Integrating a Numerical Taxonomic Method and Molecular Phylogeny for Species Delimitation of Melampsora Species (Melampsoraceae, Pucciniales) on Willows in China.

The species in genus Melampsora are the causal agents of leaf rust diseases on willows in natural habitats and plantations. However, the classification and recognition of species diversity are challenging because morphological characteristics are scant and morphological variation in Melampsora on willows has not been thoroughly evaluated. Thus, the taxonomy of Melampsora species on willows remains confused, especially in China where 31 species were reported based on either European or Japanese taxonomic systems. To clarify the species boundaries of Melampsora species on willows in China, we tested two approaches for species delimitation inferred from morphological and molecular variations. Morphological species boundaries were determined based on numerical taxonomic analyses of morphological characteristics in the uredinial and telial stages by cluster analysis and one-way analysis of variance. Phylogenetic species boundaries were delineated based on the generalized mixed Yule-coalescent (GMYC) model analysis of the sequences of the internal transcribed spacer (ITS1 and ITS2) regions including the 5.8S and D1/D2 regions of the large nuclear subunit of the ribosomal RNA gene. Numerical taxonomic analyses of 14 morphological characteristics recognized in the uredinial-telial stages revealed 22 morphological species, whereas the GMYC results recovered 29 phylogenetic species. In total, 17 morphological species were in concordance with the phylogenetic species and 5 morphological species were in concordance with 12 phylogenetic species. Both the morphological and molecular data supported 14 morphological characteristics, including 5 newly recognized characteristics and 9 traditionally emphasized characteristics, as effective for the differentiation of Melampsora species on willows in China. Based on the concordance and discordance of the two species delimitation approaches, we concluded that integrative taxonomy by using both morphological and molecular variations was an effective approach for delimitating Melampsora species on willows in China.

Association of Pneumococcal Carriage and Expression of Foxp3+ Regulatory T Cells and Th17 Cells in the Adenoids of Children.

BACKGROUND: Pneumococcal carriage in the nasopharynx is a primary means of transmission and a necessary prerequisite for pneumococcal disease. OBJECTIVES: We analyzed the relationship between expressions of Foxp3+ regulatory T (Treg) cells and Th17 cells, and pneumococcal carriage in the adenoids of children who were either positive or negative for pneumococci. METHODS: We collected adenoidal tissue and nasopharyngeal swab samples from children undergoing an adenoidectomy. Adenoidal mononuclear cells were isolated, cultured and then stimulated with culture concentrated supernatant (CCS) obtained from a D39 bacterial strain. RESULTS: Foxp3+ Treg cells were upregulated and Th17 cells were downregulated in populations of adenoidal mononuclear cells obtained from the pneumococcus-positive group. Following CCS stimulation, the increment in Foxp3+ Treg cells in the pneumococcus-positive group was significantly greater than that in the pneumococcus-negative group, while the increment in Th17 cells was less as compared to that in the pneumococcus-negative group. These results were consistent with variations in levels of Foxp3 mRNA and retinoic acid receptor-related orphan receptor-γt mRNA in adenoidal mononuclear cells. Levels of IL-17A and IL-6 in adenoid tissue were higher in the pneumococcus-negative group, and the levels of TGF-ß in adenoid tissue were lower in the pneumococcus-negative group compared to the pneumococcus-positive group. Pneumococcal carriage in children was closely associated with the expressions of Foxp3+ Treg and Th17 cells in the adenoid. CONCLUSION: Upregulation of Foxp3+ Treg cells might downregulate the production of Th17 cells in the adenoid, resulting in decreased scavenging of Streptococcus pneumoniae and chronic pneumococcal carriage.

Clinical, molecular, and T cell subset analyses in a small cohort of Chinese patients with hyper-IgM syndrome type 1.

