Degenerative changes in autonomic nervous system happen in elderly. The cardiac function reduces, the ability of regulating the target organ decreases, and the stability of hemodynamics is not easy to maintain in old people. How to maintain the hemodynamic stability of elderly patients during anesthesia has become the focus of clinical research. Dexmedetomidine, as a new type of 2- alpha adrenergic agonist, has the function of anti-anxiety, sedation, analgesia and anti-sympathy, and which is wildly used in clinic. This article describes the research progress of dexmedetomidine used in elderly patients during anesthesia.

Anti-angiogenesis mechanism plays a vital role in tumor targeting immunotherapy. Based on the amino acid sequence of an anti-VEGFR-2 scFv-Fc fusion antibody (AK404R-Fc), this article is aimed to generate an anti-VEGFR-2 human IgG1-like full length antibody (Mab-04). Firstly, the light chain (L-chain) and heavy chain (H-chain) were obtained by overlap PCR and then linked to eukaryotic expression vector pcDNA3.1, separately. The recombinant plasmids (pcDNA3.1-L-chain and pcDNA3.1-H-chain) were then co-transfected into CHO-k cells using liposome transient transfection. Subsequently, Mab-04 antibody was expressed and purified by Protein A affinity chromatography. Western blotting was applied to identify the expression of Mab-04 and its affinity was detected by ELISA assay. DNA sequencing revealed the successful construction of recombinant plasmids and Western blotting assay proved the successful expression of full-length antibody (1 microg x mL(-1)). Finally, ELISA assay illustrated that the binding of the antibody to its antigen was in a concentration-dependent manner (IC50: 50 nmol x L(-1)). These outcomes above indicated that Mab-04 was successfully expressed and assembled, which laid the foundation for further preparation and antineoplastic activity study.

OBJECTIVE:To evaluate the economics of azithromycin vs. amocillin clavulante in the treatment of lower respirato-ry tract infections. METHODS:System evaluation was adopted to retrieve the randomized controlled trials(RCT)about azithromy-cin(test group)vs. amoxicillin clavulanate(control group)in the treatment of lower respiratory tract infections. Information was col-lected and Meta-analyses were performed. On this basis and short-term decision tree model,cost factors were added to conduct the pharmacoeconomics by the principle of PICO of Treeage Pro 2011 edition software. RESULTS:Totally 18 RCT were enrolled,in-volving 3 365 patients. Results of Meta-analysis showed that there were no significant differences in the effective rate [RR=0.93, 95%CI(0.55,1.55),P=0.77] and incidence of adverse reactions [RR=0.79,95%CI(0.62,1.0),P=0.05] between 2 groups. The av-erage treatment cost in test group and control group was respectively 790.4 yuan and 884.4 yuan,and cost-effectiveness ratio was respectively 216.0 and 245.7,and the incremental cost-effectiveness ratio(ICER)was -1 392.59. CONCLUSIONS:Azithromycin has similar efficacy and safety to amoxicillin clavulanate in the treatment of lower respiratory tract infection,however,azithromy-cin has better cost-effectiveness.

Objective:To evaluate the role of DNA methyltransferase in acute lung injury in septic mice.Methods:Forty-eight healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (group Sham), sham operation+ DNA methyltransferase inhibitor group (group Sham+ 5-Aza), sepsis group (group Sepsis) and sepsis+ DNA methyltransferase inhibitor group (group Sepsis+ 5-Aza). Sepsis model was developed by cecal ligation and puncture (CLP) in anesthetized mice.Mice were sacrificed at 24 h after CLP, and lung tissues were obtained, DNA was extracted to determine the global DNA methylation by colorimetry, and RNA was extracted to detect the expression of DNA methyltransferase (DNMTl, DNMT3a, DNMT3b) mRNA by real-time fluorescent quantitative polymerase chain reaction, the wet/dry lung weight ratio (W/D ratio) was measured, the histopathological changes of lung tissues were determined by HE staining, the contents of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), high-mobility group box 1 protein (HMGB1) and malondialdehyde (MDA) and activities of superoxide dismutase (SOD) and catalase were measured by enzyme-linked immunosorbent assay. Results:Compared with group Sham, the global DNA methylation was significantly increased, the expression of DNMT1 and DNMT3a mRNA was up-regulated, the lung injury score, W/D ratio, and contents of IL-6, TNF-α, HMGB1 and MDA were increased, and activities of SOD and CAT were decreased at 24 h after CLP in group Sepsis and group Sepsis+ 5-Aza ( P<0.05), and no significant change was found in the indexes mentioned above in group Sham+ 5-Aza ( P>0.05). Compared with group Sepsis, the global DNA methylation was significantly decreased, the expression of DNMT1 and DNMT3a mRNA was down-regulated, the lung injury score, W/D ratio, contents of IL-6, TNF-α, HMGB1 and MDA were decreased, and the activities of SOD and CAT were increased in group Sepsis+ 5-Aza ( P<0.05). Conclusions:DNA hypermethylation mediated by DNMT1 and DNMT3a is involved in the process of acute lung injury in septic mice.

