Objective To study the protective effect on CCl4-injured L-02 hepatocytes of polysaccharides from Hyriopsis cumingii(HCP) and try to explain the mechanism.Methods The L-02 hepatocytes were incubated and then injured by CCl4.The levels of AST,ALT,MDA and SOD in cultural supernatant were detected by general methods.Cell viability was assayed by MTT method.Results The polysaccharides(25,250 and 1000?g?L-1)could reduce the levels of AST,ALT,MDA in cultural supernatant which increased by CCl4.It also could boost the hepatocytes viability and elevate the level of SOD which reduced by CCl4.Conclusion The results suggest that polysaccharides from Hyriopsis cumingii have direct protective activity on hepatocytes injured in vitro.It might be associated with the anti-oxidative activity of polysaccharides from Hyriopsis cumingii.

Objective A HPLC method was established to measure the contents of major nucleosides in Hyriopsis cumingii.Method Separation was performed on a Lichrosper-C_(18) column, pure water was used as mobile phase,the flow rate was 1.0mL min~(-1),the column temperature at 30℃and the detective walvelength at 254 nm.Results The contents of uracil,hypoxanthine, xanthine,uridine and thymine were 153.2～173.3?g?g~(-1),106.6～360.8?g?g~(-1),117.6～172.8?g?g~(-1),70.3～153.1?g?g~(-1) and 343.9～511.4?g?g~(-1),respectively. Conclusion This method showed good repeatability and flexibility.It could be used as a conventional method for determination of major nucleosides in H.cumingii.

Thunder-fire wonder moxibustion is one of pressing moxibustion therapies and has a very good therapeutic effect on limb pains, furuncle-carbuncle and cold syndrome. To reveal the mechanism of clinical action of ancestors’ thunder-fire wonder moxibustion and seek the physical basis of its therapeutic advantage, this study, by a series of experiments, compared heat transfer regularities of thunder-fire wonder moxibustion versus pure moxa stick in simulated biological tissues under different conditions, preliminarily revealed heat radiation and heat transfer regularities of thunder-fire wonder moxibustion, tried to find pressing strength suitable for clinical operation of pressing moxibustion and had thoughts about changes in the clinical operation of past dynasties.

Objective To investigate the expression and significance of mRNAand exon mutationof NT5C2 gene in acute lymphoblastic leukemia (ALL) bone marrow.Methods Case control study design was used in this study.Bone marrow samples were collected from ALL patients in Anhui Provincial Cancer Hospital in recent 4 years.The patientswere divided into the initial diagnosis group , the complete remission group and the recurrence group.And they could specifically be divided into 36 patients initially diagnosed, 36 patients who achievedcomplete remission and 16 patients who relapsed with children B -ALL,15 patients initially diagnosed,15 patients who achievedcomplete remission and 9 patients who relapsed with children T -ALL, 18 patients initially diagnosed,18 patients who achievedcomplete remission and 12 patients who relapsed with adult B-ALL, and 11 patients initially diagnosed,11 patients who achievedcomplete remission and 6 patients who relapsed with adult B -ALL.The initial diagnosis,complete remission and recurrence samples were matched.8 children and 8 adults without hematologic malignanciewere used as controls .Real-time PCR was performed to detect the level of NT5C2 mRNAin ALL patients.The exons of NT5C2 gene were cloned and sequenced for the common mutations in all cases .The results of NT5C2 mRNA levels in different groups were performed using non -parametric test by SPSS16.0 analytics software, and then non-parametric test together with correlation analysis was analyzed between NT 5C2 mRNA levels of different initial diagnosis groups and gender, age, leukocyte level and risk classification .Results (1)The expression of NT5C2 mRNA levels of recurrence group were higher than that of initial diagnosis group ,complete remission group and controls in children and adult B -ALL respectively(P <0.01).(2)NT5C2 mRNA expression in children and adult T-ALL showed no difference in initial diagnosis ,complete remission, recurrence group and controls (P >0.05).(3)NT5C2 mRNA expression of initial diagnosis group in children and adult B -ALL and T-ALL was not correlated with risk classification (P >0.05).(4)A newheterozygousmutation p.P414A of NT5C2 was discovered in a recurrencesample.Conclusions (1) High expression ofNT5C2 mRNA is associated with recurrence inchildren and adult B-ALL, and it may be an indicator of monitoring recurrence .(2)The incidence of exons mutation of NT5C2 gene in ALL is low in China.

