Molting is an important physiological phenomenon of many metamorphosis insects, during which the old and new epidermis are separated by enzymes present in the molting fluid. Various proteomic studies have discovered the presence of Bombyx mori carboxypeptidase A (Bm-CPA) in the molting fluid of silkworm, but its function remains unclear. In order to better understand the role of Bm-CPA in the molting process of silkworm, Bm-CPA was analyzed by bioinformatics analysis, real-time fluorescence quantitative PCR, antibody preparation, immunofluorescence staining, and expression in Pichia pastoris. The results showed that Bm-CPA had a conserved M14 zinc carboxypeptidase domain and glycosylation site. Its expression was regulated by ecdysone 20E, and large expression was observed in the epidermis of the upper cluster stage. Immunofluorescence staining showed that Bm-CPA was enriched in the epidermis during the molting stage, and the inhibitor of Bm-CPA led to the larval death due to the inability to molt. We also successfully obtained a large number of recombinant Bm-CPA proteins by Pichia pastoris expression in vitro. These results may facilitate further understanding the molting development process of silkworm.
Animals ; Molting/genetics* ; Bombyx/genetics* ; Carboxypeptidases A/metabolism* ; Proteomics ; Larva/metabolism* ; Fluorescent Antibody Technique ; Insect Proteins/metabolism*

glioma; U87 cell; U251 cell; MAGE-A1; MAGE-A3; DNA methylation; histone acetylation To detect the expressions of melanoma antigen genes MAGE-A1 and MAGE-A3 in glioma tissues and to explore their clinical significance. Methods: Seventy-eight surgically resected glioma specimens and 15 normal brain tissue samples from donors suffered traffic accidence were collected at the Department of Neurosurgery, the Fourth Hospital of Hebei Medical University between January 2006 and January 2010, and the mRNAexpressions of MAGE-A1 and MAGE-A3 in collected tissues were detected with RT-PCR; their associations with the overall survival of patients were also analyzed.The promoter methylation status of the two genes was observed with methylation specific PCR, and the relationship between the gene expressions and promoter methylation status was analyzed. The expressions of MAGE-A1 and MAGE-A3 genes in U251 and U87 glioma cell lines were detected by RT-PCR before and after the treatment with DNA methyltransferase inhibitor 5-aza-CdR and/or histone deacetylase inhibitor trichostatin A (TSA). Results:The positive expression rates of MAGE-A1 and MAGE-A3 genes in glioma tissues were 65.34% and 38.46%, respectively; however, the two genes were not detected in 15 cases of normal brain tissues.The 5-year overall survival of patients in MAGEA1 positive expression group was shorter than that of negative expression group (P<0.05). There was significant correlation between the mRNA expressions of two genes and their promoter methylation status (all P<0.01). There was no mRNA expressions of MAGEA1 and MAGE-A3 in U87 cells untreated with 5-Aza-CdR and TSA, but a small amount of MAGE-A1 mRNA and MAGE-A3 mRNA were detected in U251 cells. TSA alone could not activate the expression of MAGE-A1 and MAGE-A3 genes. 5-Aza-CdR alone or in combination with TSA could activate the expression of both genes, and the combined effect was better than that of single administration. Conclusion: There are different degrees of MAGE-A1 and-A3 expression in glioma tissues, and the expression of MAGE-A1 is a negative prognostic factor for glioma patients. DNApromoter methylation and histone acetylation are important mechanisms of the activation of MAGE-A1 and MAGE-A3 expression.

Objective? To explore the nursing cooperation of laparoscopic radical distal gastrectomy. Methods? From January 2014 to December 2017, we summarized the main points of nursing cooperation among 41 patients with laparoscopic radical distal gastrectomy at Xi'an Gaoxin Hospital. Results? Among 41 patients, all operations were successful except four of them with laparotomy because of difficult peripheral tissue isolation of stomach or uncontrolled bleeding. The operation time, intraoperative blood loss and postoperative hospital stay were 240 to 350 min, (200±42) ml and (7.0±2.3) d respectively. No patients developed surgical complications during their hospital stay. Conclusions? During operations, nurses are familiar with the surgical procedure, performance as well as methods of application of laparoscopic instruments, and cooperate reasonably and precisely in operation which is the key to carrying out laparoscopic radical distal gastrectomy successfully.