Objective To construct a lentiviral vector delivering the Nicastrin (NCT) gene-targeted short hairpin RNA (shRNA) and determine gene-silencing efficiency of the vector in the human immortalized keratinocyte cell line HaCaT,and to construct a NCT gene-silenced HaCaT cell model to lay an experimental foundation for subsequently studying effects of NCT gene silencing on biological behavior of keratinocytes.Methods Three NCT gene-targeted shRNAs were designed and inserted into the pGLV3/ H1/GFP + Puro vector to construct three recombinant plasmids,which were then confirmed by sequencing.Recombinant plasmids combined with lentivirus packaging plasmids were co-transfected into 293T cells to obtain lentivirus particles,and the virus titer was determined.Cultured HaCaT cells were divided into 3 groups:blank group receiving no treatment,negative control group infected with the empty vector LV3-shNC,interference groups infected with lentivirus NCT-shRNA1,-shRNA2,-shRNA3,respectively.Flow cytometry was performed to determine transfection efficiency,and real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were conducted to determine efficiency of target gene silencing in HaCaT cells,so as to select the most efficient interference sequence.Results Sequencing analysis indicated that recombinant lentiviral vector NCT-shRNA was constructed successfully.After co-transfection of recombinant plasmids and lentivirus packaging plasmids into 293T cells,the titer of recombinant lentivirus particles was about 109 TU/ml.Flow cytometry showed that the transfection efficiency was greater than 95％.qRT-PCR revealed that the NCT mRNA expression was obviously down-regulated in the interference group compared with the negative control group,and NCT-shRNA1 was the most efficient sequence with interference efficiency being 75％.Western blot analysis showed that the inhibition rate of NCT protein expression was 71.7％ in the shRNA1 group compared with the negative control group.Conclusion The most efficient NCT-shRNA interference sequence is screened out,and the recombinant lentiviral vector NCT-shRNA and an NCT gene-silenced HaCaT cell model are both constructed successfully.

Objective:To explore the differences of clinical laboratory indicators between Kawasaki disease (KD) and systemic juvenile idiopathic arthritis (SJIA), providing objective evidence for diagnosis and differential diagnosis of these diseases.Methods:A total of 41 children patients with KD (KD group) and 33 children patients with SJIA (SJIA group) who received treatment in Huainan Maternal and Child Health Hospital between September 2017 and January 2022 were retrospectively analyzed. An additional 50 healthy children who concurrently received physical examination in the same hospital were included in the control group. Platelet count (PLT), white blood cell count (WBC), and erythrocyte sedimentation rate (ESR) as well as C-reactive protein (CRP), serum procalcitonin (PCT), interleukin-6 (IL-6), interleukin-10 (IL-10), and serum ferritin (SF) levels were compared among groups before treatment.Results:One-way analysis of variance and pairwise q test were performed to compare laboratory indicators among KD, SJIA and control groups. CRP, ESR, SF and IL-6 levels in the KD group were significantly lower than those in the SJIA group [CRP: (57.80 ± 25.23) mg/L vs. (77.72 ± 45.64) mg/L; ESR: (67.02 ± 28.80) mm/h vs. (83.84 ± 47.64) mm/h; SF: (320.21 ± 182.53) μg/L vs. (945.58 ± 604.65) μg/L; IL-6: (50.35 ± 20.54) ng/L vs. (89.35 ± 45.54) ng/L, q = 4.34, 3.42, 11.51, 8.85, all P < 0.05]. IL-10 level in the KD group was significantly higher than that in the SJIA group [(18.52 ± 16.71) ng/L vs. (10.01 ± 3.24) ng/L, q = -5.25, P < 0.05]. WBC, CRP, ESR, PCT, PLT, IL-6, IL-10 and SF in the KD and SJIA groups were significantly higher than those in the control group (all P < 0.05). Conclusion:Detection of CRP, ESR, SF, IL-6, IL-10 in blood can provide objective evidence for the early diagnosis and differential diagnosis of KD and SJIA, thereby reducing the misjudgment of clinical diagnosis.

