Triple negative breast cancer (TNBC)is difficult to benefit from endocrine therapy or tras-tuzumab targeted drug therapy.Biological overexpression of breast cancer susceptibility gene 1,p53 gene,vas-cular endothelial growth factor and microRNA suggests that TNBC is easy to metastasis and recurrence and has a poor prognosis.Exploring the molecular subtypes of TNBC,setting out the treatment plan for subtypes and finding the corresponding monoclonal antibody targets are the research direction of TNBC in the future.

The directed molecular evolution of enzyme in vitro can not only improve the efficiency of evolution, but also evolve enzyme according to the investigator's desire. This review summed up the feasible methods of this novel technique.

Objective To screen and clone the target genes transactivated by nonstructural protein 5A (NS5A) of hepatitis C virus (HCV). Methods The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-NS5A and pcDNA3.1(-) empty vector, respectively. Suppression subtractive hybridization (SSH) method was employed to analyze the differentially expressed DNA sequence between the two groups. The obtained sequences were searched for homologous DNA sequence from GenBank. The new gene with no homology with known genes in this database was confirmed, and electric polymerase chain reaction was conducted for cloning the full-length DNA of the new gene and in conjunction with Kozak rule and the terminus of polyadenyl signal sequence. The reverse transcription PCR (RT-PCR) was used to amplify the new gene from mRNA of HepG2 cell as the template. The coding sequence of the new gene was deduced according to the nucleotide sequence. Results A new gene with unknown function was named as NS5ATP3. The nucleotide sequence of the NS5ATP3 gene and its corresponding amino acid have been determined, which contained 1 572nt and 524aa. The sequence of the NS5ATP3 gene was deposited into GenBank, with the accession number AF529364. Conclusions NS5ATP3 gene transactivated by HCV NS5A protein was cloned and identified successfully by combining molecular biological technology and bioinformatics technique. These results will pave the way for the study of the molecular mechanism of the transactivating effects of HCV NA5A protein and the development of new therapy for chronic hepatitis C.

Objective:To study the relationship between sleep quality and mental health of pilots.Method:356 pilots in their recuperation at our sanatorium were evaluated with Pittsburgh Sleep Quality Index and Mental Health Inventory. Result:Good sleepers were significantly different to the poor ones in mental health indexes. The score of sleep quality was positively correlated with that of self-confidence, energy,character,over self-concern, anxiety, optimism, sensitivity,nervousness mental bealth and self-assertion (r=0.31-0.47,P

Objective To explore the changes of expression of tight junction protein occludin in the brain microvascular endothelial cells of rats with acute necrotizing pancreatitis (ANP), and to study the relationship between the occludin expression and degree of pancreatic pathologic changes. Methods 80 rats were randomly divided into sham-operation (SO) group and ANP 3 h, 6 h, 12 h, and 24 h groups. Retrograde pancreatic duct injection of 5% choleate sodium in rats was used to establish ANP models. Pancreatic pathologic changes was evaluated. Immunohistochemistry, Western blot and RT-PCR were used to determine the expressions of the tight junction protein occludin in the brain microvascular endothelial cells. Results Occludin protein was linearly expressed along brain vascular vessels. The expression of occludin protein at 3h was 0. 35±0.09 in brain tissues from rats with ANP and 0. 49±0.08 in SO group, the expression of occludin mRNA were decreased from 1.50±0.30 to 1.01±0.18 (P < 0.05), the lowest expression rate occurred at 6 h, and the values in the two groups were 0. 26±0.07 and 0. 93±0. 19, respectively, the expressions of occludin protein and mRNA of ANP group at 12 h, 24 h were notably increased when compared with those at 6 h (P < 0. 05), but they were still lower than those in SO group (P < 0.05). The occludin protein and mRNA expressions were negatively related with the degree of pancreatic pathologic changes (Pearson correlation coefficient: -0.48 and -0.536, P < 0.05). Conclusions The expression of tight junction protein occludin decreased at both protein and mRNA level in brain microvascular endothelial cells of ANP rats. Furthermore, the expression of occludin was negatively correlated with pathological change of pancreas.

