[Effects of transient receptor potential vanilloid type 4-specific activator on human vascular endothelial cell functions and blood supply of rat perforator flap and its mechanism].

Objective: To analyze the effects of transient receptor potential vanilloid type 4 (TRPV4) activation on the function and endothelial-to-mesenchymal transition (EndMT) of human umbilical vein endothelial cells (HUVECs), as well as to explore the effects of TRPV4 activation on blood perfusion and survival of rat perforator flap and the mechanism. Methods: The experimental research methods were used. The 3rd to 6th passages of HUVECs were used for experiments and divided into 0.5 µmol/L 4α-phorbol 12, 13-didecanoate (4αPDD) group, 1.0 µmol/L 4αPDD group, 3.0 µmol/L 4αPDD group, 10.0 µmol/L 4αPDD group, and phosphate buffer solution (PBS) group, which were cultivated in corresponding final molarity of 4αPDD and PBS, respectively. The cell proliferation activity at 6 and 12 h of culture was detected using cell counting kit-8 (CCK-8). Another batch of cells was acquired and divided into PBS group, 1 µmol/L 4αPDD group, and 3 µmol/L 4αPDD group, which were treated similarly as described before and then detected for cell proliferation activity at 6, 12, 24, and 48 h of culture. The residual scratch area of cells at post scratch hour (PSH) 12, 24, and 48 was detected by scratch test, and the percentage of the residual scratch area was calculated. The number of migrated cells at 24 and 48 h of culture was detected by Transwell experiment. The tube-formation assay was used to measure the number of tubular structures at 4 and 8 h of culture. The protein expressions of E-cadherin, N-cadherin, Slug, and Snail at 24 h of culture were detected by Western blotting. All the sample numbers in each group at each time point in vitro experiments were 3. A total of 36 male Sprague-Dawley rats aged 8 to 10 weeks were divided into delayed flap group, 4αPDD group, and normal saline group according to the random number table, with 12 rats in each group, and iliolumbar artery perforator flap models on the back were constructed. The flap surgical delay procedure was only performed in the rats in delayed flap group one week before the flap transfer surgery. Neither rats in 4αPDD group nor normal saline group had flap surgical delay; instead, they were intraperitoneally injected with 4αPDD and an equivalent mass of normal saline, respectively, at 10 min before, 24 h after, and 48 h after the surgery. The general state of flap was observed on post surgery day (PSD) 0 (immediately), 1, 4, and 7. The flap survival rates were assessed on PSD 7. The flap blood perfusion was detected by laser speckle contrast imaging technique on PSD 1, 4, and 7. The microvascular density in the flap's choke vessel zone was detected by immunohistochemical staining. All the sample numbers in each group at each time point in vivo experiments were 12. Data were statistically analyzed with analysis of variance for factorial design, analysis of variance for repeated measurement, one-way analysis of variance, least significant difference t test, and Bonferroni correction. Results: At 6 and 12 h of culture, there were no statistically significant differences in cell proliferation activity in the overall comparison among PBS group, 0.5 µmol/L 4αPDD group, 1.0 µmol/L 4αPDD group, 3.0 µmol/L 4αPDD group, and 10.0 µmol/L 4αPDD group (P>0.05). At 6, 12, 24, and 48 h of culture, there were no statistically significant differences in cell proliferation activity in the overall comparison among PBS group, 1 µmol/L 4αPDD group, and 3 µmol/L 4αPDD group (P>0.05). At PSH 12, the percentages of the residual scratch area of cells in 1 µmol/L 4αPDD group and 3 µmol/L 4αPDD group were close to that in PBS group (P>0.05). At PSH 24 and 48, compared with those in PBS group, the percentages of the residual scratch area of cells in 3 µmol/L 4αPDD group were significantly decreased (with t values of 2.83 and 2.79, respectively, P<0.05), while the percentages of the residual scratch area of cells in 1 µmol/L 4αPDD group showed no significant differences (P>0.05). At 24 h of culture, the number of migrated cells in 1 µmol/L 4αPDD group