The vitamin K epoxide reductase complex subunit 1 (VKORC1), the rate-limiting enzyme for vitamin K recycling, is significantly down-regulated in the kidneys of urolithiasis patients. This study searched for direct evidence to define the inhibitory activity of VKORC1 against calcium oxalate (CaOx) crystal formation. In the experiment of VKORC1 overexpression, HK-2 cells were transfected with the pFLAG-CMV-7.1-VKORC1 plasmid as a pFLAG-CMV-7.1-VKORC1 transfection group or the pFLAG-CMV-7.1 plasmid as a pFLAG-CMV-7.1 control group. In the experiment of VKORC1 knockdown, HK-2 cells were transfected with the PGPU6/GFP/Neo-VKORC1shRNA-2 as a PGPU6/GFP/Neo-VKORC1shRNA-2 transfection group or the PGPU6/GFP/Neo-shRNA-NC plasmid as a PGPU6/GFP/Neo-shRNA-NC control group. The expression of VKORC1 in HK-2 cells was detected by real-time quantitative PCR and Western blotting. The CaOx crystal formation was observed under the laser-scanning confocal microscope. It was found that the expression levels of VKORC1 mRNA and protein were significantly higher in the pFLAG-CMV-7.1-VKORC1 transfection group than in the pFLAG-CMV-7.1 control group (P<0.01). The number of CaOx crystals in HK-2 cells incubated in fluorescently labeled CaOx monohydrate (COM) crystal medium for 48 h was 14±4 per field (100×) in the pFLAG-CMV-7.1-VKORC1 transfection group and 26±5 per field (100×) in the pFLAG-CMV-7.1 control group respectively under the laser-scanning confocal microscope. The amount of CaOx crystal aggregation and formation in the pFLAG-CMV-7.1-VKORC1 transfection group was significantly reduced as compared with the pFLAG-CMV-7.1 control group (P<0.05). The expression levels of VKORC1 mRNA and protein were significantly lower in the PGPU6/GFP/Neo-VKORC1shRNA-2 transfection group than in the PGPU6/GFP/Neo-shRNA-NC control group (P<0.05). The number of CaOx crystals in HK-2 cells incubated in fluorescently labeled COM crystal medium was 65±11 per field (100×) in the PGPU6/GFP/Neo-VKORC1shRNA-2 transfection group and 24±6 per field (100×) in the PGPU6/GFP/Neo-shRNA-NC control group respectively under the laser-scanning confocal microscope. The amount of CaOx crystal aggregation and formation in the PGPU6/GFP/Neo-VKORC1shRNA-2 transfection group was significantly increased as compared with the PGPU6/GFP/Neo-shRNA-NC control group (P<0.05). These findings suggested that the VKORC1 protein could inhibit CaOx salt crystallization, adhesion and aggregation. This research would help us to understand the mechanisms involving the interaction between crystallization and epithelial cells and the formation of CaOx.

After estrogen receptor?(ER?)in MC,63 cell was knocked down by RNA interference, expression of osteoprotegerin(OPG)induced by 17?-estradiol was assayed by RT-PCR.17?-estradiol with various concentration obviously upregulated the expression of OPG mRNA of MG63 cell with the maxima]effect at the concentration 10~(-7)mol/L,which was not inhibited by suramin(G protein inhibitor).The present result suggested that ER?was not involved in regulation of OPG mRNA expression in MC,63 cell by 17?-estradiol.

