Objective To evaluate the relative quantities of flt-1, KDR, endostatin mRNA in tissue specimens of human hemangioma and great saphfenous vein with the modified reverse transcriptase-polymerase chain reaction (RT-PCR). Methods Total RNA from human specimens by Trizol was reverse-transcribed and amplified by PCR in 4 deffirent tubes containing one of the primer pairs such as flt-1, KDR, endostatin or ?-actin. The target genes and beta-actin PCR products were about 200 bp in length. The ratio of the yield of the target gene PCR product to the beta-actin PCR product could be calculated after 35 cycles of amplification.Results These ratios were correlated positively ( P=0.014,P=0.019) in two groups, but no relationship in endostatine level was observed (P=0.436).Conclusions VEGFR may play a very important role in the pathogenesis of the congenital hemangioma in children. But the endostatin may take little effect on it.

OBJECTIVE: In order to investigate the interaction between the cytokines and keratinocyte and determine the role of cytokines in hyperproliferative of chronic otitis media with cholesteatoma, we observe the expression of matrix metalloproteinase 9 (MMP9), vascular endothelial growth factor (VEGF), keratinocyte growth factor (KGF) and its receptor (KGFR) in middle ear cholesteatoma. METHOD: We examined the expression of MMP9, VEGF, KGF, KGFR and Ki-67 by immunohistochemistry in 50 specimens from chronic otitis media with cholesteatoma and 15 specimens from the normal skin of external auditory meatus. Ki-67 as an evaluation of cholesteatoma proliferation markers were used to detect the keratinocyte proliferative activity. RESULT: (1) The expression of VEGF and MMP9 in cholesteatoma specimens was higher than normal skin, and the difference was statistically significant (t = 4.914, P < 0.01; t = 3.284, P < 0.01). (2) The expression of KGF and KGFR in middle ear tissues was higher than normal skin, and the difference was statistically significant (t = 4.814, P < 0.01; t = 3.104, P < 0.01); The expression of KGF and KGFR increased, and the expression of Ki-67 also correspondly increased in the cholesteatoma. (3) In the tissue MMP9 and VEGF were positive. Mean optical density increased as well. KGF expression also increased accordingly. CONCLUSION MMP9, VEGF, KGF and KGFR proteins played an important role in hyperproliferation of cholesteatoma tissues. VEGF, MMP9 and KGF had a synergistic effect in hyperproliferation of cholesteatoma tissues.
Cholesteatoma, Middle Ear ; pathology ; Cytokines ; metabolism ; Ear Canal ; metabolism ; Ear, Middle ; metabolism ; Fibroblast Growth Factor 7 ; metabolism ; Humans ; Immunohistochemistry ; Keratinocytes ; cytology ; Ki-67 Antigen ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Otitis Media ; pathology ; Receptor, Fibroblast Growth Factor, Type 2 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

Objective:The effect of naltrexone on function of murine peritoneal macrophage in vitro was studied in order to illuminate its immune activity futher. Methods:Peritoneal macrophages were divided in three groups:RPMI1640 blank control group, LPS positive control group and NTX treated group. Various phenotypic and functional indices were tested by MTS, flow cytometry technology,phagocytosis experiment and ELISA. Results: Compared with RPMI1640 group,at a LDN exhibits paradoxical properties;the expression of CD64 on surface increased while the expression of CD206 decreased in NTX group;the expression of tumour necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β)were increased. Conclusion:The results of experiment had proved that LDN could influence macrophage polarzation, regulate the inflamatory mediators production and affect the phagocytosis function of peritoneal macrophage.

Objective Deletion detection of Duchenne muscular dystrophy(DMD).Methods Totally 135 patients were tested through polymerase reaction of amplification with 12 dystrophin exons,polyacrylamide gel electrophoresis make gene analysis.Results The deletion of different region was found in 54 patients.Conclusion The deletion region is concentrated in 45～53 exons,the deletion of 48 exons is the peak.

Purpose To compare pharmacokineics of puerarin and crude extract in rats.Methods Rats received 500 mg/kg puerarin and puerarin crude extract by oral administration respectively.Hydroxybenzoic acid was selected as internal standard and the plasma concentration of the puerarin and crude extract was analyzed by HPLC.The pharmacokinetics parameters were calculated with DAS2.0.Results The pharmacokinetics of puerarin and puerarin crude extract was both best fitted with two-compartment models in rats after oral administration,and the pharmacokinetics main parameters of the two formulations were different:the AUC_(0-t) and C_(max) of puerarin were much greater than those of puerarin crude extract,but T_(max),t_(1/(2z)),CL/F and V_z/F were much lesser than those of puerarin crude extract.Conclusion The complex components in pueraria crude extract can affect the pharmacokinetics of puerarin in rat in vivo.

