Nonaqueous capillary electrophoresis-chemiluminescence (NACE) method was developed for the determination of five phenolic environmental estrogens including bisphenol A (BPA),4-nonylphenol(4-NP) ,4-tert-octylphenol(4-tOP),4-tert-butylphenol(4-tBP) and tetrabromobisphenol A(TBBPA) in food packaging material. Target compounds in soaking solution of food packaging material samples were separated by NACE-CL after derivatization with 4-(4,5-diphenyl-lH-imidazol-2-yl) benzoyl chloride ( DIB-Cl) ,reacted with per-oxyoxalate chemiluminescence system,and then detected through magnifying light signal with photomultiplier. With 17β-E_2 as internal standard,qualitative and quantitative analysis of the target compounds were performed by relative migration time and the ratio of relative chemiluminescence intensity,respectively. Several influence factors on separation with NACE,such as the composition and proportion of organic solvent,the concentration of electrolyte,temperature surrounding of the capillary,the concentration of acetic acid and work voltage,were investigated. And the peroxyoxalate chemiluminescence system was optimized. Under these conditions,4-tBP,BPA,4-OP,4-NP and TBBPA were all separated and showed good linearities in the ranges of 0.0095-3.0 mg/L,0.0087-3.0 mg/L,0.0085 -3.0 mg/L,0.011 -3.0 mg/L,0.009-3.0 mg/L,respectively,with correlation coefficients over 0. 9947. The RSDs of relative migration time and relative chemiluminescence intensity ratio were 0.9% -3.0% and 6.5% - 8.6%. Recoveries for the spiked samples ranged from 86.7% to 98. 8%. Five common food packaging material samples were analyzed. The proposed method is simple and sensitive for the quantitative determination of phenolic environmental estrogens in food packaging material samples.

Objective To study the clinical diagnostic value of transvaginal ultrasonic in early cervical carcinema and cervical neoplasis.Methotis The suspected patients with cervical carcinoma and cervical neoplasis were detected with transvaginal ultrasonography,liquid based cytology and cervical biopsy.The sonograms of transvaginal ultrasonic were retrospectively analyzed.Results In early cervical carcinoma and cervical neoplasis,the diagnostic sensitivity of transvaglnal ultrasonic was 90.9%and 83.3%;and the diagnostic specifity of transvaginal ultrasonic was 70.6% and 60.0%;the rate of missed diagnosis of transvaginal ultrasonic was 9.1% and 16.7%.Conclusion Transvaginal ultrasonic plays an important part in the clinical diagnosis of early cervical carcinoma.

Objective:To observe the relationship between the occurrence of symptomatic hypocalcemia (SH) and various potential influencing factors in patients after thyroidectomy, stratify according to the scope of thyroidectomy, and explore the predictive value of intact parathyroid hormone (iPTH) for postoperative SH.Methods:Among 3 379 patients with thyroidectomy who admitted into the First Affiliated Hospital of Zhengzhou University from January 2020 to February 2021, 122 patients with SH after thyroidectomy were collected retrospectively and set as SH group. 100 patients of the remaining 3 200 patients who did not suffer from SH in the same year were selected by systematic sampling method and set as control group. Pearson correlation analysis was used to analyze the potential influencing factors such as age, preoperative calcium, postoperative calcium, preoperative iPTH, postoperative iPTH, central lymph node number, blood loss, operation duration, gender, lymph node dissection method, thyroidectomy range, postoperative pathological type and other. Among them, the measurement data of normal distribution were expressed by mean±standard deviation( Mean± SD), t-test was used for the comparison between the two groups, and Chi-square test was used for count data. By drawing the receiver operating characteristic curve (ROC), the iPTH levels in patients with and without SH before/after operation (different surgical methods) were studied, and the diagnostic threshold, sensitivity and specificity of iPTH were predicted. Results:Among 3 379 patients, 122 patients suffered from SH after thyroidectomy, with the incidence rate of 3.6%. There were significant differences in gender (8 males and 114 females in SH group; 27 males and 73 females in control group), whether lateral area dissection was performed (58 cases with dissection and 64 cases without dissection in SH group; 7 cases with dissection and 93 cases without dissection in control group), thyroidectomy range (14 cases with one side and 108 cases with both sides in SH group; 73 cases with one side and 27 cases with both sides in control group), age (40.1 years old vs 43.2 years old), dissection number of central lymph nodes (8.6 vs 4.6), dissection number of cervical lymph nodes (12.3 vs 0.7), blood loss (22.8 mL vs 11.0 mL), operation duration (1.7 h vs 0.8 h), postoperative iPTH (16.4 pg/mL vs 41.9 pg/mL), preoperative iPTH (39.4 pg/mL vs 47.8 pg/mL) in SH group; and postoperative calcium level (1.9 mmol/L vs 2.2 mmol/L). There was significant differences between the two groups ( P<0.05). However, there was no significant differences between them with postoperative pathological type (4 cases with toxic goiter, 3 cases with medullary thyroid carcinoma, 1 case with thyroid follicular carcinoma, 114 cases with papillary thyroid carcinoma in SH group; 1 case with medullary thyroid carcinoma, 1 case of thyroid follicular carcinoma, 98 cases with papillary thyroid carcinoma in control group, P=0.25) and preoperative calcium (2.3 mmol/L vs 2.3 mmol/L, P=0.10). For patients with bilateral thyroidectomy, SH was easy to occur when postoperative iPTH < 20.08 pg/mL, and its sensitivity and specificity were 74.07% and 96.30%; however, for patients with unilateral thyroidectomy, SH was easy to occur when iPTH < 24.00 pg/mL after operation. Conclusions:Gender, age, postoperative calcium, preoperative iPTH, postoperative iPTH, central lymph node number, blood loss, operation duration, lymph node dissection method and thyroidectomy range are important factors affecting the occurrence of SH after thyroidectomy. With the expansion of surgical range, the postoperative iPTH level gradually decreases, which predicts the occurrence of symptomatic hypocalcemia. In order to avoid the occurrence of symptomatic hypocalcemia after operation, it is necessary to supplement calcium in time according to the range of operation and postoperative iPTH level.