Objective To study the changes of learning-memory ability and synapse of hippocampus after radiation injuries. Methods Sprague-Dawley rats were grouped according to radiation dose to 20Gy group,30Gy group radiated by linear accelerator and control group were used before radiation and 120 days after radiation. Morris maze test were taken to study the learning and memory ability of rats in each group. Average escape latency and search strategy were scaled and analyzed in each group. The parameters of synapse in CA3 area of hippocampus were studied by using electron microscope and image analyzer. Results AEL of 20Gy group was (41. 17 ±10.76 ) s and score of SS was 27.13 ± 2.34 after 120 days' radiation but AEL of 30 Gy group was (78.49 ± 9.32)s and the score of SS was (23.19 ± 7.65 ) nm. There were significant statistic differences Compared with control group and before radiation (P ＜ 0.05 ). The thickness of PSD of 20 Gy group was ( 22.03 ± 6.84 ) nm after 120days' radiation and (23.19 ± 7.65 )nm in 30 Gy group. There were significant statistic differences compared with control group and before radiation. It was observed that both in 20 Gy and 30 Gy group' s the length of synaptic activity area was shorter,the curvature of synaptic interface was smaller,the width of synaptic cleft and the thickness of PSD was narrower than that of control group. Conclusion There was close relation between the changes of learning-memory ability and synapse of hippocampus after radiation injuries.

Objective To investigate the effects of airway pressure release ventilation (APRV) in children with severe pneumonia-related acute respiratory distress syndrome(ARDS).Methods Ten children suffering severe pneumonia-related ARDS with APRV were included in Pediatric Intensive Care Unit, Beijing Children's Hospital,Capital Medical University from March 2011 to October 2014.Ventilation variables, changes of airway pressure and Ramsay scores were collected and compared with that in conventional ventilation (CV).Clinical variables were measured at CV before APRV and at 1,4,12,24 hours after transition to APRV.Results High airway pressure(Phigh) at each time point during APRV was significantly lower than peak airway pressure (Ppeak) or plateau airway pressure (Pplat) in CV[(26.00 ±2.94) cmH2O(1 cmH2O =0.098 kPa) ,(24.40 ±3.34) cmH2O,(23.30 ±3.46) cmH2O,(23.00 ± 3.80) cmH2O vs (31.80 ± 5.59) cmH2O, P ＜ 0.01].Mean airway pressure (Pmean) at each time point during APRV was significantly higher than that in CV [(23.00 ± 2.86) cmH2 O, (21.69 ± 3.12) cmH2 O, (20.89 ± 3.31) cmH2 O, (20.46 ± 3.48) cmH2 O vs (17.50 ± 2.37) cmH2 O, P ＜ 0.05].Fraction of inspired oxygen (FiO2) were significantly decreased at 4, 12 and 24 hours after APRV than that in CV [(73.00 ± 22.39) ％, (63.50 ± 20.16) ％, (63.00 ± 21.11) ％ vs (88.00 ± 15.49) ％, P ＜ 0.05].Ramsay scores were significantly decreased at each time point during after APRV than that in CV [(3.90 ± 0.74) scores, (2.90 ± 0.88) scores, (3.00 ± 1.15) scores,(3.50 ± 0.71) scores vs (4.60 ± 0.52) scores, P ＜ 0.05].Conclusions Compared with CV, APRV had a lower Phigh and FiO2 ,a higher Pmean and more shallow sedation.APRV may be an effective ventilation mode in children's severe pneumonia-related ARDS.

