Along with prolongation of overall survival and increasing of therapeutic methods in advanced non-small cell lung cancer, whole-process management has become more and more important. We reviewed the whole-process management strategy according to difference of mutation state of epidermal growth factor receptor (EGFR) gene.

Objective To study the influence of site-directed deglycosylation of the HIV-1 envelope (Env) on its immunogenicity and assembly of functional pseudovirus. Methods Site-directed deglycosylation were performed using cycling mutagenesis and selection of mutants with DpnⅠ. Single-cycle infection assay was employed to analyze the effect of the mutations on the ability of functional pseudovirus assembly. The influence of deglycosylations on the immunodeficiency of Env was evaluated using pseudovirusbased neutralization assay and ELISPOT assay. Results Mutant N197Q induced higher neutralization activities for both pseudoviruses, but lower Env-specific T-cell response. And N197Q rendered the Env to lose the ability of functional pseudovirus assembly. Mutant G2 induced higher neutralization activities for pseudovirus 74-2 but lower for pseudovirus Wt, and had almost no influence on Env-specific T-cell response and functional pseudovirus forming. Conclusion The site-directed deglycosylation of the HIV-1 Env affects the pseudovirus forming and its immunogenicity.

Objective To establish a pseudovirus-based neutralization assay. Methods The functional gp160 genes were amplified from plasmids containing HIV-1 gene. The products were ligased into pcDNA3.1 plasmid and positive clones were screened by digestion with restriction enzymes. The pseudoviruses were harvested by co-transfection of the positive clone and pSG3△env plasmid. The neutralizing activities of monoclonal antibodies and HIV-1 antibody positive plasma were measured by these pseudoviruses. Results The four strains of psedoviruses (CHB01, CHB02, CHBC03 and CHAE04) had been successfully obtained. Monoclonal antibody 4E10 could neutralize all of 4 pseudoviruses while 2G12 could not neutralize any pesudoviruses. Monoclonal antibody 2F5 could neutralize pseudovirus CHB01, CHB02 and CHAE04 but not CHBC03, while IgG1b12 could neutralize pseudovirus CHB01, CHB02 and CHBC03 but not CHAE04. The neutralizing activities of 43 of HIV-1 antibody positive plasma against different subtypes of pseudovirus were significant differences and the cross-neutralization effects for some samples exist. Conclusion The harvested pseudoviruses could be used in the neutralization assay. However, the neutralizing characteristics of different pseudoviruses may be different.

Objective To investigate the radiosensitizing effect of knock-down the expression of miR-21 on human SHG-44 glioma cells and explore the possible mechanism. Methods Antisense oligonuleotidas of miR-21, mediated by LipofectamineTM 2000, were transfected to SHG-44 cells. Three groups were: blank control group ( mock group), negative control and antisense transfected group ( AS-miR-21 gorup). Cells of each group were irradiated with 6 MeV X-rays at the doses of 0,1,2,4,6 and 8 Gy.Dose-suvivial curve was established by colony-forming assay. The influence of AS-miR-21 on cell cycle and cell apoptosis was analyzed by flow cytometry assay after 6 Gy irradiation. Results The value of D0 and Dq of AS-miR-21 group declined obviously compared with the mock group and negative control group. Flow cytometric analysis showed that cell cycle distribution changed( G0/G1 phase arrest, S phase decreased)after transfected with AS-miR-21 (t = 8.18, -4.52,P ＜ 0.05 ). The sensitization enhancement ratios of D0 and Dq were 1.32 and 2.10 respectively. Apoptosis assay showed the early apoptosis rate was signiflcantely increased in AS-miR-21 、irradition alone and combined group than mock control group( t = 20.14,11.11,50.07, P ＜ 0.05). Conclusions AS-miR-21 can enhance the radiosensitivity of human glioma cells SHG-44 by promoting cell apoptosis and faciliating cell cycle redistribution.

