Objective To screen the Doc 1R gene recombinant plasmid by use of colony PCR. Method The recombinant colonies were transfered into the PCR reaction mixture. The PCR primers were used for constructing mouse Doc 1R genomic sequence. Result Among the 5, 3 positive strips in the size of 1 500 bp were visible, which were the same as the Doc 1R gene fractions in terms of their sizes were screened as positive clones. The positive colony were further confirmed by double digestion and DNA sequencing. Conclusion Colony PCR is a simple, efficient and reliable technique for screening the recombinant.

Interdisciplinary can be acted in any stages of researching procedure.The interdisciplinary process research thinking were addressed including how to builds variables,which are depth and width,fixed discipline or not,and also to reflect the collaboration level of interdisciplinary in certain period.By this quantity evaluation mode building,it will useful for the interdisciplinary research in the future,especially in medical and life science fields.

ObjectiveTo investigate the effects of platelet derived growth factor (PDGF) on brain cell apoptosis rate and serum neuron-specific enolase (NSE) concentration after hypoxic-ischemic brain damage (HIBD) in neonatal rats. MethodsForty-eight HIBD models of 7-day old neonatal Wistar rats were established and then divided into two groups randomly:PDGF group and normal saline control group (n =24 in each).Another 24 neonatal Wistar rats were taken into the sham operation group.The treatment group received intraperitoneal injection of PDGF-BB (50 ng/kg) once,while the other two groups received normal saline at the same time.In each group,rats were randomly sacrificed immediately at 12,24 and 72 hours after injection (n=8).The serum of rats were reserved for NSE concentration determination by enzyme linked immunosorbent assay,and the right brains of the sacrificed rats were used to prepare brain cell suspension for neurocyte apoptosis rate examination by flow cytometry.Mono-variate analysis and q-test were performed for statistical analysis. Results(1) The brain cell apoptotic rates of treatment group [ (6.09 ± 0.70)％,(9.67 ± 1.52) ％ and (14.15±1.52)％] and control group [(8.00± 1.10)％,(11.45±2.42)％ and (22.90±2.03) ％] were significantly increased compared to that of sham group [(2.11 ± 0.54)％,(2.34 ±0.46)％ and (2.21±0.49)％] at all time points (all P＜0.01 or ＜0.05),the apoptotic rate of treatment group was lower than that of control group (P＜0.01 or ＜0.05).Statistical differences were found among the three groups at 12,24 and 72 hours (F =39.01,66.60 and 194.20respectively; P＜0.01).(2) Serum NSE concentration was significantly increased in the treatment group [(8.43 ± 0.17) μg/L,(6.73 ± 0.16) μg/L and (6.12 ± 0.13) μg/L] and control group [(10.04±0.19) μg/L,(9.330.15) μg/L and (8.36 ± 0.16) μg/L] than in the sham group [(4.22±0.53) μg/L,(3.96±0.60) μg/L and (3.59±0.55) μg/L] at all time points,and it was significantly lower in treatment group than in control group (P＜ 0.01).Statistical difference was found among three groups at 12,24 and 72 hours (F=371.25,245.61 and 236.22 respectively,P＜0.01). ConclusionsPDGF might have neuroprotective effect,which could inhibit apoptosis of neural cells and decrease the serum NSE concentration.

