Objective To establish a satisfactory method of isolating,culturing and determining gut neural crest stem cells,which may provide theoretical basis for clinical application.Methods The guts of embryonic mice removed and dissociated were plated into serum-free DMEM/F12 medium.The mitogen-free DMEM/F12 medium supplementd with 10% fetal bovine serum was used to induce differentiation of GNCSCs.Neurospheres and their derivations were determined with immunocytochemical and immunofluorescent staning.Results Neurospheres were generated in the simplified serum-free medium.The staining results showed that enteric neurospheres were GNCSCs and could differentiate into neurons,glial cells and smooth muscle cells by serum-induction.Conclusion GNCSCs have the capacity of self-renewal and proliferation,and give rise to neurons,glial cells and myofibroblasts.

Cause of Death ; Female ; Hospital Mortality ; Hospitalization ; statistics & numerical data ; Humans ; Infant, Newborn ; Inpatients ; statistics & numerical data ; Male

Objective To study pathogensand diagnostic value of beta2-microglobulin(?2-MG) in cerebrospinal fluid(CSF) in children with aseptic meningitis(AM).Methods Polymerase chain reaction (PCR) or reverse transcription-PCR (RT-PCR) assay,enzyme linked immunosorbent assay (ELISA) and passive particle agglutination were employed respectively to detect herpes simplex virus type-1(HSV-1) DNA,enterovirus(EV) RNA in CSF and the specific antibodies of HSV-1 and mycoplasma pneumonia(MP) in blood and CSF from 80 children with AM (AM group) and 20 cases of complete remission acute leukaemia(control group).Children with AM were divided into the EV meningitis group,HSV-1 meningitis group and MP meningitis group according to respective diagnostic criteria.The levels of ?2-MG in CSF from all groups were detected by radio immunoassay.SPSS 13.0 software was used to analyze the data.Results The infection rate of EV,HSV-1 and MP were 52.5%,11.25%and 5.0%,respectively in children with AM.The infection rate of EV was highest.The sensitivity of PCR was higher than ELISA.There could be false negative PCR results in bloody CSF specimens with HSV-1 DNA.The levels of ?2-MG in CSF was significantly higher in children with AM than that in control group(P

Objective To compare Western blot analysis and immunofluorescence method in Survivin expression. Methods By using Western blot analysis and immunofluorescence method, Survivin gene expressions of the peripheral blood mononuclear cells in 18 patients with leukemia and 10 healthy persons were analysed. Results Positive expression of Survivin gene was detected in 13 of 18 leukemic patients by Western blot analysis and in 11 of 18 leukemic patients by immunofluorescence method. Negative expression of Survivin gene was detected in 10 healthy persons. The results of Survivin expression were the same in 16 patients by Western blots analysis and immunofluorescence method (consistent rate is 88.89%), but the positive expression of Survivin by Western blot analysis was the negative expression by immunofluorescence method in another 2 patients. Conclusion Survivin gene expressed selectively in leukemic cells, but not in normal mononuclear cell in peripheral blood. It indicated that Survivin expression is effective monitoring index in diagnosis and prognosis of leukemia. Survivin expression by Western blot analysis was consistent with immunofluorescence method.

Objective To explore the relation ship between expression of survivin gene in leukemia cells and its clinical effects, and to study the mechanism of survivin resist-apoptosis function in leukemia.Methods Survivin expression was detected by Western blots analysis and expressions of Fas and Caspase were examined by immunohistochemistry in 18 leukemia patients.Results Thirteen cases in peripheral blood mononuclear cell survivin positive expression was detected in 18 leukemia patients(72.2%), but no survivin expression in 10 normal persons. There were significant difference of survivin expression in ALL/ANLL patients groups and different WBC groups(P

