Objective To study the construction of tissue engineering adipose in vitro with silk fibroin scaffold and human umbilical cord mesenchymal stem cells (hUCMSCs) transfected by recombinant human insulin gene lentivirus.Methods hUCMSCs infected with recombinant lentiviral pLenti6.3-insulin-IRES-EGFP (Group A) by the best MOI=10 were seeded in silk fibroin scaffold; hUCMSCs transfected by EGFP gene (Group B)was regarded as negative control; and then the cell-scaf-fold complexes were cultured in adipogenic differentiation medium. MTT test was used to detect whether recombinant lentiviral affected the growth and proliferation of hUCMSCs and the growth activity of hUCMSCs seeded in silk fibroin.Results After 5-7 days for adipogenic culture,the numbers of fat-like cells in group A were significantly more than those in group B (P =0.007,P＜0.01).RTPCR results showed that the expression of PPARγ-2 in group A was much stronger than that in group B.MTT test showed that there was no significant difference in optical density (A)at each time between transfected group and nontransfected group (P=0.056,P＞0.05).And there was also no significant difference in optical density (A)between cell group and cell-scaffold group (P =0.066,P＞0.05).Conclusions Insulin gene could obviously promote hUCMSCs getting into adipose,and carrying recombinant human insulin gene lentiviral vector transfection of hUCMSCs and silk fibroin scaffolds could effectively construct tissue engineering adipose in vitro.

Objective To evaluate the efficacy and safety of Prostant TM in the treatment of chronic prostatitis. Methods A multi center, randomized, double blinded, placebo controlled trial was carried out during March 2001 and May 2001. 72 cases of patients who had been diagnosed as chronic prostatitis were separated into tow groups: 36 cases in the trial group treated with Prostant TM and the other 36 cases in the controlled group using placebo. The efficacy was evaluated by the WBC in EPS and NIH Chronic Prostatitis Symptom Index after a one month follow up. The efficacy of the therapy was divided into four levels:cure,the symptom index score being reduced over 90%, and the number of WBC in the EPS less than 10/HP; effective, the symptom index score being reduced 60%～89% and the number of WBC in the EPS lowered over 50% or less than 15/HP;improved,the symptom index score being reduced 30%～59% and the number of WBC in the EPS lowered over 25%; no effect, the symptom index score being reduced less than 30% or the number of WBC in the EPS lowered no more than 25%. Results All but two cases had completed the follow up.One case (2.8%) was completely cured,remarkable effective in 7 cases (20.0%) of the trial group;and improred in 16 cases (45.7%).Only 2 cases (5.7%)seemed effective and 8 cases (22.8%) improved in the controlled group. The efficacy showed significant difference between these two groups ( P

Humans ; Lymphohistiocytosis, Hemophagocytic ; diagnosis ; therapy

Objective To establish an inductively coupled plasma‐mass spectrometry (ICP‐MS) method for analysis of elements in peripheral blood of children .Methods A total of 474 healthy children in Hunan area were enrolled in this study ,and six ele‐ments ,including Ca ,Mg ,Fe ,Cu ,Zn and Pb ,in peripheral blood specimens were detected by using ICP‐MS method .Results The levels of elements ,including Ca ,Mg ,Fe ,Cu ,Zn and Pb ,in peripheral blood of healthy children showed skewed distributions ,and no significant differences were found in levels of these elements between male and female children(P>0 .05) .The reference intervals of Ca ,Mg ,Fe ,Cu ,Zn and Pb in peripheral blood of healthy children in this area were 57 .30 -81 .40 mg/L ,30 .40 -44 .80 mg/L , 361 .20-531 .40 mg/L ,848 .10-1 469 .20 μg/L ,2 .68 -6 .54 mg/L and 0 .00 -100 .00 μg/L respectively .Conclusion The ICP‐MS method for simultaneously detecting Ca ,Mg ,Fe ,Cu ,Zn and Pb in peripheral blood of children and reference interval of each ele‐ment are successfully established .

