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With literature of professor CHEN Yinglong such as Medical Notes of Doctor CHEN Yinglongfor Taiwan Compatriots,the'experience of doctor CHEN Yinglong treating common diseases in Fujian and Tai-wan is summarized. The diseases in the paper are constipation, retention of urine, asthma, vec6rdia, bi syndromeinsomnia, thoracic obstruction, pediatric obesity and freckle of face, etc. It is discovered that professor CHENYinglong treated diseases with combination of acupuncture and medication and accurate acupoints according to thefeature of climate in Fujian and Taiwan.
Subject(s)
Humans , Acupuncture Points , Acupuncture Therapy , History , Asthma , Therapeutics , Constipation , Therapeutics , History, 20th Century , TaiwanABSTRACT
Objective To establish a high-performance induction culture system for Raw264.7 cells to differentiate into osteoclasts(OC)invitro by improving the cell culture program.Methods The Raw264.7 cells were cultured withα-MEM medium containing 50 μg · L-1 M-CSF, 100 μg · L-1 RANKL, and 1 × 10-8 mol · L-1 1α,25-(OH)2 D3 in 5% CO2 for 12 d at 37℃. The cells were digested transiently every time before the medium was changed after every three days. The morphologic changes of the Raw264.7 cells were observed by inverted microscope.The maturation and phagotrophic function of OC were identified by HE,TRAP,FITC-phalloidin staining and immunofluorescence.Results The cells remained to grow in single layers all the time in most areas of the well during the whole induction by the improved culture program. The observation results of inverted microscope and HE staining showed that the growth area of the polykaryotic OC reached to 70% of the well on day 1 2. FITC-phalloidin staining showed that in the maturation of the OC, the cluster-shaped podosomes in the pseudopodia gradually transformed into rings,which finally fused to form a large belt surrounding the periphery of the cytoplasm. The calcitionin receptor (CTR) expressed by OC was markedly enhanced compared with the precursor cells by immunofluroescence staining,and a large number of red granules appeared in the cytoplasm of OC with TRAP staining on day 1 2. These results comfirmed that the obtained OC were maturated and owned phagotrophic function. Conclusion A high-performance induction culture system for Raw264. 7 cells to differentiate into OC in vitro induced by combination of 50μg · L-1 M-CSF, 100 μg · L-1 RANKL,and 1 × 10-8 mol·L-1 1α,25-(OH)2 D3 is established by improving the cell culture program.
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Objective To probe the relationship between the expression of TL1A and the level of IFN-γ secreted by T cells in the acute stage of Guillain-Barre syndrome (GBS). Methods ① Six-week female Bal b/c mice were immunized by purified recombinant human soluble TNF-like molecular 1A (rhsTL1A) protein. The polyclonal antibody against rhsTL1A was identified by immunofluorescence using human umbilical vein epithelial cells (HUVEC). ② To detect the biologic activity of rhsTL1A, the peripheral blood mononuclear cells (PBMC) from the healthy donors were separated by Ficoll gradient centrifugation and were seeded on 96-well plates with medium containing 2 μg/ml PHA (control group), 2 μg/ml PHA + 25 ng/ml rhsTL1 A, 2 μg/ml PHA + 100 ng/ml rhsTL1A and 2 μg/ml PHA + 400 ng/ml rhsTLlA respectively. T cell proliferation assay was carried out using ~3H-TdR. ③ IFN-γ productions in the sera of the children with GBS in the acute stage were detected by ELISA. ④ The ratio of CD_3~+ TL1A~+ T cells to CD_3~+ T cells in the peripheral blood of the children with GBS in acute stage was detected with flow, cytometry. ⑤PBMC from the children in acute GBS were separated and cultured in the environment adding 2 μg/ml PHA and 400 ng/ml rhsTL1A in vitro. Then, the IFN-γ in the supernatant was determined by ELISA kit after 72 hours. Results ① hTL1A A expressed by eukaryotic HUVECs was recognized by rhsTL1 A polyclonal antiserum. ② The result of T cell proliferation assay showed that SI of 25 ng/ml rhTL1A, 100 ng/ml rhTL1A A and 400 ng/ml rhTL1A group was increased compared with control group. The SI of 2 μg/ml PHA +400 ng/ml rhsTL1 A group was the highest (2. 