Type 1 hyper-IgM syndrome (HIGM1) is a rare primary immunodeficiency disease caused by mutations in the CD40L gene. Patients often present with recurrent infections and autoimmune manifestations. We investigated the clinical and molecular characteristics of HIGM1 in thirteen patients from the Chinese mainland and examined the proportion of CD4(+)CD25(+)FoxP3(+)Treg, Th17, and Th1 cells in the peripheral blood. We identified ten distinct CD40L mutations in eleven patients: one missense mutation, one nonsense mutation, one insertion mutation (in frame), and seven deletions. Six of these mutations were novel. We observed the percentage of Tregs in the peripheral blood of HIGM1 patients decreased markedly compared with that in healthy controls, but no statistically significant differences was found in the percentages of Th17 and Th1. The identified mutations reflect the heterogeneity of the CD40L gene in HIGM1. Precise genetic diagnosis of HIGM1 will enable appropriate therapeutic interventions, reliable detection of carriers, and genetic counseling. Skewed Treg, Th17/Treg, and Th1/Treg profiles may be associated with immune responses to autoimmunity or infection, which requires replication in larger studies.

[Comorbidities and functional impairments in children with attention deficit hyperactivity disorder].

OBJECTIVE: To assess comorbidities and functional impairments in children with attention deficit hyperactivity disorder (ADHD), and to investigate their relationship with the core symptoms (attention deficit and hyperactivity) of ADHD. METHODS: A total of 319 children with suspected ADHD were included in the study. The Vanderbilt ADHD Parent Rating Scale (VADPRS) was completed by their parents. Diagnosis and classification were performed based on the Diagnostic and Statistical Manual of Mental Disorders, 4th Edition. Comorbidities and functional impairments were evaluated according to the VADPRS. Children with various types of ADHD were compared in terms of comorbidities and functional impairments, and their relationship with the core symptoms of ADHD was analyzed. RESULTS: Of the 319 children, 196 were diagnosed with ADHD, including 84 cases of predominantly inattentive type (ADHD-I), 35 cases of predominantly hyperactive-impulsive type (ADHD-HI) and 77 cases of combined type (ADHD-C); 123 did not meet the diagnostic criteria for ADHD. At least one other psychiatric disorder (oppositional defiant disorder, conduct disorder or emotional disorder) was seen in 63.8% (125/196) of the children with ADHD, versus 37.4 % (46/123) of the children without ADHD (P<0.05). The incidence of oppositional defiant disorder and conduct disorder in the ADHD-C subgroup was significantly higher than in the ADHD-I subgroup (P<0.05). The sums of oppositional defiant disorder, conduct disorder and emotional disorder symptoms were weakly correlated with the sums of hyperactive-impulsive and inattentive symptoms (P<0.01). Up to 89.8% of children with ADHD and 74.8% of children without ADHD showed functional impairments (P<0.05). The ADHD-C subgroup had a significantly higher overall incidence of functional impairments than the ADHD-I and ADHD-HI subgroups (P<0.05). The sum of inattentive symptoms was weakly correlated with the scores of learning ability, sibling relationship and participation in organized activities (P<0.01), and the sum of hyperactive-impulsive symptoms was weakly correlated with the score of sibling relationship (P<0.01). CONCLUSIONS: The incidence of comorbidities and functional impairments among children with ADHD is high, especially in those with ADHD-C. The severity of core symptoms in children with ADHD can influence the occurrence of comorbidities and functional impairments. The incidence of psychiatric disorders and functional impairments is also high in children with suspected ADHD who do not meet the diagnostic criteria for ADHD, so attention also needs to be paid to interventions among these children.

[Diagnostic value of Vanderbilt ADHD Parent Rating Scale in attention deficit hyperactivity disorder].

