OBJECTIVE: To develop a HPLC method for assaying strychnine and brucine in traditional Chinese drug Maqianzi San. METHODS: With μBondapak C18 (10 μm) column (4 mm×300 mm), MeOH-H2O-HAc-(C2H5)3N(70∶230∶2.4∶0.3) as mobile phase, we set a detector at 254 nm. RESULTS: Linear range of strychnine and brucine was 0.367 2 μg～0.061 2 μg and 0.945 0 μg～0.157 5 μg respectively. The recovery rate of strychnine and brucine was 100.36% (RSD=1.09%) and 100.01% (RSD=1.46%) respectively. CONCLUSION: This method provides a simple and feasible way to control the quality of Maqianzi San.

AIM:To examine the relationship between the gene defect,change of protein hydrophobicity,spacial structure change and clinical phenotypes of Duchenne muscular dystrophy(DMD),and to explore the molecular pathogenesis of DMD.METHODS:The gene sequences of 59 cases of DMD/BMD patients with deletion from mutation were analyzed.The relationship between the protein hydrophobicity,3D-spacial structure and clinical phenotypes was examined by biological informatic technology.RESULTS:50 cases of frameshift mutation were all DMD.In other 5 cases with codon mutation that involved the 3rd hydrophobic region,4 cases were diagnosed as DMD and the rest one was BMD.The exon 3 deletion leaded to the intortion of dystrophin N-terminal,which in turn affected the combination of dystrophin and troponin resulting in the DMD pathopoiesis.CONCLUSION:The severity of clinical phenotypes of muscular dystrophy diseases is related to whether the deletion destroys the reading frame,involves the 3rd hydrophobic region or changes the protein special structure.The biological informatic technology provides a new potential research methodology for studying the pathogenesis of DMD.

With the information section of the 187th Central Hospital of PLA as an example , the paper introduces the application of the PDCA cycle model in the quality management of its information services .As a result, it finds that the service satisfaction degree after the implementation of the PDCA cycle is significantly higher than that before the implementation .It indicates that the management method with the PDCA cycle can optimize the information service process and promote the continuous improvement of information service quality .

Objective To analyze the rules of acupoint selection in treating knee osteoarthritis with acupuncture and moxibustion therapy based on data mining; To provide references for clinical treatment.MethodsArticles about treating knee osteoarthritis with acupuncture and moxibustion were retrived from China National Knowledge Infrastructure (CNKI), Wanfang academic periodical full-text database, Chinese Journal Full-text Database (VIP) and SinoMed in China from Jan. 1st 2006 to Jan. 1st 2016. The database of acupuncture and moxibustion prescription was set up. The Apriori association rules, system cluster analysis and factor analysis were adopted in order to analyze the rules of acupoint selection for knee osteoarthritis.ResultsTotally 190 articles were included, covering 228 acupuncture- moxibustion prescriptions, 65 acupoints and the total frequency for 1736 times. The usual acupoints in the lower limbs were Dubi (ST35), Neixiyan (EX-LE 4), and Yanglingquan (GB34). The main maridians were Stomach Meridian of Foot-Yangming, Spleen Meridian of Foot-Taiyin, and Gallbladder Meridian of Foot-Shaoyang, accounting to for 1626 times (93.66%). Cluster analysis achieved other four groups of acupoint selection based on syndrome differentiation.ConclusionLocal acupoints and dialectical point selection were important principle in treating knee osteoarthritis combined with acupoint selection based on syndrome differentiation.

Objective To investigate the expression of monocyte chemoattractant protein-1(MCP-1)and CCR2 in peripheral blood and pleural fluid of tuberculous pleurisy patients.Methods Flow cytometry was used to detect the expression of CCR2 in the peripheral blood mononuclear cell and pleural fluid cell of tuberculous pleurisy patients;ELISA was used to detect the content of MCP-1 in serum and pleural fluid.Results MCP-1 in surem and pleural fluid and CCR2 in the peripheral blood mononuclear cell and pleural fluid cell of tuberculous pleurisy patients was significantly higher than that of the healthy normal controls[MCP-1:(340.8±220.8)and(9.0±3.8)ng/L,P＜0.01;CCR2(18.2±10.1)%and(6.9±3.5)%,P＜0.05];Both MCP-1 and CCR2 were detected in pleural fluid and both of them were positivley correlated(r=0.227,P＜0.05).Conclusion The expression of MCP-1 and CCR2 in peripheral blood of tuberculous pleurisy patient are significantly elevated which are significance molecule participating in the pathogenesis of tuberculous pleurisy.

