DANCR Regulates hESC Differentiation Towards Definitive Endoderm / 中山大学学报(医学科学版)

ObjectiveTo explore the function of DANCR during the differentiation of human embryonic stem cells （hESC） toward definitive endoderm （DE）. MethodsThe in vitro DE differentiation system was established and its efficiency was verified. The correlation between the expression level of DANCR and DE differentiation process was detected. Using lentivirus system， we stably knocked down DANCR in hESC. The shDANCR hESC line was applied to DE differentiation， using qPCR and Western blot to detect the expression of DE marker genes SOX17 and FOXA2， and that of primitive streak marker genes Brachyury （T）， EOMES， MIXL1 and GSC. Dual luciferase reporter assay and qPCR were used to confirm the interaction between DANCR and the WNT pathway during DE differentiation. ResultsThe in vitro differentiation system mimicked DE differentiation efficiently. And the expression of DANCR was gradually downregulated during differentiation. DANCR was efficiently knocked down in the shDANCR hESC line （P < 0.001）. Compared with those in the control group， the expression levels of primitive markers Brachyury （T）， EOMES， MIXL1 and GSC， as well as DE markers SOX17 and FOXA2， were significantly decreased in shDANCR groups （P < 0.05）. Furthermore， the transcriptional activity of the WNT pathway in shDANCR groups was lower than that in the control group （P < 0.05）. And RNA levels of downstream genes of the WNT pathway， FZD5， FZD8， SFRP1， FRZB and ANKRD6， were significantly decreased in shDANCR groups （P < 0.05）. However， differences in protein levels of the TGFβ pathway effectors SMAD2/3 and p-SMAD2 were statistically insignificant in shDANCR and control groups （P > 0.05）. Forced activation of β-CATENIN rescued DANCR knock down-induced deficiency in DE differentiation. ConclusionsThe expression of DANCR decreases during DE differentiation. DANCR may promote DE differentiation through modulating the activity of the WNT pathway.

MDM2 Inhibitor RG-7388 Promotes Apoptosis and Cell Cycle Arrest of Diffuse Large B Cell Lymphoma / 肿瘤防治研究

Objective To investigate the effect of MDM2 inhibitor RG-7388 on the proliferation, cell cycle, and apoptosis of diffuse large B-lymphoma (DLBCL) cells. Methods DLBCL cell strains SUDHL2 and HBL1 were treated with 2, 4, and 8 μmol/LRG7388, respectively. Cell proliferation was detected by CCK8 and EdU methods. Apoptosis was measured by Annexin V–FITC/PI double staining and Caspase 3/7-Glo enzyme activity methods. Cell cycle was assessed by flow cytometry. Changes in the expression of cell cycle and apoptosis-related proteins were determined by Western blot. Results The IC50 of RG7388 for inhibiting SUDHL2 and HBL1 cells were 3.36 and 3.76 μmol/L, respectively, and the inhibitory effect of RG7388 was dose dependent. The proportions of G1 phase and apoptotic cells in the SUDHL2 and HBL1 cells treated with different doses of RG7388 were significantly higher than those in the control group (all P<0.05). The activity of Caspase 3/7 increased gradually with RG7388 concentration, compared with that in the control group. The expression levels of p53, p27, p21, and PARP increased, whereas the expression of Mcl-1 and Bcl-xL was down-regulated (all P<0.05). Conclusion MDM2 inhibitor RG-7388 inhibits the proliferation of DLBCL cells, triggers cell cycle arrest in the G1 phase, and induces apoptosis through the p53 pathway.