Objective:To evaluate the value of intraoperative magnetic resonance imaging (iMRI) combined with neuronavigation for the resection of insular gliomas.Methods:From August 2014 to October 2017 in the First Hospital Affiliated to Sun Yat-sen University,clinical data of 41 patients with insular glioma,who underwent the surgery assisted with 3.0T iMRI and neuronavigation,were analyzed retrospectively,and the resection extent,complications and prognosis were evaluated.Results:Subtotal tumor resection was achieved in 21 patients and partial resection was done in 20 after iMRI scanning.After further resection,total tumor resection was achieved in 16 patients,subtotal resection in 18 and partial resection in 7.There was a statistical significant difference in tumor resection between pre-iMRI and post-iMRI according to the Fisher test (P＜0.05).In the follow-up from 3 months to 3 years,the symptoms of the 41 patients had improved.Conclusion:iMRI corrected the shift of brain.Neuronavigation can accurately and timely assess the degree of resecting tumor.The combination of neuronavigation with surgery can maximally and safely resect insular glioma.

Objective To construct a recombinant eukaryotic expression vector of human tumor suppressor p53-binding protein 2(TP53BP2)and transfect human embryonic kidney Expi293F cells.High-purity recombinant human full-length TP53BP2 protein was obtained and its biological activity was identified.Methods The TP53BP2 gene sequence was queried on the UniProt website,and the Expi293F expression system was optimized.The TP53BP2 gene was connected to pcDNA3.1(+)-P2A-eGFP vector by homologous recombination,and identified by double enzyme digestion and sequencing.Transect pcDNA3.1(+)-P2A-eGFP-TP53BP2 plasmid into Expi293F cells of Polyethylenimine(PEI),observe the transfection efficiency with a fluorescence microscope,collected cells from the experiment group and control group.The expression level of TP53BP2 recombinant protein was detected by Western blot(WB).Protein was purified by His label purification kit and Superdex 20010/300 GL chromatographic column.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis.The purified recombinant protein was identified by SDS-PAGE.Combining recombinant human full-length TP53BP2 protein with p65 protein was investigated for Co-immunoprecipitation(Co-IP)precipitation.Recombinant human full-length TP53BP2 protein was co-localized with p65 protein by Immunofluorescence(IF).The surface plasmon resonance(SPR)technique was used to detect the interaction between purified recombinant human full-length TP53BP2 protein and TP53BP2 antibody.Results The recombinant plasmid pcDNA3.1(+)-P2A-eGFP-TP53BP2 was successfully constructed by sequencing and double digestion.The fluorescence microscopy results showed that the transfection efficiency was about 60%.WB showed that the TP53BP2 protein was overexpressed in Expi293F cells,which proved that transfection was successful.SDS-PAGE results showed that the purity of the purified recombinant protein was above 90%,which proved that the purification was successful.Co-IP results showed that the TP53BP2 could interact with p65 protein.The results of IF showed that His tag protein,TP53BP2 protein,and p65 protein were co-located,indicating the interaction between the three proteins.SPR results showed that the purified TP53BP2 recombinant protein had good binding activity with the TP53BP2 antibody.These results all prove that the recombinant human full-length TP53BP2 protein has biological activity.Conclusion The eukaryotic expression vector of TP53BP2 gene was successfully constructed and the recombinant full-length human TP53BP2 protein with biological activity was successfully expressed in human embryonic kidney Expi293F cells.It lays a foundation for further study on the structure and function of TP53BP2.