65) among them. ③ IFN-γ productions in the sera of the children with GBS in the acute stage ((102. 25±22. 17) pg/ml) were increased significantly compared with healthy control ((28.75 ± 1.31) pg/ml, t = 3. 309, P < 0. 05). ④ The ratio of CD_3~+ TL1A~+ T cells to CD_3~+ T cells in the peripheral blood of the children with GBS in acute stage (18.22%± 1.83%) was enhanced significantly compared with healthy control (5. 17% ±0. 48%, t = 6. 884, P < 0. 01). ⑤ PBMC both in healthy control and the acute GBS secreted more IFN-γ markedly ((43.56± 4.41) pg/ml and (180.64 ± 38.39) pg/ml) after being incubated in 2 μg/ml PHA and 400 ng/ml rhsTL1A (t =4. 523 and 2. 600, P <0. 01 and 0. 05 respectively). Moreover, PBMC in acute GBS secreted more IFN-γ, than that of the healthy group markedly (t = 3. 545, P < 0. 05). Conclusions ① The mouse antiserum recognizing rhsTL1A is successfully obtained. ② In this study, 400 ng/ml rhsTL1A promotes the proliferation of T cells activated by 2 μg/ml PHA, indicating that rhsTL1A has biological activity. ③ The expression of hTL1A of activated T cells in the peripheral blood of the children with acute GBS is up-regulated. These TL1A proteins promote the secretion of IFN-γ through binding to their receptors DR_3.
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Objective:To probe into the long-term effect of La(NO3)3 fed orally on gast ric mucosa of rats.Methods:The gastric mucosa of rats was studied after La(NO3)3 was administe red orally for a long-term,by the use of ordinary histological technology and m orphometry.Results:In the groups of 20 mg·kg-1 and 10 mg·kg-1,the cytopl asma of more parietal cells lying in the top of the gastric gland was loose.Proporti on of cell types of the gastric gland was related to the doses. The acid mucus l evel of the mucous neck cells of male rats in the groups of 20 mg·kg-1 an d 10 m·kg-1 decreased. In the groups of 2 mg2kg-1, 0.2 mg·kg -1 and 0.1 mg*kg-1,the acid mucus level of the mucous neck cells of both male and female rats increased.Conclusion:La(NO3)3 fed orally for a long-term could injure gastric muc osa in higher dose (20 mg·kg-1,10 mg·kg-1) but promote the protect ive action of the gastric mucosa in lower dose (0.2 mg·kg-1,0.1 mg·kg- 1).
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Objective To explore the effect of Acorus gramimeus and its major components ?-asarone on N-methyl-D-asperate recepter 1 (NMDAR1) mRNA levels of hippocamp neurons in epileptic animal models induced by pentylenetetrazol (PTZ). Methods PTZ 60 mg/kg was injected to the abdominal cavity of three weeks Wistar rats to prepare epileptic animal models. The animal models were randomly divided into three groups: epilepsy control, A. gramimeus and ?-asarone as well as the normal control. Each group was administered through intraperitoneal injection. Both of two control groups were injected physiologic saline 1.0 mL/d, and the rest groups were injected A. gramimeus 2 350 mg/(kg?d) or ?-asarone 29 mg/(kg?d) for 7 d, respectively. All rats were treated by PTZ 60 mg/kg through intraperitoneal injection next day. After their behaviors were observed for 24 h, animals were sacrificed. The expressions of NMDAR1 mRNA in hippocamp CA1 and CA3 of all rats were detected by in situ hybridization and semiquantitative RT-PCR. Results The results of in situ hybridization showed that positive staining granules were located in the cyptoplasm of hippocamp neurons. The number of positive cells and average absorbance of A. gramimeus group or ?-asarone group was markedly less than that of epilepsy control (P