ObjectiveTo investigate the three-dimensional (3D) culture and morphology of human eccrine sweat gland cells. MethodsThe human eccrine sweat gland cells were isolated from normal abdominal full thickness skin by digestion of type II collagenase, and cultured in defined-keratinocyte serum free medium supplemented with 5 ng/mL recombinant human epidermal growth factor, 25 mg/mL bovine pituitary extract, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37℃ in a humidified atmosphere of 5%CO2/95% air incubator. When the cell fusion reached above 80%, the cells were harvested and the concentration was adjusted to 1×105 cells/mL. The mixture of 0.3 mL cell suspension and 0.3 mL Matrigel basement-membrane matrix was cultured in 12-well plate. The cell growth was observed under an inverted phase contrast microscope. At 14 days after culture, frozen sections were prepared and were stained with HE to observe the cells morphology, and immunohistochemical analysis was used to detect the antigen expressions of cytokeratin 7 (CK7) and CK19. ResultsInverted phase contrast microscope observation showed that many free eccrine sweat gland tissues were seen after digestion of type II collagenase; eccrine sweat gland cells grew adhering to the wall at 3-5 days and continued division for 2-3 weeks to form single ring around the block sweat glands; cellular senescence were observed after 3-4 weeks. During the process of 3D culture, the single eccrine sweat gland cell divided into 2-4 cells after 2-3 days, and these cells subsequently formed small cell clusters, tubular-like structures and finally spheric-like shapes. After cultured for about 2 weeks, there was crack in part of the gelled mixture or liquefaction occurred. HE staining of frozen sections of the 3D cultures showed some of the tubular-like structures composed of 1-2 layers of epithelial cells, which were similar to the secretion part and the duct part of the eccrine sweat gland. Immunohistochemical analysis showed that CK7 and CK19 antigens expressed positively in the cells. ConclusionHuman eccrine sweat gland cells cultured in Matrigel can form the 3D structures which simulate the morphology of eccrine sweat glands in vivo.
ObjectiveTo investigate the application of the double skin paddle arterialized venous flaps for reconstruction of soft tissue defects in the middle and proximal parts of double fingers. MethodBetween September 2011 and December 2014, 6 cases (12 fingers) of soft tissue defects in the middle and proximal parts of double fingers underwent reconstructive surgery with the double skin paddle arterialized venous flaps. There were 5 males and 1 female with an average age of 33.8 years (range, 19-52 years). The causes included cut injury in 4 cases and crush injury in 2 cases. Five index fingers, 3 middle fingers, 2 ring fingers, and 2 little fingers were involved. All defects located at proximal and middle fingers and defect did not exceed the distal interphalangeal joint. The defect area ranged from 2.5 cm×2.5 cm to 5.5 cm×4.0 cm. All cases had bone or tendon exposure, and 2 cases had phalangeal fracture. The disease duration was 1.5-7 hours (mean, 3.5 hours). The flap size was 8 cm×3 cm-14 cm×5 cm. The donor site was directly sutured (≤ 3.0 cm in width) or was repaired with skin graft (>3.0 cm in width). ResultsThe operation time was 2.5-5.0 hours (mean, 4.0 hours). All flaps survived completely. Tensive blisters occurred in 4 cases and were improved at 1 week after removal of suture around pedicle. Partial distal flap necrosis was noted in 1 case, healing was obtained after secondary debridement; other wounds healed in one stage. The patients were followed up 6-18 months (mean, 13 months). The flap had good texture, elasticity, and appearance. According to the hand function evaluation criteria issued by the Chinese Hand Society, the results were excellent in 3 cases, good in 2 cases, and fair in 1 case at last follow-up. The two-point discrimination of the flap was 8-10 mm (mean, 9 mm). ConclusionsThe double skin paddle arterialized venous flaps have the advantages of simple technique and definite effectiveness for reconstruction of soft tissue defects in the middle and proximal part of double fingers.
ObjectiveTo systematically review the correlation between acute gastrointestinal infection and IBS. MethodsLiterature search was performed in The Cochrane Library (Issue 8, 2013), PubMed, EMbase, Web of Science, CBM, CNKI, VIP and WanFang Data to collect the prospective cohort studies about association between acute gastrointestinal infection and IBS, from inception to August 2013. Two reviewers independently screened the literature according to the inclusion and exclusion criteria, extracted the data, and assessed the quality using NOS, and then Meta analysis was conducted using RevMan 5.2 software. ResultsA total of 11 cohort studies involving 6 274 patients were included. According to the different follow-up times for subgroup analysis, the results of meta-analysis showed that, compared with the healthy volunteers who did not expose the acute gastrointestinal infection, the patients with acute gastroenteritis had a increase risk of irritable bowel syndrome within 3 months, 6 months, 12 months, and 2-3 years (3 months: RR=6.46, 95%CI 1.85 to 22.58, P=0.003; 6 months: RR=4.68, 95%CI 2.07 to 10.60, P=0.000 2; 12 months: RR=4.95, 95%CI 2.90 to 8.45, P < 0.000 01; 2-3 years: RR=3.11, 95%CI 2.72 to 3.56, P < 0.000 01). However, after the fifth year of acute gastroenteritis, there was no statistical significance in the risk of irritable bowel syndrome between the two groups (RR=1.69, 95%CI 0.68 to 4.24, P=0.26). ConclusionAcute gastrointestinal infection within 3 years after onset was associated with the risk of IBS. Sex, diarrhea duration, bloody purulent stools and abdominal cramps at acute stage are important risk factors of intriguing the occurrence of post-infectious IBS. The acute gastrointestinal infection and IBS are not associated in the fifth year; however, more high-quality trials are needed for further verifying the aforementioned conclusion.
Urokinase plasminogen activator receptor (uPAR) is a membrane protein which is attached to the cellular external membrane. The uPAR expression can be observed both in tumor cells and in tumor-associated stromal cells. Thus, in the present study, the human amino-terminal fragment (hATF), as a targeting element to uPAR, is used to conjugate to the surface of superparamagnetic iron nanoparticle (SPIO). Flowcytometry was used to examine the uPAR expression in different tumor cell lines. The specificity of hATF-SPIO was verified by Prussian blue stain and cell phantom test. The imaging properties of hATF-SPIO were confirmed in vivo magnetic resonance imaging (MRI) of uPAR-elevated colon tumor. Finally, the distribution of hATF-SPIO in tumor tissue was confirmed by pathological staining. Results showed that the three cells in which we screened, presented different expression characteristics, i.e., Hela cells strongly expressed uPAR, HT29 cells moderately expressed uPAR, but Lovo cells didn't express uPAR. In vitro, after incubating with Hela cells, hATF-SPIO could specifically combined to and be subsequently internalized by uPAR positive cells, which could be observed via Prussian blue staining. Meanwhile T2WI signal intensity of Hela cells, after incubation with targeted probe, significantly decreased, and otherwise no obvious changes in Lovo cells both by Prussian blue staining and MRI scans. In vivo, hATF-SPIO could be systematically delivered to HT29 xenograft and accumulated in the tumor tissue which was confirmed by Prussian Blue stain compared to Lovo xenografts. Twenty-four hours after injection of targeting probe, the signal intensity of HT29 xenografts was lower than Lovo ones which was statistically significant. This targeting nanoparticles enabled not only in vitro specifically combining to uPAR positive cells but also in vivo imaging of uPAR moderately elevated colon cancer lesions.