Objective To obtain highly expressing cell lines by inserting the glutamine synthetase (GS) screening system and replacing the promoter of the vector.Methods The mutation of the point BamHⅠwas induced to build a new vector pIRES2-EGFP.The marker gene GS was inserted by AseⅠ and NheⅠ, and the promoter hCMV was replaced by PacⅠand NheⅠ.The new vector pHGS1.0 and the vector pIRES2-enhanced screen fluorescein protein( EGFP)-B were inserted by the recombinant protein TEM8 ( 1-227 )-VEGFR1 domain2-IgG2 ( TV-IgG2 ) gene to analyze the advantages of the expression.Results The glutamine synsthetase is successfully inserted, the human cytomegalovirus replaced, and recombinant protein is increased 5-fold by human immunoglobulin quantification kit.Conclusion The GS system is a highly protein expressing system.

Extracorporeal membrane oxygenation(ECMO)is an important cardiopulmonary support technique.It is the ultimate treatment for heart failure and lung failure after conventional treatment.ECMO enables the body to temporarily replace the pulmonary function and part of the heart function by extracorpore-al gas exchange,and creates a time window for the recovery of respiratory or cardiac function. In 1974, ECMO was first used in neonates.There are many complications in the application of ECMO in neonates, including mechanical complications,bleeding and thrombosis,hemolysis,neurological complications,renal dysfunction,gastrointestinal complications,infection,etc.Long term follow-up found that growth and develop-ment are affected by varying degrees. Timely monitoring,identification of complications and appropriate treatment will improve outcomes.

Objective To explore the behavioral characteristics of response to name in 16-30 months old infants with autism spectrum disorders (ASD),in order to provide a theoretical basis for the early identification and early diagnosis.Methods Two professionals,according to the scoring criteria and using video analysis methods,evaluated the response score (RS),reaction time (RT),duration time (DT) and the rate (RR) of response to their names among ASD infants (ASD group,61 cases),who were diagnosed at Child Developmental and Behavioral Center,the Third Affiliated Hospital of Sun Yat-Sen University from April to December 2017.Then they were compared with infants with developmental delays (DD group,32 cases) and neuro-typical (NT group,33 cases) infants.Finally,researchers predicted the diagnosis for ASD infants according to the behavioral indicators,which had significant differences compared with other groups.Results Compared with DD group [RS 2 (1) score,RT 1.32 (4.65) s,DT 2.69(1.84) s] and NT group [RS 2 (1) score,RT 1.37 (4.37) s,DT 2.90 (2.23) s],RS was significantly lower [1 (1)score],RT was significantly longer [5.87 (4.64) s],and DT was significantly shorter [0.77 (1.88) s] in ASD group,and the differences were statistically significant (H =-4.91,-5.94;5.36,5.41;-4.47,-5.78;all P ＜0.05);while the differences between DD group and NT group were not significant(all P ＞0.05).The RR was significantly lower in ASD group [0.25 (0.50)] and DD group [0.50 (0.25)],compared with NT group [0.75 (0.50)],and the differences were statistically significant (H =-6.39,-4.45,all P ＜ 0.01);while the differences between DDgroup and ASD group were not significant(P ＞0.05).When detecting ASD from ASD and NT infants,the area under the receiver operating characteristic curve (AUC) was 0.889 (P ＜0.01);when detecting ASD from ASD and DD infants,AUC was 0.924 (P ＜ 0.01);when detecting ASD from all infants,AUC was 0.868 (P ＜ 0.01),according to all indicators of response to name.Conclusions There are significant differences between ASD infants and DD and NT infants in response to name domain.Behavioral characteristics in the procedure of response to name can predict ASDwell.Response to name as an early social behavioral indicator,being tested at 2 years old,is still of importance for the early identification and early diagnosis of ASD.

OBJECTIVETo evaluate the therapeutic effect of integrative Chinese and Western medicine in treating mid-advanced crescentic nephritis (MACN).

METHODSThirty-two patients, their diagnosis was confirmed as MACN by renal biopsy, were divided, adopting randomized, controlled method, into two groups, the treated group and the control group, they were all, excepting one, treated with impact therapy of methyl-prednisolone followed with oral intake of prednisone, to part of them cyclophosphamide or mycophenolate mofetil was given in addition, to those with hypo-hemoglobin (< 90 g/L), subcutaneous injection of erythropoietin was administered. Decoction of Qingre Huoxue recipe (QHR), consisted of oldenlandia herb 30 g, honey-suckle stem 30 g, violet herb 30 g, red peony root 15 g, rehmannia root 15 g, solomonseal rhizome 15 g, asiabell root 30 g, red sage root 30 g, prepared rhubarb 12 g and giant-hyssop herb 12 g, were additionally given one dose per day to patients in the treated group. The renal function, improvement of anemia and immunosuppressive agents needed in patients were observed after 3 months treatment.

