0.05),there were more apoptosis-starting cells found in normal skin(1738.33?348.89/mm2)than in keloid(891.67?395.00/mm2)(t=-5.565,P=0.000).The number of positive stained fibroblast cells in dermis significantly increased in keloid group(911.67?323.61/mm2),compared with that in normal skin group(220.00?80.00/mm2)(t=7.188,P=0.000).This result that the protein expression level of Bid was higher in keloid(0.46?0.08)than that in normal skin tissue(0.02?0.01)(t=18.905,P=0.000)was proven by western blotting test.Conclusions The number of apoptosis-starting cells in epidermis of normal skin is more than that in keloid.Bid protein in dermis of keloid is over expressed.

Objective To investigate the isolation, purification and culture methods of human embryonic germ cells (EGCs) on feeder layer cells of human embryonic fibroblasts. Methods Primordial germ cells(PGCs) from the genital ridges of 6-11 weeks(post fertilization) human embryos were cultrued on feeder layer cells of human embryonic fibroblasts(HEFs) which were isolated from the 3-4 month embryos and routinely cultured for over 25 passages. The medium composed of growth factors and differentiation inhibitory factors. The cultures were analyzed with the expression of alkaline phosphatase, immunologic markers (SSEA-1,SSEA-4) and the transcription factor Oct-4 that have been used to routinely to characterize EGCS. Results A large-scale EG cells were obtained and meintained on feeder layers for over 8 passages. The cell surface marker showed that the EGCs possess high levels of AP activity. EGCs colonies stained strongly for SSEA-4,SSEA-1 and they expressed the transcription factor Oct-4.Conclusion EGCs have potential to maintain long term proliferation and undifferentiation state on human embryonic fibroblasts in vitro.;

Objective To evaluate the clinical safety and efficacy of focused ultrasound body contouring apparatus in abdominal fat reduction and body contouring.Methods Forty healthy subjeets were equally randomized into treatment group and control group,and received two treatments with the JCS-01 device (3H Medical Technology Co,Ltd,Beijing,China) in the abdominal region at one-week interval.During each treatment,the subject was placed on the treatment bed and the region of interest was spread with an acoustic coupling medium.Then the ultrasonic transducer was fixed,and the operator controlled the computerized system to move the bed in a designed regon.The control group would not receive any energy therapy during treatment time.Subjects were followed for 28 days after the last treatment (day 35).Abdominal circumference,regional photos and safety parameters were recorded at the time instantly before and after treatment and days 14 and 35.Subject satisfaction survey was conducted at day 35.Results One subject in control group was loss of follow-up.No local skin reactions,such as erythema edema or papules,and no changes with clinical significance in laboratory examinations occurred in all other 39 subjects.The proportion of subjects with over 0.5 cm abdominal circumference reduction in the treatment group was significantly higher than the control group at all time points (P＜0.05).Peak abdominal circumference reduction was on day 35.The abdominal circumferences of 19/20 subjects in the treatment group were reduced by 0.83-6.33 cm,and the mean abdominal circumference was reduced by 3.09 cm on day 35.The total effective rate was 95 ％,and the subject satisfactory rate was 75 ％.Conclusions The focused ultrasound treatments for body contouring is safe and tolerable and also significantly reduces abdominal circumference.

Objective To explore a more efficient surgical method to cure axillary osmidrosis through a comparative study on results from two surgical treatments of axillary osmidrosis in the armpit area.Methods 72 patients were randomized into two groups,in which 36 patients (group A) with micro-incision simple liposuction and 36 patients (group B) with dissection and excision of subcutaneous tissue from axillary folds.The clinical outcomes in the two groups were comparatively analyzed,based on surgery time,effect,complications,scarring and recovery time.Results The total efficiency of axillary osmidrosis treatment was 100％ after 6-12 months clinical follow-up towards two groups.There was no significantly statistical difference on the effect of both groups.Micro-incision simple liposuction was obviously superior to dissection and excision of subcutaneous tissue from axillary folds based on surgery time,complications,recovery time,scarring and armpit hair survival rate.The two groups were significantly different (P＜0.05).Conclusions Minimally invasive simple liposuction,as a treatment for axillary osmidrosis is a method with more effectiveness and reliability.It is a more acceptable approach to patients in view of safety and convenience,smaller and invisible scarring,fewer complications,and less recovery time.

