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 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 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 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 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 investigate the correlation between high myopia and MMP-2 in aqueous humor (AH), by comparing the protein level of MMP-2 in AH from patients with cataract combined with high myopia and with emmetropia. Design Experimental study. Participants 30 AH samples of patients with age-related cataract. Methods AH samples were collected from the patients with age-related cataract during phacoemulsification, including 15 AH samples from patients combined with high myopia (experimental group) and 15 AH samples from patients with emmetropia (control group). The expression of pro-MMP-2 and MMP-2 in AH were ana-lyzed by Western blot technique. Main Outcome Measures The gray value of MMP-2 protein band detected by Western blot. Results The level of pro MMP-2 was statistically higher than that of MMP-2 in experimental group (430.4±57.3 versus 294.5±35.2, t=10.400, P= 0.000) and control group (402.8±57.7 versus 280.3±49.7, t=8.400, P=0.000). There was no statistically difference in the level of pro-MMP-2(t=1.320, P=0.200)and MMP-2 (t=0.900,P=0.375) between the two groups. Conclusions No abnormal expression of MMP-2 was detected in AH from patients with high myopia, according to this study based on limited samples. Pro-MMP-2 was the main form of MMP-2 in AH of patients with cataract combined with high myopia or emmetropia, which possessed potential ability of transferring into form of active MMP-2. (Ophthalmol CHN, 2009, 18: 260-264)

Objective To investigate the relation between the expression of toll receptor-4 (TLR-4) and the increase of blood brain barrier (BBB) permeability in the rats with acute pancreatitis.Methods Male Sprague-Dawley rats were randomly divided into 8 groups:control group;acute edematous pancreatitis (AEP) 2 h,6 h groups;acute necrotizing pancreatitis (ANP) 2 h,6 h,12 h,24 h,48 h groups with 8 rats in each group.The BBB permeability,pathological score of pancreas,TLR-4 expression were determined and the relationships between them were analyzed.Results The BBB permeability in control group;acute edematous pancreatitis (AEP) 2 h,6 h groups;ANP 2 h,6 h,12 h,24 h,48 h groups were 1.55±0.29,1.64±0.17,1.69±0.24,1.89±0.12,2.66±0.32,2.91±0.29,2.89±0.69 and 1.84±0.07,respectively;the pathological scores of pancreas were 0,2.38±0.92,3.13±0.64,8.50±1.07,9.75±0.71,10.25±1.28,11.13±1.25 and 10.13±1.13,respectively;there was no significant difference between AEP groups and control group,while there was significant difference between AEP groups and ANP groups (P＜0.05 orP ＜0.01).BBB permeability was correlated with pancreatic injury ( r = 0.626,P ＜0.01).There was no TLR-4 mRNA and protein expression in the control and AEP group,while they were significantly expressed in ANP groups,and the expression were positively related with BBB permeability ( r =0.208,P = 0.027 ).Conclusions BBB permeability was present in the course of ANP.Activation of TLR-4 signal pathway may be involved in the BBB permeability increase caused by ANP.

Objective　To explore the effects of hypoxia on the syntheses and secretion of adrenomedullin (AM), calcitonin gene related peptide (CGRP) and c-type natriuretic peptide (CNP) and the relationship between these peptides. Methods　Rat models were established with hypoxia for 10, 20 and 30 d respectively and rats under normal altitude were served as control. Pulmonary artery pressure and the maximum increasing speed of right ventricle (RVdp/dtmax) were measured in every group. The dynamic changes of AM, CGRP and CNP concentrations in plasma were studied with radioimmunoassay. Results　During hypoxia, pulmonary artery pressure and RVdp/dtmax were enhanced. Plasma AM and CNP concentrations were increased while CGRP was decreased significantly. The plasma level of AM had positive correlation with that of CNP, but negatively correlated with that of CGRP. Conclusion　Results indicate that hypoxia may cause pulmonary artery pressure change and right ventricle has compensatory reaction to hypoxic pulmonary hypertension. Dynamic changes of plasma AM, CGRP and CNP concentrations can be regarded as indexes for condition of illness.

Objective To study the clinical features of invasive pneumococcus disease (IPD) with resistance to antimicrobial agents in children,and to improve the diagnosis and treatment of this disease.Methods The clinical data from 21 IPD patients younger than 13 years old were collected from January 2008 through December 2010 in Pediatric Intensive Care Unit in Beijing Children's Hospital for retrospective analysis. Specimens of blood,pleural effusion,cerebrospinal fluid and soft tissue aspirated were collected from these children,and 23 strains of streptococcus pneumonia (SP) were cultured,isolated and confirmed,and the antibiotics susceptibility to penicillin and other antibiotics of these strains were assayed.Results Among the 21 IPD children,the ratio of male to female was 0.9∶1,and the age was 5 months to 13 years,with 61.9％ of them under 2 years.Of them,12 patients (57.1％ ) had purulent pleurisy,and 1 (4.8％ )patient had an underlying disease diagnosed to be X - linked agammaglobulinemia (XLA).There was no seasonal difference in the occurrence rate of this disease. Eight (38.1％) patients were cured,11(52.4％ ) were improved,while 2 (9.5％ ) patients not improved without death.There was no statistically significant difference in the annual detection rate of invasive SP (x2 =3.711,P =0.156).The incidences of penicillin-intermediate susceptibility SP (PISP) and penicillin-resistant SP (PRSP) were 47.8％ and 26.1％ respectively.The rate of resistance to multiple antibiotics was 91.3％.Conclusions Children aged less than 5 years,especially younger than 2 years,are prone to IPD,and purulent pleurisy and septicemia are often seen in this disease. Some patients had the underlying diseases.The complications included hemophagocytic syndrome,acute respiratory distress syndrome,septic shock,bronchial pleural fistula and so on.The multidrug resistance rate was 91.3％.It is important to put great emphasis on the monitoring antibiotics resistance to invasive pneumococcal disease.