69, P < 0. 01 ) , but no significant difference was observed for ADM levels (F = 2. 25 , P > 0. 05 ) . Plasma levels of ADM and U- Ⅱ in acute stage showed positive correlation with symptom scores, and the regression coefficients were 0.884 (P =0. 000) for ADM and 0. 943 (P = 0. 000) for U-Ⅱ . Conclusion Plasma ADM and U-Ⅱ levels in children with capillary bronchiolitis are increased in acute stage and correlated with the symptom scores, which may serve as laboratory indicators for assessing the severity of the disease.

Objective To compare the safety and efficacy of minimally-access percutaneous nephrolithotomy (Mini-PCNL) in modified lateral position and prone position in high-risk patients with upper urinary tract stones and explore the clinical value of the modified lateral position. Methods We retrospectively analyzed 82 cases of high-risk patients underwent ultrasound-guided percutaneous nephrolithotomy from June 2010 to December 2015. 43 cases in group of modified lateral position and 39 case in group of prone position. The mean operating time, the success rate of stone fragmentation, hospitalization days, the value of postoperative hemoglobin decline and complication rate of patients were record and compared between the two groups. Results There was no statistical significance between the modified lateral position group and prone position group (P > 0.05) in gender, age, stone type, BMI, the score of ASA and preoperative complication. There was statistical significance in found access time, (7.88 ± 0.82) min in the modified lateral position group and (8.50 ± 0.80) min in the prone position group (P < 0.01). The stone free rate in the modified lateral position group (95.35 %) was higher than that in prone position group (74.36 %), the value of postoperative hemoglobin decline were (9.33 ± 2.49) g/L and (10.90 ± 3.54) g/L, respectively (P < 0.05); Two groups of the pleural damage rate had significant difference (P < 0.05); the hospitalization days and operating time were no statistical difference between the two groups (P > 0.05). Conclusions The Mini-PCNL in modified lateral position for high-risk patients had advantages of found access time, stone clearance rate, pleural injury, blood loss, comfort degree and security and worthy of clinical promoting.

Objective To investigate the correlation between the measurement methods of carotidcerebral pulse wave velocity (ccPWV) and the traditional method of brachial-ankle pulse wave velocity (baPWV).Methods A total of 136 healthy volunteers were divided into a youth group (20-39 years),a middleaged group (40-59 years),and an elderly group (more than 60 years) according to their ages.While detecting baPWV,transcranial Doppler ultrasound was used to simultaneously monitor the ipsilateral common carotid artery and the terminal segment of internal carotid artery.The time differences of the beating points of their cardiac cycles and the distanceses of the skin surfaces between the 2 probes were measured and ccPWV was calculated.Results The systolic blood pressure,pulse pressure and mean arterial pressure in the elderly group were signifificantly higher than those in the middle-aged group and the young group.The ccPWVs in the youth,middle-aged and elderly groups were 418 ± 52 cm/s,489 ±54 cm/s,and 599 ± 58 cm/s,respectively.The elderly group was significantly faster than the middle-aged group (t =7.308,P ＜0.001),and the middle-aged group was significantly faster than the youth group (t=6.758,P＜0.001).A Pearson correlation analysis showed that ccPWV was significantly positively associated with the age (r=0.847,P＜0.001) and baPWV (r =0.548,P ＜0.001).The multiple linear regression analysis showed that ccPWV was significantly positively associated with the age and diastolic blood pressure (partial correlation coefficients were 0.742 and 0.293respectively,P ＜0.001 and ＜0.010 respectively).Conchlusions ccPWV is a new measurement method for cerebrovascular stiffneas,and it has a good correlation with the traditional measurement method.

BACKGROUND: It has been believed mesenchymal stem cells (MSCs) play a role in treatment through paracrine mechanism. Various side effects such as embolism, tumorigenesis and immunological reaction caused by direct injection of MSCs can be avoided by extracting MSC lysate. However, there is a larger difference in current collection methods and standards of MSC lysate. OBJECTIVE: To compare repeated freeze-thaw and ultrasonication for the collection of lysate of MSCs. METHODS: Adipose-derived mesenchymal stem cells (ADMSCs) were isolated from the abdominal subcutaneous fat of healthy individuals, and purified with adherence screening method, followed by in vitro amplification using fetal bovine serum medium. The common surface makers of these cells were tested by flow cytometry (1×109, 2×109, 4×109/L). Repeated freeze-thaw and ultrasonication were employed for cell cytoclasis at three different densities respectively in saline and double distilled water, and a comprehensive comparison was performed on cytoclasis rate and the content of protein in cell lysate between the two methods. RESULTS AND CONCLUSION: (1) ADMSCs obtained from in vitro isolated human adipose tissue grew in a swirl or radial pattern with a homogenous size and neat arrangement. CD44, CD90, CD105 and other commonly used surface markers were highly expressed. (2) The study for optimization of lysate collection revealed that the higher cell density implicated a longer time for cell wall disruption and cytoclasis, as well as significantly increased cytoclasis rate. (3) BCA protein assay showed that the highest content of protein was obtained in saline solvent using ultrasonication method. Comprehensive analysis on the results leads to a conclusion that ultrasonication method with saline as the solvent is the optimized method for extraction of ADMSCs lysate, and the cell concentration of less than 4×109/L is recommended.