AIM: To observe the effects of overload exercise on skeletal muscles in X-linked muscular dystrophy(mdx) mice.METHODS: Mdx mice and C57 mice were carried out swimming and hanging tail movement tests (mdx mice as control did not exercise). It lasted for 13 minutes each time per day, and lasted 3 days. Evans blue was injected into tail vain. The mice were killed the next day, and the hind limbs were taken photographs after skins were flayed. The gastrocnemius muscles and diaphragms cryostat sections were made. Under a fluorescence microscope, Evans blue staining was seen. Then the sections were tested by routine HE staining, the histological change of muscles was analyzed under a light microscope.RESULTS: Many blue colored longitudinal lines were observed in skeletal muscles of mdx mice, whereas they were hardly seen in control mdx and C57 mice. Under a fluorescence microscope, some muscle fibers of mdx mice were stained with Evans blue, few muscle fibers of control mdx mice were stained, and C57 mice were not. Under a light microscope, HE staining of muscles showed some degenerated muscle fibers became round in shape and the myonuclei became condensed, or necrotic fibers had amorphous structures, most of them in the degenerated and necrotic fibers of diaphragms C57 mice did not have these changes.CONCLUSION: Overload exercise did harm to skeletal muscles of mdx mice; Vital staining with Evans blue is useful not only for distinguishing degenerating muscle fibers, but also for studying the degeneration process in dystrophin-deficient muscle.

AIM: To detect dystrophin expression in skeletal muscles of mdx mice after bone marrow transplantation (BMT), and to evaluate the effect of BMT on Duchenne muscular dystrophy (DMD). METHODS: Bone marrow cells were cultured for three days, and then transplanted into mdx mice irradiated lethally through tail veins. After 4 and 6 months, dystrophin expression on myocytes membranes in mdx mice was detected by fluorescent immunohistochemical staining. The centrally nucleated fibers (CNF) were calculated by HE staining, and the physiologic parameters measured and the motor function detected by traction test, rotating rods test and rotating wheels test were also observed. RESULTS: Until 4 and 6 months after BMT, dystrophin was expressed partly on myocytes membranes in mdx mice, and the ratio of CNF decreased, physiologic functions improved, the motor ability reinforced in treated group. CONCLUSION: After BMT, marrow stem cells settled in injured skeletal muscles and bone marrow, then differentiated into myocytes with dystrophin expression and caused the improvement of pathology, physiology and motor function in treated group finally. These results give a powerful proof for the treatment of DMD with BMT.

AIM: To study the effect of bone marrow stem cell transplantation on mdx mice at different ages. METHODS: The bone marrow stem cells of C57BL/6 mice (4 - to-weeks age) were cultured in vitro for 3 days, then injected intravenously into the 6 -week and 8-week aged mdx, which were preconditioned with 7 Gy ? ray. 12 weeks after being transplanted, the mdx mice were studied for the dystrophin protein expression on the skeletal muscle membrane. RESULTS: Three months after transplanted with bone marrow stem cells, about 16% and 7% muscles cells in 6-week and 8-week mdx mice expressed dystrophin protein, respectively. CONCLUSION: 12 weeks after transplantation with bone marrow stem cells of homologous series mice, different amounts of dystrophin protein expressed on the membrane of skeletal muscle cells were observed in different aged mdx mice. Bone marrow stem cell transplantation show more benefic effect for younger mdx mice.

【Objective】To observe the dynamic interference of cellular endotoxin specific receptors of Toll-like receptor 4(TLR4) mRNA and nuclear factor ?B p65(NF-?B p65) in rats with damp-heat syndrome of seasonal febrile disease by Ganlu Xiaodu Dan(GXD),and to explore the therapeutic mechanism of prescriptions with the actions of clearing heat and resolving dampness.【Methods】Wistar rats were randomized into 3 groups: A(normal control),B(models established by feeding of high-fat diet and colon bacillus under the high-temperature and high-humidity condition),and C(rat models treated with GXD).Reverse transcription-polymerase chain reaction(RT-PCR) analysis was used to detect the TLR4 mRNA level in hepatic macrophages and immunohistochemical method was used to detect the activation of NF-?B p65.【Results】The expression of TLR4 mRNA and NF-?B p65 was increased gradually in group B 6,12 and 24 hours after infection with colon bacillus,and the differences were significant,indicating that the expression of TLR4 mRNA and NF-?B p65 increased with the prolongation of disease course.In group C,TLR4 mRNA expression was decreased in different time points after infection,the difference of TLR4 mRNA expression 12 and 24 hours after infection was significant as compared with group B and group A(P