Objective: To investigate the coping strategies and it’s relationship with self- rated health state of military personnel in Tibet. Methods: 490 military personnel in Tibet were assessed by Simplified Coping Style Questionnaire and Self- rated Health Measurement Scale. Results: Military personnel in Tibet adopting positive strategies (23.3%) were more than those adopting negative strategies(16.2%) when facing stresses. The most common coping strategies they adopted were "restraining negative emotion(36.9%)" and "consoling themselve(s41.2%)". The scores of positive coping style had signifi- cant correlation with the scores of three subscales and the total scores of the self- rated health. Conclusion: Different cop- ing strategies have different effects on self- rated health of military personnel in Tibet, and the positive strategies could im- prove their status of self- rated health, but negative ones had contrary effect to health.

Objective To discuss the short-term clinical curative effect of laparoscopic colorectal cancer radical resection for rectal cancer. Methods Clinical data of 50 patients with rectal cancer underwent radical resection from November 2015 to November 2016 were retrospectively analyzed. Among them, 27 cases underwent laparoscopic radical resection (Laparoscopy group), the other 23 cases underwent radical resection (Laparotomy group). Then observe and record the operation time, tumor diameter, specimen length, number of lymph node cleaning, time of ambulation, postoperative anal exhaust time, postoperative defecation time, postoperative complications and postoperative eating time of the two groups. Results The tumor diameter, length of specimens and number of lymph node dissection in laparoscopic group were (3.8 ± 1.4) cm, (18.5 ± 2.1) cm and (7.2 ± 3.1), while in Laparotomy group were (3.9 ± 1.4) cm, (18.6 ± 2.3) cm, and (7.7 ± 3.4), the difference has no statistical significance (P > 0.05). The intraoperative blood loss, operation time, ambulation time, postoperative anal exhaust time, postoperative defecation time, postoperative eating liquid diet time, postoperative hospitalization time in laparoscopic group were (105.3 ± 23.8) ml, (140.2 ± 22.3) min, (4.0 ± 1.2) d, (6.0 ± 1.5) d, (3.0 ± 1.0) d, (3.5 ± 0.5) d and (4.0 ± 1.0) d, while in Laparotomy group were (210.4 ± 21.3) ml, (118.9 ± 20.7) min, (4.5 ± 1.1) d, (7.8 ± 1.2) d, (7.0 ± 1.6) d, (8.1 ± 2.0) d and (10.0 ± 3.2) d, there was significant difference between the two groups (P < 0.05). Conclusion Laparoscopic surgery for rectal cancer is safe and effective. It can achieve radical tumor resection, and intraoperative less bleeding, faster postoperative recovery, shorter hospitalization time.

Objective To observe the effects of CO2 pneumoperitoneum on the enteric motor function and acetylcholine esterase(AchE) neuron and nitric oxide synthase(NOS) neuron in the enteric nervous system,and explore the neuromechanism of the CO2 pneumoperitoneum on renteric motor function.Methods Thirty-six Sprague Dawley rats were randomly divided into experiment group(n=24) and control group(n=12).The experiment group was divided into two subgroups namely pneumoperitoneum 30min group and pneumoperitoneum 60min group(12 each) based on the maintenance time of pneumoperitoneum.Rats in each group were gavaged with medicinal carbon powder,and then the transmission of carbon powder in small intestine was determined.The spreading specimens of intestinal myenteric plexus of small intestine were prepared and the stained AchE and NOS neurons were observed and compared.Results The propellant velocity of carbon powder was slower in pneumoperitoneum 60min group than that in pneumoperitoneum 30min group and control group(28.55%?3.45% vs 45.90%?6.30%,48.25%?5.28%,P0.05).The number of positive expression of AchE neurons in intestinal myenteric plexus decreased in pneumoperitoneum 60min group compared with that in pneumoperitoneum 30min group and control group(48.00?3.16 vs 58.82?4.62,61.83?4.17,P0.05).The number of positive expression of NOS neurons in intestinal myenteric plexus increased in pneumoperitoneum 60min group compared with that in pneumoperitoneum 30min group and control group(42.17?4.45 vs 32.50?4.34,30.83?3.6,P0.05).Conclusions Prolonged CO2 pneumoperitoneum can affect or damage cholinergic neurons and nitroxidergic neurons in the enteric nervous system to some extent,and it may be the underlying mechanism of the intestinal motor dysfunction after operation.