OBJECTIVE: To study the value of the Vanderbilt ADHD Parent Rating Scale (VADPRS) in the diagnosis of attention deficit hyperactivity disorder (ADHD). METHODS: VADPRS were completed by parents of 319 children with suspected ADHD. The children were then evaluated by a specialist based on the Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV) and 196 of them were diagnosed with ADHD. The value of VADPRS in the diagnosis of attention deficit and hyperactivity was evaluated using ROC curves. Diagnostic evaluation indexes at best operating point were calculated. Kappa values were calculated to explore the consistency of items in VADPRS and corresponding items in the DSM-IV criteria. RESULTS: The area under the ROC curve for the diagnosis of attention deficit by VADPRS was 0.791. At the best operating point, its sensitivity was 0.83, specificity was 0.63, positive predictive value was 0.69 and negative predictive value was 0.79. The area under the ROC curve for the diagnosis of hyperactivity by VADPRS was 0.855. At the best operating point, its sensitivity was 0.82, specificity was 0.76, positive predictive value was 0.65, and negative predictive value was 0.88. The negative predictive value of VADPRS in general population screen was 0.99, based on the results of this study. The consistency of items in the VADPRS and corresponding items in DSM-Ⅳ criteria was poor, with the Kappa value of most items being less than 0.40. CONCLUSIONS: VADPRS is suitable for a general population screen for ADHD and it is helpful in the clinical diagnosis of ADHD, but its results can be influenced by parents' awareness and perception of children's behavior, and cannot replace the interview and judgment of professionals.

Epidemiological survey on newborns born at the obstetric departments in hospitals in mid-southern region of China in 2005.

OBJECTIVE: To investigate the situations at birth of newborns in the mid-southern region of China by performing a survey on the newborns born at urban hospitals. METHODS: A total of 23 hospitals in the mid-southern region of China were selected to participate in this survey. The data of 15,582 newborns who were born at the obstetric departments from January 1, 2005 to December 31, 2005 were retrospectively investigated. RESULTS: The male to female ratio among newborns was 1.16:1. The incidence of preterm birth was 8.11%, while very low birth weight (VLBW) infants accounted for 0.73%. The rates of spontaneous vaginal delivery and cesarean section ware 57.52% and 40.82%, respectively, while the other delivery modes accounted for 1.66%. The incidence of neonatal asphyxia was 3.78% (0.75% for heavy asphyxia). The mortality of newborns was 0.55% (5.56% for preterm infants). CONCLUSIONS: The incidences of preterm birth and neonatal asphyxia are relatively high in the mid-southern region of China. The rate of cesarean section is abnormally high, which is due mainly to social factors.

[A survey of neonatal births in maternity departments in urban China in 2005].

OBJECTIVE: To study the epidemiology of births in urban China. METHODS: A retrospective study was conducted on neonates born in 2005 in the maternity departments of 72 urban hospitals from 22 provinces in China. RESULTS: A total of 45722 infants born between January 1, 2005 and December 31, 2005 were enrolled. The male to female sex ratio was 1.13:1. Preterm births accounted for 8.1%. The incidence of very low birth weight infants was 0.7%. A total of 99.7% of mothers delivering at term had conceived naturally and 0.3% had experienced assisted reproduction. A total of 98.4% of mothers who delivered preterm had conceived naturally and 1.6% had experienced assisted reproduction. The proportion of vaginal deliveries was 50.8% compared to 49.2% delivered by cesarean sections. Many cesarean sections (38.1%) were due to social factors. Infants with an Apgar score≤7 at 1 minute accounted for 4.8%, and 1.6% of infants had an Apgar score≤7 at 5 minutes. Of all the infants included in the study, 7.14% were admitted to neonatal units for treatment. The death rate of all included infants was 0.74%. CONCLUSIONS: The proportion of preterm births was higher in 2005 than in 2002-2003. The proportion of cesarean section deliveries was much higher in urban China than in most other Asian countries and America.

Epidemiologic survey on hospitalized neonates in China.

