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ObjectiveTo observe the effect of ginsenoside Rg1 （G-Rg1） on the biological activity of cryopreserved Schwann cells （SCs） of the rat sciatic nerve and explore the feasibility of G-Rg1 in reducing the cryopreservation-induced injury in SCs. MethodBilateral sciatic nerves of SD rats were randomly divided into a fresh group， a blank group， and five G-Rg1 groups of different doses （1×10-7， 1×10-6， 1×10-5， 1×10-4， and 1×10-3 mol·L-1）. The nerves in the blank group and the G-Rg1 groups were preserved in liquid nitrogen solutions containing 0， 1×10-7， 1×10-6， 1×10-5， 1×10-4， and 1×10-3 mol·L-1 G-Rg1 for four weeks. The apoptosis of SCs was detected by TdT-mediated dUTP-biotin nick end labeling （TUNEL）/S100 immunofluorescence staining. The expression of cysteinyl aspartate-specific protease （Caspase）-9， Caspase-3， major histocompatibility complex （MHC）-Ⅰ， and MHC-Ⅱ was detected by Western blot. Subsequently， all nerves were cultured in the incubator at 37 ℃ with 5% CO2 for 7 days. The expression of glial cell line-derived neurotrophic factor （GDNF） and nerve growth factor （NGF） was detected by Western blot. In addition， the above cryopreserved nerves in the blank group and the 1×10-6， 1×10-5， and 1×10-4 mol·L-1 G-Rg1 groups were transplanted to the Wistar rats by allografting （blank transplantation group and the 1×10-6， 1×10-5， and 1×10-4 mol·L-1 G-Rg1 transplantation groups）， and fresh sciatic nerve allograft and isograft control group were set up. Sixteen weeks after transplantation， compound muscle action potential （CMAP） and nerve conduction velocity （NCV） were measured by electrophysiology. Nerve filament （NF）200 immunofluorescence staining， transmission electron microscopy， and toluidine blue staining were used to analyze the histology of the regenerated nerves. ResultCompared with the fresh group， the blank group and the G-Rg1 groups showed increased expression of Caspase-9， Caspase-3， and the apoptosis of SCs （P<0.05，P<0.01） and decreased expression of GDNF， NGF， MHC-Ⅰ， and MHC-Ⅱ （P<0.01）. Compared with the results in the blank group， the expression of Caspase-9 and Caspase-3 decreased in the 1×10-7， 1×10-6， 1×10-5，1×10-4 mol·L-1 G-Rg1 groups （P<0.01）， and the apoptosis of SCs was reduced in the 1×10-7-1×10-4 mol·L-1 G-Rg1 groups（P<0.05，P<0.01） and increased in the 1×10-3 mol·L-1 group （P<0.05）， while the expression of GDNF and NGF increased in the 1×10-7， 1×10-6， 1×10-5，1×10-4 mol·L-1 G-Rg1 groups and decreased in the 1×10-3 mol·L-1 group （P<0.05）. There was no statistical significance in the expression of MHC-Ⅰ and MHC-Ⅱ between the blank group and the G-Rg1 groups. Compared with the 1×10-7 mol·L-1 and 1×10-3 mol·L-1 G-Rg1 groups， the 1×10-6 1×10-5， 1×10-4 mol·L-1 G-Rg1 groups showed decreased expression of Caspase-3 and the apoptosis of SCs （P<0.05，P<0.01） and increased expression of GDNF and NGF （P<0.05，P<0.01）. There was no statistical significance in MHC-Ⅰ and MHC-Ⅱ expression among G-Rg1 groups. Sixteen weeks after transplantation， compared with the isograft group， the blank transplantation group and the G-Rg1 transplantation groups showed decreased CMAP， NCV， myelin sheath thickness， and number of myelinated nerve fibers （P<0.01）， and the 1×10-6 and 1×10-4 mol·L-1 G-Rg1 transplantation groups showed decreased NF200 （P<0.01）. Compared with the allograft group， the blank transplantation group and the G-Rg1 transplantation groups showed increased CMAP， NCV， NF200， myelin sheath thickness， and number of myelinated nerve fibers （P<0.05，P<0.01）. Compared with the blank transplantation group， the G-Rg1 transplantation groups showed increased CMAP， NCV， NF200， myelin sheath thickness， and number of myelinated nerve fibers （P<0.05，P<0.01）. Among all groups of G-Rg1 transplantation， each index of the 1×10-5 mol·L-1 G-Rg1 transplantation group was superior to that of the 1×10-4 and 1×10-6 mol·L-1 G-Rg1 transplantation group （P<0.05）. ConclusionG-Rg1 at a certain centration can maintain the biological activity of cryopreserved SCs of rat sciatic nerve， alleviate the cryopreservation-induced injury of rat sciatic nerve， and promote nerve regeneration after allograft.