and 3 µmol/L 4αPDD group were close to that in PBS group (P>0.05). At 48 h of culture, the number of migrated cells in 1 µmol/L 4αPDD group and 3 µmol/L 4αPDD groups were significantly greater than that in PBS group (with t values of 6.20 and 9.59, respectively, P<0.01). At 4 h of culture, the numbers of tubular structures of cells in 1 µmol/L 4αPDD group and 3 µmol/L 4αPDD group were significantly greater than that in PBS group (with t values of 4.68 and 4.95, respectively, P<0.05 or <0.01). At 8 h of culture, the numbers of tubular structures of cells in 1 µmol/L 4αPDD and 3 µmol/L 4αPDD groups were similar to that in PBS group (P>0.05). At 24 h of culture, compared with those in PBS group, the protein expression level of E-cadherin of cells in 3 µmol/L 4αPDD group was significantly decreased (t=5.13, P<0.01), whereas there was no statistically significant difference in the protein expression level of E-cadherin of cells in 1 µmol/L 4αPDD group (P>0.05); the protein expression level of N-cadherin of cells in 3 µmol/L 4αPDD group was significantly increased (t=4.93, P<0.01), whereas there was no statistically significant difference in the protein expression level of N-cadherin of cells in 1 µmol/L 4αPDD group (P>0.05); the protein expression levels of Slug of cells in 1 µmol/L 4αPDD group and 3 µmol/L 4αPDD group were significantly increased (with t values of 3.85 and 6.52, respectively, P<0.05 or P<0.01); and the protein expression level of Snail of cells in 3 µmol/L 4αPDD group was significantly increased (t=4.08, P<0.05), whereas there was no statistically significant difference in the protein expression level of Snail of cells in 1 µmol/L 4αPDD group (P>0.05). There were no statistically significant differences in the protein expression levels of E-cadherin, N-cadherin, Slug, or Snail of cells between 1 µmol/L 4αPDD group and 3 µmol/L 4αPDD group (P>0.05). The general condition of flaps of rats in the three groups was good on PSD 0. On PSD 1, the flaps of rats in the three groups were basically similar, with bruising and swelling at the distal end. On PSD 4, the swelling of flaps of rats in the three groups subsided, and the distal end turned dark brown and necrosis occurred, with the area of necrosis in flaps of rats in normal saline group being larger than the areas in 4αPDD group and delayed flap group. On PSD 7, the necrotic areas of flaps of rats in the 3 groups were fairly stable, with the area of necrosis at the distal end of flap of rats in delayed flap group being the smallest. On PSD 7, the flap survival rates of rats in 4αPDD group ((80±13)%) and delayed flap group ((87±9)%) were similar (P>0.05), and both were significantly higher than (70±11)% in normal saline group (with t values of 2.24 and 3.65, respectively, P<0.05 or P<0.01). On PSD 1, the overall blood perfusion signals of rats in the 3 groups were basically the same, and the blood perfusion signals in the choke vessel zone were relatively strong, with a certain degree of underperfusion at the distal end. On PSD 4, the boundary between the surviving and necrotic areas of flaps of rats in the 3 groups became evident, and the blood perfusion signals in the choke vessel zone were improved, with the normal saline group's distal hypoperfused area of flap being larger than the areas in delayed flap group and 4αPDD group. On PSD 7, the blood perfusion signals of overall flap of rats had generally stabilized in the 3 groups, with the intensity of blood perfusion signal in the choke vessel zone and overall flap of rats in delayed flap group and 4αPDD group being significantly greater than that in normal saline group. On PSD 7, the microvascular density in the choke vessel zone of flap of rats in 4αPDD group and delayed flap group were similar (P>0.05), and both were significantly higher than that in normal saline group (with t values of 4.11 and 5.38, respectively, P<0.01). Conclusions: After activation, TRPV4 may promote the migration and tubular formation of human vascular endothelial cells via the EndMT pathway, leading to the enhanced blood perfusion of perforator flap and microvascular density in the choke vessel zone, and therefore increase the flap survival rate.

