In order to increase the production of insecticidal crystal proteins Cry1 and Cry2, firstly, Plack-ett-Burman design was applied to evaluate the effectiveness of the related nutrition factors; it was found that the soybean powder and MnSO4?H2O were significant factors for Cry1 production, but the yield of Cry2 wasn’t effected remarkably in such medium. Then the steepest ascent experiment was adopted to approach the optimal region of the medium composition. Lastly, the optimal concentration of the soybean powder and MnSO4?H2O was 11.5 and 0.02 g/L, obtained by response surface methodology (RSM). The final yields of Cry1 and Cry2 was 0.32 mg/mL and 0.11 mg/mL, increasing twice more than that in the medium optimized before. The median lethal concentration (LC50) of optimal medium was 1.09 ?L/mL. The toxicity to Heli-coverpa armigera was significantly enhanced than the old one.

OBJECTIVETo establish a new non-radioactive method for electrophoretic mobility shift assay (EMSA) to investigate the binding between glucocorticoid induced leucine zipper (GILZ) and peroxisome proliferator-activated receptor-gamma 2 (PPARgamma2) promoter oligonucleotides.

METHODSGILZ protein prepared by prokaryotic expression was linked to PPARgamma2 promoter oligonucleotides end-labeled with IRDye 800 infrared dye. The DNA-protein complex was separated with non-denatured polyacrylamide gel and scanned with the Odyssey. Infrared Imaging System.

RESULTSOne lane of DNA-protein complex was clearly presented, and the signal intensity increased along with the increment of the protein load.

CONCLUSIONThis infrared imaging system can be used for EMSA for detecting the DNA-protein complex with high sensitivity efficiency and allows easy operation.

Binding Sites ; DNA ; chemistry ; DNA-Binding Proteins ; chemistry ; metabolism ; Electrophoretic Mobility Shift Assay ; instrumentation ; methods ; Fluorescent Dyes ; chemistry ; Gene Expression Regulation ; Humans ; Infrared Rays ; Protein Binding ; Protein Interaction Domains and Motifs ; physiology ; Proteins ; chemistry

Objective To have a better understanding of the effort-reward imbalance for learning and learning burnout of high school students and their relationship. Methods A sample of 420 high school students was selected by stratified random sampling. Scales were used to study the effort-reward imbalance for learning and learning burnout and multiple linear regression analysis was used to study their relationship. Results A total of 387 high school students were actually investigated and 42.38% of which had effort-reward imbalance for learning. There were no significant differences in the rates of effort-reward imbalance for learning between students of different grades and between students of different homeplaces (P>0.05) . Female students had higher level of excessive input than males (P<0.05) . The average score of learning burnout was 56.93±13.22. There were no statistically significant differences in the scores of learning burnout between students of different genders and between students of different homeplaces (P>0.05) . The students who had effort-reward imbalance for learning scored higher in learning burnout than that who did not (P<0.05) . Senior Three students scored higher in learning burnout than Senior Two and One students (P<0.05) . The multiple linear regression analysis showed that learning reward and excessive input was both negatively correlated with learning burnout (all P<0.05) . Conclusion The high school students in Lishui City generally had the effort-reward imbalance for learning and learning burnout. learning reward and excessive input have effects on learning burnout.

OBJECTIVETo analyze the consistency of evolution condition between HA gene and the whole genome of influenza virus subtype A/H3N2 strains isolated in Zhejiang province from 1998 to 2009, and to study the potential antigenic region on the whole genome.

METHODSThe sequences of whole genome of 19 Zhejiang influenza virus isolates circulated from 1998 to 2009, which conserved by influenza laboratory of Zhejiang Provincial Centre for Disease Prevention and Control, were amplified using RT-PCR assays. The obtained sequences were used to conduct phylogenetic analysis with 10 contemporaneous vaccine strains. Three methods, including comparison of the amino acid substitutions, calculation of the entropy value and the filtering of positive selection sites, were used to confirm the mutable sites on each gene.

RESULTSThe whole genome of influenza virus subtype A/H3N2 was 4466 amino acids in length, with 137 stable mutations. The 144, 158 aa of HA gene mutate four and three times respectively; 93, 143, 307, 370, 372 aa of NA gene and 450 aa of NP gene mutate twice, and there were 29% (12/41) and 77% (24/31) mutations of HA and NA genes occurred on the non-epitope regions respectively. Analysis of the entropy value suggest that many amino acid sites on the non-epitope regions were prone to mutation, including 3, 225, 361 aa of HA gene; 93, 143, 147, 150, 372 aa of NA gene; 113, 576, 586 aa of PB1 gene; 101,256, 382, 421, 437 aa of PA; 377, 450 aa of NP gene; 218 aa of M1 gene and 31 aa of M2 gene.

CONCLUSIONBased on the whole genome of influenza virus subtype A/H3N2 strains isolated in Zhejiang province in 1998 to 2009, there may be several unknown or new antigen sites existing on the non-epitope regions of HA and NA genes and parts of internal genes. The phylogenetic tree constructed based on the complete sequence was more comprehensive than on the HA gene to reflect the genetic relationship and law of evolution among the influenza virus strains.