Objective To study the effect of a new immunomodulator composed of muramyl dipeptide(MDP) and anti-CD10monoclonal antibody(MDP-Ab)on the dendritic cells(DC)of children with acute leukemia. Methods DC was adopted to divide the children with acute lymphoblastic leukemia into 6 groups,including the control group,unconjugated anti-CD10alone,unconjugated MDP alone,MDP-Ab alone,lipopolysaccharide(LPS)alone and MDP-Ab + LPS.The immunophenotypes,the endocytosis interleukin-12(IL-12)were detected.The stimulation index of autologous lymphocytes was assayed by adopting 5-(and 6)-carboxyfluorescein diacetate,succinimidyl es-ter(CFSE)-staining method.The supernatants of DC and autologous lymphocytes were used to detect the level of in-terferon-γ(IFN-γ)by using enzyme-linked immunosorbent assay.Results (1)DC immunophenotype:The ex-pressions of human leukocyte antigen-DR(HLA-DR),mature molecule(CD83)and co-stimulatory molecules (CD80and CD86)were increased significantly upon DC triggered with MDP-Ab,compared with the control group,un-conjugated anti-CD10group,and unconjugated MDP group,but lower than those in LPS and combination of MDP-Ab with LPS(F=629.62,P=0.000).(2)The level of IL-12:a significant increase in IL-12 level was detected in MDP-Ab group,LPS group,and combination of MDP-Ab with LPS group,compared with the control group,uncon-jugated anti-CD10group,and unconjugated MDP group(F=857.87,P=0.000). There were significant differences among the first three groups.(3)Endocytosis assay:The uptake of DCs stimulated by unconjugated anti-CD10,un-conjugated MDP,MDP-Ab immunoconjugate,LPS or combination was lower than that of immature DC in the control group which was(81.3 ± 10.1)%.(4)Mixed lymphocyte reaction and IFN-γ level:DC,treated with MDP-Ab, LPS and combination,stimulated more CFSE positive cells and higher level of IFN-γ secretion than the control group and unconjugated anti-CD10group,unconjugated MDP group. The most significance was observed in combination of MDP-Ab with LPS(F=393.36,P=0.000;F=2 497.18,P=0.000).Conclusion It is concluded that MDP-Ab could promote the proliferation and maturation of DC derived from blood of children with acute leukemia.

This paper was aimed to study the safety of moxibustion with FMEA method.Failure mode and effects analysis (FMEA) were used in every aspect of the operation process of moxibustion.And the local skin temperature was measured in 80 patients treated with moxa box moxibustion.The results showed that the skin temperature reached the highest when the moxibustion was given for 15 min,which was in consistence with the patients' chief complaints and their tolerances.It indicated that moxibustion for 15 min was the best moxibustion amount.Meanwhile,inspection should be made to avoid burning.After the application of FMEA,the RPN of the inspection activities,the temperature and distance of moxibustion were significantly decreased (P ＜ 0.05).It was concluded that the application of FMEA management mode strengthened the risk management of moxibustion treatment,standardized treatment process,provided the basis for the temperature and distance of moxibustion,and ensured the safety and efficacy of treatment.

Objective To investigate the application effect of point-factor method in constructing nursing performance management mode in Operating Room, so as to provide reference for improving quality of operating room nursing.Methods Relative indexes were collected and compared before ( December 2014) and after ( December 2015) implementation of nursing performance management mode in Operating Room basing on point-factor method, including performance assessment of nurses, professional standard test of nurses and satisfaction of nursing.Results The nursing performance management mode constructed by point-factor method included 4 first-level indexes and 20 second-level indexes, after implementation of which, Operating Room nurses′scores of theoretical level, duty work quality, equipment operating quality, round going quality and total scores were all improved ( P<0.05);scores of their three basis theory, three basis operation and professional achievements were also improved ( P<0.05);satisfaction of patients, doctors and Operating Room nurses to the nursing work were improved (P<0.05).Conclusions The nursing performance management mode constructed by point-factor method can promote management efficiency in the Operating Room and improve quality of nursing.

