Objective To introduce the outcomes of tracheal resection combined with primary end-to-end anastomosis for benign cervical tracheal stenosis,and to discuss the strategy for prevention of surgical complications.Methods The clinical data of 22 patients due to different causes benign cervical tracheal stenosis surgery were analyzed retrospectively.Results The length of cervical tracheal stenosis ranged was 2.2-4.2 cm.Grade Ⅱ stenosis was present in 6 patients.Grade Ⅲ stenosis was present in 11 patients and grade Ⅳ stenosis in 5 patients.Successful extubation was achieved in all 22 cases.After surgery,temporary hoarseness occurred to 1 case;unilateral pulmonary atelectasis with pleural effusion occurred to 1 case; subcutaneous emphysema with infection occurred to 1 case; mild dysphagia occurred to 3 cases;a slight deepening of the tone of voice in 10 patients with women occurred to 5 cases,granulation tissue growth near the suture occurred to 3 cases,and suture dehiscence did not occur in any patient.The follow-up period ranged from 6-45 months,no patient developed restenosis.Conclusions It presents a high success rate and good functional result of tracheal resection combined with primary end-to-end anastomosis.Therefore,it is an effective and reliable approach for the management of benign cervical tracheal stenosis.To avoid complications,the preoperative assessment,patients selection and postoperative management should be emphasized.

Objective To investigate the effect of small dose of dexmedetomidine on the development of postoperative nausea and vomiting in the patients undergoing laparoscopic gynecologic surgery.Methods A total of 207 patients,aged 18-60 yr,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective laparoscopic gynecologic surgery under general anesthesia,were randomly divided into either dexmedetomidine group (group D,n=103) or control group (group C,n=104) using a random number table.Before induction of anesthesia,dexmedetomidine 0.1 μg/kg was injected intravenously over 1 min in group D,while the equal volume of normal saline was given instead in group C.At 20 min before the end of surgery,dexmedetomidine 0.04 μg/kg was injected intravenously over 30 s in group D.For the patients requiring patient-controlled analgesia (PCA),PCA solution contained sufentanil 125 μg in 125 ml of normal saline.The PCA pump was set up with a 2 ml bolus dose,a 8 min lockout interval and background infusion at a rate of 1 ml/h.Pethidine was used for PCA after surgery in the patients who did not require PCA.Numeric rating scale scores were maintained ＜4 after surgery.Sufentanil 3 μg was injected intravenously as rescue analgesic in postanaesthesia care unit (PACU).The occurrence of nausea and vomiting was recorded within 0-1 h,1-6 h,6-12 h,and 12-24 h periods after surgery.The occurrence of hypotension and bradycardia during surgery,emergence time,Ramsay sedation score on admission to PACU,requirement for sufentanil as rescue analgesic in PACU,requirement for PCA and antiemetics,and occurrence of somnolence and shivering were recorded.Results Pethidine was not given in the patients who did not require PCA in the two groups.Somnolence and shivering were not found in the two groups.Compared with group C,the incidence of bradycardia was significantly increased,the requirement for antiemetics in PACU and incidence of nausea and vomiting within 1 h after surgery were significantly decreased (P＜0.05),and no significant change was found in the emergence time,Ramsay sedation score,requirement for sufentanil as rescue analgesic,requirement for PCA and total incidence of postsurgical nausea and vomiting in group D (P＞0.05).Conclusion For the patients undergoing laparoscopic gynecologic surgery,small dose of dexmedetomedine can only decrease the development of nausea and vomiting in the early postsurgical period (in PACU),and does not influence the recovery from anesthesia.

Objective To investigate the relation between lesions of medullary infarction and clinical symptoms and signs.Methods Eleven patients with medullary infarction confirmed by MRI were identified.Their clinical presentations and the relation between clinical presentation and location of lesion in MRI were assessed and analyzed.Results Among the 11 patients,5 cases suffered from lateral medullary infarction,6 medial medullary infarction(including two cases of bilateral infarction).Regarding clinical symptoms,common symptoms in lateral medullary infarction are blurred speech,dizziness,choking water,swallowing difficulties and facial numbness;in medial medullary infarction are limb weakness and blurred speech.The common signs of lateral medullary infarction are dysarthria,sensory disturbance,Horner sign and facial paralysis;in medial medullary infarction are paralysis of limbs and peripheral paralysis of the tongue.Conclusion Clinical manifestations are vailed depending on the sites of medullary infarction.MRI is helpful in diagnosing of medullary infarction.