OBJECTIVE: To carry out a nationwide epidemiologic survey on the neonates in urban hospitals with an attempt to understand the disease spectrum and treatment outcomes of hospitalized neonates in China. METHODS: The clinical data of 43,289 hospitalized neonates from 86 hospitals in 47 Chinese cities (22 provinces) between January 1, 2005 and December 31, 2005 were retrospectively analyzed. RESULTS: The male:female ratio was 1.73:1. Premature infants accounted for 26.2% of the hospitalized neonates, which was higher than that reported in 2002 (19.7%). The top three diseases during the neonatal period were jaundice, pneumonia, and hypoxic-ischemic encephalopathy. The incidences of pneumonia, meconium aspiration syndrome, and bilirubin encephalopathy in term infants were higher than those in premature infants, while the incidences of asphyxia, respiratory distress syndrome, and pulmonary hemorrhage in term infants were lower than those in premature infants. The incidences of asphyxia, small for gestational age infant, and wet lung were higher in neonates whose mother had pregnancy induced hypertension. The outcomes of these hospitalized neonates included: recovered, 63.9%; improved, 27.3%; discharged due to the family's own decisions, 7.6%, and died, 1.2%. Nearly half (46.4%) of the neonatal death occurred within 24 hrs after admission. CONCLUSION: The incidence of premature birth shows an increasing trend among hospitalized neonates. Since the neonatal deaths mainly occur within 24 hrs after admission, monitoring during this period should be enhanced.

[An epidemiologic investigation of newborns from obstetric departments in the central south region of China].

OBJECTIVE: To investigate the birth information of newborn infants from obstetric departments in the Central South Region of China. METHODS: A retrospective investigation was carried out in 15582 newborns from obstetric departments of 23 hospitals in the Central South Region of China between January 1 and December 31 of 2005. RESULTS: The sex ratio (male/female) of neonates was 1.16∶1. The proportion of preterm infants was 8.11%. The very low birth weight infants accounted for 0.73%. The neonates born by spontaneous labor accounted for 57.52%. Cesarean sections accounted for 40.82% (social factor of cesarean section: 29.91%). The incidence of neonatal asphyxia was 3.78%, in which 0.75% of the cases were severe asphyxia. The mortality of newborn infants was 0.55%, in which the mortality of preterm infants was 5.56%. CONCLUSIONS: The proportion of preterm infants and the incidence of neonatal asphyxia is high in the Central South Region of China. The proportion of births delivered by cesarean section is high, and social factors are probably responsible for the high rate.

The role of ß-catenin signaling pathway on proliferation of rats neural stem cells after hyperbaric oxygen therapy in vitro.

ß-catenin, a protein that functions in both cell adhesion and Wnt signaling, plays a key role in mammalian neural development. To investigate the role of ß-catenin in hyperbaric oxygen therapy (HBO)-induced neurogenesis after hypoxic ischemic brain damage (HIBD), we transfected ß-catenin siRNA and negative control siRNA into neural stem cells (NSCs) after HIBD. We found that HBO promoted NSCs differentiate into neurons or oligodendrocytes, and inhibited NSCs differentiate into astrocytes; HIBD brain tissue extract conditioned cultures promoted NSCs differentiate into neurons; ß-Catenin siRNA decreased the NSE-positive neurons and increased GFAP-positive astrocytes in the NSCs in vitro. Furthermore, the expression of Ngn1 protein and mRNA in NSCs was increased when HBO promoted NSCs differentiate into neurons after HIBD, and the expression of BMP-4 protein and mRNA was decreased when HBO depressed NSCs differentiate into astrocytes after HIBD. These results showed that ß-catenin-mediated transcriptional activation functions in the decision of NSCs to proliferate neurogenesis during HBO-induced after HIBD, and suggested that HBO therapy promotes the proliferation of neural stem cells in vitro, an effect that may be correlated with ß-catenin protein and HBO therapy could promote neurogenesis by ß-catenin-induced activated Ngn1 gene and repress astrocytogenesis by ß-catenin-induced down-regulated BMP-4 gene.

