Humans ; Infant, Newborn ; Infant, Premature ; Infant, Premature, Diseases

Objective To explore the effect of progesterone on the rate of neuronal apoptosis and nitric oxide(NO) level in the cortex and hippocampus tissue of newborn rats with hypoxic-ischemic encephalopathy(HIE).Methods Thirty 7-day-old neonatal rats were randomly divided into 3 groups:sham-operated group,hypoxic-ischemic(HI) group and pretreatment group.Rats in HI group and pretreatment group were subjected to left common carotid artery ligation,then were exposed to 80 mL/L oxygen and 920 mL/L nitrogen gas in 37 ℃ closed container for up to 2.5 h to establish HIE model.Progesterone was injected intraperitoneally into rats in the pretreatment group respectively for 30 minutes before hypoxia,and solution was injected into the sham-operated group and HI group.All rats were killed at 24 h after operation.The neuron apoptosis was identified and analyzed by flow cytometry.Nitrate/nitrite was assayed to represent nitric oxide content of brain tissues.Results The ratio of neuronal apoptosis and NO contents in cortex and hippocampus tissue in HI group [(10.09?0.36)%,(12.32?0.28)%,(51.36?9.71) ?mol/L,(52.34?4.26) ?mol/L] were significantly higher than those in sham-operated group [(2.49?0.23)%,(2.58?0.26)%,(18.16?6.24) ?mol/L,(19.28?3.58) ?mol/L)](P_a

Objective To observe the changes of neural stem cells(NSCs) in subventricular in developing human fetal brain at(diffe)-rent ages.Methods Thirty cases of embryoes at gestational age 24-28 weeks induced by labor with water bag were collected to determin distribution,shapes and growth modes in subventricular with hybridization in situ under light microscope.Results NSCs expressing Nestin mRNA existed in subventricular from human fetal brain at different ages,NSCs existed in subventricular of different fetal age included astro-NSCs,each had enations from 3 to 6,and all projections crowded together into a reticular plexiform,in which NSCs districuted.Nucli were round in shape,each had nucleoli from 1 to 4.NSCs had rarefaction chromatin,most NSCs existed in a single growth mode,symmetral cleavage growth mode and clony with 3 NSCs would be seen.There were no differences between positive Nestin mRNA NSCs in distribution and cell shapes,but had differences in growth mode.Conclusion NSCs exist in subventricular from different gestational ages and their growth mode are changing with difference of gestational age.

Objective To explore a precise and dependable method for determining neural stem cells(NSCs)in hippocampus from human fetal brain.Methods Ten cases of embryo at gestational age 32 weeks and induction of labor with water bag were collected,affused and sliced including frost slice and paraffin slice for determining alteration of tissue and NSCs in hippocampus from human fetal brain with HE stain and immunohistochemical staining under light microscope.Results Nestin proteins located in NSCs and the number of NSCs were less in paraffin slice of tissue than those in frost slice.Integrality of structure of tissues and cells in paraffin slice excelled in frost slice.Conclusions There is a precise and dependable method including induction of labor with water bag and allusion and frost slice,which is necessary for wide useness in study of NSCs in human fetal brain.

Cerebral Hemorrhage ; epidemiology ; etiology ; prevention & control ; Cerebral Ventricles ; Humans ; Incidence ; Infant, Newborn ; Infant, Premature ; Infant, Premature, Diseases ; epidemiology ; etiology ; prevention & control

Objective To investigate a simple and dependable approach to establish the hypoxic-ischemic encephalopathy model in neonatal rat. Methods Twenty-one neonatal rats of 7 days old were randomly divided into control group,hypoxic group,and hypxic-ischemic group.Every group was randomly divided into 3 hours,6 hours,1 day,3 days,7 days, 14 days,and 21 days group,according to the time of killing.Left common carotid artery of neonatal rats at age of 7 days in hypoxic-ischemic group were ligated.Then,the rats in hypoxic and hypoxic-ischemic group were put in a state of 8% oxygen for 2.5 hours. Brain tissues of rats in 3 groups were observed with HE staining under light microscope.Results In hypoxic-ischemic group,there was found mild brain damage after hypoxic-ischomic 3 hours,the brain lesion was most severe at 1 day,glial cell proliferation was found at 3 days,much neur were losed at 14,21 days,and colloid scar was formed in cortex,striatum and hippocampi.Conclusion The method that left common carotid ontery of neonatal rats were ligated and then put in 8% oxygen for 2.5 hours is simple, rapid and dependable, which can be applied widely.

Objective To investigate the characters of culture of neural stem cells(NSCs) from neonatal rats in different states in vitro.Methods Forty-two neonatal rats at age 7 days were divided randomly into 3 groups as control group,hypoxic group and hypoxic-ischemic group,each having 14 rats.Forteen rats of every group divided randomly into 7 small groups,each including 3 h,6 h,1 d,3 d,(7 d),14 d and 21 d,according to the time to put to death,each having 2 rats.After builting rat models of hypoxic-ischemic encephalopathy,NSCs from hippocampus in 3 groups were isolated,then cultured,passed,differentiated and differentiated with single cell clone and immunocytochemistry tecnique.Results NSCs in hippocampus from 3 groups were cultured in form of typical neuraospheres in suspension.The cells could be cloned,passed continuously and induced.There were differences among 3 groups when primary NSCs were cultured at 3 h and 6 h time points.But at 1 d,3 d,7 d,14 d and 21 d time points,clony neuraospheres from primary NSCs in hypoxic group were the most among 3 groups while clony neuraospheres from primary NSCs in hypoxic-ischemic group were the lest.Conclusions NSCs from hippocampus of neonatal rats in different states remain to be cultured,meanwhile,NSCs are decreased with increase of age,elongation of illness course and progress of state of an illness.

Objective To study the effects of matrine on accumulation of ? globin mRNA and induction of erythroid differentiation in K562 cells in vitro.Methods K562 cells were cultured for 6 days with different concentration of matrine,viable cell counts were determined by trypan-blue dye exdusion test. Erythroid differentiation was evaluated by percentage of benzidine-positive cells at different days after culture. Morphological changes were observed under microscope after Wright-Gimesa staining; ? globin mRNA was quantitative by real time quantitative reverse transcript polymerase chain reaction(RT-PCR).Results Different concentrations of matrine inhibited proliferation of K562 cells in dose-dependent manner; otherwise, K562 cells were successfully induced by erythroid differentiation with matrine. After treatment with matrine, percentage of benzidine-positive cells significantly increased from 0.7% to 15.7% and characteristic changes of erythroid differentiation in the cell morphology were observed, G? globin mRNA had a preferential increase (2.7 fold)in K562 cells. Conclusions Matrine accumulation G? globin mRNA and induced erythroid differentiation of K562 cells. The results provides an experimental evidence for the pharmacological therapy of hematological diseases associated with a failure in the expression of normal ? globin genes.

Objective To explore the therapeutic effectiveness and pathways of human umbilical cord mesenchymal stem cells (hUCMSCs) transplantation for acute hepatic failure in rats.Method hUCMSCs were isolated from umbilical cord with attachment culture method,and the surface antigens were tested by flow cytometry.Forty-eight male Sprague Dawley rats were randomly divided into four groups.The animal model of acute liver failure was induced by injecting intraperitoneally with 50％ olive oil solution of carbon tetrachloride (2.5 ml/kg).The treatment groups were injected with hUCMSCs suspension separately through the tail vein or injected into the liver 24 h post-modeling.Blood serum and liver tissues were collected at several time points to analyze the improvement of liver function and histological repair.Real-time PCR was used to detect the expression of human CK8,CK18 and AFP mRNA in liver tissues.Immunohistochemistry was used to detect the expression of human CK18 in liver tissues.Result There were statistically significant differences among liver functions after transplantation (P＜0.05).hUCMSCs improved histological status through enhancing hepatocellular regeneration and reducing inflammatory cells.Real-time PCR results showed that the expression of CK8,CK18 and AFP mRNA was obviously increased in the tail vein transplantation group and hepatic lobe injection transplantation group as compared with the model group (P＜0.05).Immunochemistry results revealed that transplanted hUCMSCs in animal liver could differentiate into functional hepatocyte-like cells that expressed human CK18 as hepatocyte-specific marker in the tail vein transplantation group and hepatic lobe injection transplantation group.No significant differences in histological repair and grade of differentiation were examined between the tail vein transplantation group and hepatic lobe injection transplantation group (P＞0.05).Conclusion hUCMSCs can prompt the repair of acute liver failure and enhance pathological repair.Transplanted cells in animal liver can differentiate into functional hepatocyte-like cells that expressing hepatocyte-specific markers.Transplantation of hUCMSCs via the tail vein or direct injection into the liver had the similar therapeutic effects.

Objective To develop a method for determining verbascoside in Mitonghua Granules by HPLC. Method Verbascoside was determined by HPLC method,Agilent ZORBAX-Extend C18 column (4.6 mm?150 mm,5?m) with Acetonitrile-1 %Acetic acid(13∶87) as the mobile phase was used for elution.The flow rate was at 1.0 mL/min and the detective wavelength was at 334 nm. The column temperature was at 30 ℃. Results There was a good linearity relationship at the range of 0.062 8～1.256 0 ?g for verbascoside (r=0.999 83) .The average recovery was 99.61 %,and RSD was 1.55 %(n=6). Conclusion The method is simple,reproducible and accurate with a good repeatability,and can be used for the quality control of Mitonghua Capsules.