[Predictive value of magnetic resonance imaging characteristics before and after radiotherapy for the occurrence of severe radiation-induced late rectal injury in patients with rectal cancer].

Objective: Severe radiation-induced late rectal injury (sRLRI) directly affects the quality of life of patients with rectal cancer. Effective prediction of sRLRI before surgery may provide important information for the selection of surgical strategies and perioperative managements. The purpose of this study is to evaluate the feasibility of predicting sRLRI based on magnetic resonance imaging (MRI) features before and after radiotherapy for rectal cancer. Methods: This was a diagnostic study. Clinical and imaging data of 90 patients with rectal cancer receiving long-term radiotherapy from June 2013 to July 2018 in the Sixth Affiliated Hospital of Sun Yat-sen University were collected retrospectively. Case inclusion criteria: (1) rectal cancer was diagnosed by pathology and age of ≥ 18 years old; (2) patients received neoadjuvant chemoradiotherapy and anterior rectal resection; (3) follow up time ≥ 3 years; (4) patients had no history of other neoplasm. Exclusion criteria: (1) patients did not receive MRI examination in our hospital within 2 weeks before and/or 8 weeks after radiotherapy; (2) images were not good enough for evaluation; (3) medical records were incomplete; (4) patients had severe gastrointestinal diseases. According to the RTOG/EORTC classification criteria for radiation reactions, severe complications of grade 3-4 requiring surgical management were defined as sRLRI. T2WI and DWI images before and after radiotherapy were evaluated. The rectal wall thickness, bladder wall thickness, rectal sacral spacing and apparent diffusion coefficient (ADC) were measured. The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of the above indicators for sRLRI. Results: Among the 90 patients with rectal cancer, 34 (37.8%) developed sRLRI. Before radiotherapy, the median rectal wall thickness of sRLRI and non-sRLRI patients was 4.530 mm and 4.355 mm, respectively; the median bladder wall thickness was 3.962 mm and 3.868 mm, respectively; the median rectal sacral spacing was 15.557 mm and 12.433 mm, respectively; the median ADC value of rectal wall was 1.620 ×10(-3) mm(2)/s and 1.653 ×10(-3) mm(2)/s, respectively. There were no significant differences in above indicators between sRLRI and non-sRLRI patients (all P>0.05). After radiotherapy, compared with non-sRLRI patients, sRLRI patients had increased rectal wall thickness (median: 8.239 mm vs. 6.223 mm, Z=-3.512, P=0.001), rectal sacral spacing (median: 17.728 mm vs. 13.885 mm, Z=-2.247, P=0.025), and change of rectal wall thickness after radiotherapy (median: 98.106% vs. 49.584%, Z=-4.169, P<0.001). After radiotherapy, there were no significant differences in the bladder wall thickness and its change value, the ADC value of rectal wall and its change rate before and after radiotherapy between the two groups (all P>0.05). The area under the curve (AUC) of the change rates of rectal wall thickness after radiotherapy, rectal wall thickness and rectal sacral spacing after radiotherapy for predicting sRLRI was 0.763, 0.722 and 0.642, respectively, while the sensitivity was 85.3%, 70.6% and 76.5%, respectively, and the specificity was 64.3%, 71.4% and 57.1%, respectively. Conclusion: Based on MRI examinations, assessments of rectal wall thickness after radiotherapy, the change rate of rectal wall thickness after radiotherapy, and rectal sacral spacing after radiotherapy are helpful for evaluating the risk of sRLRI after radiotherapy for patients with rectal cancer.

[Risk factors of blood loss during liver transplantation in children with biliary atresia and its influence on prognosis].