Objective: To observe the therapeutic efficacy of warm needling moxibustion plus intra-articular injection of sodium hyaluronate in treating chondromalacia patellae and its effect on inflammatory factors in knee joint fluid. Methods: Sixty-eight patients with chondromalacia patellae were randomized into a control group and an observation group, with 34 cases in each group. The control group was treated with intra-articular injection of sodium hyaluronate, while the observation group was given additional warm needling moxibustion treatment. Before and after treatment, the two groups were scored using Western Ontario and McMaster Universities osteoarthritis index (WOMAC) and visual analog scale (VAS), examined by magnetic resonance imaging (MRI) and determined for the levels of nuclear factor-κB (NF-κB), tumor necrosis factor (TNF)-α and interleukin (IL)-1β in knee joint fluid. Clinical efficacy was estimated after treatment. Results: The effective rate was higher in the observation group than in the control group (P<0.05). After treatment, the scores of pain, stiffness and daily activities, as well as the general score of WOMAC declined significantly in both groups (all P<0.05), and were lower in the observation group than in the control group (all P<0.05); the time cost for climbing up and down one staircase and VAS score decreased markedly in both groups (all P<0.05), and were shorter or lower in the observation group than in the control group (both P<0.05); the MRI grading showed no significant change in the control group after intervention (P>0.05), while the grading in the observation group showed notable improvement (P<0.05), and was better than that in the control group (P<0.05); the levels of NF-κB and IL-1β in knee joint fluid dropped significantly in the control group after treatment (both P<0.05), while the levels of NF-κB, TNF-α and IL-1β in knee joint fluid all decreased significantly in the observation group (all P<0.05) and were lower than those in the control group (all P<0.05). Conclusion: Warm needling moxibustion plus intra-articular injection of sodium hyaluronate can produce definite efficacy in treating chondromalacia patellae; it can mitigate the clinical symptoms, improve the lesion extent of chondromalacia and down-regulate the levels of NF-κB, TNF-α and IL-1β in knee joint fluid.

Objective To explore the high risk factors of brain injury in preterm infants,and to reduce its morbidity and improve the developmental outcome.Methods One hundred and thirty preterm infants,who were admitted to our neonatal intensive care unit(NICU)between Aug.2005 and Aug.2007,were scanned by echo in 1,3,4,7,15 days,and 1,3 and 6 months after birth,respectively.Those who had intraventricular hemorrhage(IVH)of grade Ⅰor Ⅱ were regarded as mild brain injury,whereas those who had IVH of grade Ⅲ or Ⅳ or periventricular leukomalacia(PVL)were regarded as severe brain injury.Logistic regression was adopted to analyze 17 factors:gestational age,birth weight,hypertension syndrome during pregnancy,premature rupture of membranes,modalities of delivery,fetal distress,asphy-xiate,resuscitation,surfactant,apnea,seizures,hypoxia,hypercarbia,hypocarbia,acidosis,use of oxygen,nasal constant positive airway pressure or mechanical ventilation.Results Among 130 preterm infants,88 cases(66.7%)were detected with brain injury,which included 29 cases(33%)with mild brain injury(5 cases with IVH of grade Ⅰ,24 cases with IVH of grade Ⅱ)and 59 cases(67%)with severe brain injury(53 cases with IVH of grade Ⅲ,1 case with IVH of grade Ⅳ and 5 cases with PVL).Gestational age and birth weight were the fundamental factors of brain injury in premature infants.The smaller the gestational age and the lower the birth weight,the highter the brain injury rate.Resuscitation,hypoxia,the use of auxiliary ventilation were also important high risk factors of brain injury in preterm infants.All these high risk factors could influence the autoregulation of cerebral blood and trigger or aggravate brain injury of preterm infants.Conclusions Smaller gestational age,lower birth weight,resuscitation,hypoxia,the use of auxiliary ventilation were all the high risk factors of brain injury in premature infants,which could influence the parameters of cerebral blood dynamics by influencing cerebral blood autoregulation of preterm infants and lead to the occurrence of brain injury in premature.