Objective:To analyze the clinical features, treatment and prognosis of asymptomatic patients with retinoblastoma.Methods:A retrospective series of case study. Eight asymptomatic patients (11 eyes) with the diagnosis of retinoblastoma by screening enrolled in Department of Ophthalmology of The Eye-ENT Hospital of Fudan University from January 2006 to March 2019 were included. There were 6 males and 3 females ranging from 2 days to 20 months, with a median age of 6 months. Five patients were unilateral retinoblastoma while 3 patients were bilateral. Based on the International Classification of Intraocular Retinoblastoma, 4 eyes were stage A, 3 eyes were stage B and 4 eyes were stage C. One patient had family history. Four patients were evaluated the Rb1 mutation. Routine ophthalmic examinations and ultra-wide field fundus imaging were performed on the 16 parents and 3 siblings of the 8 patients. Systemic intravenous chemotherapy was performed using the Carboplatin, Vincristine, Etoposide protocol, intra-arterial chemotherapy using Carboplatin and Melphalan, and local treatment involved cryotherapy and transpupillary thermotherapy. The mean follow-up time is 47.25 months.Results:None of the 8 children had any ocular symptoms. Six patients received intravenous chemotherapy (5-6 times), 1 patient received intra-arterial chemotherapy (3 times), and 1 patient just received local treatment. Among the 11 eyes, 9 eyes were treated with local cryotherapy and 8 eyes were treated with transpupillary thermotherapy. During the follow-up period, 2 patients had new tumor, and the average time was 6.3 months after the last chemotherapy. At the last follow-up, the tumor disappeared in 11 eyes, remained stable in 11 eyes. The eye protection rate was 100% (8/8) for patients without eyeball excision. The best corrected visual acuity was 0.1 for 3 eyes and 1.0 for 5 eyes. Three eyes were not found. One heterozygous mutation of Rb1 gene [1c.35_69del (p.T12fs)] was identified in 1 patient, and the other 3 patients were not detected. One had bilateral bulbar tuberculosis of the 16 parents, 1 had bilateral RB of the 3 siblings. They were the mother and brother of a child with bilateral RB.Conclusions:Fundus screening is helpful for the detection of early RB. The eye protection rate is high and the long-term vision prognosis is good after systemic or topical chemical drugs (IVC, IAC) and ocular topical treatment (cryopreservation and transpupillary thermotherapy).

Objective:To systematically review parents ′ experience of caring for children with chronic kidney disease (CKD) to fully understand care needs and improve the psychological state and caregiving quality of parents. Methods:The qualitative studies on parents ′ experience of caring for children with CKD were retrieved from following databases, including PubMed, Web of Science, CNKI, VIP, CBM, and WanFang Data from inception to March 2020. The quality of included researches was evaluated according to the JBI Critical Appraisal Tool for qualitative studies in Australia. The results were integrated by pooled integration methods. Results:A total of 14 studies were included. 69 results were summarized and integrated to form 10 categories. These categories extracted 4 integrated results: parents' physical and mental condition are affected, relationship between parents and their support system has changed, reconstruction of parents ′ life, unmet care needs and problems. Conclusions:Parents are crucial to the disease management of children with CKD, so clinical medical staff should not only provide medical services for children, but also pay more attention to the psychological status and needs of parents, so as to provide guidance and support to promote parents to better implement care and disease management for children.