OBJECTIVETo observe the effects of conditioned medium from keloid fibroblasts under hypoxia on angiogenesis, and to investigate the role of hypoxic microenvironment in invasive growth of keloid.

METHODSPrimary keloid fibroblasts and human umbilical endothelial cells (HUVEC) were cultured as conventional method. Keloid fibroblasts were cultured either in a hypoxic incubator (2% O2) for 48 h or in a normoxic incubator (20% O2) as control. Then those cell culture mediums were collected and mixed with endothelial cell medium by the proportion of 1:1 as conditioned medium. The mRNA and secreted protein of pro-angiogenic factors such as vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1) and periostin of keloid fibroblasts under hypoxia were detected by real time PCR and ELISA. The proliferation, migration and invasion, tube formation of HUVEC cultured with conditioned medium were evaluated by CCK-8 assay, Transwell assay and matrigel tube formation assay, respectively.

RESULTSHypoxia increased the expression of VEGF, Ang-1 and periostin in both mRNA (increased by 75%, 43% and 118% respectively, P < 0.05) and secreted protein (increased by 30.2%, 14.2% and 19.5% respectively, P < 0.05) levels; the proliferations of HUVEC in hypoxic conditioned medium in 1, 2 and 3 d were 0.67 +/- 0.07, 0.84 +/- 0.09 and 1.08 +/- 0.10 respectively, which were higher compared to those in control group (0.52 +/- 0.08, 0.72 +/- 0.10 and 0.91 + 0.14, P < 0.05); the numbers of migration, invasion and tube formation of HUVEC were (73.2 +/- 8.9), (56.3 +/- 12.5), (9.66 +/- 1.96) cells/HP, which were higher compared to those in control group [(59.0 +/- 8.0), 35.5 +/- 8.5), (6.5 +/- 1.87) cells/HP, P < 0.05].

CONCLUSIONSHypoxia increases the expression of pro-angiogenic factors of keloid fibroblasts, and its conditioned medium under hypoxia could promote angiogenesis. The results suggest hypoxic microenvironment may play a significant role in the invasive growth of keloid by inducing angiogenesis.

Objective To investigate effective approach to decrease portal venous hypertension and high perfusion of portal vein caused by small-for-size (SFS) liver graft transplantation with the aim of improving hepatocellular microcirculation.Methods Rat models with SFS liver graft (n =62) were well estab lished and divided into SFS group and trans-portal intrabepatic portosystemic shunt (TPIPSS) group.Hemodynamic parameters,histopathologically morphologic changes,postoperative complications,accumulated survival rate were recorded and analyzed.Venous filling time after liver reperfusion,hemodynamic parameters were evaluated using t test and Kruskal-Wallis test.Kaplan-Meier method was performed for survival analysis.Results Venous filling time after liver reperfusion was remarkably prolonged with the application of multihole cone-shaped tubes.Compared with SFS group,the filling time was 4-second longer in TPIPSS.At each endpoints of reperfusion within 90 mins,the portal vein pressures were lowered in the TPIPSS group than those of SFS group.Liver grafts were present with more regular structures in TPIPSS group,with no sign of hepatic sinusoid congestion or irregular clearance extension.In the aspect of postoperative complications,all the rat receivers showed ascites in the SFS group.Nevertheless,there was no ascites observed in TPIPSS rats,and 50％ rats (5/10) experienced clinical manifestations of hepatic encephalopathy.Persistent fever over 7 days was showed in 10％ rats (1/10) of SFS group and 40％ rats (4/10) of TPIPSS group,respectively.The mean survival was superior in TPIPSS group (37.2 ± 23.5) d than SFS group (17.7 ± 13.5) d,P ＜ 0.05.Conclusion TPIPSS could be a safe and feasible approach to improve portal venous hypertension caused by SFS liver graft and hepatocellular reperfusion.