Objectives: To study the risk factors for massive intraoperative blood loss in children with biliary atresia who underwent liver transplantation for the first time,and to analyze their impacts on graft survival,hospital stay and postoperative complications. Methods: The data of 613 children with biliary atresia who underwent liver transplantation at Department of Pediatric Organ Transplantation,Tianjin First Central Hospital from January 2015 to December 2018 were collected and analyzed. There were 270 males and 343 females, aged 7.4 (3.9) months (range: 3.2 to 148.4 months), the body weight of the recipients were (7.8±3.5) kg (range: 4.0 to 43.3 kg).According to the 85th quad of estimated blood loss(EBL),they were divided into two groups:massive EBL group(96 cases) and non massive EBL group(517 cases). The age,height,weight and other factors between the two groups were analyzed and compared. Univariate Logistic regression and multiple stepwise regression were used to determine the risk factors of massive EBL. Then,the postoperative complications of the two groups,including portal vein thrombosis and portal vein anastomotic stenosis etc.,were analyzed and compared by chi square test. Kaplan Meier curve and log rank test were used to analyze the recipient and graft survival rate of the two groups. Results: During the study period,713 transplants were performed and 613 patients were enrolled in the study. Ninety-six patients(15.7%) had massive EBL,and the postoperative hospital stay was 21(16) days(range:2 to 116 days),the hospital stay of non-massive EBL group was 22(12)days(range:3 to 138 days)(U=24 224.0,P=0.32). Univariate Logistic regression analysis showed that the recipient's weight,Kasai portoenterostomy,platelet count,operation time and cold ischemia time were the risk factors of massive EBL during biliary atresia transplantation. Multiple regression analysis showed that cold ischemia time ≥10 hours,prolonged operation time(≥8 hours) and body weight<5.5 kg were important independent risk factors for massive EBL.The incidence of portal vein thrombosis,hepatic vein stenosis,intestinal leakage and pulmonary infection in patients with massive EBL were significantly higher than those without massive EBL(3.1% vs. 0.8%,9.4% vs. 2.1%,6.3% vs. 0.8%,30.2% vs. 20.1%,all P<0.05). The 3-year overall graft and recipient survival rate were significantly lower in patients with massive EBL than those without massive EBL(87.5% vs. 95.7%,P=0.001;84.4% vs. 95.4%,P<0.01,respectively). Conclusions: In children with biliary atresia who underwent liver transplantation for the first time,the effective control of intraoperative bleeding should shorten the operation time and reduce the cold ischemia time as far as possible,on the premise of ensuring the safety of operation. For children without growth disorder,the weight of children should be increased to more than 5.5 kg as far as possible to receive the operation. Reducing intraoperative bleeding is of great significance to the prognosis of children.

[Clinical study of causes and outcomes in pediatric liver retransplantation].

Objective: To investigate the etiology,clinical features and prognosis of pediatric liver retransplantation. Methods: The data of 1 024 cases of pediatric liver transplantation (<18 years old) from January 2014 to December 2019 operated at Tianjin First Central Hospital were collected,retrospectively. Retransplantation was performed in 26 cases,among which 25 cases received secondary liver transplantation and 1 case received a third liver transplantation. There were 13 male and 12 female patients among the 25 patients. The median age was 12.9(20.5) months(range: 5.8 to 134.8 months), the body weight was 8.0(5.6) kg(range: 5.0 to 30.0 kg) at the time of retransplantation. The pediatric end-stage liver disease(PELD) score was 17.0(21.3) (range: 0 to 45) before retransplantation. The etiology of retransplantation was biliary complications in 7 cases,primary nonfunction of liver graft in 5 cases,antibody-mediated rejection in 4 cases,hepatic artery thrombosis in 3 cases,portal vein thrombosis in 3 cases,concomitant hepatic artery and portal vein thrombosis in 2 cases,thrombogenesis of inferior Vena Cava in 1 case and sinusoidal obstruction syndrome in 1 case. The patients were divided into two groups according to the time interval(30 days) between two liver transplantations,8 patients were classified into early-retransplantation(≤30 days) group and 18 patients were classified into late-retransplantation (>30 days) group. The etiology of liver retransplantation,pre-transplant score,time interval between two transplantations,surgical aspects,major complications and survival rates were compared between the two groups. Continuous variables with normal distribution were compared with t test,while Mann-Whitney U test was applied to compare variables without normal distribution. Categorical variables were compared with chi-square test. The survival curves were created by Kaplan-Meier method and compared by Log Rank test. Results: The median follow-up time was 26.8(30.2) months(range: 1 day to 85.7 months), and the incidence of retransplantation was 1.9%. In the early-retransplantation group,the duration of surgery was (439.8±151.0)minutes,the graft-to-recipient weight ratio was 5.0(1.8)%(range:3.6% to 6.1%),the main cause for retransplantation were primary nonfunction and vascular complications. In the late-retransplantation group,the duration of surgery was (604.4±158.0)minutes,the graft-to-recipient weight ratio was 3.4(2.1)%(range:1.4% to 5.3%),the main cause for retransplantation were biliary complications,antibody mediated rejection and vascular complications.The 3-month,1-year and 2-year recipient survival rates in the early-retransplantation group were all 62.3%,while the recipient survival rates in the late-retransplantation group were 100%,93.8% and 93.8%,respectively. The difference of recipient survival rates was significant between the early-retransplantation group and the late-retransplantation group(P=0.019). The overall 3-month,1-year and 3-year recipient survival rates after the primary liver transplantation were 97.1%,95.4%,94.1%,respectively. Conclusions: The vascular complications,biliary complications,primary nonfunction and antibody-mediated rejection are the main causes of liver retransplantation.The PELD score is higher in patients receiving early retransplantation,while the surgery is relatively more complex in patients receiving late retransplantation,which is reflected by longer duration of surgeries. Patients in the late-retransplantation group showed similar recipient survival rates with primary liver transplantation recipients,and the survival rates are superior to those of patients in the early-retransplantation group. Infection and multiple organ failure are the most common fatal causes after retransplantation.

