BACKGROUND: In previous studies on tissue-engineered adipose, the pore size of scaffolds is easily neglected. If the pore size was large, cells would flow away along the pore and could not maintain in the scaffold. If the pore size was small, cells would distribute on the surface of the scaffold, and could not enter the scaffold. Simultaneously, it would be not contribute to growth of new vessels. OBJECTIVE: To screen appropriate silk fibroin porous scaffolds to construct tissue-engineered adipose. METHODS: Under unchanged concentration of flbroin, six kinds of silk fibroin porous scaffolds in different aperture were prepared by changing freezing and drying temperature and time. Human umbilical cord mesenchymal stem cells (hUCMSCs) were harvested using attachment method. The potentiality of its osteogenic and adipogenic differentiation was detected by chemistry staining. The apertures of six kinds of silk fibroin porous scaffolds were measured by scanning electron microscope. The adhesion and proliferation of hUCMSCs on silk fibroin porous scaffolds in different aperture were observed. RESULTS AND CONCLUSION: The apertures of six silk fibroin porous scaffolds were (39.94±17.27), (53.51±16.18), (63.97±19.76), (71.08±18.07), (87.33±21.78), (121.97±44.10) μm, respectively. The strong ability of attachment of hUCMSCs appeared in the number two scaffold, and in other materials (number 1,3,4,5,6), there were not any cell in them except number one and three. Therefore, 50 μm might be a suitable aperture to construct tissue-engineered adipose with silk flbroin porous scaffolds using hUCMSCs.

To explore the role of dentritic epidermal T lymphocytes ( DETCs) in immune rejection of skin allograft in mice and its related mechanism. Methods (1) Full-thickness skin was harvested from back of one male wild type (WT) C57BL/6 mouse. Epithelial cells were isolated for detection of the expression of DETCs and their phenotype with flow cytometer. Another male WT C57BL/6 mouse was used to harvest full-thickness skin from the back. Epidermis was isolated for observation of the morphological characteristics of DETCs with immunofluorescence technology. (2) Four male green fluorescence protein (GFP)-marked C57BL/6 mice, 7 female WT C57BL/6 mice (group WT), and 7 female ybT lymphocytes 8 gene knock-out (GK) C57BL/6 mice (group GK) were used. Full-thickness skin in the size of 1.4 cm x 1.4 cm on the back of mice in groups WT and GK were excised, and the wounds were transplanted with full-thickness skin in the size of 1.2 cm x 1.2 cm obtained from male GFP-marked C57BL/6 mice. The survival time of skin grafts was affirmed with small animal in vivo imager and naked eyes and recorded. (3) Two male WT C57BL/6 mice were used to isolate epithelial cells. Cells were inoculated into 48-well plate and divided into activation group (A) and control group (C) according to the random number table, with 4 wells in each group. Cells in group A were treated with 10 pL concanavalin A in the concentration of 2 microg/mL for 24 hours, while those in group C with PBS in the same volume as that in group A. The expression of interferon y in DETCs was detected with flow cytometer. (4) Four male GFP-marked C57BL/6 mice were used as donors. Fourteen female WT C57BL/6 mice were used as receptors and divided into interferon gamma neutralizing group (IN) and control group (C) according to the random number table, with 7 mice in each group. The skin transplantation model of C57BL/6 male to C57BL/6 female was established as in part (2). Before surgery and 72 hours after, mice in group IN were intraperitoneally injected with 200 pL interferon y neutralizing antibody in the concentration of 1 mg/mL, and those in group C with normal saline in the same volume as that in group IN. The survival time of skin grafts was observed and recorded using the methods in part (2), and the result of group IN was compared with that of group GK in part (2). The survival curve of skin grafts was processed with Log-rank ( Mantel-Cox) test. Results (1) The positive expression rate of DETCs in epithelial cells of skin in mouse was 7.27%, and they were all CD3 cells. DETCs were found to be scattered in the epidermis of skin in mouse with dendritic morphology. (2) The survival time of skin grafts of mice in group GK was 22-35 d, obviously longer than that in group WT (12-16 d, y2 = 14. 10 , P < 0.001). (3) Expression of interferon gamma was detected in 22. 70% DETCs in group A, which was obviously higher than that in group C (0.51%). (4) The survival time of skin grafts of mice in group IN was 19-24 d, which was obviously longer than that in group C (12-16 d, chi 2 = 13.60, P < 0.001) but close to that in group GK as in part (2) (chi2 = 0.06, P = 0.810). Conclusions DETCs are involved in promotion of immune rejection of skin allograft probably by secretinf interferon gamma.
Allografts ; Animals ; Epidermis ; Female ; Graft Survival ; immunology ; physiology ; Interferon-gamma ; immunology ; metabolism ; Lymphocytes ; Male ; Mice ; Mice, Inbred C57BL ; Skin ; Skin Transplantation ; T-Lymphocytes ; immunology

