Objective To investigate the effect of different promoters on the expression level of transgene containing MAR expression vector in recombinant CHO cells.Methods The CMV promoter and 3-globin MAR were amplified by PCR,then CMV promoter was replaced the SV40 promoter in pCAT1 for constructing the expression vector droved by CMV promoter.The control vectors of pCAT1 and pCAT2 without containing MAR were simultaneously transfected into the CHO cells.Then the stably transfected cell line was screened by G418.The CAT gene expression level was analyzed by ELISA.Results The expression level of CAT enzyme in the cells transfected with MAR-containing vectors was increased compared with the cells transfected by pCATG and pCAT3 vectors without containing MAR,which were increased by 1.75 and 1.25 times respectively(P＜0.05);but CAT enzyme expression level in the pCAT1 transfected cells droved by SV40 promotor with the MAR-containing expression vectors was 1.4 times higher than that in the pCAT2 vector droved by the CMV promoter(P＜0.05).Conclusion MAR can enhance the transgene expression level in stably recombinant CHO cells,and the promoting efficiency of SV40 promoter and MAR combination is superior to that of CMV promoter and MAR combination.

Objective: To discuss the clinical features of basaloid squamous carcinoma(BSC)and the factors relating to its prognosis and to compare patient survival between poorly differentiated squamous cell carcinoma(PDSC)and BSC. Methods: Clinical and pathological data of BSC and PDSC cases seen in our hospital between January 2004 and December 2008 were reviewed. Results: There were no statistical differences in demographic and clinical features between PDSC and BSC patients,with the exception that a larger proportion of BSC patients were female(P=-0.001).Additionally,higher tobacco consumption was observed among BSC male patients (P=0.003).There were no significant differences in survival rate between BSC and PDSC groups(X2=0.03,P=0.5470).The median survival time of BSC and PDSC patients was 19 months and 30 months，respectively.The 4-year survival rate was 22.4%and 36.1%，respectively(u=0.740,P=0.230).No significant difference was found in survival rate between stage Ⅰ and stage Ⅱ patients(X~2=0.109，P=0.2974).The median survival time of stage Ⅰ and stage Ⅱ patients was 19 months and 46 months，respectively；and the 4-year survival rate of stage Ⅰ and stage Ⅱ patients was 47.3% and 45.2%，respectively(u=0.122，P=0.450).Using Cox proportional hazard model，we found that surgical types and clinical stages of BSC were correlated with its prognosis.Compared with that of patients who received lobectomy,the postoperative mortality hazard of patients who received pneumonectomy and segmentectomy was increased by 1.379 times(P=0.031)and 1.634 times(P=0.061)，respectively.A more advanced clinical stage was associated with an increase in the postoperative morta,ty hazard ratio(X~2=14.12,P=0.000).The postoperative mortality hazard of patients of stage Ⅲ and stage Ⅳ was 2.437 times higher than that of stage Ⅰ patients(P=0.018).There were no statistical differences in postoperative mortality risk between stage Ⅰ patients and stage Ⅱ patients(P=0.057). Conclusion: Compared with that of PDSC,the incidence of BSC is higher among females.However,there is no difference in the prognosis between BSC and PDSC.BSC can be treated with the same therapies as those for other types of non-small cell lung cancer(NSCLC).

Objective To investigate the effects of matrine on proliferation and apoptosis of human Tenon capsule fibroblasts (HTFs) in.vitro.Methods After treated with 0,0.3,0.6 and 0.9 g/L matrine in vitro,cell counting kit-8 (CCK-8) method was used to assay the proliferation of HTFs at 24,48 and 72 hours,Western blot and PCR were performed to evaluate the expression of apoptosis-associated factor caspase-3 on both protein and RNA levels.Results The activity of human Tenon capsule fibroblast at 48 hours and 72 hours after treated with 0.3,0.6,0.9 g/L matrine was significantly inhibited when compared with the 0 g/L matrine group,and the inhibitory effect was dose-dependent and time-dependent (F ion =1 019.51,P =0.00;Ftime =5 848.66,P =0.00;Fi ion =147.45,P=0.00).After treated with 0,0.3,0.6 and 0.9 g/L matrine,the early apoptosis rate of HTFs was (2.68±0.30)％,(5.08±0.47)％,(6.97±0.69)％ and (10.30±1.20)％,the grey value ofcaspase-3 protein was 1.00±0.13,1.90±0.19,2.50±0.30 and 2.67±0.30,the relative expression of caspase-3 mRNA was 0.98 ±0.12,2.01 ±0.34,6.15 ± 0.60 and 11.40 ± 1.12,respectively,with significant differences among them (F =55.74,66.01,154.50;all at P＜0.01),the early apoptosis rate of HTFs,the grey value of caspase-3 protein and the relative expression of caspase-3 mRNA were all increased significantly as the concentration of matrine increased,with significant differences between any two groups (all at P＜0.05).Conclusions Matrine can inhibit the proliferation of HTFs and induce the apoptosis of HTFs in a time-and dose-depended manner.

