UNLABELLEDOBJECTIVE To explore the effect of combined use of rehmannia (RM) and rhodiola (RD) on peripheral leukopenia and bone marrow hematopoietic function suppression induced by cyclophosphamide (CTX) in mice.

METHODSICR mice were established into bone marrow inhibition models by intraperitoneal injection of CTX, and were administered with RM, RD or its extract (RDE), singly or in mixture, via gastrogavage for 10 days. The changes of peripheral hemogram, bone marrow nucleated cell proliferation, CFU-GM colony formation, GM-CSF and erythropoietin (EPO) secretion were observed.

RESULTSCompared with the un-treated model mice, the peripheral white blood cell count was significantly higher in model mice treated with RDE and RM mixture; the bone marrow nucleated cells count, CFU-GM formation, and GM-CSF production were significant higher in model mice treated with RD and RM mixture, showing statistical significance (P < 0.01); while EPO production in the RD and RM mixture treated group was slightly elevated, but the difference showed no statistical significance.

CONCLUSIONRD and RM mixture could regulate hematopoietic system by promoting the production of bone marrow cells and colonies, as well as enhancing the synthesis of related cytokines, such as GM-CSF, so as to increase the amount of peripheral white blood cells and restore the hematopoietic function of organism.

Animals ; Bone Marrow ; drug effects ; Cyclophosphamide ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Erythropoietin ; secretion ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; secretion ; Hematopoiesis ; drug effects ; Leukopenia ; chemically induced ; drug therapy ; Male ; Mice ; Mice, Inbred ICR ; Phytotherapy ; Rehmannia ; chemistry ; Rhodiola ; chemistry

OBJECTIVE: To establish the electricity plate digestion and inductively coupled plasma-mass spectrometry (ICP-MS) method for determination of 33 inorganic elements in human hair. METHODS: Lithium (6Li), Germanium (72Ge), Yttrium (89Y), Indium (115In), and Terbium (159Tb) were used as internal standards. The electric heating board digestion in a mixture of nitric acid and hydrogen peroxide was used as the pre-treatment of the hair. Thirty-three inorganic elements in human hair were analyzed by ICP-MS method. RESULTS: The detection limit of ICP-MS was 0.0001 microg/g(Th)-10.9 microg/g (Ca) and the limit of quantitation was 0.0005 microg/g (Th)-25 microg/g (Ca). The recovery rate of this method was 86%-113%. The RSD for the intra-day and inter-day were less than 9.2%. The method was not statistically different from microwave digestion method. CONCLUSION This method is highly efficient and accurate. It can be used for analysis of 33 inorganic elements in human hair.
Electricity ; Hair/chemistry* ; Humans ; Limit of Detection ; Mass Spectrometry/methods* ; Microwaves ; Reference Standards ; Sensitivity and Specificity ; Trace Elements/analysis*

OBJECTIVE: To determine the normal reference values of 33 elements, Ag, Al, As, Au, B, Ba, Be, Ca, Cd, Co, Cr, Cs, Cu, Fe, Ga, Hg, Li, Mg, Mn, Mo, Ni, Pb, Rb, Sb, Se, Sr, Th, Ti, Tl, U, V, Zn and Zr, in the blood and urine samples from the general population in Sanmen County of Zhejiang province, a typical coastal area of eastern China. METHODS: The 33 elements in 272 blood and 300 urine samples were determined by inductively coupled plasma-mass spectrometry (ICP-MS). The normality test of data was conducted using SPSS 17.0 Statistics. The data was compared with other reports. RESULTS: The normal reference values of the 33 elements in the blood and urine samples from the general population in Sanmen County were obtained, which of some elements were found to be similar with other reports, such as Co, Cu, Mn and Sr, while As, Cd, Hg and Pb were generally found to be higher than those previously reported. There was a wide variation between the reports from different countries in blood Ba. CONCLUSION The normal reference values of the 33 elements in the blood and urine samples from the general population in Sanmen County are established, and successfully applied to two poisoning cases.
Blood Chemical Analysis ; China ; Elements ; Environmental Monitoring ; Humans ; Urinalysis

BACKGROUNDOlfactory ensheathing cells (OECs) can promote many kinds of neuron growth and axonal extension. The aim of the study was to investigate the effects of co-culturing with OECs on neuron apoptosis in vitro.