The stability of melittin in different solvents (water, deoxygenated water, physiological saline, PBS, 50% ethanol, ethanol, glycerol)was studied and the results showed that the stability of melittin is not influenced by light, temperature and pH in 50% ethanol, which melittin can be completed dissolved when compared with ethanol and glycerol, in such, 50% ethanol was chosen as solvent storage when measured content of melittin. Then the effect of different concentrations of PBS, the pH of PBS and rat skin ho- mogenates were tested, and the results showed that melittin was degraded rapidly at low concentration solution and low ionic strength. Increasing pH of PBS and rat skin homogenate can accelerate the degradation of melittin. These researches provide an experimental ba- sis for further study of melittin.
Animals ; Drug Stability ; Ethanol ; chemistry ; Hydrogen-Ion Concentration ; Melitten ; chemistry ; Rats ; Skin ; drug effects ; Solvents ; chemistry ; Temperature

Objective: To explore the correlation between moxibustion sensation and distance of moxa stick and provide reference for clinical practice. Methods: A total of 16 healthy volunteers aged 18-35 years old in college were recruited and given mild moxibustion at Shousanli (LI 10), Zusanli (ST 36), Shenshu (BL 23) and Tianshu (ST 25) with moxa stick, and the occurrence and frequency of moxibustion sensation were recorded at distances of 5 cm, 4 cm, 3 cm and 2 cm. Mild moxibustion scale was used to count the score. Results: Warm was the main moxibustion sensation, burning pain and soreness decreased with the rise of distance; for the same acupoint, score of mild moxibustion scale increased with the decrease of distance; score ranged between 5.5 and 6.5 at distance 3 cm, which was the most comfortable distance for volunteers. Conclusion: The distance of 3 cm is the most comfortable distance in mild moxibustion.

Objective: To explore the beneficial regulatory effect of mild moxibustion from different distances at Zusanli (ST 36) of healthy population on the functions of temperature-related brain regions. Methods: In 20 recruited healthy subjects, the change of the temperature-related brain regions induced by mild moxibustion from different distances at Zusanli (ST 36) was observed by functional magnetic resonance imaging (fMRI). Results: In comparison of the values in amplitude of low-frequency fluctuation (fALFF) during and before moxibustion, it has been found that in moxibustion of 2 cm distance, fALFF value increased in the brain regions of the left anterior cingulated cortex and lateral surrounding cerebral regions, and fALFF value decreased in the cerebral regions of the peripheral cortex of the calcarine fissure; in moxibustion of 3 cm distance, fALFF value increased in the brain regions of the right and medial side and paracingulated gyrus, and fALFF value decreased in the cerebral zone of the left middle temporal gyrus; in moxibustion of 4 cm distance, fALFF value increased in the brain regions of the right and medial and paracingulated gyrus; and in moxibustion of 5 cm distance, fALFF value increased in the brain regions of the left hippocampus. In comparison of the value of regional homogeneity (ReHo), it has been found that in moxibustion of 2 cm distance, ReHo value increased in the cerebral zone of the posterior lobe of the right cerebellum, and ReHo value decreased in the cerebral zone of the right occipital lobe; in moxibustion of 3 cm distance, ReHo value increased in the brain regions of the left cerebellar posterior lobe and left frontal lobe, and ReHo value decreased in the cerebral zone of the right inferior temporal gyrus; in moxibustion of 4 cm distance, ReHo value increased in the brain regions of the right superior frontal gyrus and ReHo value decreased in the brain regions of the right parietal lobe and angular gyrus; in moxibustion of 5 cm distance, ReHo value increased in the cerebral zone of the right frontal lobe and ReHo value decreased in the cerebral zone of the right brainstem. Conclusion: In moxibustion of 3 cm distance, the changes in the brain regions basically conform to the transmission route of body trunk temperature.