ObjectiveTo study the relationship between EphA2 and laryngeal carcinoma angionegesis.MethodsImmunohistochemical staining was used to determine the expression of EphA2 and CD34 in 20 normal squamous epithelium tissues and 50 laryngeal carcinoma.The relationship between EphA2 expression and Microvascular density was analyzed bg Spearman correlation and linear regression.ResultsThe MVD in laryngeal carcinoma (54.89 ± 13.67) was significantly higher than that in normal squamous epithelial tissues ( 17.15 ± 5.21 ) ; The EphA2 in laryngeal carcinoma(4.56 ± 1.38) was significantly higher than that in normal squamous epithelial tissues ( 2.49 ± 1.23 ) ( t =16.721,5.847,all P ＜ 0.05 ).There was significant positive linear correlation between EphA2 and MVD in laryngeal squamous cell carcinoma(P ＜ 0.01 ).ConclusionEphA2 may correlate with angionegesis in laryngeal squamous cell carcinoma.

Objective To investigate the expression and correlation of cell cycle protein-D1 and Ki-67 in laryngeal cancer tis-sues.Methods Immunohistochemical streptavidin-perosidase ( SP) staining method was used to detect the expression of Cyclin D 1 and Ki-67 proteins in laryngeal squamous cell carcinoma tissues and their correlation between Cyclin D 1 and Ki-67 was analyzed .Re-sults Among 56 cases of Cyclin D1 positive over-expressed laryngeal squamous cell carcinoma tissues , 41 cases of Ki-67 had positive over-expression.Among 51 cases of Ki-67 low-expressed tissues, 35 cases of Cyclin D1 had low-expression.Spearman rank correlation test showed that expressions of Cyclin D 1 and Ki-67 in laryngeal squamous cell carcinoma tissues had a significant positive correlation ( rs =0.620, P <0.01).Conclusions Cyclin D1 and Ki-67 expressions in laryngeal squamous cell carcinoma have a significantly positive correlation .

Objective To establish reference values of normal fetal heart conduction time intervals by tissue Doppler imaging(TDI)and pulsed Doppler(PD)echocardiography,and to assess their correlation with gestational-age and fetal heart rate.Methods One hundred and eighty-nine pregnant women underwent detailed echocardiographic examinations.Atrio-ventricular conduction time interval(AV)and the time interval from onset of ventricular contraction to the onset of atrial contraction of next cardiac cycle(VA)were measured by TDI and PD echocardiography.Results TDI-AV was(126.56±15.33)ms(95% CI 124.10～129.03 ms),TDI-VA was(285.22±24.53)ms(95% CI 281.27～289.16 ms),PD-AV was(127.42±12.88)ms(95% CI 125.35～129.49 ms),PD-VA was(287.42±25.19)ms(95% CI 283.37～291.47 ms).A paired t test revealed no systematic difference between the two approaches used to measure AV and VA.AV and VA were significantly positively correlated with gestational age,and significantly negatively correlated with fetal heart rate.Heart conduction time intervals were altered in fetus with paroxysmal arrhythmia.Conclusions This study established the normal values of fetal heart conduction time intervals.Prenatal determination of fetal heart conductional time intervals has important potential clinical utility in assessing fetal arrhythmia.

Objective:To analyze the level of self-management and its influencing factors in patients with systemic lupus erythematosus (SLE).Methods:A cross-sectional study. From January to December 2021, patients with SLE in the Department of Rheumatology and Immunology in the First Affiliated Hospital of Henan University of Traditional Chinese Medicine were selected as the study objects by convenience sampling method. A total of 135 questionnaires were distributed in this study, 135 were recovered, and 128 were valid. The general data of the patients were collected, and the self-management level, self-efficacy, family care and meaning in life of the patients were evaluated with the chronic disease self-management behavior scale, general self-efficacy scale (GSES), family adaption partnership growth affection resolve index (APGAR) and the meaning in life questionnaire (MLQ). Multiple linear regression analysis was applied to explore the factors affecting the self-management of SLE patients.Results:The total score of chronic disease self-management behavior scale in the 128 SLE patients was (28.77±6.58) points. Pearson correlation analysis showed strong correlation between self-efficacy, sense of life meaning and self-management ( r=0.748, 0.717, both P<0.001), there was a moderate correlation between family care and self-management ( r=0.560, P<0.001). Multiple linear regression analysis showed that educational level ( β′=0.457), disease activity ( β′=-0.211), self-efficacy ( β′=-0.372), family care ( β′=-0.510), meaning in life ( β′=-0.386) were influencing factors of self-management level in patients with SLE (all P<0.05). Conclusions:The self-management level of SLE patients needs to be improved. Disease activity, self-efficacy, meaning in life and other factors can affect the patients′ self-management level.

