The major challenge in the development of recombinant biologics lies in generating and isolating rare high-producing stable single clone in a short period of time. The selection marker is an essential component of the plasmid vector, it plays an important part in the generation and screening of producing cell lines. Engineering the selection marker to enhance the stringency of selection for high producing cells is one of the most effective approaches to improve the cell line development process. Here, using Chinese hamaster overy (CHO) cells as an example, we introduce the application of selection marker for generation of recombinant biologics producing mammalian cell lines, methods of engineering the selection markers to enhance the selection stringency, and propose considerations on cell substrate stability and selection marker safety, in order to provide references for high-efficiency development of recombinant biologics.

Objective To retrospectively evaluate different methods (laboratory tests,liver / gallbladder ultrasound and magnetic resonance cholangiopancreatography)in differentiating biliary atresia from infant hepatitis syndrome.Methods Seventy infants with cholestatic jaundice,50 cases of biliary atresia and 20 cases of infant hepatitis syndrome were studied prospectively from January 2010 to December 2012.All cases underwent abdominal ultrasound and magnetic resonance cholangiopancreatography.The accuracy,sensitivity,specificity and predictive values of these various methods were compared.Also the laboratory parameters were statistically analyzed and compared between groups.Results Patients with BA had significantly higher GGT values at presentation [(743.5 ± 564) IU/L] compared with infants with IHS [(198.8 ± 197.8) IU/L],showing statistically significant difference (P ＜ 0.05).The sensitivity,specificity,and accuracy of the ultrasound in diagnosis of BA were 84.0％ 、100.0％ 、88.6％ respectively(P ＜ 0.05).The values for magnetic resonance cholangiopancreatography were 82.0％ 、80.0％ 、81.4％ (P ＜ 0.05).Conclusion Currently,the method of ultrasound is more reliable than MRCP for differentiating biliary atresia from infant hepatitis syndrome.

Objective To evaluate the inter -list equivalency of the disyllabic materials for Mandarin speech perception test (MSP) by measuring the speech recognition score of patients with cochlear implant (CI) .Methods According to Latin -square design ,disyllabic recognition scores (quiet background) were measured for each of the 10 phonemically balanced lists in 50 Mandarin-speaking CI users(aged 30 .44 ± 12 .71 years)in sound field .RM -ANOVA was administered to confirm the list equivalency .Results The sentence recognition scores were 59 .26% ± 23 .49% ,64 .31% ± 23 .35% ,59 .97% ± 23 .07% ,62 .40% ± 25 .16% ,62 .75% ± 24 .47% ,62 .29% ± 23 .55% , 62 .85% ± 24 .60% ,61 .35% ± 23 .73% ,61 .82% ± 25 .28% ,58 .83% ± 25 .13% ,respectively for the 10 lists .There was no significance difference in sentence recognition scores across the 10 disyllabic lists [F(9 ,490)=0 .255 ,P=0 .986>0 .05] .Conclusion The good inter -list equivalency of the disyllabic materials for Mandarin speech perception test (MSP) has been proved to be useful for assessing speech recognition performance of Mandarin -speaking CI users .

Objective To evaluate the value of ultrasound (US) in assessment knee joint inflammation in patients with juvenile rheumatoid arthritis(JRA).Methods US scans of the knees obtained in 30 children at clinically active stage; JRA was compared with those obtained in 30 healthy children and 10 JRA patients in clinical remission.Results Changes in synovial membrane thickness and presence of fluid in suprapatellar bursa showed statistically significant differences between JRA patients with active disease and the other subjects.Alterations in contour of the articular cartilage were demonstrated in 10 knees of patients with JRA.Conclusion US is a simple sensitive and reliable methods for the assessment and monitoring of knee joint involvement in JRA.

Objective To investigate the changes in the expression of GABAAα1, receptors in medial prefrontal cortex (mPFC) in a rat model of neuropathic pain. Methods Nine male Wistar rats weighing 200-210 g were randomly divided into 3 groups ( n = 3 each): control group (group C) , sham operation group (group S) and neuropathic pain group (group P). Neuropathic pain was induced by chronic constrictive injury. The right sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 3-0 chromic catgut. In group S, the right sciatic nerve was exposed but not ligated. The thermal and mechanical pain threshold was measured at 1 d before and 1,4,7, 10 and 14 d after operation. The animals were then sacrificed and the mPFC was removed. The expression of GABAAα1, receptors in mPFC was determined by Western blot. Results Compared with C and S groups, thermal and mechanical pain threshold were significantly decreased and the expression of GABAAα1, receptors was up-regulated in group P ( P ＜ 0.01) . There was no significant difference was in the thermal and mechanical pain threshold and expression of GABAAα1 receptors between C and S groups (P ＞ 0.05). Conclusion Up-regulation of GABAAα1 receptor expression in mPFC may be involved in the development and maintenance of neuropathic pain in rats.

