Adolescent ; Classical Lissencephalies and Subcortical Band Heterotopias ; diagnosis ; Female ; Humans

Objective To knock out the GP73 gene in H22 cells originating in mice using CRISPR/Cas9 gene editing system and construct H22 GP73 gene knockout stable strain for identification of its functions .Methods Two pairs of sgRNAs that could specifically identify the upstream and downstream of GP 73 gene first promoter were designed before a recombinant eukaryotic expressional plasmid was constructed using pX 459 .After enzyme digestion and sequencing , two pairs of recombinant plasmids were co-transfected into H22 cells before puromycin was used to screen positive cells and monoclonal cells which stably knocked out GP 73 gene were developed .The knockout effect was measured by Western blotting.Cell Titer 96? AQueous One Solution Assay was used to detect the effect on cell reproductive capacity when the GP73 was knocked out .The transferability was detected through wound healing test .Results The result of Western blotting suggested that GP73 protein was undetected in the construction of H22 GP73 knockout gene stable strain after transfection.The transfer and reproduction slowed down .Conclusion H22 GP73 gene knockout stable strain can be successfully built using CRISPR/Cas9 gene editing system ,thus facilitating studies on the function of GP 73 in hepatocarcinogenesis .

Objective:To observe the effects of bama nectar in kidney-yang deficienct mice and on the mating ability and sperm counts of normal mice. Methods:The weight, body temperature, locomotor activity, cold-resistant,genital organ coefficient and lipid peroxidation of the hydrocortisone-induced kidney-yang deficient mice were detected. The genital organ ccoefficient, immune organ co-efficient and erectile function of the ovariectomized mice were detected. The mating ability and the sperm counts of normal mice were measured. Results:Compared with the model group, bama nectar obviously increased the weight, body temperature and locomotor ac-tivity of the hydrocortisone-induced kidney-yang deficient mice, and improved the locomotor activity, the organ coefficient of seminal vesicle, prostate gland, preputial gland and epididymis. Furthermore, bama nectar significantly increased the SOD activity in serum and reduced the level of MDA in serum when compared with the model group (P<0.05 or P<0.01). Compared with the model group, bama nectar could increase the organ coefficient of foreskin and thymus gland in the ovariectomized mice, and shorten the erec-tile latency (P<0. 05). Compared with the normal group, bama nectar improved the mating ability and the sperm counts of the male mice (P<0. 05 or P<0. 01). Conclusion:Bama nectar has beneficial effects on invigorating kidney and strengthening yang.

Objective To approach the application value of diffusion weighted imaging with background suppression (DWIBS) combined with routine MR sequence in differentiating benign and malignant lymph nodes,and assessing therapeutic effect of lympho-genic tumors.Methods 48 patients with cervical lymph node enlargement who were confirmed by pathology and follow up under-went DWIBS and routine MRI examination,malignant lymph nodes were 83,benign lymph nodes were 79.16 patients with malig-nant lymph nodes were rechecked after radiotherapy and chemotherapy,Apparent diffusion coefficient (ADC)values of the solid part for lymph nodes were compared.Results More lymph nodes can be detected in DWIBS than conventional sequence.ADC values of the solid part for malignant lymph node(0.898±0.111)×10-3 mm2/s were lower than that of benign lymph node(1.043±0.106)× 10-3 mm2/s,there was significant difference between them (P<0.05).ADC values of all malignant lymph nodes after treatment (1.205±0.121)×10-3 mm2/s were significantly higher than that of pretherapy (0.883±0.090)×10-3 mm2/s (P<0.05).Conclu-sion DWIBS could more sensitively detect lymph node than conventional MR sequence.ADC value could provide some reference values for differentiating benign and malignant lymph nodes and assessing therapeutic effect.