METHODSApoptosis was induced by treatment of cultured dorsal root ganglion neurons with 1 mmol/L hydrogen peroxide (H(2)O(2)). Cells were randomly arranged into the following treatment groups. In group 1, OECs at different density (10(4)/ml to 8 x 10(5)/ml) were added immediately after H(2)O(2) treatment and cells were co-cultured for 24 hours. In group 2, OECs were added at different time points (0, 4, 8, 12 and 24 hours) after H(2)O(2) treatment. Apoptotic cell death was determined by Hoechst 33258 staining and flow cytometry (FCM). Cell viability was determined by using methyl thiazoleterazolium (MTT) assays.

RESULTSThe results showed in the Hoechest 33258 staining, FCM and MTT that OECs have both the density-dependent protection and time-dependent protection on neuron apoptosis. The apoptosis decreased and the dorsal root ganglion neuron viability increased, when the density of OECs was increased in co-culture groups. But further increasing OEC density above 2 x 10(5)/ml (i.e. 8 x 10(5)/ml) failed to exert additional protection. As the interval between adding H(2)O(2) and adding OECs was increased, the amounts of apoptosis cells were also increased. When OECs were added 24 hours after H(2)O(2), no significant protection was observed.

CONCLUSIONThese results indicated that OECs could protect dorsal root ganglion neurons from apoptosis induced by H(2)O(2) in a density- and time-dependent manner.

Animals ; Apoptosis ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Coculture Techniques ; Ganglia, Spinal ; cytology ; drug effects ; Hydrogen Peroxide ; toxicity ; Male ; Olfactory Bulb ; cytology ; physiology ; Rats ; Rats, Sprague-Dawley ; Time Factors

Objective To investigate the isolation and culture of the neural stem cells (NSCs) derived from the embryonic mouse spinal cord and striatum, and observe the differences in their proliferation and differentiation characteristics in vitro, thereby providing evidences for identifying more suitable seed cells for repainng spinal cord injuries. Methods The spinal cord and striatum of 14-day-old embryonic mice were dissected for primary NSC culture. After 4 passages, the cells were examined for the expression ofnestin, β-tubulin Ⅲ, glial fibrillary acidic protein (GFAP) and MAG using immunohistochemistry. Results The NSCs derived from the spinal cord and the striatum both possessed in vitro proliferation capacity and expressed nestin antigen. After induced differentiation, the NSCs expressed specific antigens of neurons, astrocytes and oligodendrocytes. Conclusion Multipotent NSCs can be obtained from embryonic mouse spinal cord and striatum through different in vitro culture methods under different conditions. According to the differenees between the two kinds of NSCs in culture situation and differentiation, the striatum derived NSC is suitable for transplantation to repair spinal cord injury.

This study was purposed to detect the expression of transforming growth factor β1 (TGF-β1) and its receptors (TGF-βR) and to investigate their roles in pathogenesis of immune thrombocytopenic purpura (ITP). The expressions of TGF-β1 and their receptors TGF-βRI, TGF-βRII and TGF-βRIII in the peripheral blood of patients with ITP and healthy persons were detected by the real-time PCR, and differences of their expression levels were analysed. The results showed that the expression of TGF-β1 and TGF-βRII mRNA in ITP patients was significantly higher than that in the healthy controls, while the TGF-βRI mRNA expression in ITP patient was significantly lower than that in the controls. The expression of TGF-βRIII was not statistically different between the two groups. It is concluded that TGF-β1 and its receptors including TGF-βRI and TGF-βRII express abnormally in the peripheral blood of ITP patients, which suggests that the TGF-β signaling pathway probably play a vital role in the pathogenesis of the ITP.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; Female ; Humans ; Male ; Middle Aged ; Protein-Serine-Threonine Kinases ; metabolism ; Purpura, Thrombocytopenic, Idiopathic ; metabolism ; pathology ; Receptors, Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Young Adult

OBJECTIVETo observe the treating effect of collage-heparin sulfate after the 10 mm rat sciatic nerve defect was bridged by it.