BACKGROUND:Dapagliflozin,an inhibitor of sodium-glucose cotransporter 2,can delay the progression of atherosclerosis by regulating glucose metabolism,inhibiting inflammation and improving endothelial cell function. OBJECTIVE:To study the effect of dapagliflozin on cell pyroptosis and endothelial dysfunction induced by oxidized low-density lipoprotein. METHODS:Human umbilical vein endothelial cells were divided into a control group(no intervention),a model group(treated with oxidized low-density lipoprotein for 24 hours),and a dapagliflozin group(treated with oxidized low-density lipoprotein + dapagliflozin for 24 hours).Endothelial cell proliferation activity was measured by cell counting kit-8 assay.The levels of intercellular adhesion molecule 1,vascular cell adhesion molecule 1,and monocyte chemotactic protein-1 in cell supernatant were detected using ELISA.Nitric oxide level in the cells was detected by nitrate reductase assay.The pyroptosis rate and characteristics of endothelial cells were detected by Hoechst 33342/PI fluorescence co-staining and lactate dehydrogenase release assay.The protein expression levels of NLRP3,caspase-1,GSDMD,interleukin-1β,and interleukin-18 were detected by western blot assay. RESULTS AND CONCLUSION:(1)Oxidized low-density lipoprotein could cause pyroptosis and dysfunction of endothelial cells.(2)Compared with the control group,the level of nitric oxide and cell activity were decreased(P<0.05),while lactate dehydrogenase,intercellular adhesion molecule 1,vascular cell adhesion molecule 1,and monocyte chemotactic protein-1 levels were significantly increased in the model group(P<0.05).Compared with the model group,cell activity and nitric oxide levels significantly increased(P<0.05),but lactate dehydrogenase,intercellular adhesion molecule 1,vascular cell adhesion molecule 1,and monocyte chemotactic protein-1 levels were significantly diminished in the dapagliflozin group(P<0.05).(3)Compared with the model group,cell pyroptosis rate and the protein expression of pyroptosis factor NLRP3,caspase-1,GSDMD,interleukin-18 and interleukin-1β significantly reduced in the dapagliflozin group(P<0.05).(4)The results indicate that dapagliflozin inhibits oxidized low-density lipoprotein-induced endothelial pyroptosis and ameliorates endothelial cell dysfunction.

Fermentative production of amino acids is one of the pillars of the fermentation industry in China. Recently, with the fast development of metabolic engineering and synthetic biology technologies, the metabolic engineering for production of amino acids has been flourishing. Conventional forward metabolic engineering, reversed metabolic engineering based on omics data and in silico simulation, and evolutionary metabolic engineering mimicking the natural evolution, have shown increasingly promising applications. A series of highly efficient and robust amino acids-producing strains have been developed and applied in the industrial production of amino acids. The increasingly fierce market competition has put forward new requirements for strain breeding and selection, such as developing high value-added amino acids, dynamic regulation of cellular metabolism, and adapting to the requirements of new process. This review summarizes the advances and prospects in metabolic engineering for the production of amino acids.
Amino Acids ; China ; Corynebacterium glutamicum/genetics* ; Metabolic Engineering ; Synthetic Biology

Objective    To explore the practical feasibility of the weaving technique for pectus carinatum. Methods    From January 2011 to December 2018, a total of 51 patients with pectus carinatum, including 47 males and 4 females at age of 9-29 (13.7±2.9) years, were applied with minimally invasive waving technique for the correction. The steel plate was inserted through the subcutaneous layer, intercostal space and over the sternal surface under direct thoracoscopic vision. The number of implanted steel plates was determined by the degree of chest wall deformity. The steel plate was removed 2 years after surgery. Results    All the operations were successfully completed, the average operation time was 63.9±15.8 min, the amount of bleeding was 19.8±8.8 mL, and the duration of postoperative hospitalization was 4.6±1.6 d. The adverse events included intercostal artery injury (n=2), pneumothorax (n=4), pleural effusion (n=3) and skin rupture (n=1). And there were 29 patients of moderate pain (numerical rating scale 4-6 points) on the first day after surgery, but no patient was asked to remove the steel palate due to intolerable discomfort. All patients were followed up after plate placement. Of the 51 patients, the plates were removed in 37 patients until 2 years after placement, and the duration of postoperative hospitalization was 1.4±0.5 d. After 33 (1-48) months of routine follow-up after the removal of the plate, 22 patients achieved excellent outcomes and 9 patients with good outcomes. Besides, there were 5 patients with fair outcome and 1 patient with poor outcome. No adverse effect was found in growth and development after the steel plate placement. Conclusion    Minimally invasive weaving technique is a safe, feasible, effective and individualized operation for pectus carinatum with substantial thoracic reconstruction.

