After sequencing, we amplified and cloned foot-and-mouth disease virus (FMDV) O/QYYS/s/06 whole genome by three fragments. These three fragments were cloned into vector P43 one by one to construct recombinant plasmid P43C, which carried the full-length cDNA of FMDV O/QYYS/s/06. Then, plasmid P43C and plasmid T7 expressing T7 RNA polymerase were co-transfected into BHK-21 cells. After 48 h, we harvested the culture broth from transfected BHK-21 cells and inoculated into 2-3 day-old sucking mice. After four generation passage, the virus harvested from sucking mice was confirmed to be type O FMDV by the indirect hemagglutination test, sucking mice's neutralization test and sequencing. The results showed that we have successfully constructed the full-length cDNA clone of FMDV O/QYYS/s/06 strain.
Animals ; Animals, Newborn ; Cloning, Molecular ; DNA, Complementary ; genetics ; DNA, Viral ; biosynthesis ; genetics ; Foot-and-Mouth Disease ; virology ; Foot-and-Mouth Disease Virus ; classification ; genetics ; pathogenicity ; Mice ; Transcription, Genetic ; Transfection

Objective To investigate the curative effect of lateral position ventilation combined with vibration sputum drainage on the patients with acute respiratory distress syndrome (ARDS). Methods A prospective randomized controlled trial was conducted. The patients with ARDS undergoing mechanical ventilation (MV) admitted to intensive care unit (ICU) of Linyi Central Hospital from January 2013 to June 2017 were enrolled, and they were divided into simple ventilation group and combined treatment group according to random number table. The patients in both groups received etiological treatment, protective ventilation strategy, sensitive antibiotics for anti-infection, and calefacient and humidifying treatment. The patients in the simple ventilation group received bilateral discontinuous alternative lateral position ventilation [pressure controlled ventilation (PCV), tidal volume (VT) ≤ 6 mL/kg, the inhaled oxygen concentration (FiO2) and positive end expiratory pressure (PEEP) was adjusted to maintain the airway plateau pressure (Pplat) ≤ 30 cmH2O (1 cmH2O = 0.098 kPa)], and those in the combined treatment group received lateral position ventilation combined with vibration sputum drainage, twice a day, 15 minutes each time. The parameters of respiratory function and inflammation as well as excretion of sputum before and after treatment were compared between the two groups, and the complication and prognosis were recorded. Results A total of 200 patients with ARDS were included, 4 patients were excluded because of severe pneumothorax, massive hemorrhage of the digestive tract, or elevated intracranial pressure, so 196 patients were finally enrolled in the analysis. There were 98 patients in the simple ventilation group and the combined treatment group, respectively. There were no significant differences in parameters of respiratory function and inflammation as well as excretion of sputum before treatment between the two groups. After treatment, the above parameters in both groups were improved, and the effect of combined treatment was more significant. Compared with the simple ventilation group, white blood cell count (WBC), procalcitonin (PCT), C-reactive protein (CRP) at 24 hours of treatment in the combined treatment group were significantly decreased [WBC (×109/L):9.1±1.6 vs. 11.8±3.6, PCT (μg/L): 14.5±2.4 vs. 22.7±3.2, CRP (mg/L): 32.2±6.3 vs. 67.2±7.2, all P < 0.01], the quantity of sputum excretion was significantly increased (mL: 49.3±12.5 vs. 36.8±11.8, P < 0.01); 72 hours after treatment, the oxygenation index (PaO2/FiO2) in the combined treatment group was significantly increased [mmHg (1 mmHg = 0.133 kPa): 278±28 vs. 238±39, P < 0.01], and PEEP, FiO2, airway resistance (Raw) were significantly lowered [PEEP (cmH2O): 5±2 vs. 7±3, FiO2: 0.40±0.11 vs. 0.49±0.12, Raw (cmH2O): 12.8±0.7 vs. 16.2±0.8, all P < 0.01]. There was no significant difference in static lung compliance (Cst) or Pplat between the two groups. Compared with the simple ventilation group, the duration of MV (days: 5.5±3.0 vs. 8.6±2.1), the length of ICU stay (days: 7.5±5.4 vs. 11.3±4.6) and the extraction time of artificial airway (days: 6.6±2.8 vs. 9.4±3.3) in the combined treatment group were significantly shortened (all P < 0.01). However, there was no significant difference in the 28-day mortality rate [4.1% (4/98) vs. 3.1% (3/98)], the rate of tracheotomy [2.0% (2/98) vs. 3.1% (3/98)] or the incidence ofⅠ-Ⅱ skin pressure sore [1.0% (1/98) vs. 2.0% (2/98)] between simple ventilation group and the combined treatment group (all P > 0.05). No artificial airway disappearance, pneumothorax, grade Ⅲ or Ⅳ pressure sore was found in the two groups. Conclusions Compared with lateral position ventilation alone, the treatment of combined with vibration sputum drainage is more effective for improving the respiratory function of patients with ARDS, reducing infection action, shortening the duration of MV and the length of ICU stay, and improve the recovery, promote the rehabitation of patients.