RESULTSAfter treatment, renal function was improved in both groups, but the effect in the treated group was better than that in the control group (P < 0.05). Anemia was partially alleviated in the two groups with no significant difference. The dosage of glucocorticoids used in the treated group was obviously lesser than that used in the control group (P < 0.01).

CONCLUSIONIntegrative Chinese and western medicine could treat crescentic nephritis to obtain good effect, and reduce the quantity of glucocorticoid necessity for treatment.

Adult ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerulonephritis ; drug therapy ; physiopathology ; Humans ; Immunosuppressive Agents ; therapeutic use ; Kidney Function Tests ; Kidney Glomerulus ; physiopathology ; Kidney Tubules ; physiopathology ; Male ; Methylprednisolone ; therapeutic use ; Middle Aged ; Phytotherapy

Chinese mitten crab (Eriocheir sinensis) is an important aquaculture species in Crustacea. Functional analysis, although essential, has been hindered due to the lack of sufficient genomic or transcriptomic resources. In this study, transcriptome sequencing was conducted on 59 samples rep-resenting diverse developmental stages (fertilized eggs, zoea, megalopa, three sub-stages of larvae, juvenile crabs, and adult crabs) and different tissues (eyestalk, hepatopancreas, and muscle from juvenile crabs, and eyestalk, hepatopancreas, muscle, heart, stomach, gill, thoracic ganglia, intes-tine, ovary, and testis from adult crabs) of E. sinensis. A comprehensive reference transcriptome was assembled, including 19,023 protein-coding genes. Hierarchical clustering based on 128 differ-entially expressed cuticle-related genes revealed two distinct expression patterns during the early lar-val developmental stages, demonstrating the distinct roles of these genes in 'crab-like"cuticle formation during metamorphosis and cuticle calcification after molting. Phylogenetic analysis of 1406 one-to-one orthologous gene families identified from seven arthropod species andCaenorhabditis elegans strongly supported the hypothesis that Malacostraca and Branchiopoda do not form a monophyletic group. Furthermore, Branchiopoda is more phylogenetically closely related to Hexapoda, and the clade of Hexapoda and Branchiopoda and the clade of Malacostraca belong to the Pancrustacea. This study offers a high-quality transcriptome resource for E. sinensis and demonstrates the evolutionary relationships of major arthropod groups. The differentially expressed genes identified in this study facilitate further investigation of the cuticle-related gene expression networks which are likely associated with'crab-like"cuticle formation during metamor-phosis and cuticle calcification after molting.

PET (polyethylene terephthalate) is one of the most important petrochemicals that is widely used in mineral water bottles, food and beverage packaging and textile industry. Because of its stability under environmental conditions, the massive amount of PET wastes caused serious environmental pollution. The use of enzymes to depolymerize PET wastes and upcycling is one of the important directions for plastics pollution control, among which the key is the depolymerization efficiency of PET by PET hydrolase. BHET (bis(hydroxyethyl) terephthalate) is the main intermediate of PET hydrolysis, its accumulation can hinder the degradation efficiency of PET hydrolase significantly, and the synergistic use of PET hydrolase and BHET hydrolase can improve the PET hydrolysis efficiency. In this study, a dienolactone hydrolase from Hydrogenobacter thermophilus which can degrade BHET (HtBHETase) was identified. After heterologous expression in Escherichia coli and purification, the enzymatic properties of HtBHETase were studied. HtBHETase shows higher catalytic activity towards esters with short carbon chains such as p-nitrophenol acetate. The optimal pH and temperature of the reaction with BHET were 5.0 and 55 ℃, respectively. HtBHETase exhibited excellent thermostability, and retained over 80% residual activity after treatment at 80 ℃ for 1 hour. These results indicate that HtBHETase has potential in biological PET depolymerization, which may facilitate the enzymatic degradation of PET.
Hydrolases/metabolism* ; Bacteria/metabolism* ; Hydrolysis ; Polyethylene Terephthalates/metabolism*