Analysis of causes and treatment methods of complication of early acute kidney injury in four severely burned patients / 中华烧伤杂志

Objective@#To analyze the causes of complication of early acute kidney injury (AKI) in four severely burned patients, and to explore the related treatment methods.@*Methods@#The clinical data of 4 patients with severe burn complicated with early AKI admitted to Guangzhou Red Cross Hospital Affiliated to Medical College of Jinan University (hereinafter referred to as our hospital) from June 2014 to December 2017 were retrospectively analyzed. All the patients were male, aged 23-33 (30±5) years old, with depth of burns ranged from deep partial-thickness to full-thickness, complicated with myofascial compartment syndrome of extremities and varying degrees of striated muscle injury, and treated in other hospitals before transfer to our hospital. The patients were numbered from small to large according to the total burn area. The total burn area of patients No. 1, 2, 3, and 4 was 10%, 80%, 90%, and 95% total body surface area respectively, their occurrence time of early AKI was 48, 11, 29, and 48 hours after injury respectively, and their time of arriving our hospital was 60, 11, 29, and 144 hours after injury respectively. Hypovolemic shock occurred in patients No. 2 and 3 at admission to our hospital. All the patients received continuous renal replacement therapy (CRRT) after admission to our hospital. Under the support of hemodynamic monitoring and organ function monitoring, the limbs complicated with myofascial compartment syndrome were incised, thorough decompression exploration was performed, and necrotic muscle tissue was removed or amputation was performed. After escharectomy and decompression of limbs, fresh granulation wounds were formed by temporarily covering wounds with Jieya dressing skin or pig skin, multiple debridements, and vacuum sealing drainage. Fresh granulation wounds and other wounds underwent staged eschar excision and shaving were covered with autologous Meek skin graft, particulate skin graft, reticular skin graft and small skin graft respectively. The treatment outcome, CRRT time, operation times, time of recovery of serum creatinine and myoglobin, length of hospital stay, and follow-up were recorded.@*Results@#All the 4 patients were cured after transfer to our hospital. Among them, totally 5 limbs of patients No. 1 and No. 4 underwent amputation because of complication of myofascial compartment syndrome and a large amount of necrotic muscle which could not be preserved. Patients No. 1, 2, 3, and 4 were treated with CRRT for 19, 35, 14, and 25 days respectively and performed with operation for 5, 6, 10, 8 times respectively. Serum creatinine of patients No. 1, 2, 3, and 4 returned to normal on 22, 35, 37, and 48 days after transfer respectively, and their serum myoglobin returned to normal on 18, 28, 25, and 30 days after transfer respectively. Patients No. 1, 2, 3, and 4 were hospitalized for 52, 105, 148, and 156 days and discharged after basic wound healing. Follow-up for 1 to 36 months showed no abnormal renal function in 4 patients.@*Conclusions@#The early AKI in patients No. 1 and 4 was caused by rhabdomyolysis after severe burn complicated with myofascial compartment syndrome, while that of the other 2 cases were also related to hypovolemic shock and poor renal perfusion. The success rate of early AKI treatment in severely burned patients can be effectively improved by removing the causes of diseases at the same time of CRRT and actively treating burn wounds under the support of organ function and hemodynamic monitoring.

Drug resistance and status of infection of Acinetobacter baumannii in burn intensive care unit during 3 years / 中华烧伤杂志