METHODSA new kind of nervous tissue engineering scaffold was produced by freeze-drying technique from collagen-heparin sulfate. Thirty-two SD rats were randomly divided into A, B, C and D groups. Sciatic nerve defect in group A was bridged by collagen-heparin sulfate. In group B, sciatic nerve was bridged by auto-nerve transplantation. Group C was the blank control group. Animals in group D were normal. And 10 mm sciatic nerve defect was bridged in the experiment. Thirty-six weeks after the operation, the experimental animals were detected by HRP labeled retrograde trace, HE staining, toluidine staining, silvering staining, S100, GAP-43 and NF immunohistological staining, MBP immunofluorescence staining and transmission electron microscope to observe the nerve regeneration inducing effect of this new scaffold.

RESULTSNine months after operation, the collage-heparin sulfate scaffold was replaced by newly regenerated nerve. The number of HRP labeled spinal cord anterior horn cells and the area of sensation nerve fiber at the posterior horn were similar with that was repaired by auto-nerve. GAP-43, NF and S100 labeled regenerated nerve fiber had passed the total scaffold and entered the distal terminal. The regenerated nerve fibers were paralleled, lineage arranged, coincide with the prearranged regenerating "channel" in the collagen-heparin sulfate scaffold. MBP immunofluorescence staining also proved that the newly regenerated nerve fiber could be ensheathed. In the experimental group, the area of myelinated nerve fiber and the thickness of the myelin sheath had no obvious difference with that of the group repaired by auto-nerve, except that the density of the regenerated myelinated sheath fiber was lower than that of the control group.

CONCLUSIONNervous tissue engineering scaffold produced by collagen-heparin sulfate can guide the regeneration of nerve fibers. The nerve function recovers fine. This kind of material has great application potential.

Animals ; Biocompatible Materials ; Heparitin Sulfate ; Male ; Prosthesis Implantation ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; pathology ; surgery ; Sulfuric Acid Esters ; Tissue Engineering ; methods

The study is to investigate the effect of asiaticoside on collagen-induced arthritis (CIA). The model of CIA mice was prepared and the change of secondary paw swelling and the arthritis scores were observed. In vitro proliferation of spleen cells was examined using MTT assay. The cell-free protein extracts from the arthritic joints and nonarthritic joints were used for the analysis of protein expression of cyclooxygenase-2 (COX-2). And the level of PGE2 in joints was assayed using PGE2 express EIA kit. The tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels in the serum were measured by ELISA. Histopathological examination was performed by hematoxylin-eosin (HE) stain method. Asiaticoside (10, 20 and 40 mg x kg(-1) x d(-1), 22 d, ig) significantly reduced paw swelling, and decreased the arthritis scores. There was a significant reduction in proliferation of spleen cells of CIA mice treated with asiaticoside as compared with that of untreated CIA mice. COX-2, PGE2, TNF-alpha and IL-6 production in CIA mice were inhibited by asiaticoside. Meanwhile, the pathological examination showed that articular cartilage degeneration with synovial hyperplasia and inflammatory cells infiltration in CIA mice was suppressed by asiaticoside. Its active mechanism may be related to inhibiting proliferation of lymphocyte and reduction of expression of COX-2 and inflammatory cytokines.
Animals ; Ankle Joint ; metabolism ; pathology ; Anti-Inflammatory Agents ; isolation & purification ; pharmacology ; Arthritis, Experimental ; chemically induced ; metabolism ; pathology ; Cell Proliferation ; drug effects ; Centella ; chemistry ; Collagen Type II ; Cyclooxygenase 2 ; metabolism ; Cytokines ; blood ; metabolism ; Dinoprostone ; metabolism ; Interleukin-6 ; blood ; Lymphocytes ; pathology ; Male ; Mice ; Mice, Inbred DBA ; Plants, Medicinal ; chemistry ; Spleen ; pathology ; Triterpenes ; isolation & purification ; pharmacology ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo evaluate the efficacy of iloprost in acute vasodilatation test during cardiac catheterization and to explore a useful hemodynamic indication regarding operability in the patients with severe pulmonary hypertension (PH) related to congenital heart disease (CHD).