Objective To propose a deep learning-based retinal image quality classification network, FA-Net,to make convolutional neural network ( CNN) more suitable for image quality assessment in eye disease screening system. Methods The main network of FA-Net was composed of VGG-19. On this basis,attention mechanism was added to the CNN. By using transfer learning method in training, the weight of ImageNet was used to initialize the network. The attention net is based on foreground extraction by extracting the blood vessel and suspected regions of lesion and assigning higher weights to region of interest to enhance the learning of these important areas. Results Total of 2894 fundus images were used for training FA-Net. FA-Net achieved 97. 65％ classification accuracy on a test set containing 2170 fundus images,with the sensitivity and specificity of 0. 978 and 0. 960,respectively,and the area under curve(AUC) was 0. 995. Conclusions Compared with other CNNs,the proposed FA-Net has better classification performance and can evaluate retinal fundus image quality more accurately and efficiently. The network takes into account the human visual system ( HVS) and human attention mechanism. By adding attention module into the VGG-19 network structure, the classification results can be better interpreted as well as better classification performance.

Objective To investigate the differences of all discomfort perceived by patients and with mechanical ventilation and nursing care .Methods A total number of 220 patients admitted to ICU accepted mechanical ventilation , and 68 nurses with over 1 year experience conducting ventilation were investigated on their perception with self-designed questionnaires .Results The total score of patients′comfort was (48.956 ± 7.136), of which physiological comfort was (27.816 ±4.158), psychological comfort (14.475 ±4.335), society comfort (8.141 ±3.895), while total patients′comfort score of nurses′perception was (59.110 ± 9.002), of which physiological comfort was (30.089 ±4.637), psychological comfort (11.032 ±4.076).The perception between nurses and patients had statistically differences (t=-8.512,-3.618,-6.708,-5.165, respectively;P<0.05).Conclusions The inconformity of perception between patients and nurses affects both evaluation of patients′discomfort and nursing care .Nurses should access discomfort of patients precisely in order to provide comfortable nursing .

Objective:To assess the risk factors for mortality and clinical outcome of carbapenem-resistant Pseudomonas aeruginosa (CRPA) infections in patients with hematological disorders. Methods:The data of in-patients with hematological disorders infected by CRPA or carbapenem-susceptible Pseudomonas aeruginosa (CSPA) were recorded in a seven-year retrospective cohort study. Risk factors for CRPA infections and impact of on mortality were identified. The primary end point was 30-day all-cause mortality. Results:A total of 81 patients with PA infections were included in the study, including 58 CSPA and 23 CRPA. Most of the primary diseases were acute leukemia or lymphoma (79.0%, 64/81). The median absolute neutrophil count at infection onset was 0.24×10 9/L. Independent risk factors associated with carbapenem-resistance included longer duration of hospital stay ( P=0.013, OR=1.045) and carbapenem exposure one month prior to infections ( P=0.005, OR=8.132). The 30-day all-cause mortality of the whole cohort was 29.6%(24/81), and 30-day attributable mortality was 13.6%(11/81). Pulmonary infection was the leading cause of death, accounting for 41.7%(10/24). The adjusted 30-day mortality rate was significantly higher in patients with CRPA compared with CSPA [60.9%(14/23) vs. 17.2%(10/58), P<0.001, respectively]. CRPA infection was an independent prognostic factor for 30-day mortality( P=0.011, OR=5.427). Other factors included old age, longer duration of neutropenia and poor functional performance. Conclusions:Patients with hematological disorders have high mortality rate and poor prognosis caused by CRPA infections, which mainly develop in lungs.

Polyhydroxyalkanoates (PHAs), as a novel class of biopolymer, are attracting more attention due to their diverse material properties and environment-independent biodegradability. Here we report the preparation of PHA exhibiting efficient antibacterial activity by embedding Nisin, a food additive generally recognized as safe, into poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx), a type of PHA with high biocompatibility. We first prepared Nisin-containing PHBHHx films using solvent casting method. Confocal laser scanning microscopy analysis showed that a well-mixed integrated structure of the films with an even distribution of the Nisin particles in the PHBHHx matrices. Then the antimicrobial activity of PHBHHx/Nisin films against Micrococcus luteus was quantified on agar plate by measuring the size of inhibition zone. Cultivation in liquid media further confirmed the releasing of Nisin from the films and the long-time antibacterial activity. Results showed that the threshold of Nisin concentration for long-time and effective inhibition against bacteria growth is 25 μg/g. These results altogether establish a technological foundation for the application of PHA in biomedicine and food industry.
3-Hydroxybutyric Acid ; chemistry ; Anti-Bacterial Agents ; chemistry ; Caproates ; chemistry ; Micrococcus luteus ; drug effects ; Nisin ; chemistry ; Polyhydroxyalkanoates ; chemistry