<p><b>OBJECTIVE</b>To analyze the detection, drug resistance, and status of infection of Acinetobacter baumannii (AB) in burn ICU during 3 years.</p><p><b>METHODS</b>A total of 2 010 specimens of wound secretion, blood, venous catheter attachment, sputum, stool and urine were collected from 505 burn patients hospitalized in our burn ICU from January 2011 to December 2013, and bacterial culture was performed. Pathogens were identified by automatic microorganism identifying and drug sensitivity analyzer. Drug resistance of all the obtained AB to 16 antibiotics commonly used in clinic, including cefoperazone/sulbactam, polymyxin, etc., was tested with K-B paper disk diffusion method. Patients with AB infection were ascertained. The WHONET 5.6 software was used to analyze the distribution of pathogens during 3 years, the isolation of AB with different sources and the status of drug resistance of AB to 16 antibiotics each year, and the status of patients with AB infection, and their outcome.</p><p><b>RESULTS</b>A total of 961 strains of pathogens were isolated, among which 185 (19.25%) strains were Gram positive cocci, 728 (75.75%) strains were Gram negative bacilli, and 48 (4.99%) strains were fungi. A total of 172 strains of AB were isolated, ranking the second place among all the detected pathogens, with 67 (38.95%) strains from wound secretion, 11 (6.40%) strains from blood, 23 (13.37%) strains from venous catheter attachment, and 71 (41.28%) strains from sputum, no AB strain was isolated from feces or urine. The AB strains were found sensitive to polymyxin and with relatively low drug resistance rate to minocycline, while the drug resistance rates were over 80.0% to the other 14 antibiotics commonly used in clinic in 2013. AB culture of wound secretion was positive in 27 patients. Among them, 7 patients suffered from wound infection, and the wound infection was caused by AB in 1 out of the 7 patients. AB culture of blood was positive in 7 patients. Among them, 3 patients suffered from bloodstream infection, and the infection was due to AB invasion in 1 out of the 3 patients. AB culture of venous catheter attachment was positive in 20 patients. Among them, 8 patients suffered from bloodstream infection, and the infection was due to AB invasion in 1 out of the 8 patients. AB culture of sputum was positive in 35 patients. Among them, 13 patients suffered from ventilatory associated pneumonia, and 2 out of the 13 patients were diagnosed as AB infection. A total of 69 patients were AB culture positive, among them 64 patients were cured, 2 patients were transferred to other hospitals, and 3 patients died, with the mortality rate of 4.35%.</p><p><b>CONCLUSIONS</b>AB in our burn ICU has a high detection rate and extensive drug resistance in above-mentioned 3 years. However, AB was mainly colonized in patients with extensive burns with a low mortality rate.</p>

Effects of unified surgical scheme for wounds on the treatment outcome of patients with extensive deep burn / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the effects of unified surgical scheme for wounds on the outcome of patients with extensive deep partial-thickness to full-thickness (briefly referred to as deep) burn.</p><p><b>METHODS</b>One hundred and thirty-seven patients with extensive deep burn hospitalized from July 2007 to November 2012 underwent unified surgery according to area of deep wound (unified scheme group, US). Among them, 57 patients with deep wound area less than 51% TBSA received escharectomy or tangential excision by stages followed by autologous mesh skin grafting; 52 patients with deep wound area from 51% to 80% TBSA underwent escharectomy or tangential excision by stages followed by autologous mesh skin grafting and/or small skin grafting, or escharectomy or tangential excision followed by large sheet of allogeneic skin covering plus autologous mesh skin grafting and/or small skin grafting after the removal of allogeneic skin; 28 patients with deep wound area larger than 80% TBSA received escharectomy or tangential excision by stages followed by autologous microskin grafting plus coverage of large sheet of allogeneic skin, or escharectomy or tangential excision followed by small autologous skin grafting and/or intermingled grafting with small autologous and/or allogeneic skin. Another 120 patients with extensive deep burn hospitalized from January 2002 to June 2007 who did not receive unified surgical scheme were included as control group (C). Except for the surgical methods in group US, in 53 patients with deep wound area less than 51% TBSA in group C escharectomy or tangential excision was performed followed by autologous small skin grafting; in 40 patients with deep wound area from 51% to 80% TBSA in group C escharectomy or tangential excision was performed followed by autologous microskin grafting plus large sheet of allogeneic skin covering, or escharectomy or tangential excision followed by large sheet of allogeneic skin embedded with stamp-like autologous skin; in 27 patients with deep wound area larger than 80% TBSA in group C escharectomy or tangential excision was performed followed by covering with large sheet of allogeneic skin embedded with stamp-like autologous skin without intermingled grafting with small autologous and allogeneic skin in group US. In group US, escharectomy of full-thickness wound in extremities was performed with the use of tourniquet in every patient; saline containing adrenaline was subcutaneously injected when performing escharectomy or tangential excision over the trunk and skin excision; normal skin and healed superficial-thickness wound were used as donor sites for several times of skin excision. The baseline condition of patients and their treatment in the aspects of fluid resuscitation, nutrition support, anti-inflammation, and organ function support were similar between the two groups. The mortality and incidence of complications of all patients and wound healing time and times of surgery of healed patients were compared between the two groups. Data were processed with independent sample t test, Mann-Whitney U test, and Fisher's exact test.</p><p><b>RESULTS</b>(1) Both the mortality and the incidence of complications of patients with deep wound area less than 51% TBSA in group US were 0, which were close to those of group C (with P values above 0.05). The number of times of surgery of healed patients with deep wound area less than 51% TBSA in group US was 2.4 ± 0.9, which was obviously fewer than that of group C (3.5 ± 1.8, U=-5.085, P<0.001), but with wound healing time close to that of group C (U=-1.480, P>0.05). (2) Both the mortality and the incidence of complications of patients with deep wound area from 51% to 80% TBSA in group US were 0, which were significantly lower than those of group C [both as 20.0% (8/40), with P values below 0.01]. The number of times of surgery and wound healing time of healed patients with deep wound area from 51% to 80% TBSA in group US were respectively 3.0 ± 1.0 and (43 ± 13) d, which were obviously fewer or shorter than those in group C [4.2 ± 2.3 and (61 ± 34) d, with U values respectively -2.491 and -2.186, P values below 0.05]. (3) Both the mortality and the incidence of complications of patients with deep wound area larger than 80% TBSA in group US were 25.0% (7/28), which were close to those of group C [both as 25.9% (7/27), with P values above 0.05]. The number of times of surgery and wound healing time of healed patients with deep wound area larger than 80% TBSA in group US were close to those of group C (with U values respectively -0.276 and -0.369, P values above 0.05).</p><p><b>CONCLUSIONS</b>Unified surgical scheme can indirectly decrease the mortality and the incidence of complications of burn patients with deep wound area from 51% to 80% TBSA; it can reduce times of surgery of healed patients of this type and shorten their wound healing time.</p>