OBJECTIVE:To investigate clinical efficacy and safety of oral loratadine combined with physiological seawater na-sal irrigation in the treatment of intermittent allergic rhinitis. METHODS:Totally 300 patients with intermittent allergic rhinitis were chosen from the Second Hospital of Hebei Medical University during Jan. 2013-Jun. 2015,and then divided into group A,B,C ac-cording to lottery method,with 100 cases in each group. Group A was given Loratadine tablets 10 mg,po,qd. Group B received nasal irrigation with physiological seawater nasal spray,every morning and evening. Group C was given oral loratadine combined with physiological seawater nasal irrigation. Treatment courses of 3 groups lasted for 28 d. Clinical efficiencies of 3 groups were compared as well as symptom and sign scores,respiration function indexes and inflammatory factor levels before and after treat-ment,and the clinical recurrences were followed up for 12 months. RESULTS:The total response rates of group A,B,C were 80.00%,78.00%,96.00%,respectively,and that of group C was significantly higher than that of group A and B,with statistical significance(P<0.05). Before treatment,there was no statistical significance in symptom and sign scores,respiration function in-dexes and inflammatory factor levels among 3 groups(P>0.05). After treatment,symptom and sign scores,the rates of PEF diur-nal variation,TNF-α,INF-γ and IL-4 in 3 groups were significantly lower than before treatment,and the levels of PEF and IL-12 were significantly higher than before treatment. Above indexes of group C were significantly better than those of group A and B, with statistical significance (P<0.05). Clinical recurrence rates of group A,B,C were 21.00%,23.00%,6.00%,and group C was significantly lower than the group A and B,with statistical significance (P<0.05). There was no statistical significance in above indexes between group A and group B (P>0.05). CONCLUSIONS:Oral loratadine combined with physiological seawater nasal irrigation in treatment of intermittent allergic rhinitis can efficiently relieve the nasal symptoms and signs,improve expiratoryfunction,reduce the inflammatory response levels and be help-ful to reduce the long-term recurrence risk.

Directed evolution was used to improve the performance of beta-1,3-1,4-glucanase (designated as PtLicl6A) from Paecilomyces thermophila J18 under acidic condition. A mutant library was constructed by error-prone PCR and DNA shuffling, and positive clones were screened by Congo red staining. More than 1 500 mutants were selected. One mutant (PtLic16AM1) exhibited an optimal activity at pH 5.5, while the optimal pH of the wild-type enzyme was 7.0. The mutant PtLic16AM1 kept the high specific activity and thermotolerence of the wild-type enzyme. Sequence analysis revealed that the mutant enzyme has four sense substitutions which caused four amino acid substitutions - namely T58S, Y110N, G195E and D221G.. Homology modeling showed that among the four amino acid substitutions, Y110N was near the active site of the enzyme, while the other three was distant. T58S and G195E may play key roles in the change of optimal pH. This study provided a new perspective of obtaining applicable 3-1,3-1,4-glucanase for industrial use.
Catalysis ; Directed Molecular Evolution ; methods ; Endo-1,3(4)-beta-Glucanase ; biosynthesis ; genetics ; metabolism ; Enzyme Stability ; Hot Temperature ; Hydrogen-Ion Concentration ; Mutant Proteins ; metabolism ; Mutation ; Paecilomyces ; classification ; enzymology ; genetics ; Protein Engineering ; methods

Objective To explore the diagnosis of clinically suspicious von Willebrand disease(vWD)in a family and its pathogene-sis.Methods The pedigree information and the biological specimen were collected from the clinically suspected VWD patient and her family members(4 persons in total)in Peking University First Hospital.The levels of platelet count(PLT),activated partial thrombo-plastin time(APTT),vWF antigen(vWF:Ag),vWF activity(vWF:Ac)and FⅧ activity(FⅧ:C)were detected,and vWF risto-cetin cofactor(vWF:RCo)assay,ristocetin-induced platelet aggregation assay(RIPA)and vWF collagen binding(vWF:CB)assay were performed for phenotype diagnosis.The peripheral blood genomic DNAs were extracted from the proband and her family members to perform whole-exome sequencing for identifying the mutation of vWF gene,The mutation site was analyzed by using bioinformation tools to explore the pathogenesis of the proband.Results The APTT of proband(m 1)was slightly prolonged and her vWF:Ag,vWF:Ac,vWF:RCo and vWF:CB were significantly decreased.There was no obvious aggregation in RIPA assay(1.0 mg/mL and 1.25 mg/mL).In her father(Ⅱ3),APTT,FⅧ:C,vWF:Ag,vWF:Ac and vWF:CB were normal,but vWF:RCo was slightly decreased.In her mother(Ⅱ4),APTT,FⅧ:C,vWF:Ag,vWF:RCo and vWF:CB were all normal,but vWF:Ac significantly decreased.In her brother(Ⅲ2),APTT and FⅧ:C were normal,but vWF:Ag,vWF:Ac,vWF:RCo and vWF:CB were reduced to varying degrees.In all the family members(father,mother and brpther),no apparent aggregation in RIPA(1.0 mg/mL)was shown.Genetic analysis showed that the proband(Ⅲ1)carried a compound heterozygous mutation of vWF gene c.7288-9T＞G and c.1117C＞T,her father(Ⅱ3)carried vWF gene c.7288-9T＞G heterozygous mutation,and vWF gene c.1117C＞T heterozygous mutation was presented in both mother(Ⅱ4)and brother(Ⅲ2).Conclusion According to the results of laboratory tests,the proband was diagnosed as type 2A vWD.The hetero-zygous mutation in vWF gene c.1117C＞T and c.7288-9T＞G may be the molecular mechanism leading to type 2A vWD in the proband.