Objective To study the imaging characteristics of the ligaments in craniocervical junction (CCJ), and to optimize the examination methods and scanning sequences of the ligaments in this region. Methods Two groups of 51 healthy volunteers in each were selected to undergo CT and MRI examination respectively. The CT and MRI features of the ligaments in CCJ were assessed. Two senior imaging doctors compared the results of showing the ligaments in CCJ by several MRI sequences, including T1 WI ,T2 WI, proton density weighted imaging (PDWI), T2 * WI and short time inversion recovery (STIR) ,and proton density fat saturate (PDFSAT). Standard normal rank transformation was done according to the primary data,and then analysis of variance of repeated measurement was applied. Results CT and MRI could both demonstrate the ligaments (except the anterior atlantooccipital membrane only seen on MRI) and their adjoins in CCJ, while MRI had more advantages than CT. The display ratio of the anterior atlantooccipital membrane was 100% (51/51) by MRI. The display ratio of the apical ligament was 29.4%(15/51) by CT and 43.1% (22/51) by MRI. The posterior atlantooccipital membrane-dura complex,tectorial ligament-dura complex, transverse ligament, and alar ligament could be demonstrated clearly by CT and MRI, the display ratio was 100% respectively (51/51). The results of PDWi scored by two doctors were both 5.0, there were no significant differences between them (F = 0.000, P > 0.05), which were significantly higher than T, WI (M = 3.0), T2Wi (M = 3.0), T2 * WI (M = 1.0), STIR (M = 1.0), and PDFSAT(M = 3.0)(P < 0.01). Conclusion MRI was superior to CT in demonstrating the ligaments in CCJ. PDWI was the optimal MRI sequence. Imaging research of normal ligamentous structures in CCJ could provide valuable diagnostic and therapeutic information for evaluating the ligamentous diseases.

Objective To investigate the influence of angelica sinenisis injection on bone marrow histology and ultrastructure in immune-induced aplastic anemia(AA) mice.Methods Female Balb/c mice were divided into normal group,model group,therapy group and contrast group.All mice were killed by cutting neck on the 12 th day, ulnas and femur were taken out. The bone marrow histology section and ultrastructure of mice ulnas were observed. The quantities of hematopoietic cells were counted. The number of femur bone marrow mononucleate cells (MNC) were measured.Results The quantities of hematopoietic cells in model group were lower than those in normal group (P

OBJECTIVE:To evaluate the effects of PDCA cycle management on perioperative prophylactic application of antibiotics in typeⅠincision surgery of orthopedics department. METHODS:In retrospective analysis,512 discharge medical records, 861 ones and 1 070 ones were selected from our hospital before PDCA cycle management(Jan.-Dec. 2013,before intervention group), after first cycle of PDCA cycle management (Jan.-Dec. 2014,first intervention group) and after second cycle of PDCA cycle management(Jan.-Dec. 2015,second intervention group),respectively. The perioperative prophylactic application of antibiotics was analyzed comparatively before and after continuous intervention. RESULTS:After 2 cycles of PDCA cycle management intervention, the constituent ratio of internal fixation in orthopedics department increased significantly;utilization ratio of antibiotics,the rate of rational type,medication ratio 0.5-1 h before surgery,the rate of rational treatment course,the proportion of antibiotics use in accordance with indications increased from 50.20%,98.08%,93.77%,6.61%,82.10% to 58.41%,100%,99.04%,52.00%, 99.04%,respectively. The number of antibiotics type decreased from 4 to 2;the proportion of cephazolin increased significantly while that of clindamycin decreased significantly compared to before intervention. Postoperative prophylactic medication course decreased from(4.63 ± 2.42)d to(1.61 ± 0.75)d;the proportion of patients with medication course <24 h or ranged 24-48 h increased significantly, while those with medication course>72 h decreased significantly, with statistical significance (P<0.05). CONCLUSIONS:PDCA cycle management improves the rational rate of perioperative prophylactic application of antibiotics in typeⅠincision surgery of orthopedics department in our hospital. The prophylactic medication course of our hospital is not yet fully controlled within 24 h so it should be further intervened.

