In this study, the crude polysaccharides was extracted from Shengfupian and purified by Sevag deproteinization. Then, the purified neutral polysaccharide fragment was obtained by the DEAE-52 cellulose chromatography column and Sephadex G-100 co-lumn. The structure of polysaccharides was characterized by ultraviolet spectroscopy, infrared spectroscopy, ion chromatography, and gel permeation chromatography. To investigate the anti-inflammatory activity of Shengfupian polysaccharides, LPS was used to induce inflammation in RAW264.7 cells. The expression of the CD86 antibody on surface of M1 cells, the function of macrophages, and the content of NO and IL-6 in the supernatant were examined. An immunodepression model of H22 tumor-bearing mice was established, and the immunomodulatory activity of Shengfupian polysaccharides was evaluated based on the tumor inhibition rate, immune organ index and function, and serum cytokine levels. Research indicated that Shengfupian polysaccharides(80 251 Da) was composed of arabinose, galactose, glucose, and fructose with molar ratio of 0.004∶0.018∶0.913∶0.065. It was smooth and lumpy under the scanning electron microscope. In the concentration range of 25-200 μg·mL~(-1), Shengfupian polysaccharides exhibited little or no toxicity to RAW264.7 cells and could inhibit the polarization of cells to the M1 type and reduce the content of NO and IL-6 in the cell supernatant. It could suppress the phagocytosis of cells at the concentration of 25 μg·mL~(-1), while enhancing the phagocytosis of RAW264.7 cells within the concentration range of 100-200 μg·mL~(-1). The 200 mg·kg~(-1) Shengfupian polysaccharides could alleviate the spleen injury caused by cyclophosphamide, increase the levels of IL-1β and IL-6, and decrease the level of TNF-α in the serum of mice. In conclusion, Shengfupian polysaccharides has anti-inflammatory effect and weak immunomodulatory effect, which may the material basis of Aconm Lateralis Radix Praeparaia for dispelling cold and relieving pain.
Animals ; Mice ; Interleukin-6/genetics* ; Cytokines/metabolism* ; Polysaccharides/chemistry* ; RAW 264.7 Cells ; Anti-Inflammatory Agents/chemistry* ; Spectrophotometry, Infrared

Child ; Humans ; Inflammatory Bowel Diseases ; Organic Anion Transporters/genetics*

Morphine is a frequently used analgesic that activates the mu-opioid receptor(MOR),which has prominent side effects of tolerance.Although the inefficiency of morphine in inducing the endocytosis of MOR underlies the development of morphine tolerance,currently,there is no effective therapy to treat morphine tolerance.In the current study,we aimed to develop a monoclonal antibody(mAb)precisely targeting MOR and to determine its therapeutic efficacy on morphine tolerance and the underlying molecular mechanisms.We successfully prepared a mAb targeting MOR,named 3A5C7,by hybridoma technique using a strategy of deoxyribonucleic acid immunization combined with cell immunization,and identified it as an immunoglobulin G mAb with high specificity and affinity for MOR and binding ability to antigens with spatial conformation.Treatment of two cell lines,HEK293T and SH-SY5Y,with 3A5C7 enhanced morphine-induced MOR endocytosis via a G protein-coupled receptor kinase 2(GRK2)/β-arrestin2-dependent mechanism,as demonstrated by immunofluorescence staining,flow cytometry,Western blotting,coimmunoprecipitation,and small interfering ribonucleic acid(siRNA)-based knock-down.This mAb also allowed MOR recycling from cytoplasm to plasma membrane and attenuated morphine-induced phosphorylation of MOR.We established an in vitro morphine tolerance model using differentiated SH-SY5Y cells induced by retinoic acid.Western blot,enzyme-linked immunosorbent assays,and siRNA-based knockdown revealed that 3A5C7 mAb diminished hyperactivation of adenylate cyclase,the in vitro biomarker of morphine tolerance,via the GRK2/β-arrestin2 pathway.Furthermore,in vivo hotplate test demonstrated that chronic intrathecal administration of 3A5C7 significantly alle-viated morphine tolerance in mice,and withdrawal jumping test revealed that both chronic and acute 3A5C7 intrathecal administration attenuated morphine dependence.Finally,intrathecal electroporation of silencing short hairpin RNA illustrated that the in vivo anti-tolerance and anti-dependence efficacy of 3A5C7 was mediated by enhanced morphine-induced MOR endocytosis via GRK2/β-arrestin2 pathway.Collectively,our study provided a therapeutic mAb,3A5C7,targeting MOR to treat morphine tolerance,mediated by enhancing morphine-induced MOR endocytosis.The mAb 3A5C7 demonstrates promising translational value to treat clinical morphine tolerance.