Objective:To investigate the influence of acrylamide ( ACR) on adult neurogenesis and expression of GSK 3βin mouse.Methods:Method Adult male Kunming mice were used and divided into two groups:control group and experimental poisoning groups,that were exposured to acrylamide by intraperitoneal injection.Brdu labeling and immunohistochemistry were used to investigate the proliferation of adult neural stem cells in the subgranular zone ( SGZ).BrdU/NeuN/GFAP triple labeling to investigate the survival and differentiation of newly generated cells.Detecting GSK3βexpression and distribution in Neuro-2a cells,the expression of GSK3βwas examined by using Western blot.Results:Compared with control mice ,lower number of BrdU-positive cells and less differentiated into neurons in ACR mice.Less neural stem cells survived ,but more glia cells were generated in the subgranular zone of acrylamide mice.Moreover,higher phosphorylated GSK 3β( Ser9 ) were detected in Neuro-2a cells and mouse dentate gyrus in ACR mice respectively .Conclusion:These results suggested that acrylamide inhibits neural stem cells proliferation and influences the survival and differentiation of newly generated cells.Acrylamide inhibits neurogenesis maybe through GSK 3βsignaling pathway.

Objective]Collecting and studying Wang Fu and his book YiLing CuanYao TanYuan systematically. [Method]Literature research on Wang Fu's life and the versions ofYiLing CuanYao TanYuan, reviewing his concept of study and academic thoughts. [Result]Confucian Wang Fu wrote more than 30 manuscripts, his medical masterpiece“YiLing CuanYao TanYuan, existing versions of two Qing printed editions and one Qing transcript, guided by medical source, centered on traditional Chinese medicine and prescription, accounted for human disease, made a comprehensive and profound elaboration on the basic theory of TCM.The recipes and traditional Chinese medicine of this book have clinical significance . [Conclusion]The collecting and studying have important literature value. These are worthy of our future generations to inherit and carry forward.

Objective To summarize the therapeutic effects of combined hyperbaric oxygen on treatment of critical ill patients and the relevant problems to attention.Methods One hundred and forty-six critical ill patients, admitted to our hospital from January 2009 and December 2013, were chosen in our study;the relations of hyperbaric oxygen efficacy with different diseases and intervention times were analyzed.Methods and measures of safe and effective treatments for hyperbaric oxygen were summarized.Results The critical ill patients were treated with hyperbaric oxygen, with an effective rate of 73.97％ and a cure rate of 32.19％.The percentage of intervention time for cured patients within 30 days (73.34％) was obviously higher than that of more than 30 days (27.66％);and the hyperbaric oxygen times in patients with early treatment (less than 30 days) was obviously less than that in patients with delayed treatment (more than 30 days).There was no obvious difference between different diseases.Conclusions Hyperbaric oxygen therapy is effective for all kinds of critical ill patients.The cure rate is high and the treatment times are less when early intervention is applied.The patients with delayed intervention should insist on the long course of treatment so as to achieve satisfactory effect.And, it is the key to make the safe and effective treatments for patients with complicated diseases.

Objective To evaluate the reliability of Pain Vision method for assessment of labor pain.Methods Eighty-nine nulliparous parturients who were at full term and at the latent period of the first stage of labor,aged 19-31 yr,weighing 55-85 kg,were enrolled in this study.The degree of labor pain was assessed using Pain Vision method and VAS.Pain degree is calculated from two parameters,current perception threshold and pain compatible electrical current by using a somatosensory evoked potential stimulator.The former parameter was defined by the lowest electrical current detected; the latter parameter defined by the electrical current judged as being compatible with the intensity of ongoing pain.Linear correlation of VAS scores with pain degree was analyzed.Results There was no correlation between pain degree and VAS scores,and the correlation coefficient was 0.206 (P ＞ 0.05).Conclusion Pain Vision method can not be applied for objective assessment of labor pain.