Objective To discuss the clinical manifestations, imaging data and DSA findings of lacunar infarction (LI). Methods One hundred and thirty-three patients, admitted to our hospital from May 2002 to April 2008, were chosen in our study; these patients with first onset as LI were confirmed by Head CT or MR; the clinical manifestations and imaging data were retrospectively analyzed; DSA was also performed on these patients and DSA findings were concluded. Results One hundred and thirty-three patients were clinically manifested as pure motor hemiplegia (PMH, n=42, 31.6%) and sensorimotor stroke (SMS, n=36, 27.1%). Two hundred and eighty-three lesions were noted by CT/MR examinations, including 78 locating at the endocyst (27.6%) and 121 locating at the corona radiate+greater oval center (91.0%). Forty-four patients were noted as having 101 intracranial vessel lesions by DSA, including 38 patients with angiostenosis, 6 with Moyamoya and 1 with single intracranial aneurysm; of the patients with angiostenosis, 95 lesions (34 in the offending vessels and 61 in other vessels) were found. Among the DSA (+) patients, PMH (n=21) and SMS (n=10) were mainly noted with their lesions locating at the endocyst (n=23) and the corona radiate+greater oval center (n=31); At least 1 high-risk factor such as hypertension, diabete, hyperlipemia, coronary heart disease and arial fibrillation was found in 44 patients. Conclusion The pathogeneses of LI are various. Main artery infarction may co-exist in some cases. PMH and SMS are common with their lesions frequently locating at basal ganglia area and corona radiate of the cerebral hemisphere. High risk factor exists in most patients with cerebrovascular diseases.

Objective To explore the protective effect of astragalus injection(黄芪注射液)on cerebral cells after craniocerebral injury.Methods Sixty male Sprague-Dawley(SD)rats were randomly divided into five groups(each n=12).In the control group,only anesthesia and cranial bone surgery was performed, which did not impact on dura mater.In the model group,a freefall method was used to cause the left parietal lobe of brain limited contusion and saline was given.In the low,medium and high-dose astragalus injection (5,10,20 ml/kg)treatment groups,astragalus injection was injected intraperitoneally after injury at 30 minutes and 12 hours after injury respectively.After 24 hours of injury in each group,the rats were executed.The brain tissue from contusion was taken out and content changes of Ca~(2+),Mg~(2+),glutamate (Glu)and plasma endothelin(ET)were measured.Results In the model group,the Ca~(2+),Glu and ET contents in the rat brain tissues were significantly higher,but Mg~(2+)content was markedly lower than that of the control group(all P0.05).Conclusion Astragalus has a protective effect on nerve cells after craniocerebral injury.

Objective To identify mutations site and clinical characteristics of a familial hypercholesterolemia(FH) proband diagnosed clinically through DNA sequencing and family analysis in the proband and his family members of 3 generations.Methods Blood samples and clinical data of the kindred of total 29 from 3 generations members were collected.Proband had a physical examination electrocar-diogrom and vascular ultrasound.The proband and his family members took routine clinical exams,and genomic DNA was isolated.The promoter region and the 18 exons of low density liporotein receptor(LDLR) gene were screened by Touch down polymerase chain reaction -single strand conformation polymorphism(PCR-SSCP) and DNA sequencing.The result of sequencing were matched gene sequence published in the BLAST database.Results 1.Increased intima-media thickness and plaque were detected in the common carotid artery,right subclavian artery of the proband.Aortic valve regurgitation was found by echocardiography.2.No mutation R3500Q of ApoB100 was observed.3.Two heterozygous mutations in exon 10 and 13 of LDLR gene (W462X and A606T) were identified.The proband and 5 members of paternal relatives showed W462X heterozygosis mutation in exon 10 of LDLR gene which introduced the change from tryptophone to a new stop codon.The proband's mother and grandmother harboured A606T heterozygous mutation in exon 13 of LDLR gene due to a single base pair substitution of G for A in the codon for residue 1 879.Conclusions Disease causing mutations of proband are W462X and A606T compound heterozygosis mutation in exon 10 and 13 of LDLR gene inherited from mother and father.Proband shows homozyous phenotype though the genotype analysis indicates heterozygous mutations.