Objective:To investigate the effect of tripartite motif-containing protein 59 (TRIM59) on glucose metabolism in macrophages and its role in regulating hypoxia-inducible factor-1α (HIF-1α)/IL-10 axis in macrophages under inflammatory conditions.Methods:The differentially expressed genes between macrophages with high expression of TRIM59 and control cells transfected with empty TRIM59 plasmid were analyzed by GO and KEGG. The expression of HIF-1α by RAW264.7 macrophages with high expression of TRIM59 was detected at different time points after lipopolysaccharide (LPS) stimulation by RT-qPCR and Western blot. Bone marrow was isolated from TRIM59-cKO and TRIM59 flox/flox mice and induced to differentiate into bone marrow-derived macrophages (BMDMs). These BMDMs were stimulated with LPS and the supernatants of cell culture were collected at 3, 6, 12 and 24 h after stimulation to detect IL-10 level by ELISA. In addition, mouse models of cecal ligation and puncture (CLP) were established, and bronchoalveolar lavage fluid (BALF) samples were collected at the same time points to detect IL-10 level by ELISA. Histopathological changes in lung tissues were observed after HE staining. Results:There was a significant change in glucose metabolism-related genes in macrophages with high expression of TRIM59, and the content of lactic acid increased significantly. Compared with the control group, the expression of HIF-1α at mRNA level in BMDMs from TRIM59-cKO mice decreased after LPS stimulation ( P<0.05); the level of IL-10 increased at 3 h and 24 h in the TRIM59-cKO group, but there was no significant difference in IL-10 level at 6 h or 12 h between the two groups. In the TRIM59-cKO mouse model of CLP, the levels of IL-10 in the BALF samples increased with time, but decreased at 24 h. The level of IL-10 was higher in the TRIM59-cKO mouse model group than that in the control group at each time point ( P<0.05 or P<0.01). Conclusions:TRIM59 can inhibit inflammation and lung injury by decreasing HIF-1α-mediated lactate secretion and IL-10 expression in macrophages. This study provides a new idea for developing novel anti-sepsis drugs based on TRIM59.

Objective:To explore the mechanism of Ginseng Radix et Rhizoma- Notoginseng Radix et Rhizoma- Chuanxiong Rhizoma medicinal pair in delaying heart aging based on animal experiments, network pharmacology and molecular docking. Methods:Mice were divided into control group, aging group, metformin group and TCM group according to random number table method. All the groups were injected subcutaneously by D-galactose except the control group to build the subacute aging model. Two weeks later, the metformin group was given metformin suspension (150 mg/kg), the TCM group was given Ginseng Radix et Rhizoma- Notoginseng Radix et Rhizoma- Chuanxiong Rhizoma lyophilized powder solution (650 mg/kg), and the control group and aging group were given an equivalent volume of ultrapure water by gastric gavage, once a day, six times a week, for 10 weeks. The level of heart TERT mRNA was detected by PCR; the expression of heart p53 was observed by immunohistochemical staining; the morphology of heart tissue was observed by HE staining. TCMSP and SwissTargetPrediciton databases were used to retrieve the active components and targets of Ginseng Radix et Rhizoma- Notoginseng Radix et Rhizoma- Chuanxiong Rhizoma medicinal pair; TTD, OMIM, Gene, HAGR, DisGeNET and other data platforms were used to screen the targets of heart aging; after the drug and disease targets were intersected, the active components of them were collected; STRING database, Cytoscape 3.8.0 software, etc. were used to make PPI of the intersection targets, and screen out the key targets; FunRich was used to perform enrichment analysis of cellular components, molecular functions, biological processes, and biological signal pathways for key targets; Schr?dinger Maestro software was used to do the molecular docking of the screened active components and key targets, and docking results were visualized via PyMOL 2.1 software. Results:Experiment results showed that Ginseng Radix et Rhizoma- Notoginseng Radix et Rhizoma- Chuanxiong Rhizoma could significantly ameliorate the damage of aging heart tissues, elevate TERT mRNA level, while significantly reducing the positive expression of p53. A total of 32 active components from the medicinal pair were screened, corresponding to 637 target genes. There were 263 targets for heart aging, and 67 intersection targets of drug active component targets and heart aging targets. 31 key targets were obtained after screening. Enrichment analysis showed that molecular functions were related to transcription factor activity and protein-tyrosine kinase activity. Biological processes involved signal transduction and cell communication. Signaling pathways mainly involved PDGFR-beta, PI3K-Akt, S1P1, Glypican, TRAIL, and Glypican 1. The molecular docking results showed that kaempferol, suchilactone, and ginsenoside Rg5_qt in the medicinal pair had a strong binding ability to p53. Conclusion:Ginseng Radix et Rhizoma- Notoginseng Radix et Rhizoma- Chuanxiong Rhizoma may achieve the effect of delaying heart aging by inhibiting p53 expression, providing a foundation for further research on mechanism of invigorating qi and activating blood circulation drugs to delay heart aging.

