The Effect of Shaoyao Gancao Decoction on Sphincter of Oddi Dysfunction in Hypercholesterolemic Rabbits via Protecting the Enteric Nervous System-Interstitial Cells of Cajal-Smooth Muscle Cells Network.

OBJECTIVE: This study observes the morphological changes in the enteric nervous system (ENS) - interstitial cells of Cajal (ICC) - smooth muscle cells (SMC) network in sphincter of Oddi dysfunction (SOD) in hypercholesterolemic rabbits following treatment with Shaoyao Gancao decoction (SGD), as well as the apoptosis of the ICC. METHODS: In this study, 48 healthy adult New Zealand rabbits are randomly divided into three groups (n = 16 in each group): the control, the model, and the SGD treatment groups. The hypercholesterolemic rabbit model is established. Hematoxylin and eosin staining, transmission electron microscopy, immunofluorescence, terminal deoxynucleotidyl transferase dUTP nick end labeling staining, immunohistochemistry, Western blot analysis, and reverse transcription-polymerase chain reaction are used to detect the morphological changes in the ENS-ICC-SMC network, the expression of apoptosis-related proteins in the ICC, and to observe the curative effect of SGD after treatment. RESULTS: Compared with the control group, the morphology and the ultrastructure of the SO are destroyed in the model group. In addition, the protein gene product 9.5 (PGP9.5), nitric oxide (NO), the SMCs, and the ICC all significantly decreased while substance P (SP) significantly increased. Compared with the model group, the SO morphology and ultrastructure are repaired in the SGD group. In addition, the PGP9.5, NO, the SMCs, and the ICC significantly increased while SP decreased. In addition, SGD may activate the stem cell factor (SCF)/c-Kit signaling pathway to treat SO dysfunction by up-regulating the expression of c-Kit and SCF. Similarly, this pathway restores SO by up-regulating the expression of Bcl2 and inhibiting cleaved caspase-3, Bax, and the tumor necrosis factor. CONCLUSION: Shaoyao Gancao decoction can promote the recovery of sphincter of Oddi dysfunction in hypercholesterolemic rabbits by protecting the ENS-ICC-SMC network.

Syntheses of four porous 3-D Cd(ii) metal-organic frameworks from a new multidentate ligand 5-(imidazol-1-yl)-N'-(pyridin-4-ylmethylene) nicotinohydrazide and their characterization and adsorption properties.

Herein, a new multidentate ligand, 5-(imidazol-1-yl)-N'-(pyridin-4-ylmethylene) nicotinohydrazide (L), with an acylhydrazone group was synthesized and characterized. Subsequently, four porous Cd(ii)-MOFs, i.e. [Cd(L)(NO3)] n (1), [Cd(L)Cl] n (2), [Cd(L)Br] n (3), and [Cd(L)I] n (4), were assembled using the ligand L by a solvothermal method and characterized by single-crystal X-ray diffraction, infrared spectroscopy, thermogravimetric analysis, and powder X-ray diffraction. Structural analysis shows that the coordination environments around Cd(ii) in all the four compounds are different due to the different coordinated anions. Among them, the coordination geometries and the arrangement of five-coordinated groups of the compound 1 containing the coordinated NO3 - anions are significantly different from those of the other three compounds containing halides. However, all the four MOFs have similar one-dimensional rhombic channels. In these channels, both the nitrate ions and the halide ions are attached to the inner walls of the pores. The CO2 adsorption properties of 1-4 were studied at 273 K, and the results showed that these compounds exhibit different adsorption capacities for CO2 due to the presence of different ions in their pores.

[Ag-Ag]2+ Unit-Encapsulated Trimetallic Cages: One-Pot Syntheses and Modulation of Argentophilic Interactions by the Uncoordinated Substituents.

Four [Ag-Ag]2+ unit-encapsulated trimetallic cages 1-4 were synthesized from one new tripodal ligand L and silver salts in different solvent systems by a one-pot method. The formation of coordination cages occurred simultaneously with the condensation of amino groups and ketone. The remarkable structural feature of cages 1-4 is their spontaneous incorporation of [Ag-Ag]2+ cationic units. Moreover, the argentophilic interactions are modulated by the uncoordinated amino substituents. The study herein shows that modification and subtle changes of the cage structures could be realized by a one-pot synthetic method.

[Significance of Th17/Treg imbalance in children with primary immune thrombocytopenia].