Glutamic acid is an important amino acid with wide range of applications and huge market demand. Therefore, by performing transcriptome sequencing and re-sequencing analysis on Corynebacterium glutamicum E01 and high glutamate-producing strain C. glutamicum G01, we identified and selected genes with significant differences in transcription and gene levels in the central metabolic pathway that may have greatly influenced glutamate synthesis and further increased glutamic acid yield. The oxaloacetate node and α-ketoglutarate node play an important role in glutamate synthesis. The oxaloacetate node and α-ketoglutarate node were studied to explore effect on glutamate production. Based on the integrated strain constructed from the above experimental results, the growth rate in a 5-L fermenter was slightly lower than that of the original strain, but the glutamic acid yield after 48 h reached (136.1±5.53) g/L, higher than the original strain (93.53±4.52) g/L, an increase by 45.5%; sugar-acid conversion rate reached 58.9%, an increase of 13.7% compared to 45.2% of the original strain. The application of the above experimental strategy improved the glutamic acid yield and the sugar-acid conversion rate, and provided a theoretical basis for the metabolic engineering of Corynebacterium glutamicum.
Citric Acid Cycle ; Corynebacterium glutamicum/metabolism* ; Glutamic Acid/metabolism* ; Metabolic Engineering ; Metabolic Networks and Pathways/genetics*

L-glutamic acid is the world's largest bulk amino acid product that is widely used in the food, pharmaceutical and chemical industries. Using Corynebacterium glutamicum G01 as the starting strain, the fermentation by-product alanine content was firstly reduced by knocking out the gene encoding alanine aminotransferase (alaT), a major by-product related to alanine synthesis. Secondly, since the α-ketoglutarate node carbon flow plays an important role in glutamate synthesis, the ribosome-binding site (RBS) sequence optimization was used to reduce the activity of α-ketoglutarate dehydrogenase and enhance the glutamate anabolic flow. The endogenous conversion of α-ketoglutarate to glutamate was also enhanced by screening different glutamate dehydrogenase. Subsequently, the glutamate transporter was rationally desgined to improve the glutamate efflux capacity. Finally, the fermentation conditions of the strain constructed using the above strategy were optimized in 5 L fermenters by a gradient temperature increase combined with a batch replenishment strategy. The glutamic acid production reached (135.33±4.68) g/L, which was 41.2% higher than that of the original strain (96.53±2.32) g/L. The yield was 55.8%, which was 11.6% higher than that of the original strain (44.2%). The combined strategy improved the titer and the yield of glutamic acid, which provides a reference for the metabolic modification of glutamic acid producing strains.
Glutamic Acid ; Corynebacterium glutamicum/genetics* ; Ketoglutaric Acids ; Metabolic Engineering ; Alanine

γ-aminobutyric acid can be produced by a one-step enzymatic reaction catalyzed by glutamic acid decarboxylase. The reaction system is simple and environmentally friendly. However, the majority of GAD enzymes catalyze the reaction under acidic pH at a relatively narrow range. Thus, inorganic salts are usually needed to maintain the optimal catalytic environment, which adds additional components to the reaction system. In addition, the pH of solution will gradually rise along with the production of γ-aminobutyric acid, which is not conducive for GAD to function continuously. In this study, we cloned the glutamate decarboxylase LpGAD from a Lactobacillus plantarum capable of efficiently producing γ-aminobutyric acid, and rationally engineered the catalytic pH range of LpGAD based on surface charge. A triple point mutant LpGAD^{S24R/D88R/Y309K} was obtained from different combinations of 9 point mutations. The enzyme activity at pH 6.0 was 1.68 times of that of the wild type, suggesting the catalytic pH range of the mutant was widened, and the possible mechanism underpinning this increase was discussed through kinetic simulation. Furthermore, we overexpressed the Lpgad and Lpgad^{S24R/D88R/Y309K} genes in Corynebacterium glutamicum E01 and optimized the transformation conditions. An optimized whole cell transformation process was conducted under 40 ℃, cell mass (OD₆₀₀) 20, 100 g/L l-glutamic acid substrate and 100 μmol/L pyridoxal 5-phosphate. The γ-aminobutyric acid titer of the recombinant strain reached 402.8 g/L in a fed-batch reaction carried out in a 5 L fermenter without adjusting pH, which was 1.63 times higher than that of the control. This study expanded the catalytic pH range of and increased the enzyme activity of LpGAD. The improved production efficiency of γ-aminobutyric acid may facilitate its large-scale production.
Glutamate Decarboxylase/genetics* ; Lactobacillus plantarum/genetics* ; Catalysis ; gamma-Aminobutyric Acid ; Hydrogen-Ion Concentration ; Glutamic Acid