Objective To explore the effect and significance of clinical pathway guided teaching in the residency standard training program of respiratory medicine. Methods Total 47 resident physicians were selected and divided into clinical pathway group and control group from March 2014 to November 2014. The pathway group (n=24) was introduced into the teaching guided by clinical pathway management. The control group (n=23) was taught by traditional teaching method. All physicians were tested for the basic theory and the ability of case analysis after 4 weeks training. The ability of chemotherapy strategy ordered by residents independently and correctly was assessed each week during training. A satisfaction questionnaire survey was conducted to evaluate the effectiveness and satisfaction of teaching guided by clinical pathway. GraphPad Prism 5.0 was used and T test was done for comparison of data between groups. Results The medical records about basic theory and case analysis in the pathway group was higher than those in the control group with significant statistical difference (P<0.05). The records of resident physicians who could issue orders for chemotherapy independently and correctly were (70.75±2.79), (81.43±1.91), (85.23±1.3), (90.62±2.34) in the pathway group and (69.65±2.06), (77.11±2.21), (80.3±1.96), (87.78±2.21) in the control group at each week time point. There was statistical increase of the records in the pathway group than in the control group since the second week time point (P<0.05) The overall satisfaction of the pathway group was 95.84%(23/24), and the teaching satisfaction was higher than that of the control group(91.29%, 21/23). Pathway group doctors believe that the relevant teaching effectively improve the level of their knowledge , experience and ability. Conclusions The teaching method guided by clinical pathway is help-ful to standardize the teaching behavior, develop the standardized medical behavior of resident physicians, improve their clinical working ability efficiently, promote the relationship between teaching and studying, which is worth application in the residency standard training program of respiratory medicine.

AIM: To investigate the biocolonization of poly 2-hydroxyethy methacrylate (PHEMA) sponge with cornea tissue and evaluate the therapeutic effects of modified porous poly 2-hydroxyethy methacrylate-Polymethyl met-hacrylate (PHEMA-PMMA) Keratoprostheses (KPro) on rabbit and monkey corneas. METHODS:The KPro were made using two-stage polymerization combined with mechanical cutting. The experiment was divided into two groups. In the control group, ten normal rabbit eyes received lamellar implanta-tion of PHEMA sponges. The sponges were obtained 2 weeks, 1,2,3 and 4 months after operation. The cell proliferation and neovascularization inside the sponges were observed using light and transmission electron microscopy (TEM) and immunohistochemistry. In the experimental group, the porous PHEMA-PMMA KPros were inserted into the lamellar pockets of eight rabbit corneas and two monkey corneas (stage I operation). The healing process was investigated by slit-lamp microscopy. The anterior lamellar cornea tissues were removed 3 months after surgery, exposing the under-neath transparent core (stage II operation). The operated eyes were then followed up for 3-6 months.light microscope, fibroblasts started to grow into the cornea 2 weeks after operation; lots of cells, accompanied with new blood vessels, invaded into the cornea 2-3 months after surgery. Invading cells of sponge, as well as keratocytes, were positive for vimentin. Under the electron microscope, the invading cells looked healthy and were surrounded by extracellular matrix and collagen. In 8 rabbit eyes which received KPro implantation, anterior lamellar cornea melting happened in two eyes after the stage II operation. The remaining 6 corneas retained their central cores during observation after the stage II operation.Two monkey operated eyes were found no complication thoughout the whole follow-up.cornea. The modified PHEMA-PMMA KPros have obtained a relatively stable results after implantation into animal corneas.

Eleven env mutants were designed and generated by site-directed mutagenesis of the regions around Nab epitopes and deletions of variable regions in env.The immunogenicities of the generated mutants were evaluated using single-cycle infection neutralization assays with two pseudoviruses and IFN-γELISPOT.Overall,five mutants(dWt,M2,M5-2,M5-1 and dM7)induced highed neutralization activities for both pseudoviruses than plasmid Wt,while only two of the mutants(dWt and M5-2)showed significant differences(P<0.05).Two mutants(M2 and dM2)induced more Env-specific T cells than plasmid Wt.Statistically however,significance was only reached for mutant M2.Thus,properly modified HIV-1 Env may have the potential to induce potent cellular and humoral immune responses.

Objective To investigate the expression of thyroid cancer-1 (TC1) and β-catenin in cervical carcinoma and precancerous lesions and their significance. Methods Immunohistochemical methods were used to examine the expression of TC1 and β-catenin in80 cervical squamous cell carcinoma (CSCC) tissues, 40 high-grade squamous intraepithelial lesions (HSIL), 40 low-grade squamous intraepithelial lesions (LSIL), and 30 normal cervical tissues. Results Although TC1 expression in CSCC was significantly higher than that in LSIL (P = 0.002) and normal cervical tissues (P ＜ 0.001), it was similar to that in HSIL (P = 0.576). TC1 expression was positively correlated with poor differentiation (P = 0.005) and advanced FIGO stage (P = 0.004) in CSCC. β-catenin expression in CSCC was significantly higher than that in LSIL (P ＜ 0.001) and normal cervical tissues (P ＜ 0.001), but was similar to that in HSIL (P = 0.907). The abnormal β-catenin expression was also correlated with poor differentiation (P = 0.025) and advanced FIGO stage (P = 0.001) in CSCC. TC1 expression was positively correlated with the abnormal β-catenin expression in CSCC (r = 0.294, P = 0.008) and cervical squamous intraepithelial lesions (r = 0.549, P ＜ 0.001). Conclusion TC1 and β-catenin expression in CSCC and HSIL was significantly higher than that in LSIL and normal cervical tissues. TC1 expression correlated with the abnormal β-catenin expression, and with poor differentiation and advanced FIGO stage of CSCC.