Mean hemoglobin (Hb) concentration of about 3 500 subjects derived from 17 studies of Himalayan highlanders (Tibetans, Sherpas, and Ladakhis) was compared with lowlanders (Chinese Han, Indian Tamils) lived in the Himalayas, and European climbers during Everest expeditions as well as Andean natives. The results found that Hb concentration in Himalayan highlanders was systemically lower than those reported for Andean natives and lowland immigrants. These comparative data demonstrated that a healthy native population may successfully reside at high altitude without a significant elevation in Hb, and the lower Hb levels of Himalayan highlanders than those of migrated lowlanders and Andean natives are an example of favourable adaptation over the generations. In addition, excessive polycythemia has frequently been used as a marker of chronic mountain sickness (CMS). Altitude populations who have a higher Hb concentration also have a higher incidence of CMS. The low Hb in Himalayans suggested as showing adaptation over many generations in Tibetan stock. Recent work in Tibet, suggested that Tibetans there may have adapted to high altitude as a result of evolutionary pressure selecting for genes which give an advantage at altitude. All of the population genomic and statistical analysis indicated that EPAS1 and EGLN1 are mostly likely responsible for high altitude adaptation and closely related to low Hb concentration in Tibetans. These data supported the hypothesis that Himalayan highlanders have evolved a genetically different erythropoietic response to chronic hypoxia by virtue of their much longer exposure to high altitude.
Adaptation, Physiological ; Altitude ; Asian Continental Ancestry Group ; genetics ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Evolution, Molecular ; Hemoglobins ; genetics ; Humans ; Hypoxia-Inducible Factor-Proline Dioxygenases ; genetics ; Tibet

Hotlines ; Humans ; Influenza A Virus, H7N9 Subtype ; Influenza, Human ; prevention & control ; virology ; Internet ; Preventive Health Services ; methods

OBJECTIVE Licorice is used throughout the world as a traditional herbal remedy. Ac-cording to Chinese traditional medicine licorice alone can be used to treat inflammation.Although there have been some studies investigated the anti-inflammatory ingredients of licorice, but for the potency of flavonoid glycoside and their aglycones on inflammation are not evaluated.This study was designed to assess the contributions of licorice flavonoid glycosides and their aglycons to its anti-inflammatory and hypnotic effects. METHODS For the flavonoid aglycone's enrichment, the extract of licorice (EL) was fermented in submerged culture of the edible fungus Grifola frondosa HB0071 mycelia which can produce β-glucosidase and catalyze the flavonoid glycosides to aglycones.EL and fermented extract of licorice (FEL) were used in this study. The anti-inflammation test was carried out in arachidonic acid (AA)-induced ear edema model and the hypnotic test was performed by using electroencephalogram (EEG)analysis method in normal freely moving SD rats.The chemicals constituents were analyzed by HPLC.RESULTS During fermentation,the falvonoid glycosides of licorice were hydrolyzed by the time process.Along with fermentation time,the concentration of the major flavonoid glycosides,liquiritin and isoliquiritin were decreased obviously, and simultaneously their aglycons, liquiritigenin and isoliquiriti-genin were remarkably increased in FEL.Moreover,the content of another major constituent glycyrrhi-zic acid and glycyrrhetinic acid were not changed after the fermentation. In AA-induced mice ear ede-ma test,after topical application,FEL(effective dose range:5-20 μg·ear-1)showed more potent inhibito-ry activity than EL(effective dose range:25-100 μg·ear-1).On the other hand,oral administration of EL and FEL exhibited the same hypnotic potency and both enhanced the total sleep time including rapid eye movement (REM) sleep and non-REM sleep time. CONCLUSION These results suggested that the enrichment of flavonoid aglycons such as liquiritigenin and isoliquiritigenin enhanced the anti-inflam-matory potency of licorice extract,and this potentiation has nothing to do with glycyrrhizic acid or glycyr-rhetinic acid.In addition,enrichment of flavonoid aglycones did not alter the hypnotic effect of licorice.

Objective To explore the influence of practical teaching in community in health education on the nurses' critical thinking ability in higher vocational school.Methods The contrast group was to fulfill practical teaching according to teaching plans.On weekends,the experimental group organizeed nurses to create wall newspapers about health education,carry out health education consultation,check residents' health responsibly and create family healthy files etc.The assessment of critical thinking ability was measured with Chinese Critical Thinking Disposition Inventory,CTDI-CV.Results There was a distinct difference about their critical thinking ability between the two groups(P ＜ 0.01).5 items showed clear differences among 7 excepts the ability of analysis and desire for knowledge.Conclusions Pratical teaching in community in health education can improve nurses' ability of critical thinking effectively.[Key words ] Health education;Community; Practical teaching; Nurse of higher vocationalschool;Critical thinking ability

Objective To investigate and compare the different effects of soluble adult wornl antigen(SWA)and soluble egg antigen(SEA)of Schistosoma japonicum on the apoptosis and cell-cycle of routine CD4~+T cells.Methods Purified CD4~+T ceUs from normal C57BL/6 mice were cultured with CFSE labeled antigen presenting clls in the presence of different stimuli for 36 h.Flow cytometry(FCM)was used to detect the apoptosis of CD4~+T cells by fluorescence conjugated caspase-3 antibodie staining.The flow cytometry was used to analyze the cell-cycle of CD4~+T cells cultured as described above for 96 h by propidium iodide staining.Results Compared with the apoptosis percentage of CD4~+T cells[(1.24±0.29)%]in the SEA stimulated group,that in the SWA stimulated group[(1.52±0.38)%]did not show statistically significant difference(P>0.05).Compared with the cell percentages in G1 phase[(78.91±2.98)%],S phase[(7.39±0.85)%]and G2/M phase[(10.69±1.05)%] in the SWA stimulated group,that of the G1 phase[(59.42±1.32)%]was significantly lower,but those in the S phase[(21.07±O.88)%] and G2/M phase[(18.88±1.21)%]were significantly increased in the SEA stimulated group(P<0.01).Conclusions There is no statistically significant difference between the apoptosis levels of CD4~+T ceHs stimulated by SWA and SEA.However,SEA significantly promotes the progression of the cell-cycle of CD4~+T cells compared with SWA.