Objective Moderate autophagy helps improve the viability of neurocytes.This study aims to investigate the effect of SP600125 on the autophagy and loss of nerve cells in the hippocampus in rats with subarachnoid hemorrhage (SHA).Methods Forty healthy male SD rats were equally randomized into a sham operation, an DMSO group, an SAH model, and an SP600125 group.The SAH model was established by vascular puncture and the rats of the SP600125 group were injected with 10 μL of SP600125 (3 μg/μL) into the lateral cerebral ventricle at 30 minutes before modeling.Sham group and SAH group were injected with equal volume of normal saline, DMSO group was injected with the same amount of DMSO.The animals were sacrificed at 24 hours after modeling for observation of the changes in the morphology and the number of neurons in the hippocampus by HE staining and qualitative and quantitative determination of the expressions of the p-JNK protein and the autophagy markers beclin-1 and LC3-II by immunohistochemistry and Western blot.Results Compared with the sham operation group, the neurons exhibited a disordered arrangement and the cells were polygonal and decreased in number in the hippocampus of the SAH models, while milder neuronal injury and more cells were observed in the rats of the SP600125 group than in the SAH models.The mean optical density values of Beclin-1, LC3-II and p-JNK in the hippocampus were significantly higher in the SAH models (14.66±4.40, 12.62±3.46, and 12.82±3.68) and DMSO (13.85±3.85、11.59±4.52、13.03±3.53), and the SP600125 group (9.86±3.14, 6.78±2.56, and 5.60±2.42) than in the sham operation group (1.56±0.28, 1.60±0.30, and 1.58±0.32) (P<0.05), but markedly lower in the SP600125 than in the SAH model group (P<0.05).The expressions of Beclin-1, LC3-II and p-JNK were remarkably increased in the SAH models (0.474±0.122, 0.668±0.130, and 0.496±0.124) and DMSO (0.432±0.102、0.628±0.113、0.416±0.094) and the SP600125 group (0.264±0.106, 0.332±0.113, and 0.219±0.104) than in the sham operation group (1.56±0.28, 1.60±0.30, and 1.58±0.32) (P<0.05), but significantly decreased in the SP600125 group as compared with the SAH models (P<0.05).Conclusion SP600125 has a protective effect on the neurocytes in the hippocampus of SAH rats, which may be associated with SP600125 moderately activating neuronal autophagy by inhibiting the activity of the JNK signaling pathway.