Effects of lung protective ventilation strategy combined with lung recruitment maneuver on patients with severe burn complicated with acute respiratory distress syndrome / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the effects of lung protective ventilation strategy combined with lung recruitment maneuver on ARDS complicating patients with severe burn.</p><p><b>METHODS</b>Clinical data of 15 severely burned patients with ARDS admitted to our burn ICU from September 2011 to September 2013 and conforming to the study criteria were analyzed. Right after the diagnosis of acute lung injury/ARDS, patients received mechanical ventilation with lung protective ventilation strategy. When the oxygenation index (OI) was below or equal to 200 mmHg (1 mmHg = 0. 133 kPa), lung recruitment maneuver was performed combining incremental positive end-expiratory pressure. When OI was above 200 mmHg, lung recruitment maneuver was stopped and ventilation with lung protective ventilation strategy was continued. When OI was above 300 mmHg, mechanical ventilation was stopped. Before combining lung recruitment maneuver, 24 h after combining lung recruitment maneuver, and at the end of combining lung recruitment maneuver, variables of blood gas analysis (pH, PaO2, and PaCO2) were obtained by blood gas analyzer, and the OI values were calculated; hemodynamic parameters including heart rate, mean arterial pressure (MAP), central venous pressure (CVP) of all patients and the cardiac output (CO), extravascular lung water index (EVLWI) of 4 patients who received pulse contour cardiac output (PiCCO) monitoring were monitored. Treatment measures and outcome of patients were recorded. Data were processed with analysis of variance of repeated measurement of a single group and LSD test.</p><p><b>RESULTS</b>(1) Before combining lung recruitment maneuver, 24 h after combining lung recruitment maneuver, and at the end of combining lung recruitment maneuver, the levels of PaO2 and OI of patients were respectively (77 ± 8), (113 ± 5), (142 ± 6) mmHg, and (128 ± 12), (188 ± 8), (237 ± 10) mmHg. As a whole, levels of PaO2 and OI changed significantly at different time points (with F values respectively 860. 96 and 842. 09, P values below 0. 01); levels of pH and PaCO2 showed no obvious changes (with F values respectively 0.35 and 3.13, P values above 0.05). (2) Levels of heart rate, MAP, CVP of all patients and CO of 4 patients who received PiCCO monitoring showed no significant changes at different time points (with F values from 0. 13 to 4. 26, P values above 0.05). Before combining lung recruitment maneuver, 24 h after combining lung recruitment maneuver, and at the end of combining lung recruitment maneuver, the EVLWI values of 4 patients who received PiCCO monitoring were respectively (13.5 ± 1.3), (10.2 ± 1.0), (7.0 ± 0.8) mL/kg ( F =117.00, P <0.01). (3) The patients received mechanical ventilation at 2 to 72 h after burn, lasting for 14-32 (21 ± 13) d. At post injury day 3-14 (7 ± 5) d, lung recruitment maneuver was applied for 2-5 (3.0 ± 2.0) d. All 15 patients recovered without other complications.</p><p><b>CONCLUSIONS</b>Lung protective ventilation strategy combining lung recruitment maneuver can significantly improve the oxygenation in patients with severe burn complicated with ARDS and may therefore improve the prognosis.</p>