Induction of protective and therapeutic antitumor immunity by a DNA vaccine with C3d as a molecular adjuvant.

Although the critical role of complement component C3d as a molecular adjuvant in preventing virus infection is well established, its role in cancer therapies is unclear. In this study, we have engineered a DNA vaccine that expresses extracellular region of murine VEGFR-2 (FLK1(265-2493)) and 3 copies of C3d (C3d3), a component of complement as a molecular adjuvant, designed to increase antitumor immunity. VEGFR-2 has a more restricted expression on endothelial cells and is upregulated once these cells proliferate during angiogenesis in the tumor vasculature. Immunization of mice with vector encoding FLK1(265-2493) alone generated only background levels of anti-VEGFR-2 antibodies and slight inhibitory effect on tumor growth. However, the addition of C3d3 to the vaccine construct significantly augmented the anti-VEGFR-2 humoral immune response and inhibited the tumor growth. The antitumor activity induced by vaccination with vector encoding FLK1(265-2493)-C3d3 fusion protein was also demonstrated via growth inhibition of established tumors following passive transfer of immune serum from vaccinated mice. Our results suggest that vaccination with vector encoding FLK1(265-2493) with C3d3 as a molecular adjuvant induces adaptive humoral activity, which is directed against the murine VEGFR-2 and can significantly inhibit tumor growth, and that administration of C3d as a molecular adjuvant to increase antibodies levels to VEGFR-2 may provide an alternative treatment modality for cancer therapies.

[Effect of hyperbaric oxygen administered at different pressures and different exposure time on differentiation of neural stem cells in vitro].

OBJECTIVE: To study the effect of hyperbaric oxygen (HBO) administered at different pressures and different exposure time on the differentiation of neural stem cells (NSCs) in vitro. METHODS: The cerebral cortices from newborn rats (0-3 days old) were sterilely collected, digested, and centrifuged. After removal of the supernatant, the cells were re-suspended with DMEM/F12 medium containing B27, bFGF and EGF. The NSCs of 2-3 passages were randomly divided into seven groups: a control (untreated) and 6 HBO treatment groups that NSCs were subjected to HBO treatment of different pressures (1, 2 or 3 ATA) and different exposure time (30 or 60 minutes). The differentiated NSCs were examined by neuron-specific enolase (NSE) immunocytochemistry 24 hrs later. Percentage of NSE positive cells differentiated from NSCs was assessed by fluorescent microscopy. RESULTS: The percentage of NSE positive cells differentiated from NSCs was the highest in the HBO 2ATA-60 min group (9.17+/-0.50%) (P<0.01), followed by the HBO 3ATA-60 min (7.89+/-0.62%), HBO 2ATA-30 min (6.72+/-0.76%), HBO 3ATA-30 min (6.08+/-0.57%), HBO 1ATA-60 min (5.45+/-0.52%), HBO 1ATA 30 min (3.85+/-0.44%) and control groups (3.72+/-0.88%). In addition to the HBO 1ATA-30 min group, the other HBO treatment groups had increased significantly percentage of NSE positive cells compared with the control group (P<0.01). Under the same pressure, the 60 min treatment groups had increased significantly percentage of NSE positive cells compared with the 30 min treatment groups (P<0.01). CONCLUSIONS: HBO treatment (2 ATA, 60 minutes) produces a best effect in the differentiation of NSCs into neurons.

[Investigation of birth state and disease spectrum in 1,434 hospitalized neonates].