Objective:To investigate the risk factors of thrombocytopenia in elderly patients with sepsis-associated acute kidney injury(SA-AKI), and to further evaluate whether the degree of thrombocytopenia is related to the increased risk of death on day 28.Methods:Elderly patients with SA-AKI admitted to ICU of our hospital from June 2017 to June 2020 were selected.The patients were divided into normal platelet group(56 cases)and thrombocytopenia group(72 cases)according to the platelet(PLT)count, and according to the degree of thrombocytopenia, they were further divided into three groups: PLT<20×10 9/L(group A, 22 cases), 20×10 9/L≤ PLT<50×10 9/L(group B, 27 cases), 50×10 9/L≤ PLT<100×10 9/L(group C, 23 cases).The general data, clinical baseline indicators and prognostic indicators of each group were compared to evaluate the risk factors of thrombocytopenia.At the same time, the impact of platelet count on the prognosis of elderly patients with SA-AKI was evaluated according to the length of stay in ICU, total length of stay and whether the patient died, and the correlation between the degree of thrombocytopenia and survival was analyzed. Results:A total of 128 elderly patients with SA-AKI were enrolled, including 73 males and 55 females.59.4 % of the patients were hospitalized in the department of internal medicine.The APACHEⅡ score was(15.5 ± 3.3)points, invasive mechanical ventilation accounted for 78.9%, positive inotropic therapy accounted for 12.8%, and 56 patients had normal platelet count.Thrombocytopenia occurred in 72 patients, including 22 patients with PLT<20×10 9/L, 27 patients with platelet count in 20-50×10 9/L, and 23 patients with platelet count in 50-100×10 9/L.There were significant differences in bloodstream infection, Gram-negative bacteria, APACHEⅡ score and procalcitonin(PCT)among the four groups( P<0.05).Further multivariate logistic regression showed that PCT was an independent risk factor for thrombocytopenia in elderly patients with SA-AKI( OR=1.05, 95% CI: 1.00-1.10, P=0.042).Compared with the normal platelet group, the 28-day mortality rate of the thrombocytopenia group was significantly higher than that of the normal platelet group, while the length of stay in ICU and the total length of stay were prolonged( P<0.05).Kaplan-Meier survival curve showed that the 28-day cumulative survival rate of thrombocytopenia group was lower than that of normal platelet group, and the risk of death increased with the degree of thrombocytopenia( χ2=31.479, P<0.001).Univariate Cox regression analysis showed that thrombocytopenia, bloodstream infection, APACHEⅡ score and PCT were risk factors for 28-day death in elderly patients with SA-AKI.Multivariate Cox analysis showed that bloodstream infection and APACHEⅡ score were independent risk factors for 28-day death.After adjusting these confounding factors, thrombocytopenia was an independent risk factor for death, and the degree of thrombocytopenia was related to the increased risk of death. Conclusions:Thrombocytopenia is common in elderly patients with SA-AKI, and elevated PCT levels increase the risk of thrombocytopenia.The degree of thrombocytopenia is an independent risk factor for 28-day mortality in such patients.

Objective To analyze the current situation of unplanned reoperation in cardiac surgery and to discuss the management measures of unplanned reoperation.Methods The information of patients undergoing cardiac surgery in a class A tertiary comprehensive hospital during 2018-2022 was collected to analyze the incidence of unplanned reoperation,major ca uses,disease types,surgica l moda lities and Complications.Results A tota l of 3902 patients underwent surgery,of whom 73(1.87%)underwent unplanned reoperation.The main cause of unplanned reoperation was bleeding(50%).The disease types with the highest unplanned reoperation composition ratio were coronary heart disease(38.4%),and the disease types with the highest incidence were dilated cardiomyopathy(11.1%).The average hospitalization cost,the average length of hospitalization,mortality rate and medical dispute rate of patients who had unplanned reoperation were significantly higher than those who did not have unplanned reoperation,the difference was statistically significant(P=0.001).Conclusion The hospital should strengthen the perioperative management of cardiac surgery,focus on supervising disease types and surgical modalities with high incidence of unplanned reoperation,and strictly implement the system of operation classification and the system of reporting unplanned reoperation to ensure the quality of patient surgery.

In recent years, with in-depth study of bladder sensation related mechanisms, numerous ion channels, neurotransmitters and nerve receptors have been found to participate in the regulation of bladder sensation, including TRPV, P2X and Piezo, as well as CBR and HCN.Thanks to the relevant research on the neural signal pathway from the cerebral cortex to the bladder wall and the maturity of clinical measurement methods for bladder sensation, we can further study the abnormal bladder sensation in patients with overactive bladder (OAB), so as to explore its mechanism.Bladder hypersensitivity, as one of the current research hotspots, is receiving increasing attention from researchers.This article reviews the mechanism of bladder hypersensitivity from the aspects of clinical measurement methods of bladder sensation, ion channel, neurotransmitters and nerve receptors related to bladder sensation, in order to explore its significance in the pathogenesis of OAB.

Stress urinary incontinence (SUI) and underactive bladder (UAB) are common types of lower urinary tract dysfunction in women.As the treatment mechanisms of the two conditions are contradictory, the treatment of SUI patients complicated with UAB remains a difficult clinical problem.In order to improve the treatment rate of such patients and promote research, this paper reviews the latest domestic and overseas diagnostic criteria of UAB, summarizes the treatment experience of conventional midurethral sling (tension-free vaginal tape or outside-in transobturator tape) and adjustable sling procedures (transobturator adjustable tape or Remeex system) combined with medication or intermittent catheterization, and the application prospects of cutting-edge technologies such as stem cell injection, cytokine therapy and gene therapy, so as to provide reference for clinicians and researchers.