Analysis of altered miRNA profiling in the colon of a mouse model with ß-lactoglobulin allergy.

OBJECTIVES: The differences in the expression profiles of colonic miRNAs between ß-lactoglobulin (ß-Lg) allergic mice and normal mice were analyzed to investigate the important role of the miRNA regulation mechanism in the pathogenesis of cow's milk allergy. METHODS: The present study performed Illumina sequencing to characterize the miRNA profile changes in mouse colon responding to ß-Lg challenge. Target genes were predicted by TargetScan 50 and miRanda 3.3a algorithms and assessed by GO and KEGG analysis. The expression levels of selected miRNAs and cytokine production were verified by cell transfection and quantitative RT-PCR. RESULTS: A total of 15 miRNAs were diversely expressed between the colon of the normal and ß-Lg-sensitized mice (P < 0.05, fold change of >1.50 or <0.67), including six up-regulated miRNAs and nine down-regulated miRNAs, among which seven miRNAs were validated using qRT-PCR. GO enrichment and KEGG pathway analyses further revealed that biological process, protein binding, cytoplasm and the pathways of cancer were significantly enriched, which were closely connected to the allergic inflammation development. Additionally, six key functional interaction pairs in ß-Lg allergy were identified in miRNA prediction algorithms and verified using qRT-PCR. CONCLUSIONS: We can conclude that our results suggested that the miRNAs regulation network participated in the pathogenesis of cow's milk allergy.

[The influence of probiotics and synbiotics on intestinal inflammation and microbiota in mice with acute colitis].

Objective: To investigate the effects of probiotics and synbiotics on inflammation and microbiota of acute colitis in mice. Methods: C57BL/6J mice were divided into 4 groups randomly. Each group had 10 mice and was given 2.5% dextran sulfate sodium (DSS) drinking water for 5 days other than the blank control group. Except for model control group, other two groups were administrated with probiotics and synbiotics, respectively. Probiotics was composed of Lactobacillus acidophilus, Lactobacillus rhamnosus and Bifidobacterium lactis, while synbiotics was composed of the aforementioned probiotics, inulin and galactooligosaccharide. Feces of different periods and mucosa samples were collected to analyze the differences of enteric flora by 16s rDNA sequencing. Results: (1) Pathological scores in probiotics group and synbiotics group were 5.40±2.79 and 7.25±2.87, respectively, which were significantly lower than those in the model control group with scores 27.00±7.94. Model control group, probiotics group and synbiotics group showed lower flora diversity, increased Bacteroides and decreased Faecalibacterium than blank control group. The mucosal microbiota was different from fecal flora in abundance and species for each group, and Mucispirillum was more common in mucosa. Conclusions: Probiotics and synbiotics alleviate the inflammation of acute colitis in mice. Imbalance of beneficial genera to harmful genera is the characteristic of acute colitis. Supplementation of probiotics and synbiotics contributes to regulating the balance of intestinal microbiota.

[Effect of VSL#3 and S.Boulardii on intestinal microbiota in mice with acute colitis].

