Objective To explore effect of miR‐302a on proliferation and apoptosis in folate deficiency mouse embryonic stem cell (mESC) .Methods The cases were divided into complete culture medium group (control group) ,folate‐deficient culture medi‐um group (folate‐deficient group) ,folate‐deficient culture medium plus miR‐302a mimic group (miR‐302a group) .The expression of miR‐302a was examed by RT‐PCR in control group and folate‐deficient group .To construct miR‐302a mimic and then was trans‐fected into the folate‐deficient culture medium mESC .Effect of miR‐302a mimic on mESC viability was detected by MTT assay ,the effect of miR‐302a mimic on mESC apoptosis was examed by Annexin V‐FITC/PI flow dual‐staining method ,the effect of miR‐302a mimic on mESC cycle was examed by flow cytometry .The activation of phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) and expression of CyclinD1 ,p21 and p27 was assayed by Western blot . Results The expression of miR‐302a was lower in folate‐deficient group(P<0 .01) .Compared with control group ,the viability of mESC was lower ,the apoptosis of mESC was higher ,the cell cycle was arrested in G1 phase ,the level of phosphorylation of AKT and mTOR was lower ,the expression of CyclinD1 was lower ,the expression of p21 and p27 was higher in folate‐deficient group , with statistical significance (P<0 .01) .Compared with folate‐deficient group ,the viability of mESC was higher ,the apoptosis of mESC was lower ,G1 phase was shortened ,the level of phosphorylation of AKT and mTOR was higher ,the expression of CyclinD1 was higher ,the expression of p21 and p27 was lower in miR‐302a group with statistical significance (P<0 .01) .Conclusion These results suggested miR‐302a exerted anti‐apoptosis and promote cell proliferation in folate‐deficient culture medium ,which might be related to PI3K/AKT/mTOR signal pathway .

The directed molecular evolution of enzyme in vitro can not only improve the efficiency of evolution, but also evolve enzyme according to the investigator's desire. This review summed up the feasible methods of this novel technique.

Objiective To observe the morphological changes of rabbits' retina at different time points at which seawater was injected into the vitreous of rabbits. Methods Twenty-five New-Zealand white rabbits were randomly assigned to five groups, as 30 minutes, 2 hours, 6 hours, 12 hours and 24 hours group following the seawater injection in the vitreous. The right eyeball served as experimental eyeball and the left one as control in each group. 0.5ml artificial seawater was injected into the intermedius of rabbits right vitreous. 0.5ml balanced salt sdution was injected into the left vitreous in teh same way. At different time points following the eyeball injury, the morphological changes of retina were observed with light microscope and transmission electron microscope. Results The retina of control eyeballs had clear layers and regular cell shape in retinal structures with conventional staining. For the experimental eyeballs, the ganglion cell layer began to swelling after 30 minutes of seawater injection. The nerve fiber swelled obviously and the mitochondria vesiculated and swelled in the inner segment at 2 hours of seawater injection. The cell population of the inner nuclear layer began to reduce in 6 hours. At 12 hours of seawater injection, the retinal damages included disorganized cell nuclei, decreased cell population in ganglion cells and inner nuclear layer. Mitochondria vesiculated in photoreceptor cells, nuclear membrane shrank and sank inward. The vesiculation on inner and outer segments increased obviously. Conclusions Seawater may cause the morphological changes of rabbits' retina at early stage, and the histological damages become aggravation following the time prolongation. The damages would be unrecoverable 6 to 12 hours after seawater injury.

Objective To observe the effects of insulin pump therapy on type 1 diabetes mellitus children with diabetic keto or diabetic ketoa-cidosis(DK/DKA).Methods Forty-three children with DK/DKA were randomly divided into treatment group(n=26)and control group(n=17).The patients in treatment group were treated with insulin pump,control group were received small amount insulin injravenous injection.The changs of blood sugar,urine ketone bodies,incidence of hypoglycemia and length of stay were observed.Results 1.The effect of decreasing sugar was the same between two groups.2.There was no hypoglycemia events in treatment group.But 2 cases in control group had hypoglycemia.3.There was significant difference of length of stay between two groups(P

Animals ; Cardiovascular System ; drug effects ; Curcuma ; Plant Extracts ; pharmacology

AIM: To evaluate effects of Pinaverium Bromide on different segments and layers of colonic smooth muscle in wrap restraint stress (WRS) rats and explore its possible therapeutic mechanism on different types of irritable bowel syndrome (IBS). METHODS: Adult SD rats were randomly divided into model group (wrap restraint stress group) and control group. Colonic smooth muscle strips were made from different segments and layers in two groups. The spontaneous contraction activities of colonic longitudinal/circular muscle (LM/CM) strips of rats were observed with organ bath system before and after addition of series concentrations of pinaverium. RESULTS: Pinaverium Bromide caused concentration-dependent inhibition of colonic smooth muscle, the inhibitory effect of pinaverium in model group was significantly stronger than that in control group(proximal colon: 28.54±4.82 vs 7.48±1.65,21.75±1.00 vs 12.56±3.15; distal colon: 15.71±5.27 vs 3.89±1.16, 20.16±3.16 vs 7.56±1.96 )(P<0.05). Compared with that of distal colon, inhibitory effect of pinaverium was significantly higher of proximal colon (P<0.05). For the inhibition of pinaverium, there was no significant difference between LM and CM strips in the same intestinal segments (P>0.05). CONCLUSION: Effects of Pinaverium Bromide on different colonic muscle layers and segments in WRS rats is probably related with its therapeutic mechanism on different types of IBS.