China ; DNA Mutational Analysis ; DNA, Viral ; genetics ; Evolution, Molecular ; Genetic Variation ; Genome, Viral ; Influenza A Virus, H3N2 Subtype ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Sequence Analysis

Objective To analyze the relationship between influenza epidemic and genetic characteristic on the whole genome of influenza virus subtype A/H3N2 strains isolated in Zhejiang province during 1998 to 2009. Methods All of the eight genes from the 19 Zhejiang influenza virus isolates, circulated during 1998 to 2009, were amplified by RT-PCR and sequenced. The obtained sequences were aligned and analyzed with the vaccine strains being used in the last 10 years.Results The highest mutation happened within HA and NA genes and the amino acid divergent ratios were 13.98% and 10.00%. Amongst the six internal proteins, the amino acid divergent ratios of NP, M2 and NS1 were 6.43%, 6.19% and 3.48% respectively, and the others were lower than 3%.Other than the HA and NA genes, mutations were also observed on six internal genes of the strains isolated in those years when the influenza virus subtype A/H3N2 was widely circulating.Additionally, there had been an obvious genetic lag between vaccine strains recommended by WHO and the contemporary Zhejiang epidemic strains for many years. Conclusion Besides on HA and NA genes, surveillance programs should also be covered mutations regarding the internal genes of influenza virus subtype A/H3N2 strains, in order to provide important information for forecasting and warning of a new round of influenza epidemic.

OBJECTIVETo observe the influence of the fusion of bone graft in spine of rabbits which were treated with lower intensity ultrasound.

METHODSForty 12-month-old rabbits were made to be the models of bone graft in post-lateral between two homonymy processus transverses in lumbar, and divided into treatment group (B) and control group (A) randomly. Twenty rabbits of treatment group were treated with lower intensity ultrasound, killed after six weeks, and took radiological examination, measured indexs of biomechanics.

RESULTSAfter 6 weeks of fusion of bone graft, treatment group were higher 6%-7% (P> 0.05) in strength, rigidity, torque andantitwist, maxload than that of control group.

CONCLUSIONLower intensity ultrasound can promote the speed and strength fusion of bone graft in young rabbits.

Animals ; Biomechanical Phenomena ; Bone Transplantation ; Female ; Humans ; Male ; Rabbits ; Random Allocation ; Spinal Diseases ; physiopathology ; surgery ; therapy ; Spinal Fusion ; Spine ; physiopathology ; surgery ; Ultrasonic Therapy

Objective:To investigate the positive rate of postpartum depression/anxiety screening and its associated factors in Jinping area, Yunnan Province.Methods:This cross-sectional survey involved 761 women who delivered live, singleton infants at or after 28 gestational weeks from October 2019 to February 2021 in the People's Hospital of Jinping Miao, Yao, and Dai Autonomous County, Honghe Hani and Yi Autonomous Prefecture, Yunnan Province. A questionnaire survey on childbirth and upbringing, the Edinburgh Postnatal Depression Scale (defined as positive when ≥9 score), and the Self-rating Anxiety Scale (defined as positive when ≥50 score) were conducted at postpartum day 1 to 3. General obstetric information and medical history were also retrieved from medical records. The risk factors of maternal depression and anxiety were analyzed using Chi-square test and multivariate logistic regression. Results:All 761 parturients completed the questionnaire. The total positive rate was 7.49% (57/761) for depression screening and 8.02% (61/761) for anxiety screening. Univariate analysis showed that postpartum hemorrhage, intrapartum infection and puerperal morbidity, neonates being transferred to the pediatric ward, attendance of prenatal classes during pregnancy, whether the neonatal gender was in line with the maternal and family expectations were all associated with both postpartum depression and postpartum anxiety. In addition, an association was found between gravidity, parity, delivery mode and postpartum depression, as well as accompanied delivery, breastfeeding and postpartum anxiety (all P<0.05). Multivariate analysis showed that postpartum hemorrhage ( OR=1.934, 95% CI: 1.010-3.704), neonates being transferred to the pediatric ward ( OR=1.990, 95% CI: 1.037-3.816), and not attending prenatal classes during pregnancy ( OR=3.393, 95% CI: 1.166-9.872) were the risk factors for postpartum depression; neonates being transferred to the pediatric ward ( OR=1.972, 95% CI: 1.040-3.740) and non-breastfeeding ( OR=2.174, 95% CI: 1.077-4.389) were risk factors for postpartum anxiety (all P<0.05). Conclusions:Parturients in Jinping area of Yunnan Province were at a lower risk of postpartum depression/anxiety. Active attendance at prenatal classes and breastfeeding may help reduce the risk of postpartum depression/anxiety.

OBJECTIVETo construct and express human CD96 gene outer membrane domain (hCD96om) in prokaryotic cells and prepare rabbit polyclonal antibody of hCD96om.