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OBJECTIVE：To compare the protective effects model mice between the aboveground and underground parts of Astragalus membranaceus on immunosuppression ，and to provide reference for further utilization and development of A. membranaceus. METHODS ：A total of 240 ICR mice were divided into 4 batches，60 mice in each batch ，with half male and half female. Each batch of mice were randomly divided into blank group ，model group ，A. membranaceus aboveground part and undergroud part low-dose and high-dose groups （3，6 g/kg，by crude drug ）according to body weight and sex ，with 10 mice in each group. Blank group and model group were given normal saline intragastrically. A. Membranaceus groups were given corresponding concentration of drug intragastrically ，10 mL/kg，once a day ，for consecutive 30 days. Except for blank group ， other groups were intraperitoneally injected with cyclophosphamide 40 mg/（kg·d）for consecutive 3 days，since 24th day of treatment，to establish immunosuppression model. The levels of serum immunoglobulin （IgG，IgM，IgA），inflammation factors [nitric oxide ，interleukin-2（IL-2），IL-6，tumor necrosis factor-α] and half hemolysis value were detected in each group. Body weight ，thymus index ，spleen index ，phagocytic index ，activity of natural killer （NK）cell，splenic lymphocyte proliferation ability，dinitrofluorobenzene-induced delayed metamorphosis reaction in mice （by weight difference between left and right ears ） and the number of hemolytic plaque were determined. RESULTS ： Compared with blank group ， the serum levels of immunoglobulin，body weight ，thymus index ，spleen index ，phagocytic index ，NK cell activity ，the proliferation ability of splenic lymphocyte，the number of hemdytic plaque and half hemolysis value were decreased significantly in model group （P＜0.05）， while inflammation factor level as well as weight difference between left and right ears were increased significantly （P＜0.05）. Compared with model group ，above indexes of mice in A. membranaceus groups were improved significantly ，in dose-dependent manner（P＜0.05）. Compared with A. membranaceus undergroud part group ，above indexes of A. membranaceus aboveground part group were improved significantly （P＜0.05）. CONCLUSIONS ：Aboveground and underground part of A. membranaceus both have pretective effect on immunosuppression model mice ，and the effect of aboveground part of A. membranaceus is stronger than underground part of A. membranaceus .