Objective:To explore the monitoring value of left ventricular functional parameters obtained by bedside ultrasound combined with clinically relevant indicators in patients with veno-arterial extracorporeal membrane oxygenation (VA-ECMO).Methods:A retrospective study was conducted. A total of 24 patients receiving VA-ECMO adjuvant support in Renmin Hospital of Wuhan University from June 2018 to January 2020 were selected. The bedside ultrasound was performed on the first day of ECMO support, the day before weaning, the clinical indicators before weaning were obtained. The differences in clinical indicators and the left ventricular functional parameters between the two groups of whether weaning successfully were compared; univariate Logistic regression analysis was used to screen out the related factors affecting weaning.Results:Sixteen patients were successful weaned and 8 patients failed. Compared with the weaning failure group, patients in the weaning success group required less continuous renal replacement therapy (CRRT, cases: 4 vs. 6, P < 0.05), mean arterial pressure (MAP) before weaning was higher [mmHg (1 mmHg = 0.133 kPa): 84.64±9.55 vs. 62.30±8.79, P < 0.05], and the pulse oxygen saturation (SpO 2) was also higher (0.966±0.670 vs. 0.866±0.061, P < 0.05), while vasoactive-inotropic score (VIS), serum creatinine (SCr) and serum lactic acid (Lac) were lower [VIS score: 7.27±1.42 vs. 16.93±8.52, SCr (μmol/L): 123.60±83.64 vs. 213.10±117.39, Lac (mmol/L): 1.94±0.91 vs. 5.62±5.48, all P < 0.05]. Univariate Logistic regression analysis showed that the MAP, VIS, SCr, Lac, SpO 2 before weaning were the related factors affecting weaning [odds ratio ( OR) were 0.306, -0.740, -0.011, -0.632, -4.069; 95% confidence interval (95% CI) were 1.065-1.732, 0.235-0.899, 0.979-0.999, 0.285-0.992 and 0.001-0.208; P values were 0.014, 0.022, 0.038, 0.047, 0.002]. In the weaning success group, left ventricular ejection fraction (LVEF), velocity of mitralannulus in systolic (LatSa), maximum flow velocity of aortic valve (AV-Vmax), velocity-time integral (VTI), left ventricular global longitudinal strain (LVGLS), left ventricular global longitudinal strain rate (LVGLSr) were all increased on the day before ECMO weaning compared with the first day of ECMO support [LVEF: 0.40±0.05 vs. 0.28±0.07, LatSa (cm/s): 6.81±0.91 vs. 4.62±1.02, AV-Vmax (cm/s): 104.81±33.98 vs. 64.44±16.85, VTI (cm): 14.56±3.11 vs. 7.96±1.98, LVGLS: (-8.95±2.59)% vs. (-5.26±1.28)%, LVGLSr (1/s): -0.48±0.11 vs. -0.29±0.09], whereas the ECMO flow was significantly reduced (L/min: 1.46±0.47 vs. 2.64±0.31), the differences were statistically significant (all P < 0.05). There was no significant difference in left ventricular functional parameters between the first day of ECMO support and the day before ECMO weaning in the weaning failure group. Compared with the weaning failure group, the weaning success group had higher LVEF, LatSa, AV-Vmax, VTI, LVGLS, LVGLSr on the day before ECMO weaning [LVEF: 0.40±0.05 vs. 0.26±0.07, LatSa (cm/s): 6.81±0.91 vs. 4.31±1.03, AV-Vmax (cm/s): 104.81±33.98 vs. 67.67±18.46, VTI (cm): 14.56±3.11 vs. 7.75±2.77, LVGLS: (-8.95±2.59)% vs. (-4.81±1.81)%, LVGLSr (1/s): -0.48±0.11 vs. -0.30±0.10, all P < 0.05] and lower ECMO flow (L/min: 1.46±0.47 vs. 2.20±0.62, P < 0.05). Conclusion:Bedside echocardiographic left ventricular function parameters (LVEF, LatSa, AV-Vmax, VTI, LVGLS, LVGLSr) combined with clinical indicators (MAP, VIS, SCr, Lac, SpO 2) were helpful to evaluate the therapeutic effect of patients receiving VA-ECMO support and can provide important guiding value in the selection of VA-ECMO weaning timing and the judgment of prognosis.

BACKGROUND:Great development has been made in the research of various repair materials and tissue-engineered bone, and different materials exhibit a strong ability to repair bone defects. OBJECTIVE:To review the research status and progress of repair materials for bone defects. METHODS:PubMed, PMC, CBM, CNKI and WanFang databases were used online to retrieve the relative articles regarding repair materials for bone defects published from June 2007 to June 2017. The key words were "biomaterials, bone repair materials, bone substitute, bone tissue engineering, bone defect, bone repair" in English and Chinese, respectively. Finally, 66 eligible articles were included, while the unrelated and repeated ones were excluded. RESULTS AND CONCLUSION:The main repair materials for bone defects include autologous bone, allogeneic bone, xenogeneic bone, demineralized bone matrix, bioceramics, metal materials, polymer materials,tissue-engineered bone and relative derived composites. Different materials have shown certain advantages and disadvantages. Clinicians should make an optimal choice in view of the characteristics of materials. Although there is a great progress in thestudies of repair materials for bone defects, further investigations are warranted on how to produce highly biomimetic materials, optimize cell adhesion and migration, precisely control the expression and release of related genes and growth factors, validate clinical safety, and provide medical evidence in clinical practice.