OBJECTIVE: To investigate the birth state of neonates and the disease spectrum of hospitalized neonates from a primary hospital, and compare with the national data of the same period. METHODS: A retrospective investigation was carried out in 1,434 neonates born or hospitalized in this hospital from January 2005 to December 2005. RESULTS: During the investigation period, there were 1,100 neonates born in the department of obstetrics. The incidence of premature birth was 2.3%. The caesarean birth accounted for 54.2%, significantly higher than the national average (49.2%, p<0.01). The neonatal mortality was 0.2%. The incidences of antepartum hemorrhage, threatened abortion, and pregnancy infection in preterm infant' s mothers were significantly higher than those in full-term infant' s mothers. A total of 344 neonates were admitted to the department of pediatrics during the investigation period. Preterm infants accounted for 38.0% which was higher than the national average (26.2%; p<0.01). Beside preterm infants, asphyxia, respiratory distress syndrome (RDS), sepsis and intracranial hemorrhage were shown to have a significantly higher proportion than the national averages. The mortality of hospitalized neonates was 0.9%. CONCLUSIONS: The higher cesarean section rate should be controlled in our hospital. Prenatal health care and fetal monitoring should be strengthened to decrease the incidence of premature birth, RDS, sepsis and intracranial hemorrhage, thus reducing the mortality of neonates.

[Hyperbaric oxygen promotes the migration and differentiation of endogenous neural stem cells in neonatal rats with hypoxic-ischemic brain damage].

OBJECTIVE: To explore the effects of hyperbaric oxygen (HBO) treatment on the migration and differentiation of endogenous neural stem cells (NSCs) in neonatal rats with hypoxic-ischemic brain damage (HIBD). METHODS: Seven-day-old Sprague-Dawley rats were randomly divided into the normal control (CON), the HIBD model and the HBO groups (HBO treatment was administered at 2 ATA, once daily for 7 days within 3 hrs after HIBD). HIBD model was prepared according to the classic Rice-Vannucci method. BrdU/DCX, BrdU/beta-tubulin, BrdU/GFAP and BrdU/O4 immunofluorescence were examined by confocal microscopy in the subventricular zone (SVZ) and the cortex 7, 14 and 28 days after HBO treatment. RESULTS: The BrdU(+)DCX(+) cells in the SVZ (84 +/- 21 cells/mm2) in the HBO group were significantly higher than those in the CON group (39 +/- 14 cells/mm2) (p<0.05) and the HIBD model group (68 +/- 17 cells/mm2) (p<0.05) 7 days after HBO treatment. Fourteen days after HBO treatment, the BrdU(+) DCX(+) cells decreased in the SVZ and more cells were observed in the cortex in the HBO group as compared with the CON group (p<0.01). The BrdU(+) beta-tubulin(+), BrdU(+)GFAP(+) and BrdU(+) O4(+) cells were observed in the cortex, and more BrdU(+)beta-tubulin(+) and BrdU(+) O4(+) cells were observed in the HBO group as compared with the CON and the HIBD model groups (p<0.05) 28 days after HBO treatment. CONCLUSIONS: HBO treatment may promote endogenous NSCs to migrate to the cortex and differentiate into mature neurocytes in neonatal rats with HIBD.

[Protective effects of delayed multiple course hyperbaric oxygen treatment against hypoxic-ischemic brain damage in neonatal rats].

OBJECTIVE: To study the protective effects of multiple course hyperbaric oxygen (HBO) treatment against hypoxic-ischemic brain damage (HIBD) in neonatal rats when HBO treatment is delayed (96 hrs after the HIBD event). METHODS: Eighty-eight 7-day-old Sprague-Dawley rat pups were randomly assigned to control, HIBD and HBO groups. The HBO group was subdivided into cohorts receiving treatment 2 h, 48 h and 96 h, respectively, after HIBD was induced. The three subgroups comprising different therapeutic windows were further randomly assigned to receive 1, 2 or 3 courses of HBO treatment ("HBO-1, -2 and -3 sub-groups"). HBO was administered once daily (2 ATA), a course lasting for seven days. There was an interval of three days between the courses. All pups were sacrificed at the end of HBO treatment (31 days after HIBD). TUNEL staining was used for testing neuronal apoptosis in the cortex and the CA1 of the hippocampus, and NSE staining was used to ascertain cortical neuronal population. RESULTS: 1.There were significantly more TUNEL positive cells in the HIBD group than in the control group; NSE positive cells were significantly lower than in controls (P<0.01). 2. With the more delayed therapeutic window, the effects of apoptosis inhibition and neuronal protection of a single course of HBO were gradually reduced. 3. With increasing courses of HBO treatment, the effects of apoptosis inhibition and neuronal protection of HBO increased gradually in rats receiving treatment 48 and 96 hrs after HIBD. In the HBO group receiving treatment 2 hrs after HIBD, the number of apoptotic cells and NSE positive cells were close to that of the control group after one course of HBO treatment. CONCLUSIONS: One course of HBO administered within 2 hrs after HIBD can effectively inhibit neuron apoptosis and protect neurons. The effects of apoptosis inhibition and neuron protection of HBO can be increased through increasing the number of HBO treatment courses in neonatal rats with HIBD even if initiation of treatment is delayed after HIBD.