OBJECTIVETo learn the status quo of lifestyle among women of five occupations in six provinces of China.

METHODSA questionnaire was administered among 7416 women from five occupations (civil servants, teachers of elementary and high schools, technical staff, enterprise managers and physical laborers) in Beijing, Hebei, Jilin, Hubei, Ningxia and Gansu of China. The sample was selected by multi-stage stratified cluster random sampling method from December 2009 to June 2010. The questionnaire information included demographic characteristics, diet, sleep habit, smoking, physical exercise. The χ(2) test was used to analyze the different in life style of different occupations.

RESULTSThere were 7416 valid questionnaires received, and the valid rate of the questionnaires was 97.58% (7416/7682). About 38.00% (2818/7416) respondents preferred to bland diet and 28.44% (2109/7416) preferred to salty and oily food and 33.56% (2489/7416) had no preference. The proportion of sleep time between seven and eight hours per day was highest (accounting for 56.23%, 4154/7416), 25.27% (1867/7416) with sleep time less than seven hours. Among the population who had the sleep time less seven hours, teacher that had the highest rate accounted for 33.19% (531/1607) and technical staff had the lowest rate accounted for 21.05% (301/1401) (P < 0.01). Most of respondents were non-smokers, accounting for 93.10% (6869/7416). 22.73% (1671/7416) respondents passively smoked. The proportion of always passive smoking was highest among civil servants and lowest among teachers, accounting for 26.60% (404/1531) and 18.71% (298/1607), respectively. The proportion of having no physical exercises was highest, accounting for 62.87% (4637/7416). The proportion of having three times physical exercises per week was 12.68% (935/7416). The proportion of having no physical exercises among physical laborers (66.42%, 912/1386), enterprise managers (66.64%, 987/1491) and teachers (62.40%, 999/1607) were higher than others and the proportion of having physical exercises per week among technical staff was 40.83% (569/1401), higher than others (P < 0.01). The proportion of person who worked in sitting quietly beyond six hours per day was 42.62% (3146/7416). The technical staff had the higher rate than other occupational populations (P < 0.01), accounting for 57.83% (809/1401).

CONCLUSIONThe female occupational population had some unhealthy lifestyles, such as taking in high salt food, lacking of sleep, smoking and passive smoking, lacking of physical exercises and working in sitting quietly. Different occupational populations had different unhealthy lifestyles.

Adolescent ; Adult ; China ; epidemiology ; Female ; Humans ; Life Style ; Middle Aged ; Occupations ; Surveys and Questionnaires ; Young Adult

The chemical constituents were isolated and purified by various chromatographic techniques indluding silica gel, reverse phase silica gel, sephadex LH-20 and pre-HPLC and identified by their physicochemical properties and spectral data. Sixteen phenolic compounds had been isolated and n-butanol extracts which were fractionated from the ethanol extract of Oplopanax horridus roots bark. Their structures were identified as below, including 7 phenylpropanoid compounds, ferulic acid (1), 3-acetylcaffeic acid (2), caffeic acid (3), homovanillyl alcohol 4-O-beta-D-glucopyranoside (4), 3-hydroxyphenethyl alcohol 4-O-beta-D-glucopyranoside (5), 3, 5-dimethoxycinnamyl alcohol 4-O-beta-D-glucopyranoside (6), and 3-dimethoxycinnamyl alcohol 4-O-beta-D-glucopyranoside (7). Three coumarins, scopoletin (8), esculetin (9) and 3'-angeloyl-4'-acetyl-cis-knellactone (10). And 6 lignan compounds, (+)-isolaricires-inol-9'-O-beta-D-glucopyranoside (11), 3, 3'-dimethoxy-4, 9, 9'-trihydroxy-4', 7-epoxy-5', 8-lignan-4, 9-bis-O-beta-D-glucopyranoside (12), (+)-5, 5'-dimethoxylariciresinol 4'-O-beta-D-glucopyranoside (13), (-)-5,5'-dimethoxylariciresinol 4'-O-beta-D-glucopyranoside (14), (-)-pinoresinol 4'-O-beta-D-glucopyranoside (15), and (+)-5, 5'-dimethoxylariciresinol 9'-O-beta-D-glucopyranoside (16). All compounds were isolated and identified for the first time from this plant All the constituents except compounds 4, 6, 12 and 13 were obtained for the first time from the genus Oplopanax.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Oplopanax ; chemistry ; Phenols ; chemistry ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization