Objective: To investigate the developmental toxicity of 2,4-dinitrochlorobenzene in zebrafish embryos. Methods: AB wild-type male and female zebrafish were selected to mate and spawn, then the eggs were cultured with Holt buffer solution. Six dose groups ( 0.4, 0.8, 1.0, 1.2, 1.6, 2.0 μg/mL ), a solvent control group and a cosolvent control group, were set up with 20 embryos each. Malformations and death of embryos were observed at 48, 72 and 96 hpf ( hours post fertilization ), the mortality and 50% lethal concentration ( LC50 ) were also calculated. Results: At 48, 72 and 96 hpf, the LC50 of 2,4-dinitrochlorobenzene on zebrafish embryos were 1.668, 1.043 and 0.895 μg/mL, respectively, with a downward trend. After 72 hpf, when the concentration reached 2.0 μg/mL, all the zebrafish died. In the range of 0.4-2.0 μg/mL, the mortality of zebrafish at 48, 72 and 96 hpf increased with the increase of 2,4-dinitrochlorobenzene concentration ( all P<0.05 ); the malformation rate of zebrafish embryos at 48 hpf increased with the increase of 2,4-dinitrochlorobenzene concentration ( P<0.05 ). Zebrafish embryos exposed to 2,4-dinitrochlorobenzene led to yolk sac edema, pericardial edema and spinal curvature. Conclusion 2,4-dinitrochlorobenzene can affect the development of zebrafish embryos, which will lead to lethal and teratogenic effects.

OBJECTIVETo investigate the feasibility and safety of autologous peripheral blood hematopoietic stem cell transplantation (auto-PBHSCT) and its therapeutic effect on refractory rheumatism among preschool children.

METHODSThree boys with juvenile rheumatoid arthritis (JRA), juvenile systemic lupus erythematosus (JSLE) and juvenile dermatomyositis (JDM) respectively, 3 to 6 years old with the mean age of 5 years with 3.5 to 22 months course of disease with 14 months on average, received auto-PBHSCT. Their conditions were so severe that conventional therapy failed to control the diseases. The changes of both clinical manifestations and immunologic indexes were observed before and after transplantation with long term following up at specialty clinic of rheumatism.

RESULTThe time when neutrophil count >or= 0.5 x 10(9)/L in the 3 children was days +9, +13 and +11 respectively, that of platelet count >or= 20 x 10(9)/L was days +14, +18 and +13 respectively. The cellular immune function remained abnormal with CD4 cells at a low level and CD4/CD8 being inverted. As to the JDM child, the skin rash had disappeared and his muscle tone was improved to grade 5 within one month after the transplantation. The EMG and serum creatase level returned to normal and muscle MRI findings were improved greatly within 2 months after the transplantation. As to the JSLE child, skin rash and proteinuria had disappeared, MRI of brain showed that the pathological changes had been absorbed and EEG returned to normal 3 months after the transplantation, all the autoantibodies turned to negative within 8 months after transplantation. As to the JRA child, the arthritis had been improved remarkably within 3 weeks after auto-PBHSCT. There was no swelling of joints nor movement limitation 3 months post transplantation. The steroids and immunosuppressive drugs were discontinued post transplantation. Cushing syndrome disappeared. Their body heights increased by 10 to 15 cm in the past 18 months, and they all returned to school. There was no relapse during follow-up periods of 25 - 27 months.

CONCLUSIONThe therapy with auto-PBHSCT for refractory rheumatism among preschool children was remarkably effective in a short-term, yet the safety and long-term effect still need to be further studied.

Child ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Peripheral Blood Stem Cell Transplantation ; Rheumatic Diseases ; therapy ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVETo detect the serum specific proteins in tumor-like polypoid lesions of the gallbladder patients and establish diagnostic model.

METHODSSurface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) technique and WCX Magnetic Beads were used to detect the serum proteomic patterns of 23 patients with tumor-like PLG, 21 patients with non tumor-like PLG and 26 normal persons. Biomarker Wizard and Biomarker Patterns Software were used in combination to analyze the data.

RESULTSPreliminary screening out 22 representative specific proteins for the diagnosis of the tumor-like PLG. Analysis system under the conditions set selected 3 specific proteins to establish diagnostic model for the tumor-like PLG. The sensitivity and specificity of the model for the diagnosis of the tumor-like PLG were 100% and 89.4%, respectively.

CONCLUSIONSELDI-TOF-MS technique can select specific protein of the tumor-like PLG, and establish diagnostic model of the tumor-like PLG.

Adult ; Aged ; Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Female ; Gallbladder Neoplasms ; diagnosis ; Humans ; Male ; Middle Aged ; Polyps ; diagnosis ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

BACKGROUNDBipolar electro-coagulation has a reported efficacy in treating epilepsy involving functional cortex by pure electro-coagulation or combination with resection. However, the mechanisms of bipolar electro-coagulation are not completely known. We studied the acute cortical blood flow and histological changes after bipolar electro-coagulation in 24 patients with intractable temporal lobe epilepsy.

METHODSTwenty-four patients were consecutively enrolled, and divided into three groups according to the date of admission. The regional cortical blood flow (rCBF), electrocorticography, the depth of cortex damage, and acute histological changes (H and E staining, neuronal staining and neurofilament (NF) staining) were analyzed before and after the operation. The t-test analysis was used to compare the rCBF before and after the operation.

RESULTSThe rCBF after coagulation was significantly reduced (P < 0.05). The spikes were significantly reduced after electro-coagulation. For the temporal cortex, the depth of cortical damage with output power of 2-9 W after electro-coagulation was 0.34 ± 0.03, 0.48 ± 0.06, 0.69 ± 0.06, 0.84 ± 0.09, 0.98 ± 0.08, 1.10 ± 0.11, 1.11 ± 0.09, and 1.22 ± 0.11 mm, respectively. Coagulation with output power of 4-5 W completely damaged the neurons and NF protein in the molecular layer, external granular layer, and external pyramidal layer.

CONCLUSIONSThe electro-coagulation not only destroyed the neurons and NF protein, but also reduced the rCBF. We concluded that the injuries caused by electro-coagulation would prevent horizontal synchronization and spread of epileptic discharges, and partially destroy the epileptic focus.

Adult ; Electrocoagulation ; methods ; Epilepsy ; surgery ; Epilepsy, Temporal Lobe ; surgery ; Female ; Humans ; Male ; Temporal Lobe ; surgery ; Young Adult

OBJECTIVETo study the possible mechanism of aqueous extract of detoxified cottonseeds (CTN-W).

METHOD AND RESULTCTN-W 0.01, 0.03, 0.10, 0.30 mg.mL-1 was incubated directly with the synaptic membrane extracted from the cerebral cortex in rats, and adenylyl cyclase (AC) activity was detected by using radio-immunoassay.

RESULTShowed that CTN-W could activate AC in a dose-dependend manner. After incubation with PC12 cells in the presence of corticosterone 2 x 10(-4)mol.L-1 for 48 h, CTN-W 0.08, 0.4, 2 mg.mL-1 protected PC12 cells from the lesion induced by corticosterone.

CONCLUSIONAntidepressant and anxiolytic effects of CTN-W are related with the activation of AC-cAMP pathway in signal transduction system, thus protecting neurons from the lesion. These two aspects maybe partly form the mechanism of CTN-W's action.

Adenylyl Cyclases ; metabolism ; Animals ; Anti-Anxiety Agents ; pharmacology ; Antidepressive Agents ; pharmacology ; Corticosterone ; antagonists & inhibitors ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gossypium ; chemistry ; Neuroprotective Agents ; pharmacology ; PC12 Cells ; drug effects ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Seeds ; chemistry ; Synaptic Membranes ; enzymology

OBJECTIVETo detect the serum specific proteins in pancreatic cancer patients and establish diagnostic model by surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) technique.

METHODSTwenty-nine serum samples from patients of pancreatic cancer were collected before surgery and an additional 57 serum samples from age and sex matched individuals without cancer were used as controls, SELDI-TOF-MS technique and WCX magnetic beads were used to detect the protein fingerprint expression of all the serum samples and the resulting profiles between pancreatic cancer patients and controls were analyzed with biomarker wizard system, established the model using biomarker patterns system software. A double-blind test was used to determine the sensitivity and specificity of the classification model.

RESULTSA panel of four biomarkers (relative molecular weight are 5705, 4935, 5318 and 3243 Da) were selected to set up a decision trees as the classification model for screening pancreatic cancer effectively. The result yielded a sensitivity of 100%, specificity of 97.4%. The double-blind test challenged the model with a sensitivity of 88.9% and a specificity of 89.5%.

CONCLUSIONSSELDI-TOF-MS offers a unique platform for the proteomic detection of serum in pancreatic cancer patients. It also offers a noninvasive method to further study the proteomic changes in the development and progression of pancreatic cancer.

Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Early Detection of Cancer ; Humans ; Mass Screening ; Pancreatic Neoplasms ; blood ; diagnosis ; Proteomics ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

The epileptic eye and head movements during epileptic seizures may be much more complicated thanpeople originally understood, which can be ipsilateral or contralateral to the electroencephalographyfocus. Here, we describe a male patient with drug resistant focal seizures associated with a directionalseparation between head and eye movement before evolving into generalized tonic-clonic seizure.His contralateral head leading turning showed forced, sustained, and unnatural features companiedby ipsilateral eye staring. Stereoeletroencephalography monitoring was performed, and 4 habitualseizures were recorded over 5 days. Three seizures showed left head leading turning and generalizedtonic-clonic seizure, and only one showed dizziness and ringing in the ears. All the seizures showedthat the ictal onset contacts were located in the posterior inferior temporal sulcus which borders onthe anterolateral part of medial superior temporal area. The patient underwent a resection includingtemporooccipital region, and the histopathology showed focal cortical dysplasia type Ic. He has beenseizure free for two years after operation. The scores of the intelligence and memory quotient improvedhalf year after operation.

OBJECTIVESevere newborn hypoxic-ischemic encephalopathy (HIE) has a very high rate of disability and no effective treatment is available. The present study aimed to preliminarily evaluate the effects of human neural stem cell transplantation in treatment of severe neonatal HIE.

METHODSThe patient was a 75-day old male infant with sequelae of severe HIE who had highly delayed development of intelligence and movement and myotonia. MRI showed multiple cerebromalacia and encephalatrophy. Cells obtained from the forebrain of an 11-week old fetus were cultured and amplified for 15 days. And then the human fetal neural stem cells were injected into cerebral ventricle of this infant.

RESULTSTwenty eight days after transplantation, remarkable improvement occurred not only in his myotonia but also in his intelligence and movement, which became similar to those of the normal infants of the same age. Positron emission tomography (PET) showed significantly increased radioactivity at temporal and occipital lobes which suggested that the cellular metabolism had increased greatly.

CONCLUSIONThe short-term effect of NSCs transplantation on the infant with severe HIE sequelae was significant. PET suggested that the implanted NSCs survived. Many more studies are needed to evaluate long-term effects of NSC transplantation in treatment of HIE.

Asphyxia Neonatorum ; complications ; Brain ; pathology ; physiopathology ; Female ; Humans ; Hypoxia-Ischemia, Brain ; etiology ; pathology ; physiopathology ; therapy ; Infant ; Infant, Newborn ; Injections, Intraventricular ; Multipotent Stem Cells ; transplantation ; Neurons ; Positron-Emission Tomography ; Prognosis ; Stem Cell Transplantation ; methods ; Time Factors ; Treatment Outcome

OBJECTIVENeonatal hypoxic-ischemic encephalopathy (HIE) harms the lives and health of newborn infants and children severely. Given the absence of effective therapies for HIE, it is important to derive new strategies. Neural stem cells (NSCs) have great potential as a therapeutic tool for the repair of a number of central nervous system disorders that involve cell loss. This study was designed to transplant the neural stem cells derived from human fetal brain (hNSCs) into cerebral ventricle of neonatal rat following hypoxic-ischemic injury and to investigate their survival, migration and differentiation in rat brain.

METHODSCells obtained from the forebrain of a 12-week old fetus were cultured in the presence of epidermal growth factor, basic fibroblast growth factor and leukemia inhibitory factor for 11 days. Animal models were built in 7-day-postnatal Wistar rats, 3-days after hypoxia-ischemia (HI), 5 microl suspension containing 5.0 x 10(5) hNSCs was injected into the left cerebral ventricle of each HIE rat by using stereotactic instrument. No immunosuppression therapy was given to the animals. At 1, 2, 4 weeks and 3 months after transplantation, the rats were sacrificed and brain tissues were harvested and were then examined by H-E staining and immunohistochemical analysis.

RESULTSImplanted cells expressing human nuclear protein (hNP) migrated form the subventricular zone (SVZ) along corpus callosum to the damaged areas, especially to the injured side of cortex and hippocampus. In different areas, the implanted hNSCs differentiated into different cell types which were similar to the host cells. The 85% implanted cells in cortex consisted of hNuc-NF or hNuc-Tublin double positive cells, while in the migratory way, 60% implanted cells differentiated into hNuc-GFAP double positive cells. Compared with the 1-week time point, an increased number of hNP-positive cells were observed at 2-weeks, but the number of these cells greatly decreased at 4-weeks and 3 months.

CONCLUSIONThe implanted hNSCs could extensively survive, migrate in the brain of neonatal rat with HIE and could differentiate into neurons and astrocytes in a regionally specific manner.

Animals ; Animals, Newborn ; Brain ; pathology ; Carotid Artery, Common ; surgery ; Cell Differentiation ; Cell Movement ; Disease Models, Animal ; Fetal Stem Cells ; transplantation ; Humans ; Hypoxia ; complications ; physiopathology ; Hypoxia-Ischemia, Brain ; pathology ; physiopathology ; therapy ; Immunohistochemistry ; Injections, Intraventricular ; methods ; Ligation ; methods ; Neurons ; Nuclear Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation ; methods ; Survival Analysis ; Time Factors

OBJECTIVETo study the tissue culture and rapid-proliferation techniques of Pueraria mirifica.

METHODThe tender branch were used as explants and cultivated in different media. The optimum media for inducing buds, proliferation and rooting were selected by adjusting the kinds and doses of plant hormones and special compounds.

RESULTThe medium of MS + IBA 0.05 mg L(-1) + BA 0.5 mg L(-1) was suitable for buds inducing and could be used in the first generation cultivation; MS + IBA 0. 02 mg L(-1) + BA 0.2 mg L(-1) and MS +BA 0.1 mg L(-1) were employed by turns in subculture, 25 days propagation coefficient was 3.0; and the medium of 1/2MS + IBA 0.1 mg L(-1) + IAA 0.2 mg L(-1) + C (special compound) 10 mg L(-1) was used for roots inducing, the rooting rate was 76.9%. Rooting plantlets were transplanted in spring and summer; the surviving rate was 81.0%.

CONCLUSIONThis technique system could be employed for rapid propagation of P. mirifica.

Pueraria ; growth & development ; Tissue Culture Techniques ; methods