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.

OBJECTIVETo establish a neonatal rat model of hypoxic-ischemic encephalopathy and clarify the changing features of neural stem cells (NSCs) in episodes of hypoxic-ischemic encephalopathy (HIE) so as to provide experimental and theoretical evidences for treating HIE by applying NSCs at the appropraite time.

METHODSTotally 210 neonatal rats aged 7 d were divided randomly into three groups, normal control group, hypoxic group and hypoxic-ischemic group with 70 rats in each. According to the time of sacrefice, 70 rats of every group were further divided randomly into seven groups including third hour (3 h), the sixth hour (6 h), first day (1 d), third day (3 d), the seventh day (7 d), the fourteenth day (14 d) and the twenty-first day (21 d), with 10 rats in each subgroup. The left common carotid artery of the neonatal rats in hypoxic-ischemic group was ligated and those in the hypoxic group were subjected to inhalation of 8% oxygen for 2.5 h. NSCs from brain tissues of the rats of the three groups were determined with HE staining and immunohistochemical method under light microscope.

RESULTSMost of neonatal rats in hypoxic-ischemic group behaved turning to the left stably 1 h after normal concentration of oxygen was given. In hypoxic-ischemic group, slight brain injury occurred at 3 h, severe brain injury appeared at 1 d, glial cells proliferated at 3 d and 7 d, brain atrophy was found at 14 d and 21 d. NSCs existed in brain tissues of rats in all the three groups. NSCs in normal control and the hypoxic group mainly distributed in hippocampus, subventricular tissues, striatum and cortex. But NSCs in hippocampus located in layers of molecule, cone cell and inner granular cell. NSCs in hypoxic-ischemic group showed obvious regional distribution, less in the regions with pathological changes. At 3 h, 6 h and 14 d, there was no difference in the number of NSCs between hypoxi and hypoxic-ischemic group. At 1 d, 3 d and 7 d, there was a highly significant difference in the number of NSCs between hypoxic and hypoxic-ischemic group. At 21 d, there was a significant difference in the number of NSCs between hypoxic and hypoxic-ischemic group, meanwhile, there was a significant difference in the number of NSCs between control and hypoxic group. At 3 d, there was a very significant difference in the number of NSCs between control and hypoxic-ischemic group. At 7 d and 21 d points, there was a highly significant difference in the number of NSCs between control and hypoxic group.

CONCLUSIONThe neonatal rat model of HIE was successfully established. NSCs increased in earlier period and decreased in later period of HIE, ultimately, NSCs located in the injured regions died one after anotner. Hypoxia induces NSCs' proliferation.

Animals ; Animals, Newborn ; Atrophy ; Brain ; pathology ; Carotid Artery, Common ; surgery ; Disease Models, Animal ; Hypoxia-Ischemia, Brain ; pathology ; Immunohistochemistry ; Ligation ; Multipotent Stem Cells ; pathology ; Neuroglia ; pathology ; Neurons ; pathology ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation ; methods ; Time Factors

OBJECTIVETo study growth characteristics of neural stem cells (NSCs) from subventricular zone (SVZ) of the different human fetal brain at different gestational age and to provide experimental and theoretical evidences for clinical application of NSCs for treatment of certain diseases.

METHODSNinety human embryos at gestational age 16 - 36 weeks were collected and were divided into six groups according to gestational age: 16 w, 20 w, 24 w, 28 w, 32 w and 36 w. Each group had 15 embryos and brain tissues were taken from each embryo's SVZ. All subjects had congenital heart disease or digestive tract abnormity diagnosed with B ultrasound at antepartum, but none had abnormal development of brain. Pregnant mother and her husband desire termination of pregnancy. The morphology, existing mode and the number of neural stem cells in subventricular zone were examined with immunohistochemical method. The NSCs in subventricular zone were cultured, passaged and differentiated with cell culture technique, then were identified with immunohistochemical method.

RESULTSNSCs in SVZ from the different human fetal brain existed in a scattered manner in the network formed by stellate cells, NSCs had round, ellipse and fusiform shape, especially in stellate shape. NSCs had larger and smaller size and distributed in dense or scattered forms, each having zero to two enations, most had one or two. NSCs had less cytoplasm. The nucli of the NSCs had a round shape with loose chromatin and 1 - 4 nucleoli. Most of NSCs existed in singular scattered form, some of them showed symmetrical or asymmetrical division, some of them showed synaptic connection with other NSCs. The number of NSCs in SVZ from groups with different fetal age decreased with increasing gestational age (chi(2) = 4644.602, P < 0.01). NSCs in SVZ from the different human fetal brain cultured with serum-free medium formed typical neurospheres in suspension. The cells could be passaged continuously, and could express nestin antigen. Serum-contained medium induced neural stem cells to differentiate and express specific antigens of neuron, astrocyte and oligodendrocyte.

CONCLUSIONSNSCs existed in SVZ of human embryos at different gestational age. There are differences in morphology, existing pattern and the number of NSCs in SVZ at different gestational age. NSCs in SVZ at different gestational age may be cultured in vitro.

Age Factors ; Astrocytes ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Cerebral Ventricles ; cytology ; Female ; Fetal Stem Cells ; Fetus ; cytology ; Gestational Age ; Humans ; Immunohistochemistry ; Intermediate Filament Proteins ; metabolism ; Male ; Nerve Tissue Proteins ; metabolism ; Nestin ; Neurons ; Oligodendroglia ; Pregnancy

OBJECTIVETo investigate the expression of Rho-GDI in the decidual tissues of patients preeclampsia and explore its clinical implication.

METHODSReal-time PCR, Western blotting and immunohistochemistry were used to detect the mRNA and protein expressions of Rho-GDI in the decidual tissues from 30 normal women with full-term pregnancy, 30 patients with early-onset severe preeclampsia and 30 with late-onset severe preeclampsia.

RESULTSRho-GDI expression was found mainly on the cell membrane and in the cytoplasm and nuclei of the decidual cells, occasionally occurring in the stroma. Both the mRNA and protein expressions of Rho-GDI in the decidual tissues were significantly higher in the normal pregnancy group than in the two severe preeclampsia groups (P<0.05), and the patients with late-onset severe preeclampsia had the lowest expressions of Rho-GDI.

CONCLUSIONThe lowered expression of Rho-GDI in the deciduas might be involved in the pathogenesis and progression of preeclampsia.

Adult ; Decidua ; metabolism ; Female ; Guanine Nucleotide Dissociation Inhibitors ; genetics ; metabolism ; Humans ; Pre-Eclampsia ; metabolism ; Pregnancy ; RNA, Messenger ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; methods ; Young Adult ; rho-Specific Guanine Nucleotide Dissociation Inhibitors

BACKGROUND: Platelet-rich plasma (PRP) and naringin can both promote proliferation and induce osteogenic differentiation of mesenchymal stem cells. However, their combined use is rarely reported. OBJECTIVE: To observe the effect of PRP combined with naringin on the osteogenic differentiation of human bone marrow mesenchymal stem cells(hBMSCs)in vitro. METHODS: BMSCs at passage 3 were divided into four groups: (1) blank control group, cells were cultured in α-MEM; (2) PRP group, cells were cultured in α-MEM containing PRP; (3) naringin group, cells were cultured in α-MEM containing naringin; and (4) combined group, cells were cultured in α-MEM containing PRP and naringin. The contents of used PRP and naringin were 12.5% and 50 μg/L respectively. Cell proliferation was detected by MTT assay. Expression of related genes in hBMSCs was detected by RT-PCR. Alkaline phosphatase staining, collagen type I immunohistochemical staining, and alizarin red staining were used to analyze the osteogenic differentiation of hBMSCs. RESULTS AND CONCLUSION: The proliferation of hBMSCs was increased in each group, especially in the combined group. Cells in all the groups except the blank control group were positive for alkaline phosphatase staining, collagen type I immunohistochemical staining, and alizarin red staining, and the positive effect was more obvious in the combined group. However, negative or weakly positive response was found in the blank control group. At 7 and 14 days, the expression of alkaline phosphatase and collagen type I was significantly higher in the PRP, naringin and combined groups than the blank control group (P < 0.05); at 14 days, the expression of alkaline phosphatase and collagen type I was significantly higher in the combined group than the PRP and naringin groups (P < 0.05). To conclude, PRP combined with naringin can promote the proliferation of hBMSCs and induce the osteogenic differentiation of hBMSCs. Moreover, there is a synergistic effect between PRP and naringin.

