OBJECTIVETo investigate the effects and regulatory mechanism of ILK on angiogenesis in hypertrophic scar.

METHODSThe human scar microvascular endothelial cells (HSMECs) were isolated from 6 patients' hypertrophic scar in vitro. The HSMECs with good condition in 2nd to 4th generation were selected as experimental objectives. (1) HSMECs were divided into the blank control group (treated with routine culture), negative control group (treated with only Lipofectamine 2000), LY294002 group (incubated with 50 nmol/L LY294002), ILK siRNA group (incubated with 20 nmol/L ILK siRNA). RT-PCR and Western Blot were used to detect the expression of ILK mRNA and its protein after transfecion for 48 h. (2) The digested HSMECs of four groups were resuspended with DMEM without serum and then seeded onto the upper compartment of transwell insert which contained complete medium in its lower compartment. The cell migration experiment was stopped in 10 h and then the migrated cells were counted to analyze the effects of different interventions on the migration ability of HSMECs. (3) The thawed ECMatrix was put into each well of pre-colled 48-well tissue culture plate, and then the plate was put into the incubator at 37 degrees C to make it to become gel. The HSMECs of four groups were seeded onto the surface of the ECMatrix gel and were put into incubator. Eight random view-fields per well should be valued by the sheet of pattern recognition about angiogenesis after 8 hours to evaluate the ability of angiogenesis in vitro between four groups.

RESULTS(1) The expression of ILK mRNA (ILK mRNA = 0.829 +/- 0.109, t = 13.151, P = 0.006) and protein (ILK protein = 0.096 +/- 0.049, t = 36.656, P = 0.000) were both inhibited obviously in ILK siRNA group compared with the blank control group (ILK mRNA = 0.829 +/- 0.109, ILK protein = 1). And, the expression of ILK in LY294002 group was slightly lower than that of black control group, but there was no statistical difference. (2) The number of migrated cells in ILK siRNA group (88.111 +/- 3.079) and LY294002 group (138. 667 +/- 2.404) were respectively lower than that in blank control group (322.333 +/- 3.712, P < 0. 05) in 10th hour. (3) Compared to blank control group (4.333 +/- 0.191), the ability of angiogenesis in vitro decreased significantly ILK siRNA group (2.625 +/- 0.125) and LY294002 group (3.125 +/- 0.250), in which, the vascular network structures were not formed perfectly in 8th hour (P < 0.05).

CONCLUSIONSThe ability of HSMECs' migration and angiogenesis in vitro are inhibited significantly when the expression of ILK is down-regulated. It reveals that ILK may play an role in the regulation of scar angiogenesis.

Cell Movement ; Cell Proliferation ; Chromones ; pharmacology ; Cicatrix, Hypertrophic ; enzymology ; pathology ; Endothelial Cells ; cytology ; drug effects ; Humans ; Lipids ; pharmacology ; Morpholines ; pharmacology ; Neovascularization, Pathologic ; etiology ; pathology ; Protein-Serine-Threonine Kinases ; genetics ; physiology ; RNA, Messenger ; analysis ; RNA, Small Interfering ; metabolism

Objective To investigate the effects of the intervention,rehabilitation and speech development of children with severe hearing loss in some rural areas.Methods 61 children,including 35 males and 26 females,were diagnosed as severe hearing loss with ABR and 40 Hz-AERP from June 2004 to July 2008.All the children failed hearing screening or visited the hospital as outpatients.The ages ranged from 2 to 72 months with the average age of 17.59 months.During telephone follow-up,the questionnaire was used to gather the data regarding the usage of hearing aids,hearing and speech rehabilitation,speech development,and communication abilities.Results 33 (54.10%) children were fitted with hearing aids,and 2 (3.28%) received cochlear implants,while 26(42.62 %) neither had hearing aids nor cochlear implants.10 cases with hearing aids also had speech training,whereas 23 children with hearing aids did not receive the training.2 cases with cochlear implants and 2 cases with hearing aids were found to have good speech development and communication ability,while 31 cases with hearing aids had delayed speech development.26 cases without any devices had to rely on sign language for their commumication.Conclusion Children in rural area with severe hearing loss experience greater speech and communication difficulties because many of them have no access to intervention and speech training.The results suggest that it would be very important to increase public awareness and educate parents to have their children wear hearing aids and receive speech training.

OBJECTIVEThis study aimed to explore the impact of specimen collection and storage consumable products on trace element quantitative analysis.