Exosome is a double-layer membrane vesicle secreted by several kinds of cells. Exosome is major way of cell paracrine activity with a wide range of sources and high security feature. This paper reviews recent progress of exosomes in promoting wound healing. Studies have shown that exosomes can be applied to wound treatment by immune regulation, regulating fibroblast activity, promoting angiogenesis, inhibiting scar formation and other mechanisms. Compared with traditional stem cell therapy, exosomes have better applied prospect in clinical practise.

Background/Aims: Abdominal bloating or distension (AB/D) is a common complaint in the outpatient of gastroenterology department. Since the potential contributors are numerous and complex, a longitudinal study on the disease spectrum and natural history of patients was performed to better understand the key factors of AB/D. Methods: Consecutive patients with the chief complaint of AB/D referred to the outpatient clinic were screened. Functional gastrointestinal disorders (FGIDs) were diagnosed according to Rome IV criteria. A 3-year follow-up was performed to seek for the changes in symptoms as well as disease spectrum. Results: A total of 261 participants were enrolled and 139 completed the follow-up. Most patients suffered from moderate to severe symptoms more than 1 day per week. Common causes of AB/D were FGIDs (51.7%) and organic diseases (17.2%). The latter group was older with lower body mass index (BMI). Functional dyspepsia was the most common type of FGIDs in AB/D. The symptoms of 18.0% of participants failed to improve at the end of the 3-year follow-up, and those diagnosed with FGIDs were most likely to continue to suffer. Abdominal pain was a positive predictive factor for good prognosis in the FGIDs group. Besides, only 22.7% of participants had a consistent diagnosis of FGIDs during follow-up. Conclusions FGIDs are the most common diagnosis in patients with AB/D. Symptoms were especially hard to be improved. Classification diagnoses of FGIDs in AB/D patients fluctuated significantly over time.

Objective To study the effects and mechanism of immediate application of transformation growth factor beta 1 (TGF-β1) neutralizing antibody postoperatively to prevent the fibrous capsule formation in animal model,and to observe the different effects between two concentrations of TGF-β1 neutralizing antibody.Methods Ten ml of smooth round silicone prostheses were implanted subcutaneously in the back of rats.Then 25 μg/0.1 ml or 50 μg/0.1 ml TGF-β1 neutralizing antibody were injected around the implanted prostheses in the experimental group,while the control group was injected with PBS 0.1 ml.On day 7,14 and 28,the thickness of capsule measured;The density of collagen;and the expression of TGF-β1,alpha smooth muscle actin (α-SMA),and fibronectin (FN) was detected.Results After 28 days,the average capsule thickness of the control group was significantly higher in the experimental group,and the difference between the control group and the 25 μg experimental group had statistical significance.TGF-β1 expression and the number of α-SMA positive fibroblasts in control group were significantly higher than that in the experimental group,the difference was statistically significant.In the control group,the expression of FN and collagen density was slightly higher than the two experimental groups,but the differences among the three groups showed no statistical significance at each time point.Conclusions Immediate application of the TGF beta 1 neutralizing antibody in the the implant prosthesis can reduce the thickness of the capsule to some extent.

Objective: To understand the effect of sirolimus on the erythropoiesis of K562 cell line and bone marrow cells from pure red cell aplasia (PRCA) patients and normal controls. Methods: Different concentrations (10, 100, 1 000 nmol/L) of sirolimus were added to the K562 cell line or bone marrow cells from PRCA patients or normal controls and cultured 14 days for BFU-E formation. Meanwhile, sirolimus was also added to the serum treated PRCA bone marrow cells to cultivate for the same priod of time. Results: Neither K562 cells, bone marrow cells from PRCA patients or normal controls showed any difference when sirolimus was added to the culture system for BFU-E. However, BFU-E formation decreased after serum was added in PRCA patients (76.40±22.48 vs 136.33±12.58, t=-4.329, P=0.001) and this suppression of BFU-E was partly corrected by 1 000 nmol/L sirolimus treatment (97.14±15.83 vs 76.40±22.48, P=0.038). Conclusions Sirolimus may modulate the suppression of erythropoiesis by serum instead of directly stimulate the growth of red blood cells in PRCA patients.