METHODShCD96om was amplified by RT-PCR from the peripheral blood of patients with acute myeloid leukemia and inserted into prokaryotic expression vector pET32a(+) to construct the recombinant plasmid pET32-CD96. The expression of hCD96om was induced by IPTG in BL21(DE3) cells, and the expression product was identified by Western blotting. The anti-hCD96 polyclonal antibody was prepared by immunization of rabbits with the fusion protein. The specificity of anti-hCD96 antibody was determined by Western blotting.

RESULTShCD96om protein was expressed in E.coli BL21(DE3) cells in the form of inclusion body, with a relative molecular mass around 37 kD. Western blotting showed a specific reaction of the prepared antiserum with the 70 kD protein extracted from human leukemia cell line HL-60 cells and with the 37 kD hCD96om fusion protein.

CONCLUSIONThe CD96 gene of human has been successfully cloned and expressed in BL21(DE3) cells, and its rabbit polyclonal antibody has been obtained.

Animals ; Antibodies ; immunology ; metabolism ; Antigens, CD ; biosynthesis ; genetics ; immunology ; Base Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Humans ; Immune Sera ; biosynthesis ; Immunization ; Leukemia, Myeloid, Acute ; immunology ; Molecular Sequence Data ; Neoplastic Stem Cells ; immunology ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology

BACKGROUNDNurr1 plays an essential role in the development, survival, and function maintenance of midbrain dopaminergic (DA) neurons, and it is a potential target for Parkinson's disease (PD). Nurr1 mRNA can be detected in peripheral blood mononuclear cells (PBMCs), but whether there is any association of altered Nurr1 expression in PBMC with the disease and DA drug treatments remains elusive. This study aimed to measure the Nurr1 mRNA level in PBMC and evaluate the effect of Nurr1 expression by DA agents in vivo and in vitro.

METHODSThe mRNA levels of Nurr1 in PBMC of four subgroups of 362 PD patients and 193 healthy controls (HCs) using real-time polymerase chain reaction were measured. The nonparametric Mann-Whitney U-test and Kruskal-Wallis test were performed to evaluate the differences between PD and HC, as well as the subgroups of PD. Multivariate linear regression analysis was used to evaluate the independent association of Nurr1 expression with Hoehn and Yahr scale, age, and drug treatments. Besides, the Nurr1 expression in cultured PBMC was measured to determine whether DA agonist pramipexole affects its mRNA level.

RESULTSThe relative Nurr1 mRNA levels in DA agonists treated subgroup were significant higher than those in recent-onset cases without any anti-PD treatments (de novo) (P < 0.001) and HC groups (P < 0.010), respectively. Furthermore, the increase in Nurr1 mRNA expression was seen in DA agonist and L-dopa group. Multivariate linear regression showed DA agonists, L-dopa, and DA agonists were independent predictors correlated with Nurr1 mRNA expression level in PBMC. In vitro, in the cultured PBMC treated with 10 μmol/L pramipexole, the Nurr1 mRNA levels were significantly increased by 99.61%, 71.75%, 73.16% in 2, 4, and 8 h, respectively (P < 0.001).

CONCLUSIONSDA agonists can induce Nurr1 expression in PBMC, and such effect may contribute to DA agonists-mediated neuroprotection on DA neurons.

Adult ; Aged ; Aged, 80 and over ; Dopamine Agonists ; therapeutic use ; Female ; Humans ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Male ; Middle Aged ; Nuclear Receptor Subfamily 4, Group A, Member 2 ; genetics ; Parkinson Disease ; drug therapy ; genetics ; RNA, Messenger ; genetics ; Young Adult

OBJECTIVETo observe the effect of rhynchophylline on methamphetamine-dependent zebrafish and explore the possible mechanism.

METHODSZebrafish were divided into control group, amphetamine group, low- (50 mg/kg) and high (100 mg/kg)-dose rhynchophylline groups, and ketamine (150 mg/kg) group. Conditioned place preference (CPP) was induced in zebrafish with methamphetamine, and the staying time in the drug box and the tracking map of the zebrafish were observed with Noldus Ethovision XT system. The protein expressions of TH, NR2B and GLUR2 in the brain of zebrafish with CPP were detected with Western blotting.

RESULTSCompared with the control group, zebrafish in methamphetamine group showed significant variations in the staying time and swimming distance in the drug box after conditioning (P<0.05) with obvious alterations of NR2B, TH and GLUR2 expressions in the brain (P<0.05). Treatment of methamphetamine-dependent zebrafish with high-dose rhynchophylline significantly reduced the variations in the staying time and swimming distance in the drug box (P<0.05) and in the expressions of NR2B, TH and GLUR2 in the brain (P<0.05).

CONCLUSIONRhynchophylline can inhibit methamphetamine dependence in zebrafish, the mechanism of which may involve the expressions of TH, NR2B and GLUR2 proteins in the brain.