Histological study of dual factor inducing axial vascularization in double-layered scaffold / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To construct an double-layered tube-shaped poly (lactic-co-glycolic acid) (PLGA) scaffold composited with vascular endothelial growth factor (VEGF)/ platelet-derived growth factor (PDGF), and to evaluate in vivo the axial vascularization from femoral arteriovenous bundle encapsulated.</p><p><b>METHODS</b>Eighteen male adult New Zealand rabbits were assigned randomly into 3 groups: Experimental group (n = 8), experimental control group (n = 8), empty control group (n=2). The femoral arteriovenous bundle were separated and encapsulated in double-layered tubeshaped PLGA scaffold prepared by solution casting and particle leaching. According to the way of cell factors being composited to the prepared double-layered scaffold: 1) experimental group: VEGF in inner layer and PDGF in outer layer; 2) experimental control group: VEGF in inner layer with blank outer layer; 3) empty control group: Pure blank. Specimens were retrieved at 7, 10, 14, 21 days postoperatively (7, 10 days in empty control group). The histological evaluation was performed.</p><p><b>RESULTS</b>At 7 days postoperatively, blood vessel sprouts were observed in experimental group and experimental control group, radially from the central femoral arteriovenous bundle. At 10 days postoperatively, completely penetrations of the double-layered scaffold by abundant new generated blood vessel sprouts were observed, and density descended gradiently from inside to outside. At 14 days postoperatively, new blood vessels in experimental group showed more thickness and layered structure of the wall, while monolayer endothelial cells in experimental control group. At 21 days postoperatively, new blood vessels in experimental group showed more mature characteristics, while less density of blood vessels in experimental control group. There was no obvious blood vessel structure in empty control group.</p><p><b>CONCLUSION</b>The double-layered tube-shaped PLGA scaffold composited with VEGF/PDGF could induce early angiogenesis.</p>

Characteristics and Management of Fungal Infection in Severely Burn Patients / 中华医院感染学杂志

OBJECTIVE To study the characteristics and management of fungal infection in severely burn patients.METHODS The clinical data of 51 severely burn patients with fungal infection in our department from 2000 to 2004 were analyzed.RESULTS The infection rate in total 213 severely burn patients was 23.72%.Candidiasis was the most frequent type in fungal infection accounting for 93.98%.The risk factors for fungal infection were the(severity) of burn,tracheal intubations,body catheterizations,long-term use of broad-spectrum antibiotic and(hospitalization).CONCLUSIONS Development and the types of fungi in severely burn patients with fungal infection have their own characterizations and inducing factors.it is important to eluminate the susceptible factors and provide early diagnosis and treatment.