METHODSDevices and consumable products of different brands used in specimen collection or storage were selected and treated separately as below:urine collection and storage tubes (Brand A, B, C and D, 2 samples for each brand) were treated with 1% of HNO(3) volume fraction for 2 - 4 h; blood taking device (Brand O, P and Q, 3 samples for each brand) were used for ultra-pure water samples collecting as simulation of blood sampling;dust sampling filters (Brand X, Y and Z, 2 samples for each brand) were cold digested by nitric acid for 12 h, followed by microwave digestion. Then cadmium, cobalt, chromium, copper, iron, manganese, molybdenum, nickel, lead, selenium, stannum, titanium, vanadium and zinc concentrations in the solutions obtained during the course of collect or storage were quantified by inductively coupled plasma mass spectrometer.

RESULTSFor the urine collection and storage consumable products, background values of elements were described as mean of parellel samples. The consentration of 14 quantified elements were relatively low for 5 ml cryogenic vials (brand B) with background values range of 0.001 - 0.350 ng/ml. The background values of copper of 50 ml centrifuge tubes (brand A), chromium of 5 ml cryogenic vials (brand C) and zinc of 1.5 ml centrifuge tubes (brand D) were relatively high, which were 1.900, 1.095 and 1.368 ng/ml, respectively. Background values of elements in blood sampling devices were described as x(-) ± s. Background values of chromium for brand O, P and Q were (0.120 ± 0.017), (0.337 ± 0.093) and (0.360 ± 0.035) ng/ml; for copper were (0.050 ± 0.001), (0.017 ± 0.012) and (0.103 ± 0.015) ng/ml; for lead were (0.057 ± 0.072), (0.183 ± 0.118) and (0.347 ± 0.006) ng/ml; for titanium were (7.883 ± 0.145), (8.863 ± 0.190) and (8.613 ± 0.274) ng/ml; zinc were (2.240 ± 0.573), (42.140 ± 22.756) and (8.850 ± 3.670) ng/ml. There were statistically differences of background values for chromium, copper, lead, titanium and zinc among the above three brands of blood sampling devices (all P values < 0.05). For air sampling filters, background values of elements were described as mean of parellel samples. Background values of chromium and nickel of sampling filters (brand X) were lowest, which were 17.000 and 15.400 ng per piece, respectively; while background values for other elements were relatively high, the quantification of cadmium, cobalt, copper, iron, manganese, molybdenum, lead, selenium, stannum, titanium, vanadium and zinc were 0.250, 0.550, 48.500, 690.000, 25.500, 0.900, 6.500, 10.550, 7.950, 10.500, 0.850, 370.000 ng per piece, respectively. Background values of chromium and nickel of sampling filters (brand Z) were highest, which were 171.000 and 29.850 ng per piece.

CONCLUSIONBackground values of trace elements varied among products of different brands, and the most noticable differences were found in chromium, manganese, nickel, lead, stannum and zinc.

Environmental Monitoring ; methods ; Quality Control ; Specimen Handling ; methods ; Trace Elements ; analysis

OBJECTIVETo evaluate the germplasm of Rehmannia glutinosa on the basis of photosynthetic pigment contents (PPC).

METHOD20 cultivars were planted on the same condition. On Oct. 23 and Sept. 25, 3 leaves per cultivar were collected on different plants, and 80 mg mesophyll was collected among upper lateral veins and was ground in 96% alcohol, and the supernatant was subjected to measure on a spectrophotometer (Angilent 8453).

RESULTThe PPCs among cultivars were significantly different at a P < or = 0.01 level. The results of the measurements were similar. Chlolophyll a was the most abundant pigment, but varied to a great extent among different cultivars. 20 cultivars were divided into 9 homogeneous groups according to the contents of chlorophyll a by Duncan's multiple range test at P < or = 0.05. In addition, the content of chlorophyll a was closely related to leaf color. The cultivars with higher chlolophyll a had deep green leaves, and those with lower had yellow green or pale green leaves.

CONCLUSIONPPC was an inherent character and an important index for the germplasm evaluation of R. glutinosa.

Chlorophyll ; analysis ; Color ; Photosynthesis ; Pigments, Biological ; Plant Leaves ; chemistry ; growth & development ; Plants, Medicinal ; chemistry ; growth & development ; Rehmannia ; chemistry ; growth & development

OBJECTIVETo summarize the clinical management of abdominal compartment syndrome (ACS) in burn patients with severe burn injury.

METHODSTwelve serious burn patients with abdominal compartment syndrome hospitalized in our center from January 2001 to April 2005 were enrolled in the study. Among them 3 patients were treated with conservative method, 4 with escharectomy of abdominal wall, 5 with laparotomy for decompression. The clinical results were analyzed statistically. Bladder pressure, central venous pressure, systolic blood pressure and arterial blood oxygen partial pressure (PaO2 ) were measured and compared before and after operation.