AIM To study the anti inflammatory and immunomodulatory actions of total glucosides of Mudan cortex(TGM)on adjuvant arthritis(AA)rats and its mechanisms. METHODS To observe the change of secondary inflammation and immune function on AA rats treated with TGM and to detect the effect of TGM on immune function in normal and AA rats in vitro. RESULTS TGM (25,50,100 mg?kg -1 ig) could significantly inhibit secondary inflammatory reaction(secondary inflammatory swelling, multiple arthritis, pathologic change of AA rats ankle arthrosis)of AA rats. Among them, TGM 50 mg?kg -1 ig had the best therapeutic effect, and could make the weight of atrophied thymus of AA rats restore to normal. Treated with TGM ig, not only the lower response of splenocytes to Con A and decreased IL 2 synthesis of AA rats were restored to normal but also the elevated IL 1 synthesis and PGE 2 release of PM? were reduced. The production of IL 1 by LPS induced PM? from normal rats were enhanced by in vitro treatment with lower concentrations (0 4～10 mg?L -1 )of TGM, but diminished with higher concentrations(50～250 mg?L -1 )of it. The production of PGE 2 were increased with dose independent manner(TGM 0 4～250 mg?L -1 ). In the range of 10～250 mg?L -1 TGM in vitro could significantly decreased the production of IL 1 and PGE 2 from AA rats and enhanced Con A induced proliferative response of splenocytes and IL 2 synthesis from splenocytes of AA rats. CONCLUSION TGM has anti inflammatory and immunomodulatory actions on AA rats mainly through inhibiting cell mediated immunity.

Objective To study the effect of chronic low dose soman on learning and memory and long term potentiation(LTP) of hippocampal slices. Methods Rat model was established by consecutive subcutaneous injection of soman(6-10 ?g?kg 1 , s. c, sig?14) for 14 days for Morris water maze test. Long term potential of synaptic transmission was observed in CA1 region by tetanization of the Schaffer commissural pathway in rat hippocampal slices. Results In the Morris water maze, latency to find a hidden platform was longer and the times of crossing the situation of platform and the time percent of swimming in northeast obviously decreased. In the experiment on hippocampal slice of rats in vitro by microelectrode method, the generation of long term potentiation was inhibited. Conclusion Chronic low dose soman may cause an evident learning and memory disturbance and decrease hippocampal synaptic plasticity.

Adolescent ; Child ; Child, Preschool ; Clinical Trials as Topic ; Flow Cytometry ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; diagnosis ; pathology ; therapy ; Neoplasm, Residual ; diagnosis ; pathology ; therapy ; Polymerase Chain Reaction ; Prognosis ; Recurrence ; Sensitivity and Specificity

Objective To observe the morphology of embryonic stem cell-derived neurons after transplanted into A?-injured rat hippocampus. Methods Neural precursor cells (NPCs) were generated from mouse embryonic stem cells (ESCs) expressing EGFP with modified serum-free methods and then transplanted into the hippocampus of A?1-40-injuried rats. The morphology, neurotransmitter phenotypes and receptors of EGFP-positive donor cells were observed with immunofluorescene methods. Results The engrafted NPCs survived and differentiated into glutamatergic and GABAergic neurons and expressed both glutamatergic and GABAergic neurotransmitter receptors. Synaptophysin-positive dots were found surrounding somata and dendrites of transplanted neurons, suggesting the presence of presynaptic terminals adjacent to their membranes. Conclusion NPCs derived from ESCs can differentiate into excitatory and inhibitory neurons after grafted into Alzheimer's disease model rats, and maybe form synapse with the host neurons.

AIM: To explore the influence of different concentration midazolam on the macroscopic voltage gated potassium currents and to discuss the relationship between potassium currents and inhibitory effect of clinical relevant concentration midazolam on sympathetic nervous system. METHODS: Superior sympathetic ganglion neurons were dissociated enzymatically from 7 to10 day old rat. Experiments were performed about 5 h after plating at room temperature (20- 24 ℃ ). Appropriate solution was chosen to separate the K + current from the other transmembrane currents. 1 ?mol?L -1 TTX was applied to the extracelluar solution to block the Na + current. Midazolam was also resolved in extracelluar solution to get various concentration ( 0.1 , 0.3 ,3,10,50,100 ?mol?L -1 ). Currents were recorded with the patch clamp technique in whole cell configuration using glass electrodes with a tip resistance of 2- 4 M . Potassium currents were evoked by test pulse from -100 mV to +30 mV with holding potential -80mV. Data were analyzed using Clampfit 6.0 and Oringih 5.0 software. Whole cell current records were corrected for leakage and capacitance by using the P/5 protocol. RESULTS: Midazolam dose dependently inhibited the whole cell potassium currents. Clinical relevant concentration midazolam ( 0.3 ?mol?L -1 ) only reduced the peak currents by 3.89 %(P= 0.88 ). The concentration required to produce 50% current inhibition(IC 50 ) was 76.065 ?mol?L -1 . CONCLUSION: Midazolam inhibits the whole cell potassium current significantly and dose dependently, but clinical relevant concentration midazolam has minor effect on the potassium currents, indicating that the inhibitory effect of midazolam on potassium current is not related to the suppression of activity of sympathetic system.

Histocytochemistry ; methods ; Humans ; Pathology, Clinical ; methods ; Specimen Handling ; methods ; Staining and Labeling ; methods