Gypenosides (GP), the main medicinal ingredient of Gynostemma Pentaphyllum Makino, exhibits excellent protective effect on various neural diseases through several mechanisms, including anti-inflammation, anti-oxidative stress and pro-neural regeneration. In the current review, we have made a summary on the research progress of GP on neural disorders, like Alzheimer' s disease, Parkinson' s disease, depression, hypoxic brain injury and so on, aiming to provide an experimental basis for the further research and drug development of GP.

Objective: To observe the clinical efficacy of auricular point pricking-bloodletting plus auricular point sticking therapy for acne vulgaris. Methods: A total of 66 patients with acne vulgaris were randomized into an observation group and a control group by the random number table, with 33 cases in each group. The observation group was treated with auricular point pricking-bloodletting plus auricular point sticking therapy, and the control group was treated only with auricular point sticking therapy. The treatments of both groups were performed twice a week, 4 weeks as a course of treatment, for 3 courses in total. The scores of skin lesions and dermatology life quality index (DLQI) scores were recorded before and after treatment to assess the clinical efficacy. Results: During the trial, there were 3 cases of drop-out both in the observation group and the control group. After 3 courses of treatment, the total effective rate of the observation group was 96.7％, while that of the control group was 76.7％. The difference between the two groups was statistically significant (P<0.05). The intra-group comparison showed that the scores of skin lesion and DLQI were both decreased with the increase of treatment times, that was, the scores were lower than those at the previous time point (allP<0.05). After 1, 2, and 3 courses of treatment, the scores of skin lesion and DLQI of both groups were statistically different from those of the same group before treatment (allP<0.05). At every time point during the treatment, the scores of skin lesion and DLQI of the observation group were lower than those of the control group, and the differences between the two groups were statistically significant (all P<0.05). Conclusion: Auricular point pricking-bloodletting plus auricular point sticking has a better curative effect than auricular point sticking therapy alone in the treatment of acne vulgaris, and has a time-effect correlation.

Objective To investigate the relationship between the CD 8 +T cells results of clinical automatic analysis platform and the CD8lowT and CD8highT cell subsets.Methods A total of 1316 cases of lymphocyte and flow cytometry data were collected from the First Affiliated Hospital of China Medical University from December 2015 to September 2016 by cross-sectional study. There were 287 cases of malignant tumor , 389 cases of autoimmune disease , 320 healthy people and 320 cases of HIV infection , then to get automatic analysis platform returns result of CD 8+T cell.FlowJo software was used to analyze the CD8low T and CD8high T lymphocyte subsets in the patients , and the results were compared with the results of CD8 +T cells returned by the clinical automatic analysis platform .Results The results of clinical returns CD8 +T cells were consistent with the results of CD 8high T cells in patients with different diseases , and were not exactly the same as the results of CD8lowT cells, and the difference was as follows:the results of CD8low T cells in HIV-infected patients were significantly lower than those of healthy people (56.2 ±42.0, 68.8 ± 45.9, cells/μl P<0.001), which were different from the clinical results of CD 8 +T cells.The results of clinical report of CD8 +T cells were statistically correlated with CD8high T cells and CD8low T cells, and the correlation between CD8 +T cells and CD8highT were higher than that of CD8lowT cells.There was a positive correlation between CD8low T cell count and CD4 +T cell count ( r=0.204, P<0.001) .CD8low T was significantly higher in patients on antiviral treatment than that in untreated group (58.3 ±43.9, 42.9 ± 26.5, cells/μl, P<0.001).After treatment for more than 2 years, the CD8lowT cells in patients with CD4<500 cells/μl were significantly lower than those in patients with CD 4>500 cells/μl (50.1 ±47.0, 66.3 ±46.6, cells/μl, P<0.001).Conclusions The clinical report of CD8 +T cells was consistent with the results of CD8highT cells, and there was a great difference between the results of CD 8lowT cells and the results of CD8 +T cells.CD8low T cells were significantly reduced in HIV infected patients , and CD8low T cells could be effectively reconstructed by antiviral therapy .