Objective: To investigate the effects of probiotics(VSL#3, S. Boulardii) on intestinal flora of mice with DDS-induced acute colitis. Methods: C57BL/6J mice were administered with 2.5% dextran sulfate sodium for 5 consecutive days to develop the acute colitis model except for the blank control group. Meantime,Mice were treated with drinking water (DSS model group),VSL#3 (1.5×10(9) CFU),or S.Boulardii(5×10(7) CFU) by gavage for 7 days respectively,and mice were sacrificed 2 days after the model of colitis was established. The fecal specimens before gavage (day 0),in the middle of experiment (day 4),and the end of gavage (day 7) and the intestinal mucosa after sacrifice were collected to analyze the differences between these four groups by 16s rDNA sequencing method. Results: Compared with the DSS model group, VSL#3 group showed a decrease in disease activity index (DAI) and histological scores, and there was no significant change in the S.Boulardii group. Fecal microbiota:in the middle of experiment,the alpha diversity of DSS model group,VSL#3 group and S.Boulardii group were lower than that of the blank control group(P=0.0135,P=0.0018,P=0.0151). After the end of gavage,the diversity of the VSL#3 group was lower than that of the blank control group(P=0.025), and the difference between any other two groups was not statistically significant. Mucosa-adherent microbiota:biodiversity of DSS model group,S.Boulardii group were lower than the blank control group(P=0.031,P=0.0437),while biodiversity of VSL#3 group was higher than DSS model group and S. Boulardii group(P=0.0394, P=0.0426). Compared with the blank control group, the DSS model group showed an increase in Bacteroides and a decrease in Lactobacillus. Abundance in the genus Turicibacter and Odoribacter increased in intestinal microbiota of mice with acute colitis, while VSL#3 inhibited them. Conclusions: VSL#3 alleviates inflammation in DSS-induced colitis of mice.Both VSL#3 and S.Boulardii can affect intestinal microbiota. Compared with healthy mice,mice with colitis showed a reduced diversity of microbiota both in feces and in intestinal mucosa. VSL#3 increases biodiversity of mucosal microbiota in mice with acute colitis,while it does not increase biodiversity of fecal microbiota. Genera such as Turicibacter and Odoribacter increase in mice with acute colitis, and these genera can be inhibited by VSL#3.

Upgraded flowing liquid lithium limiter for improving Li coverage uniformity and erosion resistance in EAST device.

We report on design and technology improvements for a flowing liquid lithium (FLiLi) limiter inserted into auxiliary heated discharges in the experimental advanced superconducting tokamak device. In order to enhance Li coverage uniformity and erosion resistance, a new liquid Li distributor with homogenous channels was implemented. In addition, two independent electromagnetic pumps and a new horizontal capillary structure contributed to an improvement in the observed Li flow uniformity (from 30% in the previous FLiLi design to >80% in this FLiLi design). To improve limiter surface erosion resistance, hot isostatic press technology was applied, which improved the thermal contact between thin stainless steel protective layers covering the Cu heat sink. The thickness of the stainless steel layer was increased from 0.1 mm to 0.5 mm, which also helped macroscopic erosion resilience. Despite the high auxiliary heating power up to 4.5 MW, no Li bursts were recorded from FLiLi, underscoring the improved performance of this new design.

Budd-Chiari syndrome: current perspectives and controversies.

Budd-Chiari syndrome (BCS) is a rare disorder caused by hepatic venous outflow obstruction with a wide spectrum of etiologies. Clinical manifestations are so heterogeneous that the diagnosis should be considered in any patients with acute or chronic liver disease. Therapeutic modalities for BCS have improved dramatically during the last few years. The concept of a step-wise treatment strategy has been established, including anticoagulation, thrombolysis, percutaneous recanalization, transjugular intrahepatic portosystemic shunt, surgery and liver transplantation. However, this strategy is primarily based on experts' opinions and retrospective case series, rather than prospective randomized trials. Furthermore, an earlier use of TIPS has been proposed in selected cases because of a relatively high mortality from BCS patients who underwent medical therapy alone. Herein, we review the advances in the classification, etiology, clinical presentation, diagnosis and treatment of BCS.

Altered circulating endothelial progenitor cells in patients with keloid.

Evidence has suggested that vascular endothelial growth factor (VEGF), a crucial growth factor in regulating endothelial progenitor cells (EPCs), plays a central role in keloid formation. However, the levels of circulating EPCs in patients with keloid have not yet been explored. The aim of this study was to determine the number of circulating EPCs in patients with keloid. Circulating EPCs (defined as CD45- CD34+CD133+VEGFR2+cells) and VEGF levels from 39 patients with keloid and 22 healthy controls (HCs) were assessed by flow cytometry and ELISA, respectively. EPCs were detectable in the peripheral blood of patients with keloid. The number of circulating EPCs and the levels of plasma VEGF were significantly higher in patients with keloid than in HCs. However, no correlation was found between the number of circulating EPCs and the serum VEGF levels. This study provides the first evidence that EPCs are increased in the peripheral blood of patients with keloid. Understanding the roles of EPCs in keloid fromation may lead to the development of novel therapeutic strategies for keloid.