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@#Objective To investigate the effects of triptolide (T10) on biological activity of sciatic nerve in cold preservation and nerve regeneration after allogeneic transplantation. Methods Cell Counting Kit-8 (CCK-8) was used to test the proliferation of SCs in logarithmic phase in 1×10-6 mol/L, 1×10-7 mol/L, 1×10-8 mol/L and 1×10-9 mol/L of T10 solution. The sciatic nerves from Sprague-Dawley rats were pretreated in 0 mol/L, 1×10-6 mol/L, 1×10-7 mol/L, 1×10-8 mol/L and 1×10-9 mol/L of T10 solution at 4 ℃ or 37 ℃ for 24 hours (n = 6). The expression of nerve growth factor (NGF), glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) was detected with Western blotting. Other sciatic nerve fragments were randomly divided into fresh nerve group (group A, n = 30), DMEM preservation group (group B, n = 30), T10 preservation group (group C, n = 30), T10 pretreatment DMEM preservation group (group D, n = 30) and T10 pretreatment T10 preservation (group E, n = 30), and were stored under 4 ℃ for four weeks. Calcein-AM/PI double staining laser confocal microscope and flow cytometry were used to detect the living cells and dead cells. The expression of the major histocompatibility complex (MHC)-I, MHC-II and intercellular cell adhesion molecule-1 (ICAM-1) was detected with Western blotting. The corresponding sciatic nerves were used to repaire 10 mm defects in Wistar rats (named groups A', B', C', D' and E'), and fresh sciatic nerve from Wistar rats were also used to do it (group F'). Compound muscle action potential (CMAP) and motor nerve conduction velocity (MNCV) were tested 16 weeks after transplantation, and then the grafts were observed for the nerve regeneration. Results SCs proliferated as the controls in the T10 solution with a concentration of 1×10-9 to 1×10-7 mol/L (P > 0.05). The expression of all the neurotrophic factors was more under 37 ℃ than under 4 ℃ in all the concentrations of T10 solution, and it was the most in the concentration of 1×10-8 mol/L whenever under 37 ℃ or 4 ℃ (P < 0.05). After four weeks of cold preservation, compared with groups B, C and D, the living nerve cells were the most in group E, and the expression of MHC-I, MHC-II and ICAM-1 was the least (P < 0.05). CMAP, MNCV and the never regeneration were better in group E' than in groups A', B', C' and D' (P < 0.05). A large number of myelinated nerve fibers were observed in groups E' and F', uniformity in size, wide distribution, and with myelin sheath, compared with those in groups A', B', C' and D'. Conclusion A certain concentration of T10 can induce the sciatic nerve of rats to express neurotrophic factor in vitro, which can improve the biological activity of cold preservation nerves, reduce the immunogenicity, and promote the regeneration of recipient nerve after allogeneic transplantation. It is even better to be pretreated with T10 before cold preservation.
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OBJECTIVE:To observe clinical efficacy and safety of Bushen sanhan tongluo decoction combined with moxi-bustion and celecoxib in the treatment of knee osteoarthritis (KOA). METHODS:A total of 70 KOA patients were selected from Chongqing Kanghua Hospital during May 2014-Dec. 2015,and then divided into observation group and control group ac-cording to odd and even number,with 35 cases in each group. Control group was given Celecoxib capsule 0.2 g,qd;observa-tion group was additionally given Bushen sanhan tongluo decoction(one dose a day,300 mL,decocted with water,taking it 3 times in the morning,noon and night)and moxibustion. A treatment course lasted for 4 weeks,and both received 2 courses of treatment. Clinical efficacies as well as TCM syndrome score,VAS score,WOMAC score,lab indexes,joint condition be-fore and after treatment,the occurrence of ADR were compared between 2 groups. RESULTS:Total response rate of observa-tion group (85.71％) was significantly higher than control group (68.57％),with statistical significance (P0.05). After treatment,TCM syn-drome score,VAS score,WOMAC score,erythrocyte sedimentation rate,CRP level and knee swelling score of 2 groups were decreased significantly,compared to before treatment;those indexes of observation group were significantly lower than those of control group,with statistical significance(P0.05). There was no statistical significance in the incidence of ADR (5.71％ vs. 2.86％) between 2 groups (P>0.05). CONCLUSIONS:Bushen sanhan tongluo decoction combined with moxibustion and celecoxib can improve clinical symptoms,relieve joint pain,joint inflammation and swelling of KOA pa-tients with good safety.
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0.05). Conclusion:Certain concentration of TG can inhibit the apoptosis of Schwann’s cell and the expression of major histocompatibility antigen of cold preserved sciatic nerve in rats,and decrease rejection after nerve allograft.