ObjectiveTo explore the effect of precocious puberty on glucose metabolism and lipid metabolism in female rats. MethodsSixty two-day-old female rats were randomly divided into 2 groups. When aged 5 days， the precocious puberty group and normal group were given a single subcutaneous injection of danazol and solvent soybean oil respectively. The vaginal opening of rats was monitored from their 21 days of age. After 12 hours of fasting， all successful modeling rats were randomly executed within 3 days after vaginal opening， when aged 7 and 12 weeks. Then we measured the rats’ body weight and length， determined the concentrations of glucose， insulin， blood lipids， estradiol， leptin and adiponectin with enzyme-linked immunosorbent assay and observed the pathological changes of perirenal fat， uterus and ovary. ResultsFor body weight and length， rats in the precocious puberty group were smaller than those in the normal group within 3 days after vaginal opening， but which did not affect their subsequent growth and development， and there was no significant difference between the two groups at 7 and 12 weeks of age. Within 3 days after vaginal opening， insulin levels had significant difference between the two groups （P = 0.001）， the precocious group showed hyperinsulinemia and increased number of perirenal adipocytes. At three execution times， no significant difference was noted in estradiol， leptin and adiponectin levels between the two groups. The same was true in the ratios of ovary or uterus to body weight between the two groups. ConclusionsPrecocious puberty makes earlier onset of pubertal development and allows body maladaptation to the sudden changes of the internal environment. However， the changes due to precocious puberty are temporary and reversible， and they may become normal in adulthood.

To evaluate the efficacy and safety of aromatase inhibitor letrozole in treatment of male adolescents with idiopathic short stature (ISS). Seventy five boys with height less than 2 standard deviation (SD) below the mean who had entered puberty were enrolled in our study from 2004 to 2017, in the Pediatric Department of the First Affiliated Hospital, Sun Yat-Sen University. Among 75 patients, 28 in letrozole group received letrozole and spironolactone, 30 in gonadotrophin releasing hormone analogue (GnRHa) group received GnRHa injection and 17 had no intervention. Height velocity (HV), increment of bone age/chronological age (ΔBA/ΔCA), the final adult height (FAH) were compared among groups and the safety of letrozole treatment was evaluated. HV maintained faster during letrozole treatment when compared with other groups. HV during GnRHa treatment showed slightly decline in the first 6 months, but decreased remarkably after 6 months, and was significantly lower than that in letrozole group ( < 0.05). The maturation of BA slowed down in both letrozole and GnRHa groups. But the ΔBA/ΔCA in letrozole group during the first and the second year of treatment were significantly higher (0.67±0.09, 0.50±0.15, respectively) when compared with GnRHa group (0.59±0.16, 0.44±0.13, respectively) ( =2.78 and 2.20, all < 0.05). FAH in letrozole group and GnRHa group were (170±4) cm and (170±6)cm, there was no significant differences between the two groups ( >0.05), and both were higher than that in no intervention group (162±4 cm, < 0.01). After 6 months of letrozole treatment, testicular volumes and serum testerone levels increased; 39.2% (11/28) boys had clinical manifestations of hyperandrogenemia, and 82.1% (23/28) boys had decreased serum high-density lipoprotein (HDL) levels. Serum levels of HDL and testerone returned normal and the hyperandrogenemia disappeared after the cessation of letrozole treatment. No significant changes in serum triglyceride, serum low-density lipoprotein (LDL), fating serum levels of insulin and glucose, HOMA-IR were observed. No abnormal liver function, myalgia, scoliosis or aggravations of scoliosis was found. Long term letrozole therapy during puberty in boys with ISS can delay bone maturation without significant decrease of linear growth, and thus can improve the final adult height. No severe adverse reactions were found.
Adolescent ; Body Height ; Bone Development ; Child ; Gonadotropin-Releasing Hormone ; Growth Disorders ; Humans ; Letrozole ; therapeutic use ; Male

Objective To investigate the effects of silencing the gene of single-stranded DNA-binding protein 1 (SSB1) on proliferation and DNA repair of rat submandibular gland (SMG) cells after irradiation, and explore the relationship between SSB1 and DNA damage repair. Methods Primary rat SMG cells were obtained by mechanical-enzyme digestion and identified by immunohistochemistry. The cells were divided into three groups, including blank control, negative control and shRNA transfection group. The shRNA was transfected into cells by recombinant adenovirus vector. Real-time quantitative PCR ( qRT-PCR) was used to detect the expression of SSB1 after silencing. The cell viability was detected by CCK-8 assay. Immunofluorescence analysis was performed to observe the dynamic formation of γ-H2AX foci. Results The SMG cells were positively stained for both Pan CK and α-Amylase. The efficiency of shRNA transfection was about 90%at 72 h post-transfection. Compared with the blank control group, the expression of SSB1 was significantly decreased in the cells transfected with shRNA (t=16. 24, P<0. 05). The cell viability of shRNA transfection group without irradiation was decreased indistinctively and became lower than the blank control group significantly until 120 h(t=3. 29, P<0. 05). After radiation with 5 Gy of γ-rays, the cell viability of shRNA transfection group was lower than that of the control groups significantly (F=10. 19-30. 13, P<0. 05). Silencing the expression of SSB1 could increase the number ofγ-H2AX foci in SMG cells at different time of radiation. Conclusions After silencing of the expression of SSB1, the SMG cells could be more radiosensitive, which indicats that SSB1 may play an important role in DNA damage repair after radiation.