RESULTSAmong these 12 patients, 5 died with the overall mortality of 41.67%. But only 3 died among 9 patients undergone operation. Most of patients were oliguric,with abnormal bladder pressure, central venous pressure, and systolic blood pressure 24 hours before operation. But these parameters were significantly improved after operation ( P <0. 01).

CONCLUSIONEarly abdominal escharectomy and timely abdominal decompression are vital for the management of ACS in burn patients.

Abdomen ; pathology ; Adult ; Aged ; Burns ; complications ; therapy ; Compartment Syndromes ; etiology ; surgery ; Female ; Humans ; Male ; Middle Aged

AIMTo investigate the mechanism of protective effect of SOD (superoxide dismutase) on damage of RBCs stored at 4 degrees C, the studies of erythrocyte glucose and energy metabolism were performed.

METHODSwhole blood collected from healthy donors and stored at 4 degrees C in ACD, GMA and SOD solutions. Before and post storage, some parameters were assayed. Standard methods were used for the in vitro tests. The 24-hour in vivo recoveries were measured by FTTC (Fluorescein 5-isothiocyanate) from SIGMA Company.

RESULTSAll parameters of red blood cell glucolysis rate without oxygen condition, ATP, PK (pyruvic kinase) and 24 h recoveries level were 86.2%, 56.4%, 64.3% and 86.2% of normal respectively stored in SOD solution at 4 degrees C for 75 days, distinctly more than in ACD and GMA groups at 75 days stored. The 24 h recovery at 75d in group SOD was near the recovery at 42d in group GMA.

CONCLUSIONWhole blood in SOD solution can be stored satisfactorily for 75 days at 4 degrees C, and furnished theoretical evidence for RBCs survival.

Animals ; Blood Preservation ; methods ; Erythrocytes ; cytology ; metabolism ; Humans ; Rabbits ; Refrigeration ; Superoxide Dismutase ; pharmacology

Animals ; Cytoskeletal Proteins ; biosynthesis ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Hepatocytes ; metabolism ; ultrastructure ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the expression of integrin-linked kinase (ILK) in scar in different growth stages, as well as its relationship with angiogenesis.

METHODS(1) Fifteen burn patients with scar formation time shorter than 6 months, ranging from 6 to 12 months, and longer than 12 months were hospitalized from December 2009 to December 2010. They were divided into A, B, and C groups according to the scar formation time, with 5 patients in each group. Scar specimens were harvested for observation of ILK expression with immunohistochemistry method, and ILK mRNA expression with real time fluorescence quantitative RT-PCR. (2) Microvascular endothelial cells (MEC) were isolated from scar tissue in A group and cultured in vitro, and then they were purified by immunomagnetic beads and identified with coagulation factor VIII marked by immunofluorescence (fibroblasts from human normal skin were used as control). The cultured cells in logarithmic growth phase were divided into control group (cultured with M131 medium containing microvascular growth supplement), transfection 1 group (transfected with empty plasmid), and transfection 2 group (transfected with ILK cDNA plasmid) according to the random number table. After 24 hours, the expressions of ILK mRNA, Flt-1 mRNA, and KDR mRNA were determined with real time fluorescence quantitative RT-PCR. Data were processed with one-way analysis of variance.

RESULTSImmunohistochemical observation showed that ILK in A group mainly expressed in the basal layer cells of epidermis, cytoplasm of fibroblasts, and MEC in scar, while ILK in B group only distributed in the basal layer cells of epidermis, but ILK expression in C group was not obvious. The expression of ILK mRNA in A group (0.34 ± 0.16) was significantly higher than those in B and C groups (0.17 ± 0.06, 0.07 ± 0.13, F = 37.007, P = 0.000). MEC grew up showing cobble stone formation after purification. The expression of coagulation factor VIII was positive in cytoplasm of purified MEC, while that was negative in fibroblast of human normal skin. The expressions of ILK mRNA (57.807 ± 5.556), KDR mRNA (0.836 ± 0.014), and Flt-1 mRNA (0.162 ± 0.005) in transfection 2 group were higher than those in control and transfection 1 groups (0.018 ± 0.003, 0.028 ± 0.020, 0.023 ± 0.004 and 0.042 ± 0.005, 0.039 ± 0.007, 0.046 ± 0.003; F(ILK) = 87.110, F(KDR) = 11.241, F(Flt) = 18.199, with P values all below 0.01).