Objective:To investigate the effect of end-expiratory positive pressure of pulmonary protective ventilation strategy in overweight patients undergoing laparoscopic surgery.Methods:Forty overweight patients, 24 kg/m 2≤BMI<28 kg/m 2, aged 20-65yr, of American Society of Anesthesiologists (ASA) physical status Ⅰ or Ⅱ, scheduled for elective laparoscopic surgery to radical resection of rectal cancer under general anesthesia, were randomly divided into 2 groups ( n=20 each) using a random number table: positive end-expiratory pressure (PEEP) group (group P), control group (group C). The rest of the settings in mechanical ventilation were the same in both groups, tidal volume (Vt)=6 ml/kg, initial respiration frequency (RR)=15 bpm, oxygen inhalation 100%, inspiratory expiratory time ratio ( I∶E)=1∶2. The concentration of oxygen inhalation was 50% and respiration frequency was adjusted to maintain P ETCO 2 35-45 mmHg after endotracheal intubation. The heart rate (HR), mean arterial pressure (MAP), tidal volume (Vt), airway peak pressure (Ppeak), airway pressure platform (Pplat) were recor-ded, and the lung dynamic compliance (Cdyn) was calculated; arterial oxygen partial pressure (PaO 2) and partial pressure of carbon dioxide in arterial blood (PaCO 2) were measured by gas analyzer; oxygenation index (OI) was calculated at the time of before induction of anesthesia (T 0) , 5 min after endotracheal intubation (T 1), 5 min after laparoscopic pneumoperitoneum (T 2), 60 min after laparoscopic pneumoperitoneum (T 3), and suturing the skin (T 4). The postoperative pulmonary complications were observed 3 days after surgery. Results:There was no significant difference between the two groups in patients characteristics and operative indicators ( P>0.05). Compared with T 0, the mean arterial blood pressure of the two groups decreased at T 3 and T 4, and there was no significant difference in heart rate at each time ( P>0.05). There was no significant difference in hemodynamic parameters between the two groups ( P>0.05). Compared with T 1, VT increased at T 2 in group P and T 3 in group C, while there was no significant difference in VT between the two groups ( P>0.05); compared with T 1, Ppeak and Pplat increased at T 2 and T 3 in both groups, while there was no significant difference between the two groups ( P>0.05); compared with T 1, Cdyn of the two groups decreased at T 2 and T 3 ( P<0.05), and cdyn of the P group at each time were higher than that of the group C ( P<0.05). Compared with T 1, PaO 2 and OI decreased and PaCO 2 increased at T 2, T 3 and T 4 in the two groups ( P<0.05). PaO 2 and OI at T 3 and T 4 in the P group were higher than those in the C group ( P<0.05). There was no significant difference in PaCO 2 between the two groups ( P>0.05). There was no significant difference in the incidence of pulmonary complications between the two groups ( P>0.05). Conclusions:Positive end-expiratory pressure of protective ventilation strategy from the induction period of general anesthesia can effectively improve dynamic lung cdyniance, improve oxygenation and promote pulmonary function recovery in overweight patients undergoing laparoscopic surgery.