Accelerated healing by composites containing herb epimedium for osteoinductive regeneration.

Porous composites composed of hydroxyapatite (HA), herb epimedium (EP), and chitosan (CS) were used to improve the repair of rabbit bone defects. The in vivo implantation of the HA/CS-EP showed that homogeneous bone formation occurred after 12 weeks' implantation and possessed good osteogenesis. The osteogenic process of the HA/CS-EP group was different from that of the HA/CS group. Direct bone formation of osteoblasts with HA/CS-EP as the matrix could be observed. Compared with the group filled with HA/CS, the group filled with HA/CS-EP showed significant increases in the number of osteoblasts and the bone formation area, and the areas of new bone formation in the HA/CS-EP group after 4 or 12 weeks' implantation reached 33% and 87%, respectively. The novel repair system of HA/CS-EP can induce bone formation, increase osteoblast quantity and improve osteogenesis, for EP can significantly promote the proliferation and activity of osteoblasts in the early stage and accelerate bone remodeling in the later stage. Composites containing EP could be a promising material with multifunctions of osteoinduction, osteoconduction and medication for bone repair, and herb medicine EP could be used as an osteoinduction material for bone tissue engineering.

Rapid and direct quantitative detection of viable bifidobacteria in probiotic yogurt by combination of ethidium monoazide and real-time PCR using a molecular beacon approach.

The potential of ethidium monoazide (EMA) real-time PCR method based on molecular beacon probe for rapid detection of viable bifidobacteria present in probiotic yogurt was evaluated in this work. A real-time PCR with molecular beacon assay was developed to determine genus Bifidobacterium quantitatively in order to increase the sensitivity and specificity of assay. EMA was used to treat probiotic yogurt prior to DNA extraction and real-time PCR detection to allow detection of only viable bacteria. The primer set of Bif-F/Bif-R which is genus-specific for Bifid. was designed. The specificity of the probes ensures that no signal is generated by non-target amplicons. Linear regression analysis demonstrated a good correlation (R² = 0·9948) between the EMA real-time PCR results and the plate counting, and real-time quantitative PCR results correlated adequately with enumeration of bifidobacteria by culture for commercial probiotic yogurt. This culture-independent approach is promising for the direct and rapid detection of viable bifidobacteria in commercial probiotic yogurt, and the detection can be carried out within 4 h. The detection limit for this method is about 104 cell/ml. In conclusion, the direct quantitative EMA real-time PCR assay based on molecular beacon described in this research is a rapid and quantitative method.

[Salvia miltiorrhizae in the treatment of the viral myocarditis].

In Order to evaluate the effect of Salvia miltiorrhizae (SM) on the acute viral myocarditis (AVM), 60 children with AVM were studied. The patients were divided in random into two groups, group 1 treated with vit. C, ATP, CoA (n = 30), group II with SM plus vit. C, ATP, CoA (n = 30). The levels of plasma lipid peroxide (LPO), erythrocyte membrane microviscosity (EMMV), LDH, CPK, GOT and ECG in each patient were determined before and after one course of treatments respectively. The results revealed that before treatment the levels of plasma LPO and EMMV in both groups increased significantly compared with those of normal controls (n = 30, P < 0.01) respectively. There was a close correlation between LPO and EMMV (r = 0.6774, P < 0.01) and a close correlation between LPO and LDH (r = 0.5703, P < 0.01). After one course, the levels of plasma LPO and EMMV in both groups decreased significantly (compared with those before treatment, P < 0.01, respectively). But the LPO level and EMMV in group I were much higher than those in normal controls yet (P < 0.05, respectively). And LDH, GOT and ECG in nearly half of the patients in group I did not recover after one course while most patients in group II recovered. The results suggested that free radical plays an important role in the pathogenesis of AVM. SM as a good antioxidant, could protect myocardium from repairing membrane damage and clearing away free radical. This provided a new approach to treatment of viral myocarditis.