CONCLUSIONSILK mainly expressed in scar tissue with formation time shorter than 6 months, and it may affect vascularization of scar by regulating gene expressions of KDR and Flt-1 in MEC, which plays an important role in early scar formation.

Adolescent ; Adult ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Endothelial Cells ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Neovascularization, Pathologic ; metabolism ; pathology ; Protein-Serine-Threonine Kinases ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism ; Young Adult

OBJECTIVETo investigate the expression of Yin Yang 1 (YY1) protein in human insulinoma and explore its clinical significance.

METHODSNineteen pancreatic neuroendocrine tumor tissue were collected from patients treated in Nanfang Hospital between 2000 and 2014. The protein expression of YY1 in benign and malignant insulinoma tissues were detected by immunohistochemistry.

RESULTSPositive expression for YY1 protein was detected in both benign and malignant tumor tissues, but the malignant tissues had a significantly greater intensity of YY1 expression than the benign tissues (P=0.042). The intensity of YY1 expression was positively correlated with the nature of the tumor, and the insulinomas with high expressions of YY1 had significantly greater malignant potentials (P=0.037).

CONCLUSIONThe high expression of YY1 protein is associated with the development of insulinima. YY1 may serve as a new tumor marker for detecting the malignant transformation of insulinoma.

Biomarkers, Tumor ; metabolism ; Cell Transformation, Neoplastic ; Humans ; Immunohistochemistry ; Insulinoma ; genetics ; metabolism ; Pancreatic Neoplasms ; genetics ; metabolism ; YY1 Transcription Factor ; genetics ; metabolism

OBJECTIVETo explore the expression of integrin-linked kinase (ILK) in fibroblasts (Fbs) of scar induced by cobalt chloride (CoCl2) and its effect on cell proliferation.

METHODSThe human hypertrophic scar Fbs of seven patients were isolated and cultured in vitro. Cells from the 5th to the 6th passages were used in the experiment. Six bottles of Fbs were obtained from each of the seven patients, and they were respectively cultured with DMEM nutrient solution containing CoCl2 in the concentration of 0, 50, 100, 150, 200, and 250 µmol/L for 24 h. The expression of ILK mRNA was determined with real-time fluorescence quantitative PCR. Fbs were stimulated by CoCl2 in the most suitable concentration (100 µmol/L) and the protein expression of ILK was determined 0, 1, 2, 4, 12, and 24 h after the stimulation. Then the Fbs were divided into control group (cultured with nutrient solution), negative control group (transfected with con-siRNA), and ILK siRNA group (transfected with ILK siRNA). They were cultured with nutrient solution containing CoCl2 in different concentrations 24 h after transfection, with 4 wells for each concentration in each group. The cell proliferation was detected by XTT assay. Data were processed with one-way analysis of variance (ANOVA) and ANOVA for repeated measurement, and LSD method was used in multiple comparisons.

RESULTSThe expression level of ILK mRNA was highest in Fbs cultured with 100 µmol/L CoCl2 for 24 h, with significant difference compared with those of Fbs cultured with other concentrations of CoCl2 (F = 50.958, P < 0.001). The expression of ILK protein in Fbs cultured with 100 µmol/L CoCl2 for 1 h (0.243 ± 0.009) was lower than that cultured for 0 h (0.387 ± 0.017), and it started to increase from 2 h (0.361 ± 0.010), and exaggerated at 4 h (0.584 ± 0.028), 12 h (0.730 ± 0.029), and 24 h (0.785 ± 0.031). The expression levels of ILK protein at 1, 4, 12, 24 h were statistically different from that at 0 h (P values all below 0.05). XTT showed that cell proliferation level was highest in control group when cultured with 100 µmol/L CoCl2 (F = 488.026, P < 0.001), which decreased from 150 µmol/L. The cell proliferation level in control group cultured with 250 µmol/L CoCl2 was significantly lower than that with 0 µmol/L (P values all below 0.05). There was no significant change in cell proliferation in ILK siRNA group among different concentrations of CoCl2 (F = 2.542, P = 0.056). The cell proliferation level in ILK siRNA group was significantly lower than that in control group and negative control group (F = 2519.542, P < 0.001).

CONCLUSIONSILK may be a key protein in response of hypoxia in Fbs. The mild hypoxia can stimulate the expression of ILK and promote the proliferation of Fbs, while severe hypoxia can reduce the expression of ILK and inhibit cell proliferation.

Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix ; metabolism ; pathology ; Cobalt ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; pathology ; Humans ; Protein-Serine-Threonine Kinases ; metabolism