To prepare recombinant human retinol binding protein 4 (RBP4) by using the baculovirus expression system and to detect its immunogenicity, the fusion DNA fragment of secretory signal peptide SS64 and human RBP4 gene was subcloned into a baculovirus transfer vector pFastBac-dual(pFBd), and the corresponding recombinant transfer plasmid was transformed into E. coli strain DH10bac, after transposition recombinant shuttle bacmid was screened out. The logarithmic phase Sf9 cells were transfected with the recombinant bacmid and then the recombinant baculovirus containing hRBP4 expression box were generated. After amplification of recombinant baculovirus, the recombinant baculovirus seeds were obtained. To express human RBP4, logarithmic phase Sf9 cells were infected with the virus seeds and SDS-PAGE and Western blotting were used to detect and identify the expression. Finally, to prepare a batch of RBP4 protein, logarithmic phase Sf9 cells in suspension culture were infected with recombinant baculovirus seeds and the supernatant was harvested after 120 hours post-infection for purification. Finally for preparation of polyclonal antibody and evaluation of immunogenicity, the recombinant hRBP4 from insect cells and from E. coli were immunized rabbits. Restriction enzyme digestion and sequencing confirmed that the recombinant baculovirus transfer plasmid was constructed correctly, and subsequently recombinant RBP4-bacmid was generated successfully. SDS-PAGE and Western blotting analysis suggested that human RBP4 protein was highly expressed in Sf9 cells with the molecular weight of approximately 23 kDa. The recombinant RBP4 protein could be secreted into the medium efficiently, and the expression level was calculated amount of 100 mg/L. Finally the rabbit antiserum was harvested after recombinant RBP4 immunization, therein the titer of antiserum against baculovirus recombinant RBP4 is 1:100 000 whereas the titer of antiserum against E. coli recombinant RBP4 is only 1:10 000. Overall, human RBP4 was high efficiently expressed successfully with good antigenicity in baculovirus system, and high affinity antiserum was obtained. A solid foundation was laid for the next step of the preparation of human serum RBP4 detection kit.
Animals ; Baculoviridae ; genetics ; Blotting, Western ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Genetic Vectors ; Humans ; Immune Sera ; Insecta ; Rabbits ; Recombinant Proteins ; biosynthesis ; immunology ; Retinol-Binding Proteins, Plasma ; biosynthesis ; immunology ; Sf9 Cells ; metabolism ; Transfection

Objective:To explore the effects of phased goal directed fluid therapy (GDFT) during anesthesia surgery on tissue perfusion and cognitive function in patients undergoing radical lung cancer surgery.Methods:A total of 108 lung cancer patients were prospectively selected and randomly divided into a control group and a study group using a random number table method. The control group received classical restrictive liquid therapy, while the study group received staged GDFT. We compared the surgical time, intraoperative blood loss, colloid fluid dosage, crystalloid fluid dosage, total output, and urine volume between two groups of patients; Two groups of patients were compared in terms of oxygenation index (OI), respiratory index (RI), central venous oxygen saturation (ScvO 2), lactate (Lac), central venous arterial carbon dioxide partial pressure difference (Pcv-aCO 2), oxygen supply index (DO 2I), and oxygen uptake rate (O 2ERe) before anesthesia induction (T 0), before single lung ventilation (T 1), 1 hour of single lung ventilation (T 2), immediate resumption of dual lung ventilation (T 3), 30 minutes of dual lung ventilation (T 4), and after surgery (T 5); The Mini Mental State Examination (MMSE) was used to evaluate the cognitive function scores of two groups of patients 1 day before surgery and 1 and 3 days after surgery, while recording the incidence of cognitive dysfunction (POCD) and pulmonary complications (including pulmonary infection, acute lung injury, pulmonary embolism, pulmonary edema, atelectasis, etc.) within 3 days after surgery. Results:The amount of crystal fluid and urine output in the research group was significantly lower than that in the control group, while the amount of colloidal fluid was significantly higher than that in the control group (all P<0.05). The OI of the study group T 1-T 5 was significantly higher than that of the control group, while the RI of T 2-T 5 was significantly lower than that of the control group (all P<0.05). The ScvO 2 of the study group T 1 to T 5 was significantly higher than that of the control group, and the Lac was significantly lower than that of the control group (all P<0.05); The MMSE scores of both groups of patients were significantly lower than those before surgery on day 1 and 3 after surgery, and the MMSE scores of the study group were significantly higher than those of the control group on day 1 and 3 after surgery (all P<0.05). The incidence of POCD within 3 days after surgery in the study group was 16.67%(9/54), lower than 37.04%(20/54) in the control group (χ 2=5.704, P=0.017); The incidence of pulmonary complications in the study group was lower than that in the control group (5.56% vs 22.22%, χ 2=4.955, P=0.026). Conclusions:The application of staged GDFT during anesthesia in patients undergoing radical lung cancer surgery can further improve tissue perfusion, improve microcirculation and oxygen supply-demand balance of systemic organs and tissues, including the brain, alleviate perioperative brain function damage, and reduce the occurrence of postoperative POCD compared to conventional liquid therapy.