Objective To study the risk factors of hyperthyroid heart diseases(HHD) by analyzing clinical features of patients in order to provide a scientific basis for prevention and treatment of HHD. Methods Nine hundred and eighty two cases were selected as objective from in-patient data of Thyroid Disease Treatment Centre of Shandong Province. The cases were divided into hyperthyroidism group and HHD group. The variables of etiology,sex, age, duration of disease, TSH, FT3, FT4 and TRAb were analyzed by comparative analysis. The risk factors were analyzed by logistic regression. Results The prevalence of hyperthyroidism complicated hyperthyroid heart disease was 7.7%(76/982), age, duration of diseases, FT3, TRAb in the HHD group were [(51.4 ± 11.5), (6.3 ±2.1) years, 21.6 pmol/L, 71.6 U/L], in hyperthyroidism group were [(37.9 ± 9.8), (2.6 ± 1.3) years, 14.9pmol/L, 49.6 U/L]. The differences were statistically significant(u = 9.93,15.23, T = 44954,48792.5, P ＜ 0.05)between the two groups. The factors of the older, higher FT3 and TRAb, longer duration, Graves disease (OR =1.751,1.470,1.483,1.445,1.234) increased the risk of HHD. Conclusions Graves disease, longer duration, old age, higher FT3 and TRAb are the risk factors of HHD. Timely prevention and control of risk factors is necessary to reduce the incidence of HHD.

Objective To observe toxic effect of deoxynivalenol(DON)on articular cartilage and synovium of New Zealand rabbits's knee ioints.Methods Fifteen male rabbits were divided randomly into 3 groups:control, high-dosage,and low-dosage group.In high-dosage and low-dosage group,saline solution of DON was injected with a dose of 0.10 and 0.05 ms/kg every 48 h into ear vein of rabbits.Specimen of articular cartilage and synovium were through pathologY methods,and IL-1β,TNF-α,NO levels were assayed in joint liquid,after 20 days. Results Morphological changes were observed, such as synovium inflammative infiltration, chondrocytes deformation and necrosis under light microscope.The levels of IL-1β,TNF-α and NO had statistical significance in comDarison between 3 grouPs(F=19.396,18.195,22.136,P＜0.05).The levels of IL-1β,TNF-α and NO were significantly higher(all P＜0.05),high-dosage[(0.451±0.091),(0.575±0.122)μg/L;(70.27±11.53)μmol/L] and low-dosage group[(0.295±0.107),(0.387±0.131)μg/L;(45.32±12.24)μmol/L]compared with control ((0.1 13±0.049),(0.138±0.087)μg/L;(23.56±9.35)μmoL/L],and high-dosage compared with low-dosage group Conclusions DON results in articular and synovial impairment,which has the symptom similar to osteoarthritis. DON probably causes osteoarthritis.

OBJECTIVETo describe the distribution of cerebral vascular hemodynamic indexes (CVHI).

METHODSA number of 25,355 age 35 and over were selected in the Northeast China by cluster sampling. CVHI were checked during baseline survey and were followed to see the occurrence of stroke. Distribution of CVHI among non-stroke population, individuals prior to the onset of stroke and patients with stroke were described.

RESULTSThe CVHI accumulative score, V(mean), V(max) and V(min) were dramatically decreasing, but RV, Zcv, WV and DR were significantly increasing as age increased. V(max), RV and CP were significantly higher in males but WV was lower than that of females. The CVHI accumulative score, V(min) and RV were 95.0, 10.23 and 75.8 in non-stroke population, 51.25, 6.71 and 122.72 pre stroke group, and 55.0, 6.78 and 115.89 in patients with stroke respectively. There were significant differences among three groups after controlling of age and sex (P < 0.01).

CONCLUSIONVariance of CVHI was closely related to age, and there appeared a significant abnormal of CVHI before and after stroke.

Age Factors ; Aged ; Cluster Analysis ; Female ; Hemodynamics ; Humans ; Male ; Middle Aged ; Risk Factors ; Sex Factors ; Stroke ; physiopathology

OBJECTIVETo investigate the outcome and relative problems of patients over 60 years old underwent orthotopic liver transplantation (OLT).

METHODSData of patients over 60 years old (>or= 60 years old group, n = 59) patients recipients who were 18 to 59 years old (< 60 years old group, n = 500) were reviewed retrospectively.

RESULTSOverall patients survival at 1 year was not significantly different among >or= 60 years old group (66%) and < 60 years group (76%). There were no differences in the operation time, the quantity of blood lost during operation, the days of hospitalization and the incidence of hepatic artery thrombosis between the two groups. The incidence rate of acute rejection reaction in >or= 60 years old group was lower. Both the duration of staying in intensive care unit and the time of using ventilator in >or= 60 years old group were longer than the other group. Moreover, the incidence rates of infection and intracerebral hemorrhage were higher in >or= 60 years old group, which were the primary causes of death in this group.

CONCLUSIONEven though the complications were higher, recipients over 60 years old underwent OLT have more excellent 1 year survival.

Aged ; Female ; Follow-Up Studies ; Humans ; Intraoperative Complications ; Length of Stay ; Liver Transplantation ; adverse effects ; methods ; mortality ; Male ; Middle Aged ; Postoperative Complications ; Retrospective Studies ; Survival Rate ; Treatment Outcome