Objective To observe the effect of sustained release type Ⅰ collagen-vascular endothelial growth factor (VEGF) on healing of bone-tendon junction injuries.Methods Partial patellectomy was conducted in 72 rabbits divided equally into control group,type Ⅰ collagen group,and collagen type Ⅰ-VEGF group.The scaffold was planted into the bone-tendon interface.Animals were sacrificed at 4,8 and 12 weeks.New bone formation into the patella-patella tendon surface was detected using X-ray films and histological observations.Quality of bone healing was assayed using biomechanical testing.Results At postoperative 4,8 and 12 weeks,X-ray films showed bone formation of type Ⅰ collagen group [(4.1 ± 0.4) mm2,(12.1 ± 0.5) mm2,(13.0 ± 1.2) mm2 respectively] and of collagen type Ⅰ-VEGF group [(3.8 ± 0.4) mm2,(11.0 ± 0.5) mm2,(13.1 ± 1.0) mm2 respectively] were more than that of control group [(2.1 ± 0.6) mm2,(4.1 ± 0.3) mm2,(6.6 ± 0.6) mm2 respectively] (P ＜ 0.05).Histology identified few new bone,massive fibrocyte accumulation and disrupted alignment of tendon fiber in control group,massive new bone formation,neat and orderly alignment of collagen fiber tissues and massive aggrecan expression at postoperative 4 and 8 weeks (fibrous cartage repair in largely) in collagen type Ⅰ-VEGF group,and massive new bone formation but worse alignment of tendon collagen fibers and less aggrecan expression (fibrous repair in largely) in type Ⅰ collagen group.Biomechanical test showed the ultimate tensile strength increased over time in all groups,with significantly higher value at 12 weeks than that at 4 and 8 weeks.At the same time point,ultimate tensile strength ranged in an order as follows:collagen type Ⅰ-VEGF group ＞ collagen type Ⅰ group ＞ control group (P ＜ 0.05).Conclusion Sustained release type Ⅰ collagen-VEGF can accelerate early healing of bone-tendon junction injury and improve the histological and mechanical properties.

Objective To study the diffusion tensor imaging (DTI)features in acute cervical spinal cord injury (CSCI)and evaluate its clinical value.Methods Eight patients with acute CSCI (within 72 hours after onset)were performed conventional MRI and fast DTI scans (112 seconds)and diffusion tensor tractography (DTT)at 3.0T Siemens Trio Tim system.Meanwhile,the fractional anisotropy (FA) values and apparent diffusion coefficient (ADC)values were calculated separately in the site of lesions,the upper and lower sections to the lesions.Then the data were analyzed by paired-samples t test analysis with SPSS 13.0 software.Results Cervical spinal cord injury occurred likely in the sites of C5-C6 (account for 4/8)and C4-C5 (account for 3/8).All MRI and DTI images were satisfied for clinical diagnosis.The FA value and ADC value of injury lesions were markedly lower than that of the normal cord.Accordingly,the injury lesions on FA map and ADC map presented low signals.There were no significant differences of FA values and ADC values between the upper and lower sections to the lesions.DTT could help in displaying the disruption of spinal fiber tract in lancination case and distortion fibers in closed cervical spinal cord injury.Conclusion Fast DTI sequence at 3.0 Tesla may obtain the qualified spinal cord images.By calculation of FA values and ADC values in CSCI patients,DTI may play an important role in detecting the changes of anisotropy and water diffusion caused by myelin sheath injury and cytotoxic edema and vasogenic edema respectively.