Objective : To investigate the role of dapagliflozin ( DAPA) in ox⁃LDL⁃induced pyroptosis of human myeloid leukemia monocytes (THP⁃1) derived foam cells . Methods : THP⁃1 ⁃derived foam cell pyroptosis model was constructed by ox⁃LDL⁃induced THP⁃1derived macrophages . The experimental groups were set as follows : the blank control group(NC) , the ox⁃LDL group(ox⁃LDL) , and the drug intervention group(ox⁃LDL + DAPA) . Oil Red Ostaining was used to detect the foam cell levels of macrophages . The cell proliferation and toxicity assay kit was used to detect the effect of DAPA on foam cell viability . Hoechst 33342/propidium iodide(PI) double staining was used to detect THP⁃1 derived foam cell pyroptosis . Cell immunofluorescence double staining was used to detect the effect of DAPA on the expression of pyroptosis key factor Caspase⁃1 in foam cells . The activity of lactate dehydrogenase (LDH) in the cell culture medium was detected using a microplate enzyme⁃linked immunosorbent assay. qRT⁃PCR and Western blot were used to detect the mRNA and protein expression levels of Nod⁃like receptor pyrindomain containing 3 (NLRP3) , cystein⁃containing aspartate⁃specific protease⁃1( Caspase⁃1 ) , apoptosis⁃associated⁃speck⁃like protein containing CARD(ASC) ,gasdermin⁃D (GSDMD) , interleukin(IL) Ⅳ18 and IL⁃1β , respectively . Results : The CCK⁃8 assay indicated that the optimal intervention concentration of DAPA was 10 μmol/L. Oil Red O staining confirmed the successful construction of the THP⁃1 ⁃derived foam cell pyroptosis model . Compared with the blank control group , the expression levels of NLRP3 , Caspase⁃1 , ASC , GSDMD , IL⁃18 , IL⁃1β mRNA and protein significantly increased in ox⁃LDL group(P < 0. 05) , as well as the number of PI⁃positive cells and LDH activity(P < 0. 05) , the fluorescence intensity of Caspase⁃1 and the number of redlipid droplets in the cytoplasm of the cells . However , these effects were significantly reversed after DAPA intervention in the ox⁃LDL + DAPA group(P < 0. 05) . Conclusion DAPA inhibits ox⁃LDL⁃induced pyroptosis in THP⁃1 ⁃derived foam cells .

Objective    To evaluate the clinical efficacy of fistula repair by stapler technique in patients with cervical tracheoesophageal fistula. Methods    Retrospective analysis of 8 patients with cervical tracheoesophageal fistula who accepted operative treatment in the Department of Thoracic Surgery, Lanzhou University Second Hospital from October 2014 to October 2016 was conducted. There were 5 males and 3 females at a mean age of 46.4±13.9 years ranging from 23 to 67 years. The fistula was induced by tracheal intubation in 4 patients, by esophageal foreign bodies in 2, by tracheal stent in 1 and by esophageal diverticulum in 1. The fistula was closed by stapler technique. The surgical effects were evaluated through Karnofsky performance score (KPS), image assessment, patient satisfaction score and assessment of improvement in feeding-induced bucking. Results    The operations were performed successfully with time of 117.5±6.6 min and intraoperative blood loss of 60.0±7.0 ml. After the operations, the patients did not suffer incision bleeding and infection, hoarseness, dyspnea, drinking-induced bucking, fistula relapse, tracheoesophageal stenosis or any other complications, and no death occurred during the perioperative period. The chest X-ray test was performed 1 week later showed that the pulmonary infection disappeared, and only 1 patient suffered from esophageal stenosis 1 year later. The postoperative KPS score was 90.0±7.0 points, which significantly improved in contrast to preoperation (P<0.01).   Postoperative pulmonary infection area reduced significantly (P<0.05), tracheoesophageal fistula disappeared, postoperative patients satisfaction rate was 90%, and assessment of feeding-induced bucking was excellent. Conclusion    Using stapler technique to repair cervical tracheoesophageal fistula is safe, easy and useful, with less operation time and postoperative complications.