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

This study was performed to explore the effect of flagellin+rapamycin on the growth and metastasis of 4T1 breast cancer in tumor-bearing mice,and their regulatory effect on several immunocytes.4T1 cell line was applied to establish an breast cancer model in Balb/c mice,which then injected with flagellin+rapamycin,and the volume and inhibition rate of tumor were recorded.MTT assay was used to detect the inhibitory effect of flagellin+rapamycin against 4T1 cells.Data showed that the combination of flagellin and rapamycin had the best inhibition effect on 4T1 cells,and the same results were found in animal model experiments.Flow cytometry result indicated that flagellin+rapamycin significantly down-regulated the levels of CD11b+Gr-1+myeloid-derived suppressor cells(MDSCs)and CD11b+F4/80+tumor-associated macrophages(TAMs)of tumor-bearing mice.Furthermore,flagellin+rapamycin alleviates the metastasis of 4T1 cancer cells into liver and lung.Taken together,flagellin+rapamycin can inhibit proliferation and metastasis of 4T1 cells,thus exert antitumor effects in mice,which may related with its regulatory effects on immunocytes in tumor microenvironment.

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

BACKGROUNDThe extended thymectomy for myasthenia gravis (MG) is currently available, but in 20% - 40% of the patients the results were not satisfactory. There are no ideal indicators forecasting surgical results before operation. The surface enhanced laser desorption ionization-time of flight-mass spectroscopy (SELDI-TOF-MS) is a currently new technique for detection of protein profiles, and some progresses have been made in cancer diagnosis and efficacy evaluation, but there is no report on efficacy forecasting of MG surgery. This study aimed to establish an efficacy prognosis model for forecasting the efficacy of surgery for MG by analysis of serum protein profiles of MG patients before surgery.

METHODSFifty-six MG patients 6 months after extended thymectomy were enrolled in the study. They were classified into effective or non-effective groups according to symptoms and medication. Their pre-operative blood samples were analyzed for protein profiles by the SELDI-TOF MS technique, and protein peaks were identified for establishment of the efficacy prognosis model of MG surgery. Additional 100 MG patients were subjected to model validation and their pre-operation protein profiles reviewed for post-operative results. The results were compared with those of the post-operative follow-up so as to validate the prognosis model.

RESULTSFor the model establishment, symptoms were improved in 33 patients and not improved in 18 patients, with an effective rate of 64.7%. Five (8.9%) patients were lost to follow-up. Within the molecular weight range of 1 000 to 20 000, 3 specific protein peaks were found to be significantly different between the effective and non-effective groups, ie M4110-76, M3394-58, and M1258-55. Using the efficacy prognosis model constructed with these data, the accuracy rate of classification was 87.9% for the effective group, and 83.3% for the non-effective group, with a total accuracy rate of 86.3%. For the model evaluation, 2 (8.9%) patients were lost to follow-up, 62 patients were effective and 36 were non- effective. By comparing with the real results of follow-up with 65 effective and 33 non-effective patients with an effective rate of 66.3%, the accuracy rate of prediction by the prognosis model was 86.2% for the effective group, and was 81.8% for the non-effective group with a total accuracy rate of 84.5%.

CONCLUSIONSBy protein profiles analysis of pre-operative blood samples taken from MG patients with the SELDI-TOF MS technique, protein peaks correlated with surgery efficacy in MG patients can be found for primary forecasting short-term efficacy of surgery for MG patients.

Adolescent ; Adult ; Blood Proteins ; analysis ; Child ; Female ; Humans ; Male ; Middle Aged ; Myasthenia Gravis ; diagnosis ; surgery ; Prognosis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Thymectomy ; 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.