Objective:To prepare recombinant protein of group 36 allergen of Dermatophagoides farinae(Der f 36),to deter-mine its immunogenicity and bioinformatics analysis was performed.Methods:Nucleic acid sequence coding for Der f 36 was obtained and artificially synthesized,pET-28a(+)was inserted to construct pET-28a(+)-Der f 36 plasmid and transformed into BL21(DE3)receptor cells.After expressed and purified in BL21(DE3),recombinant allergen rDer f 36 was obtained,recombinant protein rDer f 36 was identified by SDS-PAGE and Western blot.Serum IgE binding rates of rDer f 36 was determined by IgE-ELISA.Cross-reactivity between rDer f 36 and rDer p 36 was detected by IgE-ELISA inhibition assay.HNEpC cells were cultured with rDer f 36 for 24 h and cytokines were detected.Bioinformatics softwares were used to analyze physicochemical properties and structures of Der f 36 and Der p 36.Results:Coding gene for Der f 36 was obtained with a total length of 690 bp,whose molecular weight was 25.6 kD;serum IgE binding rate of rDer f 36 was 42.1%by IgE-ELISA;IgE-ELISA inhibition assay showed that rDer p 36 had 40%(8/20)inhibition rate(>50%)for rDer f 36,with an average inhibition rate of 52.98%.Compared with control group,HNEpC cells cultured with rDer f 36 showed that IL-6,IL-8,IL-33,IL-25 and TSLP expressions had increased;bioinformatics analyses show that sequence consistency of Der f 36 and Der p 36 was 77.63%,with similar physicochemical properties.Secondary structure analysis showed that both of them contained α helix,β-turn and random coils,and content of random coils was the highest;structure of C2 domains was also highly overlapping(RMSD=0.046).Conclusion:Recombinant allergen rDer f 36 is successfully prepared with good immunogenicity,laying foundation for diagnosis and treatment of dust mite allergen single component.High similarity in physical and chemical properties,secondary structure,and tertiary structure between Der f 36 and Der p 36 determines their cross reactivity.

Objective:To prepare recombinant protein of group 27 allergen of Dermatophagoides farinae(Der f 27),and to determine its immunoactivity.Methods:pET-28a(+)-Der f 27 plasmid was constructed and inserted into E.coli BL21(DE3)cells.After being expressed and purified,recombinant allergen rDer f 27 was obtained.IgE binding rates of rDer f 27 with sera from patients with allergic rhinitis induced by Dermatophagoides farinae was determined by ELISA and Western blot.PBMC from patients with allergic rhinitis and BESA-2B cells were cultured with rDer f 27 for 24 h,respectively,and cytokine expression was measured.Bioinformatics softwares were used to analyze physicochemical properties and structures of Der f 27.Results:pET-28a(+)-Der f 27 plasmids were prepared successfully and transformed into BL21(DE3)cells.After expressed and purified with PTG,SDS-PAGE and Western blot identified a band about 48 kD.IgE binding rates of rDer f 27 were 39.5%and 45.5%with sera from patients with allergic rhinitis allergic to Dermatophagoides farinae by IgE-ELISA and IgE-Western blot,respectively.Compared with control group,IL-6 and IL-8 expressions were increased in PBMC from patients with allergic rhinitis being cultured with rDer f 27(P<0.05);expressions of IL-10 and TGF-β were decreased in BESA-2B cells being cultured with rDer f 27,while IL-17A and IL-23A expressions were increased.Bioinformatics analysis showed that Der f 27 belong to serine protease inhibitor family and had universal structure and func-tion of this family.Secondary structure of Der f 27 was mainly composed of α-helix(42.62%)and random coil(35.60%).Conclusion:Recombinant allergen rDer f 27 has been prepared successfully with good immunoreactivity and immunogenicity,becoming one of important allergens of allergic rhinitis.