Objective To evaluate the relationship between serum matrix metalloproteinase-9(MMP-9), white blood cell(WBC)count and type 2 diabetes mellitus with macroangiopathy and to investigate the mechanism of the protective effects of rusiglitazone(RSG)on blood vessel.Methods Serum MMP-9 was determined by ELISA in 30 normal controls and 80 type 2 diabetic patients(including 40 cases with macroangiopathy and 40 without maeroangiopathy).WBC count and other clinical parameters were also determined.32 type 2 diabetic patients received RSG(4rag qd)for 12 weeks.All parameters were determined after 4 weeks(17 patients)and after 12 weeks(all patients)to observe the changes in MMP-9,WBC and other parameters.Results In the diabetic patients,serum MMP-9 and WBC were markedly higher as compared with normal controls;and MMP-9 and WBC in patients with macroangiopathy[579(440-949)?g/L,(7.51?1.47)?10~9/L]were higher than those [324(275-423)?g/L,(6.22?0.79)?10~9/L]in the cases without macroangiopathy(P

Objective To detect the expressions of nm23-H1 and heat shock protein 27 (HSP27) and their clinical significance on development and metastasis in non-small cell lung carcinoma (NSCLC).Methods 75 tumor tissues from patients with NSCLC were included as experimental group and 28 pulmonary benign lesion tissues were as control group.The expressions of nm23-H1 and HSP27 in patients with different clinical and pathological characters were detected by immunohistochemistry.Results nm23-H1 and HSP27 were mainly expressed in cytoplasm,the positive rates of nm23-H1 and HSP27 were significantly higher in the experimental group than that in control group [41.3 ％ (31/75) vs 7.1％ (2/28),x2 =10.946,P =0.001,80.0 ％ (60/75) vs 46.4 ％ (13/28),x2 =11.131,P =0.001].Compared with control group,the positive rate of HSP27 was correlated with the degree of tumor differentiation (x2 =4.191,P =0.041).nm23-H1 was related with HSP27 in lung cancer (r =0.284,P =0.013).Conclusion nm23-H1 and HSP27 are related to the occurrence and development of NSCLC.The joint detection of nm23-H1 and HSP27 should be helpful to the diagnosis and judge the biological behavior of NSCLC.

Objective To detect the expression of human epithelial growth factor receptor 2 (HER-2) in advanced lung adenocarcinoma with epithelial growth factor receptor (EGFR) mutation, and to explore the potential of HER-2 as a therapeutic target for drug resistance in patients with EGFR mutations. Methods HER-2 is commonly expressed in the advanced lung adenocarcinoma with EGFR mutations, mainly in the cell membrane. Results The overexpression rate of HER-2 protein in tissues of advanced lung adenocarcinoma with EGFR mutations was 33.3%(28/84). The overexpression rate of HER-2 protein in patients>50 years of age was 40.3%(27/67), which was significantly higher than that of patients ≤50 years of age [5.9 % (1/17)], the difference was statistically significant (χ2=7.227, P=0.007). The overexpression rate of HER-2 protein in patients with high pathological differentiation [44.4 % (8/18)] was higher than that in patients with poor pathological differentiation [30.3%(20/66)], but the difference was not statistically significant (χ2=1.273, P=0.259). The overexpression of HER-2 protein in patients with EGFR 21 exon mutation [40.5 % (17/42)] was significantly higher than that of EGFR19 exon mutation [25.0%(10/40)], but the difference was not statistical significance (χ2=2.222, P=0.136). Conclusions The overexpression rate of HER-2 protein in advanced lung adenocarcinoma patients with EGFR mutation is high, which is related to the age and tumor differentiation. HER-2 is expected to be a potential therapeutic target for drug resistance patients with EGFR mutations.