Exogenous R-Spondin1 Induces Precocious Telogen-to-Anagen Transition in Mouse Hair Follicles.

R-spondin proteins are novel Wnt/ß-catenin agonists, which signal through their receptors leucine-rich repeat-containing G-protein coupled receptor (LGR) 4/5/6 and substantially enhance Wnt/ß-catenin activity. R-spondins are reported to function in embryonic development. They also play important roles in stem cell functions in adult tissues, such as the intestine and mammary glands, which largely rely on Wnt/ß-catenin signaling. However, in the skin epithelium and hair follicles, the information about R-spondins is deficient, although the expressions and functions of their receptors, LGR4/5/6, have already been studied in detail. In the present study, highly-enriched expression of the R-spondin family genes (Rspo1/2/3/4) in the hair follicle dermal papilla is revealed. Expression of Rspo1 in the dermal papilla is specifically and prominently upregulated before anagen entry, and exogenous recombinant R-spondin1 protein injection in mid-telogen leads to precocious anagen entry. Moreover, R-spondin1 activates Wnt/ß-catenin signaling in cultured bulge stem cells in vitro, changing their fate determination without altering the cell proliferation. Our pioneering study uncovers a role of R-spondin1 in the activation of cultured hair follicle stem cells and the regulation of hair cycle progression, shedding new light on the governance of Wnt/ß-catenin signaling in skin biology and providing helpful clues for future treatment of hair follicle disorders.

[Observation on Therapeutic Effects of Acupuncture Combined with Cutaneous Electrical Stimulation for Dysphagia in Patients with Cerebral 1nfarction].

OBJECTIVE: To observe the clinical effect of acupuncture combined with neck-skin electrical stimulation (NSES) on dysphagia in patients with cerebral infarction (CI). METHODS: A total of 120 CI patients with dysphagia were randomly divided into acupuncture group, NSES group and acupuncture + NSES group (combined treatment group, n = 40 in each group). Acupuncture stimulation of Fengchi (GB20), Yifeng (TE 17), etc., and blood-letting of Jinjin (EX-HN 12) and Yuye (EX-HN 13) were administrated. NSES was applied to the bilateral sites of the neck-median line. The treatment was given once daily for two weeks. The swallow function and swallow dysfunction degree of the dysphasia patients were evaluated by water swallow test and food-intake scale, respectively. RESULTS: After one week's and two weeks' treatment, the water swallow score and swallow dysfunction score were significantly improved in the acupuncture, NSES and combined treatment groups (P<0. 01), and the difference values between pre- and post-treatment of the water swallow score and swallow ability score in the combined treatment group were obviously higher than those of the acupuncture and NSES groups (P<0. 01, P<0. 05). No significant differences were found between the acupuncture and NSES groups in both the water swallow score and swallow ability score after one and two weeks' treatment (P>0. 05). Of the three 40 cases in the acupuncture, NSES and combined treatment groups, 16, 18 and 27 were basically cured, 2, 3 and 5 experienced marked improvement, 15, 13 and 7 were improved, and 7, 6 and 1 failed in the treatment, with the effective rates being 82.5%, 85.0% and 97. 5%, respectively. The therapeutic effect of the combined treatment group was apparently superior to that of the simple acupuncture and simple NSES groups (P<0. 01). CONCLUSION: Acupuncture and NSES intervention is effective in improving dysphasia in CI patients and the effect of combined treatment of acupuncture and NSES is obviously better than that of the simple acupuncture and simple NSES.

[The protective effects of sacral nerve electrostimulation on intestinal mucosal mechanical barrier in rats with spinal cord injury].

OBJECTIVE: To study the protective effects of sacral nerve root electrostimulation on intestinal mechanical barrier in rats with spinal cord injury (SCI). METHODS: Fifty six Wistar rats were divided into normal group, SCI control group and SCI group with sacral nerve root electrostimulation (8 rats in each subgroup at 24, 48, 72 h after spinal cord injury). The following experiments were performed respectively in rats from the 3 groups: bacteria culture from intestinal mesentery lymph nodes, liver, spleen, intestinal morphology observation and detection the protein expression level of ZO-1. RESULTS: The intestinal mucosa appeared different degree of damage in SCI control group; cell-cell connections between intestinal epithelial cells were destroyed; Endotoxin levels in blood and the number of bacterial translocation increased obviously. Sacral nerve stimulation was found toimprove the intestinal mucosal, reduce the endotoxin content in the blood to normal level and the decrease the incidences of bacterial translocation of the gut origin. The expression of tight junction protein ZO-1 of rat intestinal tissue had no statistical differences among the 3 groups. On the other hand, the distribution of tight junction protein ZO-1 appeared different degrees of scattered and irregular in the control group while that in the experimental group appeared different degree of improvement as determined by the immunohistochemistry of rat intestinal tissue. CONCLUSION: sacral nerve root electrostimulation can rehabilitate the peristalsis of denervated colon, promote defeacation and decrease bacterial amount, protection of the intestinal mechanical barrier between intestinal epithelial cells and tight junction, reducing the endotoxin content in the blood and suppressing bacterial translocation from the gut.

NASA-approved rotary bioreactor enhances proliferation of human epidermal stem cells and supports formation of 3D epidermis-like structure.

The skin is susceptible to different injuries and diseases. One major obstacle in skin tissue engineering is how to develop functional three-dimensional (3D) substitute for damaged skin. Previous studies have proved a 3D dynamic simulated microgravity (SMG) culture system as a "stimulatory" environment for the proliferation and differentiation of stem cells. Here, we employed the NASA-approved rotary bioreactor to investigate the proliferation and differentiation of human epidermal stem cells (hEpSCs). hEpSCs were isolated from children foreskins and enriched by collecting epidermal stem cell colonies. Cytodex-3 micro-carriers and hEpSCs were co-cultured in the rotary bioreactor and 6-well dish for 15 days. The result showed that hEpSCs cultured in rotary bioreactor exhibited enhanced proliferation and viability surpassing those cultured in static conditions. Additionally, immunostaining analysis confirmed higher percentage of ki67 positive cells in rotary bioreactor compared with the static culture. In contrast, comparing with static culture, cells in the rotary bioreactor displayed a low expression of involucrin at day 10. Histological analysis revealed that cells cultured in rotary bioreactor aggregated on the micro-carriers and formed multilayer 3D epidermis structures. In conclusion, our research suggests that NASA-approved rotary bioreactor can support the proliferation of hEpSCs and provide a strategy to form multilayer epidermis structure.