OBJECTIVE: To investigate the significance of Th17/Treg imbalance in the development and treatment of primary immune thrombocytopenia (ITP) in children. METHODS: Thirty-two children diagnosed with ITP between May and August, 2015 and 22 healthy children were enrolled. Flow cytometry was used to determine the Th17/Treg ratio in peripheral blood of healthy children and children with ITP before and after treatment with immunoglobulin. RESULTS: Compared with the patients with ITP before treatment, the healthy children and the patients treated with immunoglobulin had a significantly lower percentage of Th17 cells in CD4+ T cells, a significantly lower Th17/Treg ratio, and a significantly higher percentage of Treg cells in CD4+ T cells in peripheral blood (P<0.05). In the 32 ITP children treated with immunoglobulin, 20 had complete response, 4 had response, and 8 had no response. The patients with complete response had a significantly lower percentage of Th17 cells in CD4+ T cells and a significantly lower Th17/Treg ratio in peripheral blood than the patients without response (P<0.05). CONCLUSIONS: The Th17/Treg imbalance can be found in children with ITP. Immunoglobulin can improve the cellular immune function by regulation of the Th17/Treg ratio. The Th17/Treg ratio may serve as an indicator for assessing the therapeutic effects of ITP.

[Functions of exogenous application of connective tissue growth factor in stimulating human dermal papilla cells and human hair follicle outer root sheath cells for reconstructive tissue-engineering hair follicles].

OBJECTIVE: To explore the functions of connective tissue growth factor (CTGF) in the restoration of hair follicles with a mixture of human dermal papilla cells and human hair follicle outer root sheath cells in vitro in nude mice. METHODS: Human hair follicle outer root sheath cells (hfORS) and human hair dermal papilla cells (hDP) were cultured in vitro and mixed in a fixed ratio (hfORS: hDP = 5:1). Flow cytometry was used to detect the content of CD200(+) cells in human hair follicle outer root sheath cells.And 8 nude mice were divided randomly into 2 groups according to a random number table and back wounds produced. Group A was transplanted with cell mixture plus 20 µg/L CTGF. Group B was transplanted with cell mixture alone. After 8 weeks of transplantation, the development of hair follicle formation was observed histologically.PCR was used to detect the expression of human specific DNA and mice DNA in transplants. RESULTS: The portion of CD200(+) cells in cultured hfORS was 19.65%. At 8 weeks after implantation, hair follicle formation could be observed in Group A (268 ± 96) more than Group B (62 ± 20). The difference was statistically significant (P < 0.05). And PCR showed that there was human composition in transplant. CONCLUSION: CTGF can induce the formation of hair follicle by promoting the interference between hDP and hfORS.

[Expressions of endoplasmic reticulum stress associated proteins in livers of severely burned rats].

OBJECTIVE: To explore the expression of endoplasmic reticulum stress (ERS) associated proteins in livers of severely burned rats and examine its potential significance. METHODS: Sixty-four Wistar rats were randomly divided into the control and burn groups (30% total body surface area full-thickness thermal injury) (n = 32 each). Livers were harvested at Day 1, 4, 7, 14 post-burn. Western blot was used to detect the expressions of endoplasmic reticulum stress associated proteins glucose regulated protein 78 (GRP78), C/EBP-homologous protein (CHOP), active caspase-12 and active caspase-3. Hepatic apoptosis was assessed by the assay of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL). RESULTS: Compared with the control group, the expression of GRP78 became elevated at Day 1, 4, 7, 14 post-burn (1.29 ± 0.11 vs 1.00 ± 0.00, 1.28 ± 0.12 vs 0.95 ± 0.16, 1.29 ± 0.14 vs 0.93 ± 0.06, 1.41 ± 0.17 vs 1.02 ± 0.13 respectively); the expression of CHOP was higher at Day 1, 4 (1.72 ± 0.07 vs 1.00 ± 0.00, 1.82 ± 0.18 vs 1.46 ± 0.08 respectively) while active caspase-12 and active caspase-3 increased at Day 1, 4, 7 post-burn (2.05 ± 0.65 vs 1.00 ± 0.00, 2.16 ± 0.69 vs 0.95 ± 0.21, 1.98 ± 0.56 vs 0.90 ± 0.22; 1.96 ± 0.15 vs 1.00 ± 0.00, 1.40 ± 0.14 vs 1.07 ± 0.12, 1.77 ± 0.17 vs 1.15 ± 0.21 respectively); the apoptotic index(%) of hepatocytes was higher at Day 1, 4, 7, 14 post-burn (27.20 ± 3.63 vs 5.00 ± 0.71, 16.40 ± 1.52 vs 5.40 ± 1.14, 27.60 ± 1.82 vs 7.40 ± 1.14, 10.20 ± 1.92 vs 5.20 ± 1.64 respectively). All results were statistically significant (all P < 0.05). CONCLUSION: ERS activates and expressions of associated proteins GRP78, CHOP, active caspase-12 and active caspase-3 increase in livers of severely burned rats.