Objective To evaluate the clinical performance of CLINITEK Atlas urine dry chemistry analyzer and its supplementary strips,which could be used for other hospitals as reference.Methods Five hundred and one samples of random fresh urine were collected and analyzed by CLINITEK Atlas urine dry chemistry analyzer.10 parameters were reported for each sample,including SG,pH,BLD,LEU,PRO,GLU,KET,UBG,BIL and NIT.According to the medicals standard of the People's Republic of China,General Technical Requirements for Urine Analyzer(YY/T 0475-2004),General Technical Requirements for Chemical Reagent Strips for Urinalysis(YY/T 0478-2004)and physical,Chemical and Microscopic Examination of Urine(WS/T 229-2002),the precision,accuracy,carryover,stability,sensitivity and consistency of each parameter were evaluated.The agreement was assessed between the results for BLD and LEU obtained from CLINITEK Atlas analyzer and phase contrast microscope,and calculated the sensitivity and specificity of CLINITEK Atlas analyzer for BLD and LEU using phase contrast microscope as the gold standard.SG and pH test was performed among 200 specimens by CLINITEK Atlas analyzer,and then compared with the results obtained from MASTER-SUR-NM specific gravity refractometer and pH precision test strips respectively.In addition to SG and pH,the other eight parameters were compared with the results obtained from CLINITEK 500 urine analyzer,and Kappa value and consistency were calculated.Results The accuracy,precision,sensitivity,carryover and stability of 10 parameters could meet all the requirement of standards.SG and pH had good correlation with urine specific gravity refractometer (r =0.9838,P ＜0.001)and pH meter (r =0.8884,P ＜0.001),respectively.Compared with phase contrast microscope,BLD and LEU had coincidence rates of 90.4％ and 90.8％,respectively; Sensitivities were 90.7％ (301/332) and 83.3％ (200/240) ; Specificities were 89.9％ (152/169) and 97.1％ (255/261).Compared with CLINITEK 500,all the parameters,except for SG and pH,had good coincidence rates of ＞ 87.6％.Conclusion The performance of CLINITEK Atlas urine dry chemistry analyzer can meet the clinical requirements of all standards.

Objective To report the clinical and magnetic resonance imaging(MRI)features of 5 eases with idiopathic orbital myositis.Methods Four females and one male,aged 27 to 57 years,presented department of neurology in the First Hospital of Peking University in October 2008 to September 2009.The duration of disease Was between 3 months and 4 years.Recurrent course appeared in 3 of them.0rbital MRI Was performed in all of them.After diagnosis they underwent long.term corticosteroid treatment.Results All patients presented ocular pain,asymmetrical and incomplete ophthalmoplegia and mild proptosis.EMG revealed no significant decline in repetitive stimulation.Muscle biopsies of limb muscle were unremarkable.Creatine kinase and thyroid function test were in normal limits.MRI revealed unilateral.focal or difluse enlargement and enhancement of extraocular muscles,involving 1 extraocular muscle in 2 cases,2extraocular muscles in 2 cases,more extraocular muscles in 1 case.No evidence indicated bone destruction or cavernous sinus abnormalities.Five Cases showed improvement and remission after long-term administration of steroids.Conclusion Persistent and asymmetrical ophthalmoplegia is connnon in orbital myositis.Extraocular muscle swelling characterized the MRI changes.

Angina, Unstable ; diagnosis ; Humans ; Medicine, Chinese Traditional ; Severity of Illness Index

Objective To observe the effects of soybean isoflavone in attenuating trans fatty acids-induced athero-sclerosis in rats .Methods The rats were randomly divided into normal diet group、trans fatty acids group、soybean isoflavone group and trans fatty acids +soybean isoflavone group .Research kit was used to test the amounts of TG , HDL,LDL and IL-6,TNF-αin rats'blood.Bovine aortic endothelial cells were cultured and divided into control group,trans fatty acids group and trans fatty acids group +soybean isoflavone group .Western blot was performed to test the NF-κB phosphorylation ,as well as to determine ICAM-1 and VCAM-1 protein contents in bovine aortic endo-thelial cells.Results Compared with normal diet group ,the amounts of TG,LDL,IL-6,TNF-αincreased evidently (P<0.05), whereas the amounts of HDL decreased evidently (P<0.05) in the rats'blood of trans fatty acids group . After the soybean isoflavone intervened ,the amounts of TG,LDL,IL-6,TNF-αall decreased (P<0.01), meanwhile the amounts of HDL increased significantly (P<0.01).Soybean isoflavone inhibited NF-κB phosphorylation induced by trans fatty acids ( P<0.01) .The high expression of ICAM-1 and VCAM-1 induced by trans fatty acids was de-creased by soybean isoflavone when comparing with those in the control group ( P<0.01 ) .Conclusions Soybean isoflavone has a role in attenuating trans fatty acids trans fatty acids-induced atherosclerosis in rats .