[Indexes of forensic identification by the digital orthopantomogram of the normal teeth].

OBJECTIVE: To explore more concise and unified forensic identification indexes for people with none dental disease in digital orthopantomogram. METHODS: To select randomly 170 digital orthopantomogram with none dental disease. Then to select indexes for full dentition patterns and dental alignment patterns according to the dental physiological variations and the characters of dental alignment respectively. Finally diversity of the indexes would be evaluated by statistical analysis. RESULTS: The group with none dental disease had 74 kinds of full dentition pattern in 170 samples, thus its diversity was 43.53%. The group had 129 kinds of dental alignment pattern, thus its diversity was 75.88%. The group had 150 kinds of full dentition/dental alignment pattern, thus its diversity was 88.24%. CONCLUSION: The diversity of the full dentition pattern was not very good. So the full dentition coding was not very effective when it was used solely. The diversity of dental alignment pattern was good. So the method of dental alignment coding could be used in the maxillofacial forensic identification. If the group was coded by the full dentition and dental alignment pattern at the same time, its diversity was better than any single pattern. So the method would be valuable in forensic identification.

[Effect of intracerebral transplantation of rat bone marrow stromal cells on brain white matter of neonatal rats with hypoxic-ischemic brain damage].

OBJECTIVE: To study the effect of intracerebral transplantation of bone marrow stromal cells (BMSCs) on brain white matter of neonatal rats with hypoxic-ischemic brain damage (HIBD). METHODS: Thirty-four 7-day-old neonatal rats were randomly assigned to three groups: normal control (n=10), HIBD (n=12) and HIBD+BMSCs transplantation (n=12). The HIBD and the HIBD+BMSCs transplantation group rats were subjected to left carotid artery ligation, followed by hypoxia exposure for 2 hrs, in order to induce HIBD. The rats in the HIBD+BMSCs transplantation group received transplantation of BMSCs labeled nucleus with Hochest 33324 into the left hippocampus 24 hrs after HIBD induction. Myelin basic protein (MBP) expression in the left corpus callosum and the subcortical white matter and the number of oligodendrocyte precursors positively stained O4 in the left periventricular area and the subcortical white matter were detected by immunohistochemistry at ages of 45 days. RESULTS: The labeled BMSCs survived and were found mainly in the left hemisphere 37 days after transplantation. The positive rate of O4 expressed by the transplanted BMSCs was 3.70+/-1.09%. More hypomyelination in the left corpus callosum and the subcortical white matter, and less number of O4 positive oligodendrocytes in the left periventricular area and the subcortical white matter were found in the HIBD group compared with the normal control group (P<0.01). The HIBD rats receiving BMSCs transplantation had increased O4 positive oligodendrocytes in the left periventricular area and the subcortical white matter and improved MBP immunoreactivity in the left corpus callosum and the subcortical white matter compared with the HIBD group (P<0.01). CONCLUSIONS: Intracerebral transplantation of BMSCs can improve brain white matter damage in neonatal rats with HIBD.