[Experimental research of correlation between anatomy structure of rabbit ear and creating hypertrophic scar animal model].

OBJECTIVE: To observe the anatomy structure of rabbit ear and the effect of different operation methods and post-operative treatments on the formation of hypertrophic scar. METHODS: The experimental animals were 25 New Zealand white rabbits. 6 pieces of full skin specimens were obtained from each of the ears in 5 rabbits for histological examination. 6 full-thickness skin wounds (d = 8 mm) were made on different sites of ventral side of each ear in the other 20 rabbits. The total number of the wounds was 240. 120 wounds in 10 rabbits were divided into 4 groups randomly to receive different treatments on day 7 postoperatively. No treatment was performed in the other 120 wounds. The wounds healing and the scar formation were observed for six months. The scars were harvested 4 weeks and 8 weeks after operation for pathologic examination and measurement of scar elevation index (SEI). RESULTS: Histological analysis showed that the anatomy structure was different in different sites of the rabbit ear. The best sites for creating hypertrophic scar model were on the medial margin of the middle- and inferior part of ear. The depth of the wound should reach the cartilage membrane of the ear to facilitate the formation of hypertrophic scar. The second strip crust on day 7 postoperatively enhanced the wounds healing and minimized the scar proliferation and hypertrophy. CONCLUSIONS: There is a close correlation between the anatomy structure of the ear and the creation of hypertrophic scar animal model. The wound site, the depth of wound and the post-operative treatment will affect the formation of hypertrophic scar. The study can help to improve the successful rate of creating hypertrophic scar animal model.

Effect of heparin on production of basic fibroblast growth factor and transforming growth factor-beta1 by human normal skin and hyperplastic scar fibroblasts.

Heparin affects both dermal fibroblast proliferation and collagen and may mediate these effects by altering the levels of basic fibroblast growth factor (bFGF) and transforming growth factor-beta1 (TGF-beta1) production as a wound healing modulator. The purpose of this study is to probe the effect of heparin on bFGF and TGF-beta1 production by human normal skin and hyperplastic scar fibroblasts. This research investigates the effect of heparin on bFGF and TGF-beta1 production by human normal skin and hyperplastic scar fibroblasts with exposure to 0, 100, 300, or 600 microg/ml heparin for 24, 48, 72, or 96 hours in a serum-free in vitro model. Levels of bFGF and TGF-beta1 in the supernatants were determined by enzyme-linked immunosorbant assay. All doses of heparin significantly stimulated production of bFGF by normal skin (393% to 1019% increase) and hyperplastic scar fibroblasts (405% to 899% increase) at all time points (P < .05). Heparin (300 and 600 microg/ml) also stimulated TGF-beta1 production by normal skin (26% to 83%) and hyperplastic scar fibroblasts (63% to 85%) with statistical significance (P < .05) at various time points. These effects of heparin on normal skin and hyperplastic scar fibroblasts may have implications for hyperplastic scar formation and wound healing in vivo.

[Study on poly A site gene mutation of transforming growth factor-beta1 receptor type II between multiple and single site keloid].

OBJECTIVE: To investigate the pathogenesy deference between multiple and single site keloid by detecting gene mutation of Poly A site of transforming growth factor-beta1 receptor type II (TbetaR II). METHODS: Collecting 20 keloid samples (6 multiple sites keloid samples and 14 single site keloid samples) and extracting DNA from them; designing and synthesizing the primers of Poly A site, then amplifying T1beta II DNA by PCR, analyzing the single strand conformation polymorphism about the products of PCR. After purifying the product of PCR, the site and type of the mutation rate of Poly A site was sequenced directly on the automatic sequencing equipment. RESULTS: It had been found that the Poly A site of TbetaR II in keloid has deletion mutation, its mutation rate in multiple sites keloid was 50% (3/6), in single site keloid 7.1% (1/14). The mutation rate of Poly A site in multiple sites keloid was significant higher than that in single site keloid (P < 0.05) CONCLUSION: It has been supposed that there are some deference in pathogenesy between the multiple and the single site keloid.