Objective:To construct a chronic obstructive pulmonary disease (COPD) assessment test (CAT) score prediction model based on a deep network fused with air data, and to explore its significance.Methods:From February 2015 to December 2017, the outdoor environmental monitoring air data near the residential area of the patients with COPD from the Respiratory Outpatient Clinics of Peking University Third Hospital, Peking University People′s Hospital and Beijing Jishuitan Hospital were collected and the daily air pollution exposure of patients was calculated. The daily CAT scores were recorded continuously. The CAT score of the patients in the next week was predicted by fusing the time series algorithm and neural network to establish a model, and the prediction accuracy of the model was compared with that of the long short-term memory model (LSTM), the LSTM-attention model and the autoregressive integrated moving average model (ARIMA).Results:A total of 47 patients with COPD were enrolled and followed up for an average of 381.60 days. The LSTM-convolutional neural networks (CNN)-autoregression (AR) model was constructed by using the collected air data and CAT score, and the root mean square error of the model was 0.85, and the mean absolute error was 0.71. Compared with LSTM, LSTM-attention and ARIMA, the average prediction accuracy was improved by 21.69%.Conclusion:Based on the air data in the environment of COPD patients, the fusion deep network model can predict the CAT score of COPD patients more accurately.

Objectives: To explore the differences in signal pathway and gene expression related to the pathogenesis of congenital microtia by the in-depth analysis of DNA methylation profiling of auricular chondrocytes from congenital microtia patients. Methods: Genome wide methylation profile of congenital microtia was obtained by MeDIP chip technology, and analyzed by Gene ontology (GO) and Pathway analysis. The gene expression levels of Wnt1 and Wnt11 were evaluated by Real-time PCR in the auricular cartilage from the healthy side and affected side of the congenital microtia patients , and healthy controls. Results: The GO and Pathway assay showed that Wnt signal pathway was enriched in differential methylated levels. The Wnt1 and Wnt11 genes were with higher methylation in the promoter region and CpG islands in healthy control group than that in microtia group, in addition the methylation level in the affected side auricular cartilage was lower than that in the healthy side. There was no difference in Wnt1 and Wnt11 gene expression in microtia patients and healthy controls. The higher Wnt11 gene expression was detected in the affected side residual cartilage tissues than in the healthy side cartilage tissues of the same congenital microtia patient. Conclusions The over expression of Wnt11 during embryonic development might be associated with the pathogenesis of congenital microtia. The mechanism of the difference in methylation levles of Wnt11 affecting pathogenesis of congenital microtia needs further research.

Objective:To evaluate the proliferative activity of and changes in the expression of related differentiation proteins in a stably NCSTN gene-silenced human immortalized keratinocyte cell line HaCaT, and to preliminarily explore the possible mechanism underlying the occurrence of acne inversa.Methods:By lentivirus-mediated short hairpin RNA (shRNA) , a NCSTN gene-silenced HaCaT cell model was established (shRNA group) , and other HaCaT cells transfected with empty lentivirus served as a negative control group. Real-time quantitative PCR and Western blot analysis were performed to determine the NCSTN gene-silencing efficiency. Cell counting kit-8 (CCK8) assay was conducted to evaluate the proliferative activity of HaCaT cells, and real-time quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of cytokeratins (CK1, CK5, CK7, CK10, CK14, CK16, CK17, CK18, CK19 and CK20) and other differentiation molecules (involucrin and loricrin) respectively in HaCaT cells. Two-independent-sample t test was used to compare the measurement data between two groups. Results:NCSTN mRNA and protein expression were significantly lower in the shRNA group (0.42 ± 0.19, 0.30 ± 0.07 respectively) than in the negative control group (1.00 ± 0.34, 1.00 ± 0.26; t = 5.196, 2.637, P < 0.001, < 0.05, respectively) , and the gene-silencing efficiency was 70%. Compared with the negative control group, the shRNA group showed higher cellular proliferative activity, but decreased protein expression of CK16, CK19 and terminal differentiation molecule involucrin ( t = 3.787, 3.817, 2.904, P < 0.01, < 0.05, < 0.05, respectively) . Conclusion:Stable silencing of NCSTN gene can lead to abnormal proliferation and differentiation of HaCaT cells, which provides new ideas for subsequent exploration of acne inversa caused by NCSTN gene mutation.