Objective To investigate the application value of diffusion-weighted imaging with background suppression (DWIBS) in lymph node metastasis of rectal cancer.Methods 30 patients with rectal cancer diagnosed by colonoscopy underwent routine MRI and DWIBS examinations.Metastatic and non-metastatic lymph nodes were confirmed by postoperative pathology.Apparent diffu-sion coefficient (ADC)values of the solid part of metastatic and non-metastatic lymph nodes were measured and compared.ROC curve analysis was used to define the threshold between the metastatic and non-metastatic lymph nodes.Results In all 73 lymph nodes,there were 52 metastatic lymph nodes and the other 21 were non-metastatic.ADC values of the solid part of metastatic and non-metastatic lymph nodes were (0.881 ± 0.094)× 10 -3 mm2/s and (1.072 ± 0.108)×10 -3 mm2/s respectively,the difference was statistically significant (P <0.05).With the threshold ADC value of 0.957×10 -3 mm2/s,the sensitivity and specificity for dif-ferentiating metastatic from non-metastatic lymph nodes were 90.5% and 80.8% respectively.The diagnostic value was excellent (Az value = 0.91 9,P <0.05).Conclusion ADC values may be used to differentiate lymph node metastasis of rectal cancer,and DWIBS has higher application value in assessing lymph node metastasis of rectal cancer.

Objective Evaluate the effects of venous-arterial modified ultrafiltration on hemodynamics compared to arterial-venous in children undergoing cardiopulmonary bypass (CPB) for repair of congenital heart defects. Methods Forty patients underwent MUF randomly divided into two groups,group V-A MUF (n =20) and group A-V MUF (n =20) for 10 min after CPB. They were studied before CPB, after CPB, 10 min after CPB, and 30 min after CPB. Haemodynamic data including heart rate, blood pressure, central venous pressure and hematocrit were recorded. Transoesophaegeal echocardiography determined left ventricular posterior wall thickness in end-systole ( LVPWs) and end-diastole (LVPWd) , end diastolic volume (EDV) , end systolic volume (ESV) and ejection fraction (EF) were measured and compared in two groups. Results Patients in V-A MUF maintained better systolic arterial blood pressure at 10 min and 30 min compared with 0 min values after CPB. A significant decrease in EF were observed in both groups immediately after CPB ( P < 0.05 ). Significant increase in EF was observed at 10 min (60% ) and 30 min (46% ) after CPB compared with 0 min value after bypass in V-A MUF (P <0.001 ). In A-V MUF, no such increase in EF was observed. EF were significantly higher at 10 min and 30 min in V-A MUF as compared with A-V MUF (P < 0. 001). There was also significant improvement in posterior wall thickness in V-A MUF (P <0.05). Haematocrit values were not different in duration of postoperative between two groups. Conclusion Veno-arterial modified ultrafiltration is a safe and effective method of improving hemodynamics in children following cardiac surgery.

Objective To establish a TaqMan real-time fluorescent quantitative PCR to detect Vibrio vulnificus based on hemolysin gene(vvhA)that coding cytolysin.Method By using software Primer Express, the PCR primers and TaqMan probe,which located in the conserved region of vvhA gene sequence,were designed for establishment of a TaqMan real-time fluorescent quantitative PCR to detect 100 bp amplicon from V.vulnificus DNA.A recombinant plasmid pMD19-vvhA100 as a positive control during detection was constructed using gene cloning technique.Minimal amplification cycles(Ct value)and fluorescence intensity enhancement (△Rn value)were used as observing index to optimize the reaction conditions of the TaqMan real-time fluorescent quantitative PCR.The DNAs with different concentrations from V.vulnificus and other eight bacteria and pMD19- vvhA100 were applied as templates to determine the specificity,sensitivity and reappearance of the TaqMan real- time fluorescent quantitative PCR.ICR mice were intraperitoneally,subcutaneously and orally infected with V. vulnificus,respectively.The detection effect of the TaqMan real-time fluorescent quantitative PCR was measured using the specimens of peripheral blood,subcutaneous tissue and intestinal content collected from the infected mice.Results The established TaqMan real-time fluorescent quantitative PCR showed positive results only for V. vulnificus DNA and pMD19-vvhA100.The detection effectiveness of the TaqMan real-time fluorescent quantitative PCR was as high as 0.01 ng of V.vulnificus DNA or 103 copies of pMD19-vvhA100.The SD values of the detection results repeated for three times using pMD19-vvhA100 with different concentrations were lease than 0.79. The detection results of TaqMan real-time fluorescent quantitative PCR were positive for all the specimens of peripheral blood and subcutaneous tissue.Conclusions The TaqMan real-time fluorescent quantitative PCR established in this study for V.vulnificus vvhA gene detection has advantages such as quickness,stability, sensitivity and specificity,indicating this method can be used for clinical laboratory diagnosis of septicemia and wound infection caused by V.vulnificus.