[Differentiation of rat bone marrow stromal cells into neural cells induced by hypoxic-ischemic brain tissue extracts in neonate rats].

OBJECTIVE: To investigate the effect of brain tissue extracts in neonate rats with hypoxic-ischemic brain damage (HIBD) on the differentiation of bone marrow stromal cells (BMSCs) into neural cells. METHODS: Fifteen 7-day-old neonate rats were induced HIBD by left carotid artery ligation and hypoxia exposure, and another 15-day-old neonate rats were served as normal rats. The left and right brain tissue extracts of the normal and HIBD rats were prepared 24 h after the HIBD (8-day old), 72 h after the HIBD (10-day old), and 7 d after the HIBD (14-day old), respectively (n=5). The rat BMSCs of passage 3-5 were cultured in the medium with or without previous brain tissue extracts. The expressions of neuron specific enolase (NSE), glial fibrillary acidic protein (GFAP) and O(4) marked oligodendrocyte were detected after 3 days by immunocytochemistry. RESULTS: The expressions of NSE, GFAP and O(4) of BMSCs cultured in the medium with left or right brain tissue extracts of different day old normal rats were higher than those of BMSCs cultured without the extracts, respectively (P<0.01), and the expressions of NSE, GFAP and O(4) of BMSCs cultured in the medium with left brain tissue extracts of 8 day old and 10 day old HIBD rats were higher than those of BMSCs cultured with right brain tissue extracts of the same day HIBD rats and BMSCs cultured with left or right brain tissue extracts of the same day normal rats (P<0.01 or P<0.05). The expressions of NSE, GFAP and O(4) of BMSCs cultured in the medium with left brain tissue extracts of 8-day-old HIBD rats were higher than those of BMSCs cultured with left brain tissue extracts of 10-day-old and 14-day-old HIBD rats (P<0.01 or P<0.05). CONCLUSION: The brain tissue extracts of normal and HIBD rats can induce BMSCS into neural cells, and the damaged brain tissue extracts of 8-day-old HIBD rats is the best inductor.

[Effect of hyperbaric oxygen therapy administered at different time on white matter damage following hypoxic-ischemic brain damage in neonatal rats].

OBJECTIVE: A recent study has suggested that hyperbaric oxygen (HBO) therapy administered within 3 hrs following hypoxic-ischemic brain damage (HIBD) may alleviate brain white matter damage (WMD) in neonatal rats. However it is unclear whether a delayed HBO therapy (more than 3 hrs following HIBD) has neuroprotective effects in neonatal rats. This study aimed to explore the effect of HBO therapy administered at different time points following HIBD on WMD in neonatal rats. METHODS: The HIBD model was prepared according to the Rice-Vannucci procedure in 7-day-old Sprague-Dawley rats. HBO therapy was administered at 3, 6, 12, 24 or 72 hrs after HIBD, once daily for consecutive 7 days. T-maze test, the foot-fault test and the radial arm maze test were performed after 14 days of HIBD. Myelin basic protein (MBP) in the callositas and corpora striata was examined by immunohistochemical method 28 days after HIBD. RESULTS: The rats receiving HBO therapy at 3, 6 and 12 hrs after HIBD performed significantly better in the T-maze test, the radial arm maze test and the foot-fault test than the untreated HIBD rats. There were no significant differences in the behavioral test results between the HBO-treated groups administered HBO at 24 and 72 hrs after HIBD and the untreated HIBD group. The MBP expression in the HBO-treated groups treated within 12 hrs after HIBD was significantly higher than that in the untreated HIBD group (P < 0.05). When the HBO therapeutic window was delayed to 24 hrs after HIBD, there were no significant differences in the MBP expression between the HBO-treated and the untreated HIBD groups. CONCLUSIONS: HBO therapy administered within 12 hrs following HIBD can alleviate brain WMD in neonatal rats, but the efficacy of HBO therapy administered 24 hrs after HIBD does not appear to be satisfactory.