According to folk usage of Aconitum carmichaelii Debx., the present study was designed to determine the feasibility of the stems and leaves of Aconitum carmichaelii Debx. as a new medicinal resource. Fourteen alkaloids in mother roots, fibrous roots, stems, and leaves of Aconitum carmichaelii Debx. were measured by HPLC-MS/MS. And multivariate analysis methods, such as clustering analysis and principal component analysis, were applied to analyze the difference among various parts. In addition, the acute toxicity, analgesia, and anti-inflammatory tests were carried out. The results suggested that the contents of alkaloids in mother roots and fibrous roots were approximate, but those of leaves and stems were different from mother roots and fibrous roots. The results of the acute toxicity testing demonstrated the toxicity of fibrous root was strongest, and mother roots were slightly less toxic than fibrous roots. The stems and leaves were far less toxic than mother and fibrous roots. In addition, the analgesia and inflammatory tests showed the effects of the various tissues had no difference each other. These results provided a basis for developing new complementary and alternative treatments for rheumatoid arthritis patients. Simultaneously, the approach may also turn wastes into treasure and promote the development of circular economy.
Aconitum ; chemistry ; Alkaloids ; administration & dosage ; chemistry ; toxicity ; Animals ; Anti-Inflammatory Agents ; administration & dosage ; chemistry ; toxicity ; Arthritis, Rheumatoid ; drug therapy ; Chromatography, High Pressure Liquid ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; toxicity ; Female ; Humans ; Male ; Mice ; Plant Leaves ; chemistry ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Tandem Mass Spectrometry

OBJECTIVETo study the distribution of polymorphism of c.212-37insC (rs3832879) in intron 1 of fibroblast growth factor 23 (FGF23) gene and its association with Kawasaki disease (KD) and coronary artery lesions (CAL).

METHODSForty children with KD were enrolled in this study, among whom 16 children had concurrent CAL. Twenty-six age-matched healthy children were enrolled as controls. PCR and gene sequencing were applied to explore the distribution of polymorphism of c.212-37insC (rs3832879) in FGF23 gene in KD patients and controls.

RESULTSAmong 40 children with KD, 14 (35%) carried the polymorphism of c.212-37insC (rs3832879) in FGF23 gene; among 26 controls, 6 (23%) carried such polymorphism. There was no significant difference in genotype distribution at this locus between the two groups (P=0.30). Among 16 children with CAL, 9 (56%) carried the polymorphism at this locus; among 24 children without CAL, 5 (21%) carried such polymorphism. As for the comparison of two subgroups with and without CAL, the difference in genotype distribution at this locus had statistical significance (P=0.02, OR=4.89, 95% CI: 1.21-19.71).

CONCLUSIONSThe polymorphism of c.212-37insC (rs3832879) in FGF23 gene may not be associated with the pathogenesis of childhood KD, but it may be associated with the development of CAL in children with KD.

Child ; Child, Preschool ; Coronary Artery Disease ; etiology ; genetics ; Female ; Fibroblast Growth Factors ; genetics ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; etiology ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic

OBJECTIVETo investigate the safety and feasibility of laparoscopy-assisted total proctocolectomy using medial-to-lateral approach.

METHODSBetween October 2005 and January 2012, 21 consecutive patients underwent laparoscopic-assisted total proctocolectomy by medial-to-lateral approach in Department of General Surgery in Nanfang Hospital. The clinical data and follow-up results were reviewed.

RESULTSTwenty cases underwent laparoscopic resection successfully, and 1 case (4.8%) was converted to open surgery because of serious peritoneal cavity adhesion. The mean operative time was (237.1±64.2) min and intraoperative blood loss was (90.0±77.7) ml. The mean time to first flatus was (2.7±0.8) days. The mean postoperative hospital stay was (11.8±5.7) days. Three patients (14.3%) experienced postoperative complications, including anastomotic leakage (n=1), lymphatic leakage (n=1), and anastomotic stricture (n=1). The median follow-up was 22 months (4-60 months). There were two deaths including one patient died of progressive disease 5 months after surgery and the other died of multiple organ failure.

CONCLUSIONSThe advantages of laparoscopy-assisted total proctocolectomy using medial-to-lateral approach include simplified surgical procedure, clearly revealed surgical plane, and shortened operative time. This procedure is safe and feasible in the experienced department of laparoscopic colorectal surgery.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Feasibility Studies ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Proctocolectomy, Restorative ; methods ; Retrospective Studies ; Treatment Outcome ; Young Adult