Objective: To examine the status of bullying behaviors and psychological resilience of primary school students and the relationship between the two, and to provide references for related interventions. Methods: Olweus bullying questionnaire and adolescent psychological resilience scale were used to select students in grades 4-6 for questionnaire survey. Results: Primary school students who were bullied, bullied others, double-involved and uninvolved accounted 33.3%, 48.7%, 15.3%, and 2.7%. There were statistically significant differences in bullying in different dimensions of different genders, grades, and previous suicidal thoughts (P<0.05); there were statistically significant differences in the bullied status of only children, divorced parents, and different caregivers (t/F=3.64，2.65，6.62，P<0.05); The status of bullying behavior of students in the class was better than that of ordinary students (t=-2.18, P<0.05); senior students have a higher level of positive cognition (F=8.71, P<0.05); mental toughness of urban students better than rural areas (t=-2.20, P<0.05); students with divorced parents had poor levels of emotional control, family support, and psychological toughness (t=-4.60，-3.92，-3.44，P<0.05); class cadres focused on focus, positive cognition, family support, interpersonal assistance, and psychological resilience were better than ordinary students (t=5.19，6.43,5.64,4.13，6.58，P<0.05); students with parental care had better levels of target concentration, emotional control, family support, interpersonal assistance, and psychological resilience (t=4.59，8.68，8.76，12.35，13.45，P<0.05); the level of psychological resilience of students was low (t=15.90，P<0.05). Correlation analysis shows that there was a negative correlation between bullying behavior and psychological resilience of all pupils (P<0.01), regression analysis showed that bullying behavior may had a negative prediction of mental resilience (F=128.37, R2=0.12, P<0.01). Conclusion The current situation of bullying behavior and mental resilience of grade 4-6 students is worrisome, and it is urgent to improve the current situation of bullying and psychological resilience of primary school students.

Twenty-five pediatric patients (10 males,15 females) with inherited epidermolysis bullosa,aged 3-16 yr,weighing 11-29 kg,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,underwent lysis and reconstructive surgery of both hands adhesions from July 2015 to April 2017 in our hospital.Ketamine 4-6 mg/kg and atropine 0.01 mg/kg were intramuscularly injected at 20 min before admission to the operating room.Oxygen 2 L/min was inhaled by mask after admission to the operating room.Anesthesia was induced by Ⅳ injection of midazolam 0.08 mg/kg and ketamine 2 mg/kg.Pediatric patients kept spontaneous breathing.Anesthesia was maintained by Ⅳ infusion of propofol 5 mg · kg-1 · min-1 and remifentanil 0.05-0.08 μg · kg 1 · min-1.Mepitel was used to protect skin during surgery,and sodium potassium magnesium calcium and glucose injection was intravenously infused to replace the physiological requirement,blood loss and continued loss.Anesthesia time was (130±43) min,surgery time (107±42) min,the amount of intraoperative fluid infused (352± 120) ml,blood loss (29± 10) ml and emergence time (15±6) min.Intraoperative respiratory depression was found in 5 cases (20％);postoperative new bullae occured in 3 eases,and among the 3 cases,one developed on the back and the other two on the buttocks.No cardiovascular events occurred during and after surgery in this study.

Objective To analyze the clinical and molecular genetic characterizations of X-linked adrenal dysplasia congenita(AHC) onset in infant.Methods Seven children (from 7 families) with X-linked AHC who were admitted to the Department of Endocrinology,Genetics and Metabolism,Children's Hospital Affiliated to Zhengzhou University,from July 2012 to June 2017 were selected.All patients were screened for dosage-sensitive-sex reversal-adrenal hypoplasia congenital critical region on the X chromosome gene1 (DAX1/NR0B1) mutations.The clinical manifestation and laboratory examination were analyzed,their clinical characterizations were summarized.Results Seven patients were all male,the onset age of the patients were from after birth to 7 months old,and 4 patients (4 families) had a family history of X-linked recessive inheritance.The clinical manifestations were skin pigmentation [100.0％ (7/7 cases)],vomiting [71.4％ (5/7 cases)],no weight gain [57.1％ (4/7 cases)] and poor spirit [28.6％ (2/7 cases)].Laboratory tests showed that hyperkalemia and hyponatremia,increased coricotrophin,normal or decreased cortisol,17α-hydroxyprogesterone,progesterone,aldosterone and dehydroepiandrosterone.Testosterone levels increased in 5 patients.The abnormalities of adrenal glands imaging could be seen in 2 patients.Two patients were misdiagnosed as congenital adrenal cortical hyperplasia.Then the definitive diagnosis were made by genetic test.DAX1/ NR0B1 gene mutations were found in all patients.Five patients were novel mutations (c.114_126del,c.872G ＞ A,c.56delG,c.884T ＞ G,c.1217delG).Conclusions The clinical manifestations of X-linked AHC with infant onset include pigmentation,poor spirit and growth retardation,which should be differentiated from congenital adrenal cortical hyperplasia.Hormone levels such as elevated blood 17α-hydroxyprogesterone and family history are the main identification points,and AHC cannot be excluded when testosterone level increases.Five novel mutations are found in this study,which enrich the gene database.