METHODSThe clinical data of 46 patients [mean age (12 ± 9) years] with severe PH related to CHD from June 2006 to December 2008 was retrospectively analyzed. All patients underwent standard right and left cardiac catheterization and a trial of inhaled iloprost test during cardiac catheterization. The mean pulmonary arterial pressure was (80 ± 13) mm Hg (1 mm Hg = 0.133 kPa) and pulmonary vascular resistance index was (17 ± 10) wood.m². A positive response to inhaled iloprost was defined as a decrease of at least 20% in pulmonary vascular resistance index (PVRI) without changes on systemic artery pressure. Patients with positive response to iloprost underwent cardiac surgical repair. The pulmonary artery pressure and PVRI was monitored by Swan-Ganz catheter postoperatively.

RESULTSOf the 46 patients, 29 (63.1%) showed a positive response after iloprost inhalation, defined by a significant reduction in PVRI from (15 ± 6) wood.m(2) at baseline to (9 ± 4) wood.m² in response to iloprost inhalation therapy (P < 0.05). The ratio of pulmonary to systemic resistance (Rp/Rs) decreased from 0.7 ± 0.2 to 0.4 ± 0.2 (P < 0.05). Seventeen patients (36.9%) didn't respond to iloprost displayed only little changes in PVRI [from (21 ± 10) wood.m(2) to (19 ± 9) wood.m²] and Rp/Rs (from 1.0 ± 0.5 to 0.9 ± 0.5). Out of 29 positive patients, 21 (72%) underwent successful cardiac surgical repair with a reduction of mean pulmonary arterial pressure (mPAP) to an average of (27 ± 10) mm Hg after the operation. Only 2 patients out of the 17 patients from the negative group were referred to surgery. Their mPAP was greater than 45 mm Hg.

CONCLUSIONSA significant reduction in pulmonary artery pressure after cardiac surgery was observed in patients with positive response to inhaled iloprost. Inhaled iloprost may be a valuable tool in the preoperative evaluation of patients with severe PH related to CHD.

Administration, Inhalation ; Adolescent ; Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; physiopathology ; surgery ; Hemodynamics ; drug effects ; Humans ; Hypertension, Pulmonary ; physiopathology ; surgery ; Iloprost ; pharmacology ; Infant ; Lung ; blood supply ; Male ; Preoperative Care ; Retrospective Studies ; Vasodilator Agents ; pharmacology ; Young Adult

Objective To investigate the efficient method which can culture and induce embryonic stem cells to neurocyte in vitro. Methods Isolate the blastula of 3.5 d from BALB/c species mouse. Culture the cells from inner cell mass (ICM) which were isolated by mechanical method on the mouse embryonic fibroblaste cell (MEF) feeder layer or 0.1% gelatin coated dishes. The stem cells were identified by characterized morphology, alkaline phosphatase stain, differential potency in vivo and immunochemistry stain. The isolated cells were differentiated by serial induction method that mimicking the intrinsic developmental process of the neural system. Results The isolated cells were positive for alkaline phosphatatse and SSEA-1 (stage specific embryonic antigen 1). Moreover they were identified pluripotent by differentiation in vivo. Therefore the isolated cells presented the characters of ESCs. Then the isolated cells were able to differentiate into neurocytes in vitro. Conclusion Mouse embryonic stem cells isolation, culture and differentiation system has been established.