Objective Tubulointerstitial fibrosis(TIF) is the most important marker reflecting the degree of renal function decline and prognosis and hydrogen sulfide ( H2 S) is crucial in maintaining normal renal function and many diseases of renal injury. The aim of the article was to investigate the effects of exogenous H2 S on the expressions of angiotensinⅡ ( AngⅡ) , proliferating cell nuclear antigen (PCNA) and transforming growth factor beta-1 (TGF-β1) in rats with unilateral ureteral obstruction (UUO). Methods TIF rat model was built with UUO. Ninety-six SD rats were randomly divided into four groups:sham operation group, modelgroup, UUO+low-dose NaHS treatment group ( low dose group) and UUO+high-dose NaHS treatment group ( high dose group) ( n=24, respectively) . Rats in model group were treated with left-side ureteral obstruction and ureteral separation without obstruction was done in sham operation group. UUO rats in two treatment groups were injected intraperitoneally with two different doses of sodium hydrosulfide (NaHS, donor of endogenous H2 S), respectively. HE and Massonstaining and immunohistochemical staining were performed at the 7 d, 14 d and 21 d, respectively. Results In sham operation group, the expressions of AngⅡ, PCNA, and TGF-β1 were found in microamount in tubulointerstitium at each time points. Compared with sham operation group, the expressions of AngⅡ, PCNA and TGF-β1 in model group increased( P<0.01) . While in comparison to model group, the expressions of AngⅡ, PCNA and TGF-β1 decreased in low dose group and high dose group, but no significant differ-ence was found between low dose group and high dose group. Conclusion Exogenous H2 S supplementation can attenuate TIF partly via downregulating the expressions of AngⅡ, PCNA and TGF-β1 .

Objective To investigate the clinical characteristics and treatment of external auditory canal chol‐esteatoma (EACC) .Methods The clinical data of 38 cases(39 ears)with external auditory canal cholesteatoma from August 2006 to December 2014 were retrospectively analyzed .Results All the cases of EACC in this study had the external ear canal full of impacted squamous material or granulation tissue .The Holt and CT imaging examinations disclosed that 10 ears were phase I ,lesions were confined to the external auditory with no bone destruction .There were 23 ears were phase II ,the lesions were located in external auditory meatus with destruction of bone ,but with no involvement of the middle ear .There were 6 ears were phase III ,showing the lesions with disrupt external audi‐tory meatus and involvement of the middle ear ( mastoid and/or tympanic sinus) .Hearing impairment and aural fullness were the most common symptoms .The phase I cases were treated by removing cholesteatomas from the ex‐ternal auditory canal .Canaloplasty and/or tympanoplasty were performed in phase II cases .The phase III cases were successfully managed by modified radical mastoidectomy and/or tympanoplasty .There were 30 ears of tympan‐ic membrane were perfect and invaginate .There were 4 ears of ossicular chain were disrupted with one ear of facial nerve exposed .All surgeries were performed at once .No recurrence except in one patient was found during the fol‐low -up period .Conclusion The clinical stages can help identify the primary lesions and determine the choice of the best surgical approach .

Objective To investigate the prevalence of vitamin D deficiency and to examine its relationship with the severity and prognosis in the critically ill children. Methods A total of 83 critically ill children admitted from November 1,2010 to December 9,2010 to Pediatric Intensive Care Unit in Beijing Children's Hospital,Capital Medical University were enrolled in the study. Serum 1,25 - Dihydroxyvitamin D concentration was measured by using an en-zyme - linked immunosorbent assay(ELISA). Anthropometric parameters such as height/ length and weight of the chil-dren were measured. Data collection also included primary disease,Pediatric Critical Illness Score(PCIS),the Pediatric Risk of Mortality Ⅲ(PRISM Ⅲ)score,multiple organ dysfunction syndrome( MODS)rate,mechanical ventilation rate,time of hospital of stay and the 28 - day survival rate. Results There were 32 cases with vitamin D deficiency on admission,vitamin D deficiency rate on admission was 38. 6% ,and there was no statistically significant difference among different primary disease groups(P = 0. 815). Vitamin D deficiency rate of malnutrition group was lower than that of the normal group[60. 0%(12 / 20 cases)vs 40. 0%(8 / 20 cases),χ2 = 5. 989,P = 0. 014]. PCIS scores of those with a normal vitamin D status was higher than those of the vitamin D deficiency group,showing a significant difference [(80. 47 ± 6. 18)scores vs(77. 16 ± 7. 59)scores,P = 0. 022]. PCIS score was positively correlated with the vitamin D level(r = 0. 267,P = 0. 015). There was no statistically significant difference among the PRISM score,MODS rate, mechanical ventilation rate,hospital stay length and the 28th day survival rate between the normal vitamin D group and the vitamin D deficiency group(all P ﹥ 0. 05). Conclusions A high prevalence of vitamin D deficiency is found in the critically ill children. The prevalence of vitamin D deficiency in children with malnutrition is higher. Vitamin D status may be correlated to the severity of the critically ill children,but the association with the prognosis is not obvious.