Objective:To evaluate the effects of Xiongzhi Dilong decoction(XZDLD)and its wind medicine on calcitonin gene-related peptide(CGRP)-induced migraine and explore the mechanism through the CGRP/inducible nitric oxide synthase(iNOS)pathway.Methods:Rats were divided into control,model,XZDLD,XZDLD(external wind),XZDLD(internal wind),and olcegepant groups.CGRP was injected into the dura mater to induce a migraine.The frequency of head scratching,cage climbing,and facial grooming was observed.The pain threshold,the levels of CGRP,pituitary adenylate cyclase activating polypeptide(PACAP),substance P(SP),and the plasma protein extravasation(PPE)ratio were measured.The phosphorylation levels of p38,extracellular signal-regulated kinase 1/2(ERK1/2),and expression of iNOS were detected by western blot.Results:Compared with the model group,the three modified XZDLD groups showed reduced frequency of head scratching and cage climbing in the first 30 min(all P＜.05).Facial grooming frequency was reduced in XZDLD and XZDLD(external wind)groups(P=.0003 and P=.0131,respectively).External wind medicine played a more important role in increasing mechanical pain threshold than internal wind medicine.Moreover,compared with the model group,the three modified XZDLD groups demonstrated reduced plasma levels of CGRP and PACAP(all P＜.05).No difference in the SP level was observed among the six groups.XZDLD reduced PPE ratio.XZDLD and XZDLD(external wind)groups suppressed the CGRP/iNOS pathway by inhibiting the p-p38/p38 ratio and the expression of iNOS.No difference in pERK1/2/ERK1/2 ratio was detected among the six groups.Conclusion:XZDLD increases pain threshold,downregulates the expression of CGRP and PACAP,and reduces PPE ratio by inhibiting the CGRP/iNOS pathway.External wind medicine is more effective than internal one on improving facial grooming and head scratching,increasing the mechanical pain threshold,and inhibiting the expression of iNOS.

Objective By analyzing the perioperative management in our hospital to explore the clinical effect and safety of single kidney transplantation from deceased juveniles' donors.Methods We retrospectively analyze 86 cases of kidney transplantations from deceased juveniles' donors in our hospital from 2007 December to 2015 August.Results The success rate of the operations was 100％.The postoperative complications occurred as fellows:7 cases of acute rejection (8.14％);10 cases of drug intoxication (11.62％);21 cases of DGF (24.44％),4 cases of leakage of urine (4.65％),7 cases of lung infection (8.14％).Two cases (2.32％) died after the operation because of serious lung infection,and by corresponding treatment 47 cases recovered after 2-4 weeks.The creatinine level in 37 cases without any complications was 131.88 ± 44.20 μmol/L during discharge.Conclusion With strict selection,the organ from a deceased juvenile donor is safe and practicable.

Migraine is a common neurovascular disorder manifested by recurrent severe headaches on one or both sides， occasionally accompanied by nausea， vomiting， photophobia， and phonophobia. It has the characteristics of recurrent attacks and family inheritance. Traditional Chinese medicine （TCM） believes that migraine belongs to the category of "head wind"， which is mostly caused by external wind and is related to the internal stirring of liver wind. Sanpiantang comes from the Record of Syndorme Differentiation·Headache （Bianzhenglu·Toutongmen） created by the physician CHEN Shiduo of the Qing Dynasty. It is composed of Chuanxiong Rhizoma， Angelicae Dahuricae Radix， Pruni Semen， Cyperi Rhizoma， Bupleuri Radix， White Mustard Seed， and Glycyrrhizae Radix et Rhizoma， with the functions of moving Qi to release pain， activating blood and resolving stasis， which is commonly used for the treatment of migraine in clinic. Current clinical studies on the application of Sanpiantang to the treatment of migraine mostly used modified Sanpiantang， either alone or in combination with western medicine/acupuncture. The results of these clinical trials showed that Sanpiantang could significantly lower migraine score， pain visual analog scale and endothelin level， reduce the frequency of painkiller use， and remarkably alleviate migraine symptoms， with few side effects. The animal experiments focused on exploring the mechanism of action of modified Sanpiantang from different anatomical levels of migraine， which mainly included reducing nitric oxide （NO） and nitric oxide synthase （NOS）， reduceing the release of neurotransmitters such as 5 -hydroxyline (5-HT) and neurotipides (NPY)， suppressing neuronal excitation， and blocking the transmission of nociceptive pathways， thereby promoting cerebral blood flow， regulating neurotransmitters and preventing migraine. Based on the pathogenesis of migraine， this paper systematically reviewed the latest progress in clinical application and experimental research of modified Sanpiantang， and summarized its mechanism of action of preventing and treating migraine， which provided new ideas for clinical treatment of migraine.