Objective:To evaluate the accuracy of pRIFLE criterion and KDIGO criterion for the diagnosis of acute kidney injury (AKI) after radical operations for tetralogy of Fallot in children from the perspective of postoperative outcomes.Methods:A total of 375 children, aged<8 yr, undergoing radical operations for tetralogy of Fallot, were selected continuously and retrospectively. According to the pRIFLE and KDIGO diagnostic criteria, postoperative AKI was diagnosed, and the children were classified into different AKI grades. The prognostic parameters (postoperative mechanical ventilation time, duration of intensive care unit (ICU) stay, postoperative length of hospital stay, total costs of hospitalization, and in-hospital mortality, etc.) were collected, and the differences among different AKI grades were compared. Logistic regression method was used to analyze the risk factors for prolonged postoperative length of hospital stay (≥14 days) when two different criteria were used to diagnose AKI. The children diagnosed as non-AKI by KDIGO criterion were further confirmed using pRIFLE criterion, and the prognostic parameters in the children who were diagnosed as AKI and non-AKI were compared.Results:When two different criteria were used to diagnose AKI after radical resection for tetralogy of Fallot, the incidence was 56.8% (pRIFLE criterion) and 40.0% (KDIGO criterion). AKI diagnosed according to the two criteria was the independent risk factor for prolonged postoperative length of hospital stay, and the levels of all the prognostic parameters (postoperative mechanical ventilation time, duration of ICU stay, postoperative length of hospital stay, total costs of hospitalization, and in-hospital mortality) were significantly higher in AKI children than in non-AKI (AKI grade 0) children ( P<0.01). Among the 225 children diagnosed as non-AKI according to the KDIGO criterion, 63 cases were diagnosed as AKI and 162 cases as non-AKI according to the pRIFLE criterion, however, there was no significant difference in each prognostic parameter between children with AKI and non-AKI ( P>0.05). Conclusion:The pRIFLE criterion has a higher sensitivity, while the KDIGO criterion produces better accuracy when used to evaluate the diagnosis of AKI after radical operation for tetralogy of Fallot in children from the perspective of postoperative outcomes.

Objective: To analyze the gene characterization on the first imported D8 genotype measles virus in Liaoning province. Methods: In this study, Vero/Slam cells were used to isolate measles viruses from throat swabs. Fragments of the H gene (1854 nucleotides) and N gene (450 nucleotides) were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the PCR products were sequenced and analyzed. Results: The measles virus isolates and World Health Organization (WHO) D8 genotype reference strain (MVi/Manchester.GBR/30.94) belonged to the same branch in the genetic relationship tree. The nucleotide homology of the N and H gene was 98.9%. Phylogenetic trees were constructed with reference strains of the genotype D8 measles virus of China downloaded from GenBank. The result showed that the nucleotide similarities between the measles virus isolated in this study and the D8 genotype measles virus prevalent in Hong Kong from 2012 to 2016 and MVi/LosPatios.COL/11.18/D8 in Columbia was 100%. Conclusions It is the first time to do surveillance for the D8 genotype measles virus since measles virus surveillance was carried out. It was of great significance to accumulate the bases of measles virus molecular epidemiology in Liaoning province, and helpful to analyze and trace the transmission of measles virus in the whole country and the world.